[go: up one dir, main page]

US20110119791A1 - Isolated polypeptides, polynucleotides useful for modifying water user efficiency, fertilizer use efficiency, biotic/abiotic stress tolerance, yield and biomass in plants - Google Patents

Isolated polypeptides, polynucleotides useful for modifying water user efficiency, fertilizer use efficiency, biotic/abiotic stress tolerance, yield and biomass in plants Download PDF

Info

Publication number
US20110119791A1
US20110119791A1 US12/810,855 US81085508A US2011119791A1 US 20110119791 A1 US20110119791 A1 US 20110119791A1 US 81085508 A US81085508 A US 81085508A US 2011119791 A1 US2011119791 A1 US 2011119791A1
Authority
US
United States
Prior art keywords
blastp
plant
seq
acid sequence
plants
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Abandoned
Application number
US12/810,855
Other languages
English (en)
Inventor
Gil Ronen
Basia J. Vinocur
Alex Diber
Sharon Ayal
Hagai Karchi
Yoav Herschkovitz
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Evogene Ltd
Original Assignee
Evogene Ltd
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Evogene Ltd filed Critical Evogene Ltd
Priority to US12/810,855 priority Critical patent/US20110119791A1/en
Assigned to EVOGENE LTD. reassignment EVOGENE LTD. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: KARCHI, HAGAI, RONEN, GIL, AYAL, SHARON, DIBER, ALEX, VINOCUR, BASIA JUDITH, HERSCHKOVITZ, YOAV
Publication of US20110119791A1 publication Critical patent/US20110119791A1/en
Abandoned legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8273—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for drought, cold, salt resistance
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
    • C12N15/8245—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine involving modified carbohydrate or sugar alcohol metabolism, e.g. starch biosynthesis
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8242—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits
    • C12N15/8243—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine
    • C12N15/8247—Phenotypically and genetically modified plants via recombinant DNA technology with non-agronomic quality (output) traits, e.g. for industrial processing; Value added, non-agronomic traits involving biosynthetic or metabolic pathways, i.e. metabolic engineering, e.g. nicotine, caffeine involving modified lipid metabolism, e.g. seed oil composition
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
    • Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
    • Y02A40/146—Genetically Modified [GMO] plants, e.g. transgenic plants

Definitions

  • the present invention in some embodiments thereof, relates to novel aquaporin polynucleotides and polypeptides, and more particularly, but not exclusively, to methods of using same for increasing abiotic stress tolerance, water use efficiency (WUE), fertilizer use efficiency (FUE), biomass, vigor and/or yield of a plant.
  • WUE water use efficiency
  • FUE fertilizer use efficiency
  • biomass vigor and/or yield of a plant.
  • Abiotic stress conditions such as salinity, drought, flood, suboptimal temperature and toxic chemical pollution, cause substantial damage to agricultural plants. Most plants have evolved strategies to protect themselves against these conditions. However, if the severity and duration of the stress conditions are too great, the effects on plant development, growth and yield of most crop plants are profound. Furthermore, most of the crop plants are highly susceptible to abiotic stress (ABS) and thus necessitate optimal growth conditions for commercial crop yields. Continuous exposure to stress causes major alterations in the plant metabolism which ultimately leads to cell death and consequently yield losses.
  • ABS abiotic stress
  • Aquaporins the water channel proteins, are involved in transport of water through the membranes, maintenance of cell water balance and homeostasis under changing environmental and developmental conditions [Maurel C. Plant aquaporins: Novel functions and regulation properties. FEBS Lett. 2007, 581(12):2227-36]. These proteins are considered to be the main passage enabling transport of water and small neutral solutes such as urea and CO 2 through the membrane [Maurel C. Plant aquaporins: Novel functions and regulation properties. FEBS Lett. 2007 Jun. 12; 581(12):2227-36].
  • AQPs are present as four subfamilies of intrinsic proteins: plasma membrane (PIP), tonoplast (TIP), small and basic (SIP) and NOD26-like (NIP).
  • PIP plasma membrane
  • TIP tonoplast
  • SIP small and basic
  • NIP NOD26-like
  • 35 AQP genes have been identified in the Arabidopsis genome [Quigley F, et al., “From genome to function: the Arabidopsis aquaporins”. Genome Biol.
  • WO2004/104162 to the present inventors teaches polynucleotide sequences and methods of utilizing same for increasing the tolerance of a plant to abiotic stresses and/or increasing the biomass of a plant.
  • WO2007/020638 to the present inventors teaches polynucleotide sequences and methods of utilizing same for increasing the tolerance of a plant to abiotic stresses and/or increasing the biomass, vigor and/or yield of a plant.
  • a method of increasing abiotic stress tolerance of a plant comprising expressing within the plant an exogenous polynucleotide encoding a polypeptide comprising an amino acid sequence at least 80% homologous to the amino acid sequence selected from the group consisting of SEQ ID NOs: 33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065 and 3067-3259, thereby increasing the abiotic stress tolerance of the plant.
  • a method of increasing water use efficiency (WUE), fertilizer use efficiency (FUE), biomass, vigor and/or yield of a plant comprising expressing within the plant an exogenous polynucleotide encoding a polypeptide comprising an amino acid sequence at least 80% homologous to the amino acid sequence selected from the group consisting of SEQ ID NOs:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065 and 3067-3259, thereby increasing the water use efficiency (WUE), the fertilizer use efficiency (FUE), the biomass, the vigor and/or the yield of the plant.
  • WUE water use efficiency
  • FUE fertilizer use efficiency
  • an isolated polynucleotide comprising a nucleic acid sequence at least 80% identical to the nucleic acid sequence selected from the group consisting of SEQ ID NOs:7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2764, 2843-2857 and 2859-3051.
  • nucleic acid construct comprising the isolated polynucleotide of the invention and a promoter for directing transcription of the nucleic acid sequence.
  • an isolated polypeptide comprising an amino acid sequence at least 80% homologous to the amino acid sequence selected from the group consisting of SEQ ID NOs:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065 and 3067-3259.
  • a plant cell comprising an exogenous polypeptide having an amino acid sequence at least 80% homologous to the amino acid sequence selected from the group consisting of SEQ ID NOs:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065 and 3067-3259.
  • a plant cell comprising an exogenous polynucleotide comprising a nucleic acid sequence at least 80% homologous to the nucleic acid sequence selected from the group consisting of SEQ ID NOs:7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2764, 2843-2857 and 2859-3051.
  • a method of increasing abiotic stress tolerance of a plant comprising expressing within the plant an exogenous polynucleotide encoding a polypeptide comprising the amino acid sequence set forth by SEQ ID NO:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065, 3067-3258 or 3259, thereby increasing the abiotic stress tolerance of the plant.
  • a method of increasing water use efficiency (WUE), fertilizer use efficiency (FUE), biomass, vigor and/or yield of a plant comprising expressing within the plant an exogenous polynucleotide encoding a polypeptide comprising the amino acid sequence set forth by SEQ ID NO:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065, 3067-3258 or 3259, thereby increasing the water use efficiency (WUE), the fertilizer use efficiency (FUE), the biomass, the vigor and/or the yield of the plant.
  • WUE water use efficiency
  • FUE fertilizer use efficiency
  • an isolated polynucleotide comprising the nucleic acid sequence set forth by SEQ ID NO:7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2764, 2843-2857, 2859-3050 or 3051.
  • nucleic acid construct comprising the isolated polynucleotide of the invention and a promoter for directing transcription of the nucleic acid sequence.
  • an isolated polypeptide comprising the amino acid sequence set forth by SEQ ID NO:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065, 3067-3258 or 3259.
  • a plant cell comprising an exogenous polypeptide having the amino acid sequence set forth by SEQ ID NO:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065, 3067-3258 or 3259.
  • a plant cell comprising an exogenous polynucleotide comprising the nucleic acid sequence set forth by SEQ ID NO:7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2764, 2843-2857, 2859-3050 or 3051.
  • the polynucleotide is selected from the group consisting of SEQ ID NOs:7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2764, 2843-2857 and 2859-3051.
  • the amino acid sequence is selected from the group consisting of SEQ ID Nos:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065 and 3067-3259.
  • the polypeptide is selected from the group consisting of SEQ ID Nos:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065 and 3067-3259.
  • the abiotic stress is selected from the group consisting of salinity, water deprivation, low temperature, high temperature, heavy metal toxicity, anaerobiosis, nutrient deficiency, nutrient excess, atmospheric pollution and UV irradiation.
  • the method further comprising growing the plant expressing the exogenous polynucleotide under the abiotic stress.
  • the promoter is a constitutive promoter.
  • the plant cell forms a part of a plant.
  • FIG. 1 is a schematic illustration of the pGI binary plasmid used for expressing the isolated polynucleotide sequences of the invention.
  • RB T-DNA right border
  • LB T-DNA left border
  • H HindIII restriction enzyme
  • X XbaI restriction enzyme
  • B BamHI restriction enzyme
  • S SalI restriction enzyme
  • Sm SmaI restriction enzyme
  • R-I EcoRI restriction enzyme
  • Sc SacI/SstI/Ecl136II
  • NOS pro nopaline synthase promoter
  • NPT-II neomycin phosphotransferase gene
  • NOS ter nopaline synthase terminator
  • Poly-A signal polyadenylation signal
  • GUSintron the GUS reporter gene (coding sequence and intron).
  • FIGS. 2A-B are images depicting root development of plants grown in transparent agar plates. The different transgenes were grown in transparent agar plates for 10-15 days and the plates were photographed every 2-5 days starting at day 1.
  • FIG. 2 A An exemplary image of plants taken following 12 days on agar plates.
  • FIG. 2 B An exemplary image of root analysis in which the length of the root measured is represented by a red arrow.
  • FIGS. 3A-F are histograms depicting the total economic fruit yield, plant biomass and harvest index for TOM-ABST36 (black bar) vs. control (white bar) plants growing in the commercial greenhouse under a 200 mM sodium chloride (NaCl) irrigation regime ( FIG. 3A-C , respectively), or under two different water-stress regimes (WLI-1 and WLI-2; FIG. 3D-F , respectively). Yield performance was compared to plants growing under standard irrigation conditions (0 mM NaCl and WLI-0). Results are the average of the four independent events. *Significantly different at P ⁇ 0.05.
  • FIGS. 3G-J are photographs of transgenic tomato plants or control plants grown under various conditions.
  • FIG. 3 G TOM-ABST36 plants growing under regular irrigation conditions
  • FIG. 3 H control plants growing under regular irrigation conditions
  • FIG. 3 I TOM-ABST36 plants after growing under a 200-mM NaCl-irrigation regime during the entire growing season
  • FIG. 3 J control plants after growing under a 200-mM NaCl-irrigation regime during the entire growing season.
  • the present invention in some embodiments thereof, relates to novel aquaporin polynucleotides and polypeptides, and more particularly, but not exclusively, to methods of using same for increasing abiotic stress tolerance, water use efficiency, fertilizer use efficiency, biomass, vigor and/or yield of a plant.
  • AQP aquaporin
  • the present inventors have employed a bioinformatics approach which combines digital expression analysis and cross-species comparative genomics and screened 7.2 million expressed sequence tags (ESTs) from 1,195 relevant EST's libraries of both monocot and dicot plant species. Using this approach 1,114 different AQP genes have been identified and were further classified to 11 subgroups (Table 1).
  • ESTs of the TIP2 subgroup are significantly over-represented in both plants' roots and in plants exposed to abiotic stress (ABS), and that polypeptides (e.g., SEQ ID NOs: 27-28, 45-48, Table 2) encoded by polynucleotides of the TIP2 subgroup (e.g., SEQ ID NOs:1, 2, 19-22, Table 2) share a common consensus sequence TLXFXFAGVGS (SEQ ID NO:2826).
  • ABS abiotic stress
  • polypeptides or polynucleotides which affect (e.g., increase) plant metabolism, growth, reproduction and/or viability under stress can also affect the plant growth, biomass, yield and/or vigor under optimal conditions.
  • a method of increasing abiotic stress tolerance, water use efficiency, fertilizer use efficiency, growth, biomass, yield and/or vigor of a plant is effected by expressing within the plant an exogenous polynucleotide encoding a polypeptide comprising the amino acid consensus sequence TLXFXFAGVGS as set forth by SEQ ID NO:2826, wherein expression of the polypeptide promotes plants' biomass/vigor and/or yield under normal or stress conditions.
  • the polypeptide's activity is structurally associated with the integrity of the above consensus sequence (SEQ ID NO:2826).
  • the activity is a water channel activity which typically resides in the vacuaolar membrane (tonoplast) and/or the plasma membrane of the plant cell and enables the transport of water and/or small neutral solutes such as urea, nitrates and carbon dioxide (CO 2 ) through the membrane.
  • abiotic stress refers to any adverse effect on metabolism, growth, reproduction and/or viability of a plant. Accordingly, abiotic stress can be induced by suboptimal environmental growth conditions such as, for example, salinity, water deprivation, flooding, freezing, low or high temperature, heavy metal toxicity, anaerobiosis, nutrient deficiency, atmospheric pollution or UV irradiation.
  • suboptimal environmental growth conditions such as, for example, salinity, water deprivation, flooding, freezing, low or high temperature, heavy metal toxicity, anaerobiosis, nutrient deficiency, atmospheric pollution or UV irradiation.
  • abiotic stress tolerance refers to the ability of a plant to endure an abiotic stress without suffering a substantial alteration in metabolism, growth, productivity and/or viability.
  • water use efficiency refers to the level of organic matter produced per unit of water consumed by the plant, i.e., the dry weight of a plant in relation to the plant's water use, e.g., the biomass produced per unit transpiration.
  • fertilizer use efficiency refers to the uptake, spread, absorbent, accumulation, relocation (within the plant) and use of one or more of the minerals and organic moieties absorbed from the soil, such as nitrogen, phosphates and/or potassium.
  • plant biomass refers to the amount (measured in grams of air-dry tissue) of a tissue produced from the plant in a growing season, which could also determine or affect the plant yield or the yield per growing area.
  • plant yield refers to the amount (as determined by weight/size) or quantity (numbers) of tissue produced per plant or per growing season. Hence increased yield could affect the economic benefit one can obtain from the plant in a certain growing area and/or growing time.
  • plant vigor refers to the amount (measured by weight) of tissue produced by the plant in a given time. Hence increase vigor could determine or affect the plant yield or the yield per growing time or growing area.
  • the term “increasing” refers to at least about 2%, at least about 3%, at least about 4%, at least about 5%, at least about 10%, at least about 15%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, increase in plant abiotic stress tolerance, water use efficiency, fertilizer use efficiency, growth, biomass, yield and/or vigor as compared to a native plant [i.e., a plant not modified with the biomolecules (polynucleotide or polypeptides) of the invention, e.g., a non-transformed plant of the same species which is grown under the same growth conditions).
  • a native plant i.e., a plant not modified with the biomolecules (polynucleotide or polypeptides) of the invention, e.g., a non-transformed plant of the same species which is grown under the same growth conditions.
  • exogenous polynucleotide refers to a heterologous nucleic acid sequence which may not be naturally expressed within the plant or which overexpression in the plant is desired.
  • the exogenous polynucleotide may be introduced into the plant in a stable or transient manner, so as to produce a ribonucleic acid (RNA) molecule and/or a polypeptide molecule.
  • RNA ribonucleic acid
  • exogenous polynucleotide may comprise a nucleic acid sequence which is identical or partially homologous to an endogenous nucleic acid sequence of the plant.
  • the exogenous polynucleotide of the invention encodes a polypeptide having an amino acid sequence at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more say 100% homologous to the amino acid sequence selected from the group consisting of SEQ ID NOs: 27-28, 45-48, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561, 2449-2450, 2453-2458, 2460-24
  • Homology can be determined using any homology comparison software, including for example, the BlastP or TBLASTN software of the National Center of Biotechnology Information (NCBI) such as by using default parameters, when starting from a polypeptide sequence; or the tBLASTX algorithm (available via the NCBI) such as by using default parameters, which compares the six-frame conceptual translation products of a nucleotide query sequence (both strands) against a protein sequence database.
  • NCBI National Center of Biotechnology Information
  • Homologous sequences include both orthologous and paralogous sequences.
  • paralogous relates to gene-duplications within the genome of a species leading to paralogous genes.
  • orthologous relates to homologous genes in different organisms due to ancestral relationship.
  • One option to identify orthologues in monocot plant species is by performing a reciprocal blast search. This may be done by a first blast involving blasting the sequence-of-interest against any sequence database, such as the publicly available NCBI database which may be found at: Hypertext Transfer Protocol://World Wide Web (dot) ncbi (dot) nlm (dot) nih (dot) gov. If orthologues in rice were sought, the sequence-of-interest would be blasted against, for example, the 28,469 full-length cDNA clones from Oryza sativa Nipponbare available at NCBI. The blast results may be filtered.
  • the ClustalW program may be used [Hypertext Transfer Protocol://World Wide Web (dot) ebi (dot) ac (dot) uk/Tools/clustalw2/index (dot) html], followed by a neighbor-joining tree (Hypertext Transfer Protocol://en (dot) wikipedia (dot) org/wiki/Neighbor-joining) which helps visualizing the clustering.
  • the exogenous polynucleotide encodes a polypeptide consisting of the amino acid sequence set forth by SEQ ID NO:27-28, 45-48, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561, 2449-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2484 or 2765.
  • the exogenous polynucleotide comprises a nucleic acid sequence which is at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, e.g., 100% identical to the nucleic acid sequence selected from the group consisting of SEQ ID NOs:1, 2, 19, 20-22, 53-55, 57-87, 89-141, 143-147, 149-195, 197-206, 208-212, 214, 1102-1103, 1106-1111
  • Identity e.g., percent homology
  • NCBI National Center of Biotechnology Information
  • the exogenous polynucleotide is set forth by SEQ ID NO:1, 2, 19, 20-22, 53-55, 57-87, 89-141, 143-147, 149-195, 197-206, 208-212, 214, 1102-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 2751-2752, 2748-2749, or 2750.
  • the exogenous polynucleotide encodes a polypeptide having an amino acid sequence at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, e.g., 100% homologous to SEQ ID NO:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769
  • the exogenous polynucleotide encodes a polypeptide consisting of the amino acid sequence set forth by SEQ ID NO:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065, 3067-3258 or 3259.
  • the exogenous polynucleotide does not encode a polypeptide having the amino acid sequence selected from the group consisting of SEQ ID NOs: 1828, 1867, 1404, 1436, 1495, 1543, 1554, 1560, 2451, 2452, 2459, 2464, 2482, 2484 and 3066.
  • the exogenous polynucleotide is at least at least about 60%, least at least about 65%, least at least about 70%, least at least about 75%least at least about 80%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, e.g., 100% identical to SEQ ID NO:7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2764, 2843-2857, 2859-3050 or
  • the polynucleotide is set forth by SEQ ID NO:7, 8, 4, 1-3, 5, 6, 9-26, 53-55, 57-87, 89-147, 149-195, 197-206, 208-212, 214-480, 482-519, 521-1103, 1106-1111, 1113-1116, 1118-1134, 1136, 1138-1400, 2748-2764, 2843-2857, 2859-3050 or 3051.
  • the exogenous polynucleotide is not the polynucleotide set forth by SEQ ID NO: 481, 520, 56, 88, 148, 196, 207, 213, 1104, 1105, 1112, 1117, 1135, 1137 or 2858.
  • polynucleotide refers to a single or double stranded nucleic acid sequence which is isolated and provided in the form of an RNA sequence, a complementary polynucleotide sequence (cDNA), a genomic polynucleotide sequence and/or a composite polynucleotide sequences (e.g., a combination of the above).
  • complementary polynucleotide sequence refers to a sequence, which results from reverse transcription of messenger RNA using a reverse transcriptase or any other RNA dependent DNA polymerase. Such a sequence can be subsequently amplified in vivo or in vitro using a DNA dependent DNA polymerase.
  • genomic polynucleotide sequence refers to a sequence derived (isolated) from a chromosome and thus it represents a contiguous portion of a chromosome.
  • composite polynucleotide sequence refers to a sequence, which is at least partially complementary and at least partially genomic.
  • a composite sequence can include some exonal sequences required to encode the polypeptide of the present invention, as well as some intronic sequences interposing therebetween.
  • the intronic sequences can be of any source, including of other genes, and typically will include conserved splicing signal sequences. Such intronic sequences may further include cis acting expression regulatory elements.
  • the polynucleotide of the invention comprises no more than 5000 nucleic acids in length. According to some embodiments of the invention, the polynucleotide of the invention comprises no more than 4000 nucleic acids in length, e.g., no more than 3000 nucleic acids, e.g., no more than 2500 nucleic acids.
  • Nucleic acid sequences encoding the polypeptides of the present invention may be optimized for expression.
  • a non-limiting example of an optimized nucleic acid sequence is provided in SEQ ID NO:2751, which encodes an optimized polypeptide comprising the amino acid sequence set forth by SEQ ID NO:27.
  • sequence modifications include, but are not limited to, an altered G/C content to more closely approach that typically found in the plant species of interest, and the removal of codons atypically found in the plant species commonly referred to as codon optimization.
  • an optimized gene or nucleic acid sequence refers to a gene in which the nucleotide sequence of a native or naturally occurring gene has been modified in order to utilize statistically-preferred or statistically-favored codons within the plant.
  • the nucleotide sequence typically is examined at the DNA level and the coding region optimized for expression in the plant species determined using any suitable procedure, for example as described in Sardana et al. (1996, Plant Cell Reports 15:677-681).
  • the standard deviation of codon usage may be calculated by first finding the squared proportional deviation of usage of each codon of the native gene relative to that of highly expressed plant genes, followed by a calculation of the average squared deviation.
  • a Table of codon usage from highly expressed genes of dicotyledonous plants is compiled using the data of Murray et al. (1989, Nuc Acids Res. 17:477-498).
  • Codon Usage Database contains codon usage tables for a number of different species, with each codon usage Table having been statistically determined based on the data present in Genbank.
  • a naturally-occurring nucleotide sequence encoding a protein of interest can be codon optimized for that particular plant species. This is effected by replacing codons that may have a low statistical incidence in the particular species genome with corresponding codons, in regard to an amino acid, that are statistically more favored.
  • one or more less-favored codons may be selected to delete existing restriction sites, to create new ones at potentially useful junctions (5′ and 3′ ends to add signal peptide or termination cassettes, internal sites that might be used to cut and splice segments together to produce a correct full-length sequence), or to eliminate nucleotide sequences that may negatively effect mRNA stability or expression.
  • codon optimization of the native nucleotide sequence may comprise determining which codons, within the native nucleotide sequence, are not statistically-favored with regards to a particular plant, and modifying these codons in accordance with a codon usage table of the particular plant to produce a codon optimized derivative.
  • a modified nucleotide sequence may be fully or partially optimized for plant codon usage provided that the protein encoded by the modified nucleotide sequence is produced at a level higher than the protein encoded by the corresponding naturally occurring or native gene. Construction of synthetic genes by altering the codon usage is described in for example PCT Patent Application 93/07278.
  • the invention encompasses nucleic acid sequences described hereinabove; fragments thereof, sequences hybridizable therewith, sequences homologous thereto, sequences encoding similar polypeptides with different codon usage, altered sequences characterized by mutations, such as deletion, insertion or substitution of one or more nucleotides, either naturally occurring or man induced, either randomly or in a targeted fashion.
  • the invention provides an isolated polypeptide having an amino acid sequence at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more say 100% homologous to an amino acid sequence selected from the group consisting of SEQ ID NO: 27-28, 45-48, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561, 2449-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2484 and 2765
  • the invention provides an isolated polypeptide having an amino acid sequence at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 81%, at least about 82%, at least about 83%, at least about 84%, at least about 85%, at least about 86%, at least about 87%, at least about 88%, at least about 89%, at least about 90%, at least about 91%, at least about 92%, at least about 93%, at least about 93%, at least about 94%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, at least about 99%, or more say 100% homologous to an amino acid sequence selected from the group consisting of SEQ ID NOs:33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450,
  • the polypeptide is set forth by SEQ ID NO: 33, 34, 30, 27-29, 31, 32, 35-52, 1401-1403, 1405-1435, 1437-1494, 1496-1542, 1544-1553, 1555-1559, 1561-1827, 1829-1866, 1868-2450, 2453-2458, 2460-2463, 2465-2481, 2483, 2485-2746, 2765-2769, 3052-3065, 3067-3258 or 3259.
  • polypeptide is not the polypeptide set forth by SEQ ID NO: 1828, 1867, 1404, 1436, 1495, 1543, 1554, 1560, 2451, 2452, 2459, 2464, 2482, 2484 or 3066.
  • the invention also encompasses fragments of the above described polypeptides and polypeptides having mutations, such as deletions, insertions or substitutions of one or more amino acids, either naturally occurring or man induced, either randomly or in a targeted fashion.
  • plant encompasses whole plants, ancestors and progeny of the plants and plant parts, including seeds, shoots, stems, roots (including tubers), and plant cells, tissues and organs.
  • the plant may be in any form including suspension cultures, embryos, meristematic regions, callus tissue, leaves, gametophytes, sporophytes, pollen, and microspores.
  • Plants that are particularly useful in the methods of the invention include all plants which belong to the superfamily Viridiplantae, in particular monocotyledonous and dicotyledonous plants including a fodder or forage legume, ornamental plant, food crop, tree, or shrub selected from the list comprising Acacia spp., Acer spp., Actinidia spp., Aesculus spp., Agathis australis, Albizia amara, Alsophila tricolor, Andropogon spp., Arachis spp, Areca catechu, Astelia fragrans, Astragalus cicer, Baikiaea plurijuga, Betula spp., Brassica spp., Bruguiera gymnorrhiza, Burkea africana, Butea frondosa, Cadaba farinosa, Calliandra spp, Camellia sinensis, Canna indica, Capsicum spp., Cassia spp., Centroe
  • the plant used by the method of the invention is a crop plant such as rice, maize, wheat, barley, peanut, potato, sesame, olive tree, palm oil, banana, soybean, sunflower, canola, sugarcane, alfalfa, millet, leguminosae (bean, pea), flax, lupinus, rapeseed, tobacco, poplar and cotton.
  • a crop plant such as rice, maize, wheat, barley, peanut, potato, sesame, olive tree, palm oil, banana, soybean, sunflower, canola, sugarcane, alfalfa, millet, leguminosae (bean, pea), flax, lupinus, rapeseed, tobacco, poplar and cotton.
  • Expressing the exogenous polynucleotide of the invention within the plant can be effected by transforming one or more cells of the plant with the exogenous polynucleotide, followed by generating a mature plant from the transformed cells and cultivating the mature plant under conditions suitable for expressing the exogenous polynucleotide within the mature plant.
  • the transformation is effected by introducing to the plant cell a nucleic acid construct which includes the exogenous polynucleotide of some embodiments of the invention and at least one promoter capable of directing transcription of the exogenous polynucleotide in the plant cell. Further details of suitable transformation approaches are provided hereinbelow.
  • promoter refers to a region of DNA which lies upstream of the transcriptional initiation site of a gene to which RNA polymerase binds to initiate transcription of RNA.
  • the promoter controls where (e.g., which portion of a plant) and/or when (e.g., at which stage or condition in the lifetime of an organism) the gene is expressed.
  • any suitable promoter sequence can be used by the nucleic acid construct of the present invention.
  • the promoter is a constitutive promoter, a tissue-specific, or an abiotic stress-inducible promoter.
  • Suitable constitutive promoters include, for example, CaMV 35S promoter (SEQ ID NO:2825; Odell et al., Nature 313:810-812, 1985); Arabidopsis At6669 promoter (SEQ ID NO:2823; see PCT Publication No. WO04081173A2); maize Ubi 1 (Christensen et al., Plant Sol. Biol. 18:675-689, 1992); rice actin (McElroy et al., Plant Cell 2:163-171, 1990); pEMU (Last et al., Theor. Appl. Genet. 81:581-588, 1991); CaMV 19S (Nilsson et al., Physiol.
  • tissue-specific promoters include, but not limited to, leaf-specific promoters [such as described, for example, by Yamamoto et al., Plant J. 12:255-265, 1997; Kwon et al., Plant Physiol. 105:357-67, 1994; Yamamoto et al., Plant Cell Physiol. 35:773-778, 1994; Gotor et al., Plant J. 3:509-18, 1993; Orozco et al., Plant Mol. Biol. 23:1129-1138, 1993; and Matsuoka et al., Proc. Natl. Acad. Sci.
  • seed-preferred promoters e.g., from seed specific genes (Simon, et al., Plant Mol. Biol. 5. 191, 1985; Scofield, et al., J. Biol. Chem. 262: 12202, 1987; Baszczynski, et al., Plant Mol. Biol. 14: 633, 1990), Brazil Nut albumin (Pearson' et al., Plant Mol. Biol. 18: 235-245, 1992), legumin (Ellis, et al. Plant Mol. Biol. 10: 203-214, 1988), Glutelin (rice) (Takaiwa, et al., Mol. Gen. Genet.
  • endosperm specific promoters e.g., wheat LMW and HMW, glutenin-1 (Mol Gen Genet 216:81-90, 1989; NAR 17:461-2), wheat a, b and g gliadins (EMBO3:1409-15, 1984), Barley ltrl promoter, barley B1, C, D hordein (Theor Appl Gen 98:1253-62, 1999; Plant J 4:343-55, 1993; Mol Gen Genet 250:750-60, 1996), Barley DOF (Mena et al, The Plant Journal, 116(1): 53-62, 1998), Biz2 (EP99106056.7), Synthetic promoter (Vicente-Carbajosa et al., Plant J.
  • Suitable abiotic stress-inducible promoters include, but not limited to, salt-inducible promoters such as RD29A (Yamaguchi-Shinozalei et al., Mol. Gen. Genet. 236:331-340, 1993); drought-inducible promoters such as maize rab17 gene promoter (Pla et. al., Plant Mol. Biol. 21:259-266, 1993), maize rab28 gene promoter (Busk et. al., Plant J. 11:1285-1295, 1997) and maize Ivr2 gene promoter (Pelleschi et. al., Plant Mol. Biol. 39:373-380, 1999); heat-inducible promoters such as heat tomato hsp80-promoter from tomato (U.S. Pat. No. 5,187,267).
  • salt-inducible promoters such as RD29A (Yamaguchi-Shinozalei et al., Mol. Gen. Genet
  • the nucleic acid construct of some embodiments of the invention can further include an appropriate selectable marker and/or an origin of replication.
  • the nucleic acid construct utilized is a shuttle vector, which can propagate both in E. coli (wherein the construct comprises an appropriate selectable marker and origin of replication) and be compatible with propagation in cells.
  • the construct according to the present invention can be, for example, a plasmid, a bacmid, a phagemid, a cosmid, a phage, a virus or an artificial chromosome.
  • the nucleic acid construct of some embodiments of the invention can be utilized to stably or transiently transform plant cells.
  • stable transformation the exogenous polynucleotide is integrated into the plant genome and as such it represents a stable and inherited trait.
  • transient transformation the exogenous polynucleotide is expressed by the cell transformed but it is not integrated into the genome and as such it represents a transient trait.
  • the Agrobacterium system includes the use of plasmid vectors that contain defined DNA segments that integrate into the plant genomic DNA. Methods of inoculation of the plant tissue vary depending upon the plant species and the Agrobacterium delivery system. A widely used approach is the leaf disc procedure which can be performed with any tissue explant that provides a good source for initiation of whole plant differentiation. See, e.g., Horsch et al. in Plant Molecular Biology Manual A5, Kluwer Academic Publishers, Dordrecht (1988) p. 1-9. A supplementary approach employs the Agrobacterium delivery system in combination with vacuum infiltration. The Agrobacterium system is especially viable in the creation of transgenic dicotyledonous plants.
  • DNA transfer into plant cells There are various methods of direct DNA transfer into plant cells.
  • electroporation the protoplasts are briefly exposed to a strong electric field.
  • microinjection the DNA is mechanically injected directly into the cells using very small micropipettes.
  • microparticle bombardment the DNA is adsorbed on microprojectiles such as magnesium sulfate crystals or tungsten particles, and the microprojectiles are physically accelerated into cells or plant tissues.
  • Micropropagation is a process of growing new generation plants from a single piece of tissue that has been excised from a selected parent plant or cultivar. This process permits the mass reproduction of plants having the preferred tissue expressing the fusion protein.
  • the new generation plants which are produced are genetically identical to, and have all of the characteristics of, the original plant.
  • Micropropagation allows mass production of quality plant material in a short period of time and offers a rapid multiplication of selected cultivars in the preservation of the characteristics of the original transgenic or transformed plant.
  • the advantages of cloning plants are the speed of plant multiplication and the quality and uniformity of plants produced.
  • Micropropagation is a multi-stage procedure that requires alteration of culture medium or growth conditions between stages.
  • the micropropagation process involves four basic stages: Stage one, initial tissue culturing; stage two, tissue culture multiplication; stage three, differentiation and plant formation; and stage four, greenhouse culturing and hardening.
  • stage one initial tissue culturing
  • stage two tissue culture multiplication
  • stage three differentiation and plant formation
  • stage four greenhouse culturing and hardening.
  • stage one initial tissue culturing
  • the tissue culture is established and certified contaminant-free.
  • stage two the initial tissue culture is multiplied until a sufficient number of tissue samples are produced to meet production goals.
  • stage three the tissue samples grown in stage two are divided and grown into individual plantlets.
  • the transformed plantlets are transferred to a greenhouse for hardening where the plants' tolerance to light is gradually increased so that it can be grown in the natural environment.
  • the transgenic plants are generated by transient transformation of leaf cells, meristematic cells or the whole plant.
  • Transient transformation can be effected by any of the direct DNA transfer methods described above or by viral infection using modified plant viruses.
  • Viruses that have been shown to be useful for the transformation of plant hosts include CaMV, Tobacco mosaic virus (TMV), brome mosaic virus (BMV) and Bean Common Mosaic Virus (BV or BCMV). Transformation of plants using plant viruses is described in U.S. Pat. No. 4,855,237 (bean golden mosaic virus; BGV), EP-A 67,553 (TMV), Japanese Published Application No. 63-14693 (TMV), EPA 194,809 (BV), EPA 278,667 (BV); and Gluzman, Y. et al., Communications in Molecular Biology: Viral Vectors, Cold Spring Harbor Laboratory, New York, pp. 172-189 (1988). Pseudovirus particles for use in expressing foreign DNA in many hosts, including plants are described in WO 87/06261.
  • the virus used for transient transformations is avirulent and thus is incapable of causing severe symptoms such as reduced growth rate, mosaic, ring spots, leaf roll, yellowing, streaking, pox formation, tumor formation and pitting.
  • a suitable avirulent virus may be a naturally occurring avirulent virus or an artificially attenuated virus.
  • Virus attenuation may be effected by using methods well known in the art including, but not limited to, sub-lethal heating, chemical treatment or by directed mutagenesis techniques such as described, for example, by Kurihara and Watanabe (Molecular Plant Pathology 4:259-269, 2003), Gal-on et al. (1992), Atreya et al. (1992) and Huet et al. (1994).
  • Suitable virus strains can be obtained from available sources such as, for example, the American Type culture Collection (ATCC) or by isolation from infected plants. Isolation of viruses from infected plant tissues can be effected by techniques well known in the art such as described, for example by Foster and Tatlor, Eds. “Plant Virology Protocols: From Virus Isolation to Transgenic Resistance (Methods in Molecular Biology (Humana Pr), Vol 81)”, Humana Press, 1998. Briefly, tissues of an infected plant believed to contain a high concentration of a suitable virus, preferably young leaves and flower petals, are ground in a buffer solution (e.g., phosphate buffer solution) to produce a virus infected sap which can be used in subsequent inoculations.
  • a buffer solution e.g., phosphate buffer solution
  • the virus When the virus is a DNA virus, suitable modifications can be made to the virus itself. Alternatively, the virus can first be cloned into a bacterial plasmid for ease of constructing the desired viral vector with the foreign DNA. The virus can then be excised from the plasmid. If the virus is a DNA virus, a bacterial origin of replication can be attached to the viral DNA, which is then replicated by the bacteria. Transcription and translation of this DNA will produce the coat protein which will encapsidate the viral DNA. If the virus is an RNA virus, the virus is generally cloned as a cDNA and inserted into a plasmid. The plasmid is then used to make all of the constructions. The RNA virus is then produced by transcribing the viral sequence of the plasmid and translation of the viral genes to produce the coat protein(s) which encapsidate the viral RNA.
  • a plant viral polynucleotide in which the native coat protein coding sequence has been deleted from a viral polynucleotide, a non-native plant viral coat protein coding sequence and a non-native promoter, preferably the subgenomic promoter of the non-native coat protein coding sequence, capable of expression in the plant host, packaging of the recombinant plant viral polynucleotide, and ensuring a systemic infection of the host by the recombinant plant viral polynucleotide, has been inserted.
  • the coat protein gene may be inactivated by insertion of the non-native polynucleotide sequence within it, such that a protein is produced.
  • the recombinant plant viral polynucleotide may contain one or more additional non-native subgenomic promoters.
  • Each non-native subgenomic promoter is capable of transcribing or expressing adjacent genes or polynucleotide sequences in the plant host and incapable of recombination with each other and with native subgenomic promoters.
  • Non-native (foreign) polynucleotide sequences may be inserted adjacent the native plant viral subgenomic promoter or the native and a non-native plant viral subgenomic promoters if more than one polynucleotide sequence is included.
  • the non-native polynucleotide sequences are transcribed or expressed in the host plant under control of the subgenomic promoter to produce the desired products.
  • a recombinant plant viral polynucleotide is provided as in the first embodiment except that the native coat protein coding sequence is placed adjacent one of the non-native coat protein subgenomic promoters instead of a non-native coat protein coding sequence.
  • a recombinant plant viral polynucleotide in which the native coat protein gene is adjacent its subgenomic promoter and one or more non-native subgenomic promoters have been inserted into the viral polynucleotide.
  • the inserted non-native subgenomic promoters are capable of transcribing or expressing adjacent genes in a plant host and are incapable of recombination with each other and with native subgenomic promoters.
  • Non-native polynucleotide sequences may be inserted adjacent the non-native subgenomic plant viral promoters such that the sequences are transcribed or expressed in the host plant under control of the subgenomic promoters to produce the desired product.
  • a recombinant plant viral polynucleotide is provided as in the third embodiment except that the native coat protein coding sequence is replaced by a non-native coat protein coding sequence.
  • the viral vectors are encapsidated by the coat proteins encoded by the recombinant plant viral polynucleotide to produce a recombinant plant virus.
  • the recombinant plant viral polynucleotide or recombinant plant virus is used to infect appropriate host plants.
  • the recombinant plant viral polynucleotide is capable of replication in the host, systemic spread in the host, and transcription or expression of foreign gene(s) (exogenous polynucleotide) in the host to produce the desired protein.
  • polynucleotide of the present invention can also be introduced into a chloroplast genome thereby enabling chloroplast expression.
  • a technique for introducing exogenous polynucleotide sequences to the genome of the chloroplasts involves the following procedures. First, plant cells are chemically treated so as to reduce the number of chloroplasts per cell to about one. Then, the exogenous polynucleotide is introduced via particle bombardment into the cells with the aim of introducing at least one exogenous polynucleotide molecule into the chloroplasts.
  • the exogenous polynucleotides selected such that it is integratable into the chloroplast's genome via homologous recombination which is readily effected by enzymes inherent to the chloroplast.
  • the exogenous polynucleotide includes, in addition to a gene of interest, at least one polynucleotide stretch which is derived from the chloroplast's genome.
  • the exogenous polynucleotide includes a selectable marker, which serves by sequential selection procedures to ascertain that all or substantially all of the copies of the chloroplast genomes following such selection will include the exogenous polynucleotide. Further details relating to this technique are found in U.S. Pat. Nos. 4,945,050; and 5,693,507 which are incorporated herein by reference.
  • a polypeptide can thus be produced by the protein expression system of the chloroplast and become integrated into the chloroplast's inner membrane.
  • the present invention also envisages expressing a plurality of exogenous polynucleotides in a single host plant to thereby achieve superior effect on abiotic stress tolerance, water use efficiency, fertilizer use efficiency, growth, biomass, yield and/or vigor.
  • Expressing a plurality of exogenous polynucleotides in a single host plant can be effected by co-introducing multiple nucleic acid constructs, each including a different exogenous polynucleotide, into a single plant cell.
  • the transformed cell can than be regenerated into a mature plant using the methods described hereinabove.
  • expressing a plurality of exogenous polynucleotides in a single host plant can be effected by co-introducing into a single plant-cell a single nucleic-acid construct including a plurality of different exogenous polynucleotides.
  • a construct can be designed with a single promoter sequence which can transcribe a polycistronic messenger RNA including all the different exogenous polynucleotide sequences.
  • the polynucleotide sequences can be inter-linked via an internal ribosome entry site (IRES) sequence which facilitates translation of polynucleotide sequences positioned downstream of the IRES sequence.
  • IRES internal ribosome entry site
  • a transcribed polycistronic RNA molecule encoding the different polypeptides described above will be translated from both the capped 5′ end and the two internal IRES sequences of the polycistronic RNA molecule to thereby produce in the cell all different polypeptides.
  • the construct can include several promoter sequences each linked to a different exogenous polynucleotide sequence.
  • the plant cell transformed with the construct including a plurality of different exogenous polynucleotides can be regenerated into a mature plant, using the methods described hereinabove.
  • expressing a plurality of exogenous polynucleotides in a single host plant can be effected by introducing different nucleic acid constructs, including different exogenous polynucleotides, into a plurality of plants.
  • the regenerated transformed plants can then be cross-bred and resultant progeny selected for superior abiotic stress tolerance, water use efficiency, fertilizer use efficiency, growth, biomass, yield and/or vigor traits, using conventional plant breeding techniques.
  • the invention encompasses plants exogenously expressing (as described above) the polynucleotide(s) and/or polypeptide(s) of the invention.
  • the level of the polypeptide encoded by the exogenous polynucleotide can be determined by methods well known in the art such as, activity assays, Western blots using antibodies capable of specifically binding the polypeptide, Enzyme-Linked ImmunoSorbent Assay (ELISA), radio-immuno-assays (RIA), immunohistochemistry, immunocytochemistry, immunofluorescence and the like.
  • RNA-in situ hybridization Methods of determining the level in the plant of the RNA transcribed from the exogenous polynucleotide are well known in the art and include, for example, Northern blot analysis, reverse transcription polymerase chain reaction (RT-PCR) analysis (including quantitative, semi-quantitative or real-time RT-PCR) and RNA-in situ hybridization.
  • RT-PCR reverse transcription polymerase chain reaction
  • the polypeptide according to some embodiments of the invention functions as a water channel.
  • the invention according to some embodiments encompasses functional equivalents of the polypeptide (e.g., polypeptides capable of the biological activity of a water channel) which can be identified by functional assays (e.g., being capable of transporting water in a plant) using e.g., a cell-swelling assay (Meng, Q. X. et al. 2008. Cell Physiol Biochem, 21. pp. 123-128).
  • polynucleotides and polypeptides described hereinabove can be used in a wide range of economical plants, in a safe and cost effective manner.
  • transgene the exogenous polynucleotide encoding the polypeptide
  • water use efficiency the exogenous polynucleotide encoding the polypeptide
  • fertilizer use efficiency the growth, biomass, yield and/or vigor
  • Abiotic stress tolerance Transformed (i.e., expressing the transgene) and non-transformed (wild type) plants are exposed to an abiotic stress condition, such as water deprivation, suboptimal temperature (low temperature, high temperature), nutrient deficiency, nutrient excess, a salt stress condition, osmotic stress, heavy metal toxicity, anaerobiosis, atmospheric pollution and UV irradiation.
  • an abiotic stress condition such as water deprivation, suboptimal temperature (low temperature, high temperature), nutrient deficiency, nutrient excess, a salt stress condition, osmotic stress, heavy metal toxicity, anaerobiosis, atmospheric pollution and UV irradiation.
  • Salinity tolerance assay Transgenic plants with tolerance to high salt concentrations are expected to exhibit better germination, seedling vigor or growth in high salt.
  • Salt stress can be effected in many ways such as, for example, by irrigating the plants with a hyperosmotic solution, by cultivating the plants hydroponically in a hyperosmotic growth solution (e.g., Hoagland solution), or by culturing the plants in a hyperosmotic growth medium [e.g., 50% Murashige-Skoog medium (MS medium)].
  • a hyperosmotic growth medium e.g. 50% Murashige-Skoog medium (MS medium)
  • the salt concentration in the irrigation water, growth solution, or growth medium can be adjusted according to the specific characteristics of the specific plant cultivar or variety, so as to inflict a mild or moderate effect on the physiology and/or morphology of the plants (for guidelines as to appropriate concentration see, Bernstein and Kafkafi, Root Growth Under Salinity Stress In: Plant Roots, The Hidden Half 3rd ed. Waisel Y, Eshel A and Kafkafi U. (editors) Marcel Dekker Inc., New York, 2002, and reference therein).
  • a salinity tolerance test can be performed by irrigating plants at different developmental stages with increasing concentrations of sodium chloride (for example 50 mM, 100 mM, 200 mM, 400 mM NaCl) applied from the bottom and from above to ensure even dispersal of salt. Following exposure to the stress condition the plants are frequently monitored until substantial physiological and/or morphological effects appear in wild type plants. Thus, the external phenotypic appearance, degree of wilting and overall success to reach maturity and yield progeny are compared between control and transgenic plants. Quantitative parameters of tolerance measured include, but are not limited to, the average wet and dry weight, the weight of the seeds yielded, the average seed size and the number of seeds produced per plant. Transformed plants not exhibiting substantial physiological and/or morphological effects, or exhibiting higher biomass than wild-type plants, are identified as abiotic stress tolerant plants.
  • sodium chloride for example 50 mM, 100 mM, 200 mM, 400 mM NaCl
  • Osmotic tolerance test Osmotic stress assays (including sodium chloride and mannitol assays) are conducted to determine if an osmotic stress phenotype was sodium chloride-specific or if it was a general osmotic stress related phenotype. Plants which are tolerant to osmotic stress may have more tolerance to drought and/or freezing. For salt and osmotic stress germination experiments, the medium is supplemented for example with 50 mM, 100 mM, 200 mM NaCl or 100 mM, 200 mM NaCl, 400 mM mannitol. See also Example 5 of the Examples section which follows.
  • Drought tolerance assay/Osmoticum assay Tolerance to drought is performed to identify the genes conferring better plant survival after acute water deprivation. To analyze whether the transgenic plants are more tolerant to drought, an osmotic stress produced by the non-ionic osmolyte sorbitol in the medium can be performed. Control and transgenic plants are germinated and grown in plant-agar plates for 4 days, after which they are transferred to plates containing 500 mM sorbitol. The treatment causes growth retardation, then both control and transgenic plants are compared, by measuring plant weight (wet and dry), yield, and by growth rates measured as time to flowering.
  • soil-based drought screens are performed with plants overexpressing the polynucleotides detailed above. Seeds from control Arabidopsis plants, or other transgenic plants overexpressing the polypeptide of the invention are germinated and transferred to pots. Drought stress is obtained after irrigation is ceased accompanied by placing the pots on absorbent paper to enhance the soil-drying rate. Transgenic and control plants are compared to each other when the majority of the control plants develop severe wilting. Plants are re-watered after obtaining a significant fraction of the control plants displaying a severe wilting. Plants are ranked comparing to controls for each of two criteria: tolerance to the drought conditions and recovery (survival) following re-watering.
  • Cold stress tolerance To analyze cold stress, mature (25 day old) plants are transferred to 4° C. chambers for 1 or 2 weeks, with constitutive light. Later on plants are moved back to greenhouse. Two weeks later damages from chilling period, resulting in growth retardation and other phenotypes, are compared between both control and transgenic plants, by measuring plant weight (wet and dry), and by comparing growth rates measured as time to flowering, plant size, yield, and the like.
  • Heat stress tolerance is achieved by exposing the plants to temperatures above 34° C. for a certain period. Plant tolerance is examined after transferring the plants back to 22° C. for recovery and evaluation after 5 days relative to internal controls (non-transgenic plants) or plants not exposed to neither cold or heat stress.
  • Germination tests compare the percentage of seeds from transgenic plants that could complete the germination process to the percentage of seeds from control plants that are treated in the same manner. Normal conditions are considered for example, incubations at 22° C. under 22-hour light 2-hour dark daily cycles. Evaluation of germination and seedling vigor is conducted between 4 and 14 days after planting. The basal media is 50% MS medium (Murashige and Skoog, 1962 Plant Physiology 15, 473-497).
  • Germination is checked also at unfavorable conditions such as cold (incubating at temperatures lower than 10° C. instead of 22° C.) or using seed inhibition solutions that contain high concentrations of an osmolyte such as sorbitol (at concentrations of 50 mM, 100 mM, 200 mM, 300 mM, 500 mM, and up to 1000 mM) or applying increasing concentrations of salt (of 50 mM, 100 mM, 200 mM, 300 mM, 500 mM NaCl).
  • an osmolyte such as sorbitol
  • salt of 50 mM, 100 mM, 200 mM, 300 mM, 500 mM NaCl
  • Water use efficiency can be determined as the biomass produced per unit transpiration. To analyze WUE, leaf relative water content can be measured in control and transgenic plants. Fresh weight (FW) is immediately recorded; then leaves are soaked for 8 hours in distilled water at room temperature in the dark, and the turgid weight (TW) is recorded. Total dry weight (DW) is recorded after drying the leaves at 60° C. to a constant weight. Relative water content (RWC) is calculated according to the following Formula I:
  • Fertilizer use efficiency To analyze whether the transgenic plants are more responsive to fertilizers, plants are grown in agar plates or pots with a limited amount of fertilizer, as described, for example, in Example 6, hereinbelow and in Yanagisawa et al (Proc Natl Acad Sci USA. 2004; 101:7833-8). The plants are analyzed for their overall size, time to flowering, yield, protein content of shoot and/or grain. The parameters checked are the overall size of the mature plant, its wet and dry weight, the weight of the seeds yielded, the average seed size and the number of seeds produced per plant.
  • NUE nitrogen use efficiency
  • PUE phosphate use efficiency
  • KUE potassium use efficiency
  • N (nitrogen) concentration determination in the structural parts of the plants involves the potassium persulfate digestion method to convert organic N to NO 3 ⁇ (Purcell and King 1996 Argon. J. 88:111-113, the modified Cd ⁇ mediated reduction of NO 3 ⁇ to NO2 ⁇ (Vodovotz 1996 Biotechniques 20:390-394) and the measurement of nitrite by the Griess assay (Vodovotz 1996, supra). The absorbance values are measured at 550 nm against a standard curve of NaNO 2 . The procedure is described in details in Samonte et al. 2006 Agron. J. 98:168-176.
  • Grain protein concentration Grain protein content (g grain protein m ⁇ 2 ) is estimated as the product of the mass of grain N (g grain N m ⁇ 2 ) multiplied by the N/protein conversion ratio of k-5.13 (Mosse 1990, supra). The grain protein concentration is estimated as the ratio of grain protein content per unit mass of the grain (g grain protein kg ⁇ 1 grain).
  • Oil content The oil content of a plant can be determined by extraction of the oil from the seed or the vegetative portion of the plant. Briefly, lipids (oil) can be removed from the plant (e.g., seed) by grinding the plant tissue in the presence of specific solvents (e.g., hexane or petroleum ether) and extracting the oil in a continuous extractor. Indirect oil content analysis can be carried out using various known methods such as Nuclear Magnetic Resonance (NMR) Spectroscopy, which measures the resonance energy absorbed by hydrogen atoms in the liquid state of the sample [See for example, Conway T F.
  • NMR Nuclear Magnetic Resonance
  • NI Near Infrared
  • the plant vigor can be calculated by the increase in growth parameters such as leaf area, fiber length, rosette diameter, plant fresh weight and the like per time.
  • the growth rate can be measured using digital analysis of growing plants. For example, images of plants growing in greenhouse on plot basis can be captured every 3 days and the rosette area can be calculated by digital analysis. Rosette area growth is calculated using the difference of rosette area between days of sampling divided by the difference in days between samples.
  • Measurements of seed yield can be done by collecting the total seeds from 8-16 plants together, weighting them using analytical balance and dividing the total weight by the number of plants. Seed per growing area can be calculated in the same manner while taking into account the growing area given to a single plant. Increase seed yield per growing area could be achieved by increasing seed yield per plant, and/or by increasing number of plants capable of growing in a given area.
  • Evaluation of the seed yield per plant can be done by measuring the amount (weight or size) or quantity (i.e., number) of dry seeds produced and harvested from 8-16 plants and divided by the number of plants.
  • Evaluation of growth rate can be done by measuring plant biomass produced, rosette area, leaf size or root length per time (can be measured in cm 2 per day of leaf area).
  • Fiber length can be measured using fibrograph.
  • the fibrograph system was used to compute length in terms of “Upper Half Mean” length.
  • the upper half mean (UHM) is the average length of longer half of the fiber distribution.
  • the present invention is of high agricultural value for promoting the yield of commercially desired crops (e.g., biomass of vegetative organ such as poplar wood, or reproductive organ such as number of seeds or seed biomass).
  • crops e.g., biomass of vegetative organ such as poplar wood, or reproductive organ such as number of seeds or seed biomass.
  • compositions, method or structure may include additional ingredients, steps and/or parts, but only if the additional ingredients, steps and/or parts do not materially alter the basic and novel characteristics of the claimed composition, method or structure.
  • a compound or “at least one compound” may include a plurality of compounds, including mixtures thereof.
  • range format is merely for convenience and brevity and should not be construed as an inflexible limitation on the scope of the invention. Accordingly, the description of a range should be considered to have specifically disclosed all the possible subranges as well as individual numerical values within that range. For example, description of a range such as from 1 to 6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6 etc., as well as individual numbers within that range, for example, 1, 2, 3, 4, 5, and 6. This applies regardless of the breadth of the range.
  • a numerical range is indicated herein, it is meant to include any cited numeral (fractional or integral) within the indicated range.
  • the phrases “ranging/ranges between” a first indicate number and a second indicate number and “ranging/ranges from” a first indicate number “to” a second indicate number are used herein interchangeably and are meant to include the first and second indicated numbers and all the fractional and integral numerals therebetween.
  • method refers to manners, means, techniques and procedures for accomplishing a given task including, but not limited to, those manners, means, techniques and procedures either known to, or readily developed from known manners, means, techniques and procedures by practitioners of the chemical, pharmacological, biological, biochemical and medical arts.
  • ESTs expressed sequence tags from 1,195 relevant EST's libraries originated from 15 different species, including both monocot and dicot species, namely: Arabidopsis, barley, Brassica rapa, cotton, grape, maize, medicago, poplar, potato, rice, sorghum, soybean, sugarcane, tomato and wheat.
  • Tomato plants were selected as a model plant based on the high quality tomato database from several tomato species which can be used for data-mining and the present inventors' experience in using the tomato genome as a model plant.
  • the relatively high salt tolerance exhibited by various tomato species makes the tomato genome an excellent candidate for identifying new stress tolerance mechanisms.
  • tomato is not only used as a model plant for genetic studies, it is also used as an important crop with well-defined yield parameters, which can be used to distinguish between genes affecting abiotic-stress tolerance and genes preventing yield loss under abiotic-stress conditions.
  • Clustering and assembly EST and mRNA sequences of each of the 15 species were extracted from GenBank versions 157, 160, 161, 162, 164, 165, 166, clustered and assembled using Compugen's LEADS clustering and assembly platform (Compugen Ltd., Tel Aviv, Israel; Yelin et. al. 2003, Nature Biotechnology 21, 379-85). Automatically extracted EST library annotations were manually accurated and classified by anatomy, developmental stage, abiotic/biotic stress treatment and cultivars. The results were loaded into Oracle database. The predicted proteins were then annotated using InterPro(2) (Hypertext Transfer Protocol://World Wide Web (dot) ebi (dot) ac (dot) uk/interpro/).
  • the present inventors used a combination of two approaches: selection of AQP clusters showing significant over expression (EST distribution versus normal is more than two folds; statistical significance of over-expression ⁇ p Value ⁇ 0.05) either in roots compared to shoots or under various abiotic stresses (including drought, cold, salinity, heat, chemical treatments, etc.), compared to non stress control. It was found that ESTs of about 9% of the AQP genes were significantly overrepresented in roots and 3.5% of them were induced under different abiotic stresses. AQP genes which are highly overrepresented in roots were selected since plants with an efficient root system are expected to capture more water from a drying soil.
  • AQP genes which are overrepresented in various abiotic stresses such as nutrient deficiency, heat, salinity and heavy metal stresses and biotic stresses such as application of elicitors and pathogens were selected considering that they can provide high tolerance to a wide spectrum of stresses.
  • the TIP2 subgroup (or subfamily) is highly expressed in roots and in abiotic stresses.
  • the TIP2 subgroup is found in 38 plant species and other organisms (nucleic acid SEQ ID NOs: 1, 2, 19, 20-22; Table 2), available in public databases [Hypertext Transfer Protocol://World Wide Web (dot) ncbi (dot) nlm (dot) nih (dot) gov/dbEST/].
  • the TIP2 gene was highly expressed in roots (6 fold, p ⁇ 1.01 E-24) and in both biotic (2 fold, p ⁇ 4.6 E-02) and abiotic stresses (4.5 fold, p ⁇ E-02) (data not shown).
  • Sequences which are homologous [showing at least 80% protein sequence identity on 80% of the global hit or query length, as calculated using BlastP and tBlastN algorithms of the National Center of Biotechnology Information (NCBI)] or orthologues of the AQP genes described in Table 2, and are expected to possess the same role in ABST and yield improvement in plants, are disclosed in Table 3 hereinbelow (SEQ ID NOs:6, 215-1101 and 1138-1400; Table 3).
  • Table 3 also includes homologous and orthologues of the AQP TIP2 subfamily (SEQ ID NOs:21, 53-214, 1102-1137) and additional homologous and orthologues (SEQ ID NOs:2844-3051).
  • RNA levels were determined using reverse transcription assay followed by quantitative Real-Time PCR (qRT-PCR) analysis. RNA levels were compared between leaves of 20 days old seedlings of tomato plants grown under salinity water. A correlation analysis between mRNA levels in different experimental conditions/genetic backgrounds was performed in order to determine the role of the gene in the plant.
  • qRT-PCR quantitative Real-Time PCR
  • Quantitative Real Time RT-PCR (qRT-PCR)—To verify the level of expression, specificity and trait-association, Reverse Transcription followed by quantitative Real-Time PCR (qRTPCR) was performed on total RNA extracted from leaves of 2 tomato varieties namely YO361 (salt tolerant variety) and FA191 (salt sensitive variety). Messenger RNA (mRNA) levels were determined for AQP genes, expressed under normal and stressed conditions.
  • Reverse transcription was performed using 1 ⁇ g total RNA, using 200 U Super Script II Reverse Transcriptase enzyme (Invitrogen), 150 ng random deoxynucleotide hexamers (Invitrogen), 500 ⁇ M deoxynucleotide tri-phosphates (dNTPs) mix (Takara, Japan), 0.2 volume of ⁇ 5 reverse transcriptase (RT) buffer (Invitrogen), 0.01 M dithiothreitol (DTT), 40 U RNAsin (Promega), diethylpyrocarbonate (DEPC) treated double distilled water (DDW) was added up to 24 ⁇ l.
  • RT reverse transcriptase
  • DTT dithiothreitol
  • DEPC diethylpyrocarbonate
  • RNA, random deoxynucleotide hexamers, dNTPs mix and DEPC treated DDW was incubated at 65° C. for 5 minutes, followed by 4° C. for 5 minutes.
  • Mix of reverse transcriptase (RT) buffer, dithiothreitol (DTT) and RNAsin was added to the RT reactions followed by incubation at 25° C. for 10 minutes and at 42° C. for 2 minutes afterwards. Finally, Super Script II Reverse Transcriptase enzyme was added to the RT reactions that were further incubated for 50 minutes at 42° C., followed by 70° C. for 15 minutes.
  • MAB58 and MAB59 cDNA was diluted 1:2 due to very weak expression and consequently for housekeeping genes cDNA was diluted 1:8 in order to insert the Ct values in calibration curve range. 5 ⁇ L of the diluted cDNA was used for qPCR.
  • primers of the AQP genes were designed, as summarized in Table 4 below.
  • the expression level of the housekeeping genes: Actin (SEQ ID NO: 2841), GAPDH (SEQ ID NO: 2842) and RPL19 (SEQ ID NO: 2747) was determined in order to normalize the expression level between the different tissues.
  • ORFs open reading frames
  • EST clusters and in some cases mRNA sequences were analyzed to identify the entire open reading frame by comparing the results of several translation algorithms to known proteins from other plant species.
  • RNA extraction was performed using standard protocols described elsewhere (Sambrook J., E. F. Fritsch, and T. Maniatis. 1989. Molecular Cloning. A Laboratory Manual., 2nd Ed. Cold Spring Harbor Laboratory Press, New York.), and are basic for those skilled in the art.
  • PCR products were purified using PCR purification kit (Qiagen) and sequencing of the amplified PCR products was performed, using ABI 377 sequencer (Applied Biosystems).
  • 2 sets of primers were ordered for the amplification of each gene, via nested PCR (meaning first amplifying the gene using external primers and then using the produced PCR product as a template for a second PCR reaction, where the internal set of primers are used).
  • one or two of the internal primers were used for gene amplification, both in the first and the second PCR reactions (meaning only 2-3 primers were designed for a gene).
  • a 8-12 bp extension is added to the 5′ primer end of each internal primer.
  • the primer extension includes an endonuclease restriction site.
  • restriction sites are selected using two parameters: (a) The restriction site does not exist in the cDNA sequence; and (b) The restriction sites in the forward and reverse primers are designed so the digested cDNA is inserted in the sense formation into the binary vector utilized for transformation.
  • Table 6 primers used for cloning tomato and barley AQPs are provided.
  • PCR products were purified (PCR Purification Kit, Qiagen, Germany) and digested with the restriction endonucleases (Roche, Switzerland) according to the sites design in the primers (Tables 8 and 9 below).
  • Each of the digested PCR products was cloned first into high copy plasmid pBlue-script KS [Hypertext Transfer Protocol://World Wide Web (dot) stratagene (dot) com/manuals/212205 (dot) pdf] which was digested with the same restriction enzymes.
  • NOS Nopaline Synthase terminator originated from the binary vector pBI101.3 [nucleotide coordinates 4417-4693 in GenBank Accession No.
  • U12640 (SEQ ID NO:2824)] was already cloned into the pBlue-script KS, between the restriction endonuclease sites SacI and EcoRI, so the gene is introduced upstream of the terminator.
  • the At6669 promoter (SEQ ID NO: 2823) is already cloned into the pBlue-script KS, so the gene is introduced downstream of the promoter.
  • the digested PCR products and the linearized plasmid vector were ligated using T4 DNA ligase enzyme (Roche, Switzerland). Sequencing of the inserted genes was conducted, using ABI 377 sequencer (Applied Biosystems). Sequences of few of the cloned AQP genes, as well as their encoded proteins are listed in Table 7, below.
  • the genes were digested again and ligated into pPI or pGI binary plasmids, harboring the At6669 promoter (between the HindIII and SalI restriction endonucleases site) (Table 8).
  • the At6669 promoter together with the gene are digested out of the pBlue-script KS plasmid and ligated into pPI or pGI binary plasmids, using restriction endonucleases as given in Table 8.
  • the pPI plasmid vector was constructed by inserting a synthetic poly-(A) signal sequence, originating from pGL3 basic plasmid vector (Promega, Acc. No. U47295; by 4658-4811) into the HindIII restriction site of the binary vector pBI101.3 (Clontech, Acc. No. U12640).
  • pGI FIG. 1
  • the original gene in the back bone is GUS-Intron, rather than GUS.
  • the cloned genes were sequenced.
  • Synthetic sequences (such as of MAB54, nucleotide SEQ ID NO: 2751, which encodes protein SEQ ID NO: 27; or MAB72 SEQ ID NO:2843, which encodes SEQ ID NO:2769) of some of the cloned polynucleotides were ordered from a commercial supplier (GeneArt, GmbH). To optimize the coding sequence, codon-usage Tables calculated from plant transcriptomes were used [example of such Tables can be found in the Codon Usage Database available online at Hypertext Transfer Protocol://World Wide Web (dot) kazusa (dot) or (dot) jp/codon/].
  • the optimized coding sequences were designed in a way that no changes were introduced in the encoded amino acid sequence while using codons preferred for expression in dicotyledonous plants mainly tomato and Arabidopsis; and monocotyledonous plants such as maize. Such optimized sequences promote better translation rate and therefore higher protein expression levels. To the optimized sequences flanking additional unique restriction enzymes sites were added to facilitate cloning genes in binary vectors.
  • Promoters used CaMV 35S promoter (SEQ ID NO: 2825) and Arabidopsis At6669 promoter (SEQ ID NO: 2823; which is SEQ ID NO:61 of WO04081173 to Evogene Ltd.).
  • Arabidopsis transformation Arabidopsis transformation of the following MAB genes and orthologues: MAB115, MAB54, MAB55, MAB56, MAB57, MAB58, MAB59 (ortholog of MAB58), MAB69, MAB70, MAB71, MAB72, MAB74, MAB76, MAB77, MAB79, MAB116 (ortholog of MAB115 and MAB55), and MAB117 (the sequence identifiers of the cloned polynucleotides and their expressed polypeptides are provided in Table 7 above) was performed according to Clough S J, Bent A F. (1998) “Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana.
  • Single colonies of Agrobacterium carrying the binary vectors harboring the AQP genes are cultured in LB medium supplemented with kanamycin (50 mg/L) and gentamycin (50 mg/L). The cultures were incubated at 28° C. for 48 hours under vigorous shaking and centrifuged at 4000 rpm for 5 minutes.
  • the pellets comprising Agrobacterium cells were resuspended in a transformation medium which contained half-strength (2.15 g/L) Murashige-Skoog (Duchefa); 0.044 ⁇ M benzylamino purine (Sigma); 112 ⁇ g/L B5 Gam strig vitamins (Sigma); 5% sucrose; and 0.2 ml/L Silwet L-77 (OSI Specialists, CT) in double-distilled water, at pH of 5.7.
  • half-strength (2.15 g/L) Murashige-Skoog (Duchefa); 0.044 ⁇ M benzylamino purine (Sigma); 112 ⁇ g/L B5 Gambourg vitamins (Sigma); 5% sucrose; and 0.2 ml/L Silwet L-77 (OSI Specialists, CT) in double-distilled water, at pH of 5.7.
  • Transformation of T 0 plants was performed by inverting each plant into an Agrobacterium suspension such that the flowering stem was submerged for 3-5 seconds. Each inoculated T 0 plant was immediately placed in a plastic tray, then covered with clear plastic dome to maintain humidity and kept in the dark at room temperature for 18 hours to facilitate infection and transformation. Transformed (transgenic) plants were then uncovered and transferred to a greenhouse for recovery and maturation. The transgenic T 0 plants were grown in the greenhouse for 3-5 weeks until siliques maturation, and then seeds were harvested and kept at room temperature until sowing.
  • seeds collected from transgenic T 0 plants are surface-sterilized by soaking in 70% ethanol for 1 minute, followed by soaking in 5% sodium hypochlorite and 0.05% Triton X-100 for 5 minutes.
  • the surface-sterilized seeds are thoroughly washed in sterile distilled water then placed on culture plates containing half-strength Murashige-Skoog (Duchefa); 2% sucrose; 0.8% plant agar; 50 mM kanamycin; and 200 mM carbenicylin (Duchefa).
  • the culture plates are incubated at 4° C. for 48 hours then transferred to a growth room at 25° C. for an additional week of incubation.
  • T 1 Arabidopsis plants are transferred to fresh culture plates for another week of incubation. Following incubation the T 1 plants are removed from culture plates and planted in growth mix contained in 250 ml pots. The transgenic plants were allowed to grow in a greenhouse to maturity. Seeds harvested from T 1 plants are cultured and grown to maturity as T 2 plants under the same conditions as used for culturing and growing the T 1 plants. At least 10 independent transformation events are created from each construct for which T2 seeds are collected. The introduction of the gene is determined for each event by PCR performed on genomic DNA extracted from each event produced.
  • Transformation of tomato (Var M82) plants with putative cotton genes—Tomato (Solanum esculentum, var M82) transformation and cultivation of transgenic plants is effected according to: “Curtis I. S, Davey M. R, and Power J. B. 1995. “Leaf disk transformation”. Methods Mol. Biol. 44, 59-70 and Meissner R, Chague V, Zhu Q, Emmanuel E, Elkind Y, Levy A. A. 2000. “Technical advance: a high throughput system for transposon tagging and promoter trapping in tomato”. Plant J. 22, 265-74; with slight modifications.
  • Assay I plant growth under osmotic stress [poly (ethylene glycol) (PEG)] in tissue culture conditions—One of the consequences of drought is the induction of osmotic stress in the area surrounding the roots; therefore, in many scientific studies, PEG (e.g., 25% PEG8000) is used to simulate the osmotic stress conditions resembling the high osmolarity found during drought stress.
  • PEG poly (ethylene glycol)
  • Digital imaging A laboratory image acquisition system, which consists of a digital reflex camera (Canon EOS 300D) attached with a 55 mm focal length lens (Canon EF-S series), mounted on a reproduction device (Kaiser RS), which included 4 light units (4 ⁇ 150 Watts light bulb) and located in a darkroom, was used for capturing images of plantlets sawn in agar plates.
  • the image capturing process was repeated every 2-5 days starting at day 1 till day 10-15 (see for example the images in FIGS. 2A-B )
  • An image analysis system was used, which consists of a personal desktop computer (Intel P4 3.0 GHz processor) and a public domain program—ImageJ 1.39 (Java based image processing program which was developed at the U.S. National Institutes of Health and freely available on the internet at Hypertext Transfer Protocol://rsbweb (dot) nih (dot) gov/). Images were captured in resolution of 10 Mega Pixels (3888 ⁇ 2592 pixels) and stored in a low compression JPEG (Joint Photographic Experts Group standard) format. Next, analyzed data was saved to text files and processed using the JMP statistical analysis software (SAS institute).
  • SAS institute JMP statistical analysis software
  • Seedling analysis Using the digital analysis seedling data was calculated, including leaf area, root coverage and root length.
  • the relative growth rate for the various seedling parameters was calculated according to the following formulas II, III and IV.
  • ⁇ rosette area is the interval between the current rosette area (measured at t 2 ) and the rosette area measured at the previous day (Area t 1 )
  • ⁇ t is the time interval (t 2 -t 1 , in days) between the current analyzed image day (t 2 ) and the previous day (t 1 ).
  • the relative growth rate of leaf area is in units of 1/day.
  • ⁇ root coverage area is the interval between the current root coverage area (measured at t 2 ) and the root coverage area measured at the previous day (Area t 1 )
  • ⁇ t is the time interval (t 2 -t 1 , in days) between the current analyzed image day (t 2 ) and the previous day (t 1 ).
  • the relative growth rate of root coverage area is in units of 1/day.
  • ⁇ root length is the interval between the current root length (measured at t 2 ) and the root length measured at the previous day (Area t 1 )
  • ⁇ t is the time interval (t 2 -t 1 , in days) between the current analyzed image day (t 2 ) and the previous day (t 1 ).
  • the relative growth rate of root length is in units of 1/day.
  • plantlets were removed from the media and weighed for the determination of plant fresh weight. Plantlets were then dried for 24 hours at 60° C., and weighed again to measure plant dry weight for later statistical analysis. Growth rate was determined by comparing the leaf area coverage, root coverage and root length, between each couple of sequential photographs, and results were used to resolve the effect of the gene introduced on plant vigor, under osmotic stress, as well as under optimal conditions. Similarly, the effect of the gene introduced on biomass accumulation, under osmotic stress as well as under optimal conditions, was determined by comparing the plants' fresh and dry weight to that of control plants (containing an empty vector or the GUS reporter gene under the same promoter). From every construct created, 3-5 independent transformation events were examined in replicates.
  • Tables 10-14 depict analyses of the above mentioned growth parameters of seedlings overexpres sing the polynucleotides of the invention under the regulation of the At6669 promoter (SEQ ID NO:2823) when grown under osmotic stress (25% PEG) conditions.
  • Tables 15-29 depict analyses of the above mentioned growth parameters of seedlings overexpres sing the polynucleotides of the invention under the regulation of the At6669 promoter in Normal Growth conditions (0.5 MS medium).
  • ABS tolerance Yield and plant growth rate at high salinity concentration under greenhouse conditions—This assay followed the rosette area growth of plants grown in the greenhouse as well as seed yield at high salinity irrigation. Seeds were sown in agar media supplemented only with a selection agent (Kanamycin) and Hoagland solution under nursery conditions. The T 2 transgenic seedlings were then transplanted to 1.7 trays filled with peat and perlite. The trays were irrigated with tap water (provided from the pots' bottom). Half of the plants were irrigated with a salt solution (40-80 mM NaCl and 5 mM CaCl 2 ) so as to induce salinity stress (stress conditions).
  • a salt solution 40-80 mM NaCl and 5 mM CaCl 2
  • the plants were analyzed for their overall size, growth rate, flowering, seed yield, weight of 1,000 seeds, dry matter and harvest index (HI—seed yield/dry matter).
  • Transgenic plants performance was compared to control plants grown in parallel under the same conditions.
  • Mock transgenic plants expressing the uidA reporter gene (GUS-Intron) or with no gene at all, under the same promoter were used as control.
  • the experiment was planned in nested randomized plot distribution. For each gene of the invention three to five independent transformation events were analyzed from each construct.
  • Digital imaging A laboratory image acquisition system, which consists of a digital reflex camera (Canon EOS 300D) attached with a 55 mm focal length lens (Canon EF-S series), mounted on a reproduction device (Kaiser RS), which included 4 light units (4 ⁇ 150 Watts light bulb) was used for capturing images of plant samples.
  • the image capturing process was repeated every 2 days starting from day 1 after transplanting till day 16. Same camera, placed in a custom made iron mount, was used for capturing images of larger plants sawn in white tubs in an environmental controlled greenhouse.
  • the tubs were square shape include 1.7 liter trays. During the capture process, the tubs were placed beneath the iron mount, while avoiding direct sun light and casting of shadows.
  • An image analysis system was used, which consists of a personal desktop computer (Intel P4 3.0 GHz processor) and a public domain program—ImageJ 1.39 (Java based image processing program which was developed at the U.S. National Institutes of Health and freely available on the internet at Hypertext Transfer Protocol://rsbweb (dot) nih (dot) gov/). Images were captured in resolution of 10 Mega Pixels (3888 ⁇ 2592 pixels) and stored in a low compression JPEG (Joint Photographic Experts Group standard) format. Next, analyzed data was saved to text files and processed using the JMP statistical analysis software (SAS institute).
  • SAS institute JMP statistical analysis software
  • Leaf analysis Using the digital analysis leaves data was calculated, including leaf number, area, perimeter, length and width.
  • Vegetative growth rate the relative growth rate (RGR) of leaf number and rosette area were calculated formulas V and VI, respectively.
  • ⁇ leaf number is the interval between the current leaf number (measured at t 2 ) and the leaf number measured at the previous day (Area t 1 )
  • ⁇ t is the time interval (t 2 -t 1 , in days) between the current analyzed image day (t 2 ) and the previous day (t 1 ).
  • the relative growth rate of leaf number is in units of 1/day.
  • ⁇ rosette area is the interval between the current rosette area (measured at t 2 ) and the rosette area measured at the previous day (Area t 1 )
  • ⁇ t is the time interval (t 2 -t 1 , in days) between the current analyzed image day (t 2 ) and the previous day (t 1 ).
  • the relative growth rate of rosette area is in units of 1/day.
  • Tables 30-45 depict analyses of plant parameters as describe above overexpressing the polynucleotides of the invention under the regulation of the At6669 promoter under salinity irrigation conditions [NaCl 40-80 mM; NaCl Electrical conductivity (E.C.) of 7-10].
  • Tables 46-58 depict analyses of plant parameters (as describe above) overexpressing the polynucleotides of the invention under the regulation of the 6669 promoter under Normal Growth conditions [Normal irrigation included NaCl Electrical conductivity (E.C.) of 1-2].
  • MAB115 Plants transgenic to the following MAB genes were assayed for fertilizer use efficiency in a tissue culture assay: MAB115, MAB54, MAB55, MAB56, MAB57, MAB58, MAB59, MAB69, MAB70, MAB71, MAB72, MAB74, MAB76, MAB77, MAB79, MAB116, and MAB117 (the sequence identifiers of the cloned polynucleotides and their expressed polypeptides are provided in Table 7 above).
  • NUE nitrogen use efficiency
  • Each plate contains 5 seedlings of same event, and 3-4 different plates (replicates) for each event.
  • For each polynucleotide of the invention at least four independent transformation events were analyzed from each construct. Plants expressing the polynucleotides of the invention were compared to the average measurement of the control plants (empty vector or GUS reporter under the same promoter) used in the same experiment.
  • Tables 59-68 depict analyses of seedling parameters (as describe above) overexpressing the polynucleotides of the invention under the regulation of At6669 promoter under Nitrogene Deficiency conditions.
  • AQP TIP2 genes on plant's stress tolerance
  • the present inventors have previously cloned and overexpressed a polynucleotide which comprises the nucleic acid sequence set forth by SEQ ID NO:2827 (also known as ABST36 set forth by SEQ ID NO:13 in WO2004/104162; or S1TIP2;2) and which encodes the TIP2 polypeptide set forth by SEQ ID NO:2828 (which comprises the consensus sequence TLXFXFAGVGS; SEQ ID NO:2826).
  • the nucleic acid constructs which comprises the ABST36 polynucleotide under the regulation of the constitutive Arabidopsis At6669 promoter (SEQ ID NO: 2823) (further referred to as the At6669::ABST36 construct) was further transformed into tomato ( Solanum lycopersicum ) as a model crop plant (Tom-ABST36), as well as into Arabidopsis thaliana.
  • tomato Solanum lycopersicum
  • Tom-ABST36 model crop plant
  • Transgenic genotypes in each field trial were compared to their null-segregant counterparts as controls.
  • Tomato Salt-stress field trial All field trials were performed in a light soil, in an open field (net-house) near Rehovot, Israel. The F1 hybrids of four independent events of the cross between ABST36-transgenic MicroTom plants and M82 tomato plants were grown for the first 3 weeks in a nursery under normal irrigation conditions. The seedlings were then transplanted into rows and grown in a commercial greenhouse. The salt-stress trial was divided into four blocks. In each block, two different irrigation systems were established: a normal water regime for tomato cultivation and a continuous irrigation with saline water (addition of 180 to 200 mM NaCl).
  • Each block consisted of a total of 60 plants divided as follows: six plants per event and six seedling null segregants were planted in the control row and a similar number of plants were planted in the salt-stressed row. At the stage of about 80% red fruits in planta, fruit yield, plant fresh weight, and harvest index were calculated. Harvest index was calculated as yield/plant biomass.
  • Tomato Water-deficiency-stress field trial All field trials were performed in a light soil, in an open field (net-house) near Rehovot, Israel.
  • the F1 hybrids of the four independent events were initially grown as described above.
  • Three-week-old seedlings were transplanted to a net-greenhouse.
  • the experiment was structured in four blocks containing three rows irrigated with different water levels and intervals (WLI-0, WLI-1, WLI-2). In each block, six transgenic plants per event analyzed and six non transgenic plants were transplanted in each row. Seedlings were transplanted after 4 weeks into wet soil.
  • the amount of water used to uniformly irrigate before transplanting reached maximum water capacity [20% weight per weight (w/w)] at 60 cm depth, but without the creation of water overload.
  • Each plant was transplanted near a dripper, with a 30-cm distance between plants, giving a total density of 2,600 plants per 1,000 m 2 , according to a commercial growth protocol.
  • Soil water capacity was measured using the standard procedures by sampling soil from the following three depths: 0 to 20 cm, 20 to 40 cm, and 40 to 60 cm. The water content in these soil layers was measured routinely every week. The soil contained 5% hygroscopic water while the maximum water capacity of the soil was 20%. All fertilizers were applied in the soil prior to plant transplantation.
  • WLI-1 plants received the amount of water required to reach maximum soil water capacity.
  • WLI-2 plants were irrigated only once a week, at the beginning of the week.
  • the water-stress experiment lasted throughout the flowering period (23 days), corresponding to four cycles of the above-described stresses. Afterwards, all treatments received the recommended amount of water.
  • the calculated water amount was equal to the difference between the water contents in dry soil and in soil with maximum water capacity.
  • the water amounts were compared between treatments according to actual water content in the soil (S3).
  • treatments WLI-1 and WLI-2 received a total of 75% less water than the controls (WLI-0).
  • Transgenic plants exhibit increased tolerance to salt stress—To induce salt-stress, transgenic and control tomato plants were continuously irrigated in field trials with 180 to 200 mM NaCl. As shown in FIGS. 3 a - c, 3 g - j and Table 69 below, Tom-ABST36 plants appeared to be more vigorous in all of the experiments than the control plants, which were smaller and showed severe symptoms of leaf and shoot necrosis (see for example, FIG. 3 j ). This was also associated with higher fruit yield in Tom-ABST36 plants relative to controls ( FIG. 3 a ).
  • Transgenic plants exhibit increased tolerance to water-deficiency stress—Transgenic plants subjected to water-deficiency stress exhibited a significantly higher (26%, p ⁇ 0.05) plant biomass compared to control plants ( FIG. 3 e ). Moreover the Tom-ABST36 plants showed a significant (up to 21%, p ⁇ 0.05) increment of fruit yield under water-deficient regimes (water level intervals WLI-1), while under normal irrigation, the yield improvement was even higher (27%, p ⁇ 0.05; FIG. 3 d ). The harvest index of the Tom-ABST36 plants was also higher when plants grew under regular and WLI-1 conditions while it remained similar to control when the water-deficient regime consisted of once-a-week irrigation (WLI-2) ( FIG. 3 f ).
  • WLI-2 once-a-week irrigation
  • Arabidopsis Salt-stress green house trial A complementary experiment with transgenic Arabidopsis plants expressing the ABST36 construct showed increased tolerance to a salt stress of 150 mM NaCl compared to control plants, as reflected in 42% higher fresh biomass and 60% higher dry biomass (Table 70 below).
  • In-vitro salt-stress assay Seeds of transgenic Arabidopsis plants harboring the At6669::ABST36 construct or 35S::GUS construct (which was used as control) were sown in 1/2 MS media containing 40 mg/l kanamycin for selection. Selected seedlings were sub-cultured to 1/2 MS media with 0 or 150 mM NaCl. Plants were grown for a period of 3 weeks. Results are the average of four independent events that were analyzed in four repeats. For the determination of shoot dry weight, shoot plants were collected and dried for 24 hours at 60° C. and then weighed.
  • File information is provided as: File name/byte size/date of creation/operating system/machine format.
  • CD-ROM1 (1 file of SEQUENCE LISTING):

Landscapes

  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Cell Biology (AREA)
  • Nutrition Science (AREA)
  • Botany (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Oil, Petroleum & Natural Gas (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Fertilizers (AREA)
US12/810,855 2007-12-27 2008-12-23 Isolated polypeptides, polynucleotides useful for modifying water user efficiency, fertilizer use efficiency, biotic/abiotic stress tolerance, yield and biomass in plants Abandoned US20110119791A1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US12/810,855 US20110119791A1 (en) 2007-12-27 2008-12-23 Isolated polypeptides, polynucleotides useful for modifying water user efficiency, fertilizer use efficiency, biotic/abiotic stress tolerance, yield and biomass in plants

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
US916607P 2007-12-27 2007-12-27
US13623808P 2008-08-20 2008-08-20
PCT/IL2008/001657 WO2009083958A2 (en) 2007-12-27 2008-12-23 Isolated polypeptides, polynucleotides useful for modifying water user efficiency, fertilizer use efficiency, biotic/abiotic stress tolerance, yield and biomass in plants
US12/810,855 US20110119791A1 (en) 2007-12-27 2008-12-23 Isolated polypeptides, polynucleotides useful for modifying water user efficiency, fertilizer use efficiency, biotic/abiotic stress tolerance, yield and biomass in plants

Related Parent Applications (1)

Application Number Title Priority Date Filing Date
PCT/IL2008/001657 A-371-Of-International WO2009083958A2 (en) 2007-12-27 2008-12-23 Isolated polypeptides, polynucleotides useful for modifying water user efficiency, fertilizer use efficiency, biotic/abiotic stress tolerance, yield and biomass in plants

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US13/450,547 Continuation US8426682B2 (en) 2007-12-27 2012-04-19 Isolated polypeptides, polynucleotides useful for modifying water user efficiency, fertilizer use efficiency, biotic/abiotic stress tolerance, yield and biomass in plants

Publications (1)

Publication Number Publication Date
US20110119791A1 true US20110119791A1 (en) 2011-05-19

Family

ID=40635469

Family Applications (4)

Application Number Title Priority Date Filing Date
US12/810,855 Abandoned US20110119791A1 (en) 2007-12-27 2008-12-23 Isolated polypeptides, polynucleotides useful for modifying water user efficiency, fertilizer use efficiency, biotic/abiotic stress tolerance, yield and biomass in plants
US13/450,547 Expired - Fee Related US8426682B2 (en) 2007-12-27 2012-04-19 Isolated polypeptides, polynucleotides useful for modifying water user efficiency, fertilizer use efficiency, biotic/abiotic stress tolerance, yield and biomass in plants
US13/867,185 Expired - Fee Related US9670501B2 (en) 2007-12-27 2013-04-22 Isolated polypeptides, polynucleotides useful for modifying water user efficiency, fertilizer use efficiency, biotic/abiotic stress tolerance, yield and biomass in plants
US15/478,253 Active US10407690B2 (en) 2007-12-27 2017-04-04 Isolated polypeptides, polynucleotides useful for modifying water user efficiency, fertilizer use efficiency, biotic/abiotic stress tolerance, yield and biomass in plants

Family Applications After (3)

Application Number Title Priority Date Filing Date
US13/450,547 Expired - Fee Related US8426682B2 (en) 2007-12-27 2012-04-19 Isolated polypeptides, polynucleotides useful for modifying water user efficiency, fertilizer use efficiency, biotic/abiotic stress tolerance, yield and biomass in plants
US13/867,185 Expired - Fee Related US9670501B2 (en) 2007-12-27 2013-04-22 Isolated polypeptides, polynucleotides useful for modifying water user efficiency, fertilizer use efficiency, biotic/abiotic stress tolerance, yield and biomass in plants
US15/478,253 Active US10407690B2 (en) 2007-12-27 2017-04-04 Isolated polypeptides, polynucleotides useful for modifying water user efficiency, fertilizer use efficiency, biotic/abiotic stress tolerance, yield and biomass in plants

Country Status (9)

Country Link
US (4) US20110119791A1 (pt)
EP (1) EP2240510B1 (pt)
CN (1) CN101977928B (pt)
AU (1) AU2008344935C1 (pt)
BR (1) BRPI0819476A2 (pt)
CA (1) CA2709517C (pt)
MX (2) MX357387B (pt)
WO (1) WO2009083958A2 (pt)
ZA (1) ZA201004460B (pt)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9101100B1 (en) 2014-04-30 2015-08-11 Ceres, Inc. Methods and materials for high throughput testing of transgene combinations
US10953581B2 (en) 2013-05-29 2021-03-23 Synventive Molding Solutions, Inc. Actuator cooling apparatus and method

Families Citing this family (48)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7554007B2 (en) 2003-05-22 2009-06-30 Evogene Ltd. Methods of increasing abiotic stress tolerance and/or biomass in plants
AU2005234725B2 (en) 2003-05-22 2012-02-23 Evogene Ltd. Methods of Increasing Abiotic Stress Tolerance and/or Biomass in Plants and Plants Generated Thereby
EP2186899A3 (en) 2004-06-14 2010-09-08 Evogene Ltd. Polynucleotides and polypeptides involved in plant fiber development and methods of using same
RU2487166C2 (ru) 2005-10-24 2013-07-10 Эвоген Лтд Изолированные полипептиды, кодирующие их полинуклеотиды, экспрессирующие их трансгенные растения и способы их использования
EP2383345B1 (en) 2006-12-20 2017-11-22 Evogene Ltd. Polynucleotides and polypeptides involved in plant fiber development and methods of using same
MX341624B (es) 2007-04-09 2016-08-26 Evogene Ltd Polinucleotidos, polipeptidos y metodos para aumentar el contenido de aceite, la velocidad de crecimiento y biomasa de las plantas.
BRPI0812742B1 (pt) 2007-07-24 2021-04-20 Evogene Ltd método de aumento da biomassa, da taxa de crescimento, da produtividade de semente, da eficiência do uso de nitrogênio, do estresse abiótico de uma planta, do comprimento de raiz, da cobertura de raiz, da taxa de crescimento da área de roseta, e da taxa de crescimento do diâmetro da roseta de uma planta
MX357387B (es) 2007-12-27 2018-07-06 Evogene Ltd Polipeptidos, polinucleotidos aislados utiles para modificar la eficacia en el uso de agua, eficacia en el uso del fertilizante, la tolerancia al estres biotico/abiotico, el rendimiento y biomasa en plantas.
CA3148194A1 (en) 2008-05-22 2009-11-26 Evogene Ltd. Isolated polynucleotides and peptides and methods of using same for increasing plant yield, biomass, growth rate, vigor, oil content, abiotic stress tolerance of plants and nitrogen use efficiency
BR122021014170B1 (pt) 2008-08-18 2022-08-09 Evogene Ltd Método para aumentar a eficiência de uso de nitrogênio, eficiência de uso de fertilizantes, rendimento, taxa de crescimento, vigor, biomassa e/ou tolerância ao estresse abiótico de uma planta
MX345298B (es) 2008-10-30 2017-01-24 Evogene Ltd Polinucleótidos y polipéptidos aislados y métodos para utilizarlos para aumentar el rendimiento de la planta, biomasa, tasa de crecimiento, vigor, contenido de aceite, tolerancia al estrés abiótico de las plantas y eficacia en el uso de nitrógeno.
AR074925A1 (es) 2008-12-29 2011-02-23 Evogene Ltd Polinucleotidos, polipeptidos codificados, y metodos para utilizarlos para modular la tolerancia al estres abiotico, biomasa y/o rendimiento en plantas que los expresan
CA2753616C (en) 2009-03-02 2018-07-17 Evogene Ltd. Isolated polynucleotides and polypeptides, and methods of using same for increasing plant yield and/or agricultural characteristics
EP3238534B1 (en) 2009-06-10 2019-11-13 Evogene Ltd. Isolated polynucleotides and polypeptides, and methods of using same for increasing nitrogen use efficiency, yield, growth rate, vigor, biomass, oil content, and/or abiotic stress tolerance
BR112012002543B1 (pt) 2009-08-04 2021-03-09 Evogene Ltd. método para aumentar a tolerância ao estresse abiótico, o rendimento, a biomassa, taxa de crescimento, e/ou vigor de uma planta, e, construto de ácido nucléico isolado
WO2011021190A1 (en) * 2009-08-17 2011-02-24 Yissum Research Development Company Of The Hebrew University Of Jerusalem Ltd. Plants producing high crop yield
EP2519097B1 (en) 2009-12-28 2016-03-02 Evogene Ltd. Isolated polynucleotides and polypeptides and methods of using same for increasing plant yield, biomass, growth rate, vigor, oil content, abiotic stress tolerance of plants and nitrogen use efficiency
MA33933B1 (fr) 2010-01-22 2013-01-02 Bayer Ip Gmbh Combinaisons de principes actifs acaricides et/ou insecticides
EP2563112A4 (en) 2010-04-28 2014-03-05 Evogene Ltd INSULATED POLYNUCLEOTIDES AND POLYPEPTIDES AND METHODS OF USING THE SAME FOR INCREASING PLANT YIELD AND / OR AGRICULTURAL CHARACTERISTICS
AR082530A1 (es) 2010-08-30 2012-12-12 Evogene Ltd Polinucleotidos y polipeptidos aislados, y metodos para utilizarlos para aumentar la eficacia en el uso de nitrogeno, rendimiento, tasa de crecimiento, vigor, biomasa, contenido de aceite y/o tolerancia al estres abiotico
WO2012040851A1 (en) * 2010-09-30 2012-04-05 Universite Laval Horsetail silicon transporter genes and uses thereof
MX354193B (es) 2010-12-22 2018-02-16 Evogene Ltd Polinucleótidos y polipéptidos aislados, y métodos para utilizarlos para aumentar la tolerancia al estrés abiótico, rendimiento, tasa de crecimiento, vigor, biomasa, contenido de aceite y/o eficacia en el uso de nitrógeno de las plantas.
CA3139245A1 (en) 2011-05-03 2012-11-08 Evogene Ltd. Isolated polynucleotides and polypeptides and methods of using same for increasing plant yield, biomass, growth rate, vigor, oil content, abiotic stress tolerance of plants and nitrogen use efficiency
CN103717076B (zh) 2011-08-10 2016-04-13 拜耳知识产权股份有限公司 含有特定特特拉姆酸衍生物的活性化合物组合物
CA3177736A1 (en) 2011-08-23 2013-02-28 Evogene Ltd. Isolated polynucleotides and polypeptides, and methods of using same for increasing plant yield and/or agricultural characteristics
CN104254242A (zh) 2011-11-21 2014-12-31 先正达参股股份有限公司 用于增强植物中的线虫抗性的组合物和方法
BR122019023079B1 (pt) 2011-11-28 2021-05-25 Evogene Ltd método para aumento da eficiência no uso de nitrogênio, produção, biomassa, taxa de crescimento, vigor, e/ou tolerância ao estresse abiótico de uma planta, e, construto de ácido nucleico isolado
BR122020022832B1 (pt) 2011-12-28 2021-08-17 Evogene Ltd Método para aumentar a produção, taxa de crescimento, biomassa, energia e/ou produção de semente de uma planta em comparação com uma planta nativa, e, construção de ácido nucleico isolado
MX390593B (es) 2012-02-29 2025-03-20 Evogene Ltd Polinucleótidos y polipéptidos aislados y métodos para utilizarlos para mejorar el rendimiento de la planta, biomasa, tasa de crecimiento, vigor, contenido de aceite, tolerancia al estrés abiótico de las plantas y eficacia en el uso de nitrógeno.
EA201491673A1 (ru) 2012-03-13 2015-07-30 Пайонир Хай-Бред Интернэшнл, Инк. Генетическое снижение мужской репродуктивной функции у растений
MX2014011037A (es) * 2012-03-13 2015-05-15 Pioneer Hi Bred Int Reduccion genetica de la fertilidad masculina en plantas.
AR092839A1 (es) 2012-05-28 2015-05-06 Evogene Ltd Polinucleotidos y polipeptidos aislados y metodos para utilizarlos para mejorar el rendimiento y/o caracteristicas agricolas de las plantas
BR122019028113B1 (pt) 2012-08-27 2022-08-30 Evogene Ltd Métodos para aumentar o rendimento, taxa de crescimento, biomassa, vigor, capacidade fotossintética, eficiência do uso de nitrogênio, tolerância ao estresse abiótico e/ou redução do tempo de floração ou o tempo de emergência da inflorescência de uma planta, e de cultivar uma colheita, e, construto de ácido nucleico
WO2014102773A1 (en) 2012-12-25 2014-07-03 Evogene Ltd. Isolated polynucleotides and polypeptides, and methods of using same for increasing nitrogen use efficiency of plants
BR122020005657B1 (pt) 2012-12-26 2022-04-05 Evogene Ltd Método para aumentar a eficiência do uso de nitrogênio, rendimento, taxa de crescimento, biomassa, vigor, rendimento de sementes, capacidade fotossintética e/ou tolerância ao estresse abiótico de uma planta, e, construção de ácido nucleico
WO2014188428A1 (en) 2013-05-22 2014-11-27 Evogene Ltd. Isolated polynucleotides and polypeptides, and methods of using same for increasing plant yield and/or agricultural characteristics
CA2916060A1 (en) 2013-08-27 2015-03-05 Evogene Ltd. Isolated polynucleotides and polypeptides, and methods of using same for increasing plant yield and/or agricultural characteristics
WO2015181823A1 (en) 2014-05-28 2015-12-03 Evogene Ltd. Isolated polynucleotides, polypeptides and methods of using same for increasing abiotic stress tolerance, biomass and yield of plants
CA3176207A1 (en) 2014-08-27 2016-03-03 Evogene Ltd. Constructs and methods for increasing plant yield or agricultural characteristics or both
CN104650202A (zh) * 2015-02-06 2015-05-27 中国农业大学 珠美海棠水通道蛋白MzPIP2;1及其编码基因与应用
ES2895223T3 (es) * 2015-06-23 2022-02-18 Iowa Corn Promotion Board Plantas que tienen características de rendimiento mejoradas y procedimientos para fabricarlas
US10766935B2 (en) 2015-12-28 2020-09-08 Evogene Ltd. Plant traits conferred by isolated polynucleotides and polypeptides
CN109956998B (zh) * 2017-12-26 2021-12-10 南京农业大学 水稻水通道蛋白编码基因OsNIP1；1的应用
CN111825755B (zh) * 2020-07-27 2021-09-10 江苏省农业科学院 吸收转运新烟碱类杀虫剂的质膜内在水通道蛋白及其编码基因与应用
CN111777673B (zh) * 2020-07-27 2021-08-31 江苏省农业科学院 一种吸收转运新烟碱类杀虫剂的质膜内在水通道蛋白及其编码基因与应用
CN113218899A (zh) * 2021-05-06 2021-08-06 塔里木大学 基于光谱特征响应的黄萎病和缺氮胁迫的鉴别区分方法
CN114853859B (zh) * 2022-04-29 2023-04-21 安徽农业大学 茶树水通道蛋白基因CsAQP95及其应用
CN117821471A (zh) * 2023-08-17 2024-04-05 石河子大学 一种天山雪莲的SiPIP1；5B抗逆基因片段、表达载体及应用

Citations (60)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4943674A (en) * 1987-05-26 1990-07-24 Calgene, Inc. Fruit specific transcriptional factors
US5296462A (en) * 1992-11-19 1994-03-22 Board Of Trustees Operating Michigan State University Method and compositions using polypeptides of arabidopsis thaliana
US5356816A (en) * 1991-11-19 1994-10-18 Board Of Trustees Operating Michigan State University Method and compositions using polypeptides of arabidopsis thaliana
US5495070A (en) * 1988-10-04 1996-02-27 Agracetus, Inc. Genetically engineering cotton plants for altered fiber
US5504200A (en) * 1983-04-15 1996-04-02 Mycogen Plant Science, Inc. Plant gene expression
US5521708A (en) * 1992-11-25 1996-05-28 Canon Information & Systems, Inc. Correlated color temperature detector
US5589330A (en) * 1994-07-28 1996-12-31 Genzyme Corporation High-throughput screening method for sequence or genetic alterations in nucleic acids using elution and sequencing of complementary oligonucleotides
US5597718A (en) * 1988-10-04 1997-01-28 Agracetus Genetically engineering cotton plants for altered fiber
US5608152A (en) * 1986-07-31 1997-03-04 Calgene, Inc. Seed-specific transcriptional regulation
US5880100A (en) * 1995-08-31 1999-03-09 Research Development Corporation Of Japan Method for inhibiting binding of human lens cells
US5961466A (en) * 1995-01-03 1999-10-05 Omnicorder Technologies, Inc. Method of detection of cancerous lesions by their effect on the spatial distribution of modulation of temperature and homogeneity of tissue
US6080914A (en) * 1997-01-21 2000-06-27 Monsanto Company Strawberry promoters and genes
US6084153A (en) * 1996-02-14 2000-07-04 The Governors Of The University Of Alberta Plants having enhanced nitrogen assimilation/metabolism
US6094198A (en) * 1994-01-10 2000-07-25 Cognitens, Ltd. System and method for reconstructing surface elements of solid objects in a three-dimensional scene from a plurality of two dimensional images of the scene
US6167151A (en) * 1996-12-15 2000-12-26 Cognitens, Ltd. Apparatus and method for 3-dimensional surface geometry reconstruction
US6201541B1 (en) * 1997-12-11 2001-03-13 Cognitens, Ltd. System and method for “Stitching” a plurality of reconstructions of three-dimensional surface features of object(s) in a scene defined relative to respective coordinate systems to relate them to a common coordinate system
US20010046316A1 (en) * 2000-02-21 2001-11-29 Naoki Miyano Image synthesis apparatus
US6359196B1 (en) * 1999-09-23 2002-03-19 Finn Lok Germination-specific plant promoters
US20020046419A1 (en) * 1998-08-25 2002-04-18 Yen Choo Regulated gene expression in plants
US20020049999A1 (en) * 1999-07-05 2002-04-25 Allen Randy Dale Cotton plants with improved cotton fiber characteristics and method for producing cotton fibers from these cotton plants
US6392122B1 (en) * 1999-04-30 2002-05-21 Agritope, Inc. Apple promoters for expression of transgenes in plants
US6403862B1 (en) * 1999-09-24 2002-06-11 Pioneer Hi-Bred International, Inc. Seed-preferred promoter from maize
US6442419B1 (en) * 2000-09-20 2002-08-27 Industrial Technology Research Institute Infrared 3D scanning system
US6472588B1 (en) * 1999-09-10 2002-10-29 Texas Tech University Transgenic cotton plants with altered fiber characteristics transformed with a sucrose phosphate synthase nucleic acid
US20020160378A1 (en) * 2000-08-24 2002-10-31 Harper Jeffrey F. Stress-regulated genes of plants, transgenic plants containing same, and methods of use
US20030005485A1 (en) * 2000-12-01 2003-01-02 Ohlrogge John B. Plant seed specific promoters
US20030084485A1 (en) * 2001-09-06 2003-05-01 The Arizona Board Of Regents Method for increasing stress tolerance in plants
US20030163839A1 (en) * 1997-08-27 2003-08-28 Pioneer Hi-Bred International, Inc. Genes encoding enzymes for lignin biosynthesis and uses thereof
US20030162294A1 (en) * 1998-08-04 2003-08-28 Cropdesign N.V. Genes involved in tolerance to environmental stress
US20030233670A1 (en) * 2001-12-04 2003-12-18 Edgerton Michael D. Gene sequences and uses thereof in plants
US6670528B1 (en) * 1998-10-14 2003-12-30 Independent Administrative Institute, Japan International Research Center For Agricultural Sciences Environmental stress-tolerant plants
US6701081B1 (en) * 2000-06-06 2004-03-02 Air Controls, Inc. Dual camera mount for stereo imaging
US6720477B2 (en) * 2000-04-07 2004-04-13 Basf Plant Science Gmbh Signal transduction stress-related proteins and methods of use in plants
US20040123343A1 (en) * 2000-04-19 2004-06-24 La Rosa Thomas J. Rice nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US6765607B2 (en) * 2001-04-09 2004-07-20 Matsushita Electric Industrial Co., Ltd. Driving aiding system
US6801257B2 (en) * 2001-01-12 2004-10-05 Cognitens Ltd. Optical three-dimensional digital imaging and mensuration system for industrial applications
US20040236225A1 (en) * 2001-04-16 2004-11-25 Murphy John C. Method for imaging and spectroscopy of tumors and determination of the efficacy of anti-tumor drug therapies
US6850862B1 (en) * 1999-10-22 2005-02-01 Meta Instruments Srl Methods and equipment for the measurement of the three-dimensional distribution of the temperatures within dielectric means
US20050096515A1 (en) * 2003-10-23 2005-05-05 Geng Z. J. Three-dimensional surface image guided adaptive therapy system
US20050108791A1 (en) * 2001-12-04 2005-05-19 Edgerton Michael D. Transgenic plants with improved phenotypes
US6965690B2 (en) * 2000-11-22 2005-11-15 Sanyo Electric Co., Ltd. Three-dimensional modeling apparatus, method, and medium, and three-dimensional shape data recording apparatus, method, and medium
US20060048240A1 (en) * 2004-04-01 2006-03-02 Nickolai Alexandrov Sequence-determined DNA fragments and corresponding polypeptides encoded thereby
US20060101543A1 (en) * 1994-09-26 2006-05-11 Monsanto Company, Inc. Production of hydroxylated fatty acids in genetically modified plants
US20060107345A1 (en) * 2003-09-30 2006-05-18 Nickolai Alexandrov Sequence-determined DNA fragments and corresponding polypeptides encoded thereby
US20060123505A1 (en) * 2002-05-30 2006-06-08 National Institute Of Agrobiological Sciences Full-length plant cDNA and uses thereof
US20060137043A1 (en) * 2003-04-15 2006-06-22 Basf Plant Science Gmbh Nucleic acid sequences encoding proteins associated with abiotic stress response and plant cells and plants with increased tolerance to environmental stress
US20060168684A1 (en) * 2002-05-08 2006-07-27 Basf Plant Science Gmbh Methods for increasing oil content in plants
US20060174373A1 (en) * 2002-07-10 2006-08-03 Basf Plant Science Gmbh Use of a gene for increasing the oil content in plants
US20060179511A1 (en) * 2004-12-21 2006-08-10 Chomet Paul S Transgenic plants with enhanced agronomic traits
US20060195943A1 (en) * 2004-12-08 2006-08-31 Ceres, Inc. Nucleotide sequences and corresponding polypeptides conferring modulated plant size and biomass in plants
US20060206961A1 (en) * 2003-04-16 2006-09-14 Basf Plant Science Gmbh Use of genes for increasing the oil content in plants
US20070006345A1 (en) * 2004-06-30 2007-01-04 Ceres, Inc. Nucleotide sequences and corresponding polypeptides conferring modulated plant growth rate and biomass in plants
US20070006346A1 (en) * 2004-06-30 2007-01-04 Ceres, Inc. Nucleotide sequences and corresponding polypeptides conferring modulated plant growth rate and biomass in plants
US20070044171A1 (en) * 2000-12-14 2007-02-22 Kovalic David K Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US20070061916A1 (en) * 2001-05-07 2007-03-15 Kovalic David K Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US20070169219A1 (en) * 2006-01-13 2007-07-19 Greg Nadzan Nucleotide sequences and corresponding polypeptides conferring improved nitrogen use efficiency characteristics in plants
US7292719B2 (en) * 2004-07-07 2007-11-06 Real Imaging Ltd. System and method for imaging
US20080076179A1 (en) * 2001-03-16 2008-03-27 Basf Plant Science Gmbh Sugar and lipid metabolism regulators in plants
US20100319088A1 (en) * 2007-07-24 2010-12-16 Gil Ronen Polynucleotides, polypeptides encoded thereby, and methods of using same for increasing abiotic stress tolerance and/or biomass and/or yield in plants expressing same
US20110080674A1 (en) * 2009-10-02 2011-04-07 Joel Durand Magnetic azimuth adjustment for tonearm

Family Cites Families (138)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
NL154600B (nl) 1971-02-10 1977-09-15 Organon Nv Werkwijze voor het aantonen en bepalen van specifiek bindende eiwitten en hun corresponderende bindbare stoffen.
NL154598B (nl) 1970-11-10 1977-09-15 Organon Nv Werkwijze voor het aantonen en bepalen van laagmoleculire verbindingen en van eiwitten die deze verbindingen specifiek kunnen binden, alsmede testverpakking.
NL154599B (nl) 1970-12-28 1977-09-15 Organon Nv Werkwijze voor het aantonen en bepalen van specifiek bindende eiwitten en hun corresponderende bindbare stoffen, alsmede testverpakking.
US3901654A (en) 1971-06-21 1975-08-26 Biological Developments Receptor assays of biologically active compounds employing biologically specific receptors
US3853987A (en) 1971-09-01 1974-12-10 W Dreyer Immunological reagent and radioimmuno assay
US3867517A (en) 1971-12-21 1975-02-18 Abbott Lab Direct radioimmunoassay for antigens and their antibodies
NL171930C (nl) 1972-05-11 1983-06-01 Akzo Nv Werkwijze voor het aantonen en bepalen van haptenen, alsmede testverpakkingen.
US3850578A (en) 1973-03-12 1974-11-26 H Mcconnell Process for assaying for biologically active molecules
US3935074A (en) 1973-12-17 1976-01-27 Syva Company Antibody steric hindrance immunoassay with two antibodies
US3996345A (en) 1974-08-12 1976-12-07 Syva Company Fluorescence quenching with immunological pairs in immunoassays
US4034074A (en) 1974-09-19 1977-07-05 The Board Of Trustees Of Leland Stanford Junior University Universal reagent 2-site immunoradiometric assay using labelled anti (IgG)
US3984533A (en) 1975-11-13 1976-10-05 General Electric Company Electrophoretic method of detecting antigen-antibody reaction
US4098876A (en) 1976-10-26 1978-07-04 Corning Glass Works Reverse sandwich immunoassay
US4879219A (en) 1980-09-19 1989-11-07 General Hospital Corporation Immunoassay utilizing monoclonal high affinity IgM antibodies
CA1192510A (en) 1981-05-27 1985-08-27 Lawrence E. Pelcher Rna plant virus vector or portion thereof, a method of construction thereof, and a method of producing a gene derived product therefrom
JPS6054684A (ja) 1983-09-05 1985-03-29 Teijin Ltd 新規ｄνａ及びハイブリツドｄνａ
US5011771A (en) 1984-04-12 1991-04-30 The General Hospital Corporation Multiepitopic immunometric assay
US4666828A (en) 1984-08-15 1987-05-19 The General Hospital Corporation Test for Huntington's disease
US4945050A (en) 1984-11-13 1990-07-31 Cornell Research Foundation, Inc. Method for transporting substances into living cells and tissues and apparatus therefor
CA1288073C (en) 1985-03-07 1991-08-27 Paul G. Ahlquist Rna transformation vector
US4683202A (en) 1985-03-28 1987-07-28 Cetus Corporation Process for amplifying nucleic acid sequences
US4801531A (en) 1985-04-17 1989-01-31 Biotechnology Research Partners, Ltd. Apo AI/CIII genomic polymorphisms predictive of atherosclerosis
US5569597A (en) 1985-05-13 1996-10-29 Ciba Geigy Corp. Methods of inserting viral DNA into plant material
GB8608850D0 (en) 1986-04-11 1986-05-14 Diatech Ltd Packaging system
JPS6314693A (ja) 1986-07-04 1988-01-21 Sumitomo Chem Co Ltd 植物ウイルスｒｎａベクタ−
US5268463A (en) 1986-11-11 1993-12-07 Jefferson Richard A Plant promoter α-glucuronidase gene construct
US5608142A (en) 1986-12-03 1997-03-04 Agracetus, Inc. Insecticidal cotton plants
DE3850683T2 (de) 1987-02-09 1994-10-27 Lubrizol Genetics Inc Hybrides RNS-Virus.
US5316931A (en) 1988-02-26 1994-05-31 Biosource Genetics Corp. Plant viral vectors having heterologous subgenomic promoters for systemic expression of foreign genes
US5693507A (en) 1988-09-26 1997-12-02 Auburn University Genetic engineering of plant chloroplasts
US5272057A (en) 1988-10-14 1993-12-21 Georgetown University Method of detecting a predisposition to cancer by the use of restriction fragment length polymorphism of the gene for human poly (ADP-ribose) polymerase
US5302523A (en) 1989-06-21 1994-04-12 Zeneca Limited Transformation of plant cells
US6329570B1 (en) 1989-07-19 2001-12-11 Calgene, Llc Cotton modification using ovary-tissue transcriptional factors
US5192659A (en) 1989-08-25 1993-03-09 Genetype Ag Intron sequence analysis method for detection of adjacent and remote locus alleles as haplotypes
US5859330A (en) 1989-12-12 1999-01-12 Epitope, Inc. Regulated expression of heterologous genes in plants and transgenic fruit with a modified ripening phenotype
ES2187497T3 (es) 1990-04-12 2003-06-16 Syngenta Participations Ag Promotores preferentemente en tejidos.
US5498830A (en) 1990-06-18 1996-03-12 Monsanto Company Decreased oil content in plant seeds
US5187267A (en) 1990-06-19 1993-02-16 Calgene, Inc. Plant proteins, promoters, coding sequences and use
US5399680A (en) 1991-05-22 1995-03-21 The Salk Institute For Biological Studies Rice chitinase promoter
US5604121A (en) 1991-08-27 1997-02-18 Agricultural Genetics Company Limited Proteins with insecticidal properties against homopteran insects and their use in plant protection
ZA927576B (en) 1991-10-04 1993-04-16 Univ North Carolina State Pathogen-resistant transgenic plants.
UA48104C2 (uk) 1991-10-04 2002-08-15 Новартіс Аг Фрагмент днк, який містить послідовність,що кодує інсектицидний протеїн, оптимізовану для кукурудзи,фрагмент днк, який забезпечує направлену бажану для серцевини стебла експресію зв'язаного з нею структурного гена в рослині, фрагмент днк, який забезпечує специфічну для пилку експресію зв`язаного з нею структурного гена в рослині, рекомбінантна молекула днк, спосіб одержання оптимізованої для кукурудзи кодуючої послідовності інсектицидного протеїну, спосіб захисту рослин кукурудзи щонайменше від однієї комахи-шкідника
US5281521A (en) 1992-07-20 1994-01-25 The Trustees Of The University Of Pennsylvania Modified avidin-biotin technique
ZA939767B (en) 1993-01-21 1994-09-14 Univ North Carolina State Nematode-resistant transgenic plants
EP0670670A4 (en) 1993-09-30 1996-04-24 Agracetus HETEROLOGICAL PEROXIDASE PROCESSING TRANSGENIC COTTON PLANTS.
US5608144A (en) 1994-08-12 1997-03-04 Dna Plant Technology Corp. Plant group 2 promoters and uses thereof
US5659026A (en) 1995-03-24 1997-08-19 Pioneer Hi-Bred International ALS3 promoter
EP0835311A2 (en) 1995-06-07 1998-04-15 Calgene, Inc. Cotton fiber transcriptional factors
JPH1094392A (ja) 1996-09-20 1998-04-14 Nisshinbo Ind Inc ワタ遺伝子
KR19990076740A (ko) 1996-10-24 1999-10-15 미즈노 마사루 식물체 수분함량 조절 방법
JPH10229883A (ja) * 1997-02-24 1998-09-02 Japan Tobacco Inc 植物水ポテンシャル維持機能の改良法およびこれに用いる水チャンネルタンパク質遺伝子
US20090093620A1 (en) 2000-09-05 2009-04-09 David Kovalic Annotated Plant Genes
US6313375B1 (en) 1998-08-13 2001-11-06 Pioneer Hi-Bred International, Inc. Maize aquaporins and uses thereof
US6313376B1 (en) 1998-08-14 2001-11-06 Pioneer Hi-Bred International, Inc. Maize aquaporins and uses thereof
US7511190B2 (en) 1999-11-17 2009-03-31 Mendel Biotechnology, Inc. Polynucleotides and polypeptides in plants
US6717034B2 (en) 2001-03-30 2004-04-06 Mendel Biotechnology, Inc. Method for modifying plant biomass
EP1033405A3 (en) 1999-02-25 2001-08-01 Ceres Incorporated Sequence-determined DNA fragments and corresponding polypeptides encoded thereby
US20100293669A2 (en) 1999-05-06 2010-11-18 Jingdong Liu Nucleic Acid Molecules and Other Molecules Associated with Plants and Uses Thereof for Plant Improvement
US20110214206A1 (en) 1999-05-06 2011-09-01 La Rosa Thomas J Nucleic acid molecules and other molecules associated with plants
US20040031072A1 (en) * 1999-05-06 2004-02-12 La Rosa Thomas J. Soy nucleic acid molecules and other molecules associated with transcription plants and uses thereof for plant improvement
US8877916B2 (en) 2000-04-26 2014-11-04 Ceres, Inc. Promoter, promoter control elements, and combinations, and uses thereof
WO2001006006A1 (fr) 1999-07-19 2001-01-25 Japan Science And Technology Corporation Gene de resistance au stress ambiant
CA2361679A1 (en) 1999-09-27 2001-04-05 Dutt V. Vinjamoori Methods for determining oils in seeds
US6407315B1 (en) 1999-11-02 2002-06-18 Pioneer Hi-Bred International, Inc. Seed-preferred promoter from barley
US6828476B1 (en) 1999-12-02 2004-12-07 The Regents Of The University Of California Cotton transcription factors and their uses
US7834146B2 (en) 2000-05-08 2010-11-16 Monsanto Technology Llc Recombinant polypeptides associated with plants
US20020170088A1 (en) 2000-11-03 2002-11-14 The Regents Of The University Of California Novel auxin binding proteins and uses thereof
BR0115782A (pt) 2000-12-08 2004-01-20 Commonwealh Scient And Ind Res Modificação de expressão de gene de sacarose sintase em tecido de planta e usos
WO2002090557A2 (en) 2001-05-03 2002-11-14 Vlaams Interuniversitair Instituut Voor Biotechnologie Vzw Freeze-tolerant eukaryotic cells
AU2002332795A1 (en) 2001-08-31 2003-03-18 The Dow Chemical Company Nucleic acid compositions conferring insect control in plants
CA2481504C (en) 2002-04-08 2011-08-23 Pioneer Hi-Bred International, Inc. Enhanced silk exsertion under stress
US20030221218A1 (en) 2002-05-17 2003-11-27 The Regents Of The University Of California Bioengineering cotton fiber properties
US7847156B2 (en) 2003-10-20 2010-12-07 Cropdesign N.V. Plants having improved growth characteristics and methods for making the same
AU2002351325A1 (en) 2002-12-04 2004-06-30 G. Banu Transgenic maize with enhanced phenotype
WO2004058963A2 (en) 2002-12-31 2004-07-15 University Of Delhi A novel gene osisap1 of rice confers tolerance to stresses and a method thereof
WO2004077010A2 (en) 2003-02-03 2004-09-10 The Scripps Research Institute Water channel assays
BRPI0408735A (pt) 2003-03-12 2006-03-07 Evogene Ltd polinucleotìdeo isolado, construção de ácido nucléico, célula transgênica, organismo transgênico, vegetal transgênico, método para produzir um vegetal transgênico, método para expressar um polinucleotìdeo de interesse em uma célula, e método para co-expressar dois polinucleotìdeos de interesse em uma célula
BR122016024209B1 (pt) 2003-05-22 2019-08-20 Evogene Ltd. Método para aumentar a biomassa e/ou tolerância de uma planta ao estresse salino e construto de ácido nucleico
US7554007B2 (en) 2003-05-22 2009-06-30 Evogene Ltd. Methods of increasing abiotic stress tolerance and/or biomass in plants
AU2005234725B2 (en) 2003-05-22 2012-02-23 Evogene Ltd. Methods of Increasing Abiotic Stress Tolerance and/or Biomass in Plants and Plants Generated Thereby
JP3819377B2 (ja) 2003-06-18 2006-09-06 株式会社東芝 半導体集積回路の静電放電の解析方法
WO2004111183A2 (en) 2003-06-19 2004-12-23 Evogene Ltd. Plant trichome-specific promoter and leucoplast signal sequence
JP4452876B2 (ja) 2003-08-06 2010-04-21 国立大学法人 香川大学 ＬＫＰ２部分ｃＤＮＡを用いた遺伝子導入による植物体の種子収量、乾燥重量の制御
US7989676B2 (en) 2006-08-31 2011-08-02 Ceres, Inc. Nucleotide sequences and corresponding polypeptides conferring modulated plant characteristics
US20060150283A1 (en) 2004-02-13 2006-07-06 Nickolai Alexandrov Sequence-determined DNA fragments and corresponding polypeptides encoded thereby
US20060143729A1 (en) 2004-06-30 2006-06-29 Ceres, Inc. Nucleotide sequences and polypeptides encoded thereby useful for modifying plant characteristics
WO2005084331A2 (en) 2004-03-01 2005-09-15 Syngenta Participations Ag Sorghum gene expression profiling
AU2005229157B2 (en) 2004-03-31 2011-07-21 Commonwealth Scientific And Industrial Research Organisation Genes involved in plant fibre development
US8049069B2 (en) 2004-03-31 2011-11-01 Commonwealth Scientific And Industrial Research Organisation Genes involved in plant fibre development
CA2563943A1 (en) 2004-04-23 2005-11-03 Ceres, Inc. Nucleotide sequences and polypeptides encoded thereby useful for modifying nitrogen use efficiency characteristics in plants
CA2564202A1 (en) 2004-05-05 2005-11-17 The Royal Veterinary And Agricultural University Ammonium/ammonia transporter
EP2186899A3 (en) 2004-06-14 2010-09-08 Evogene Ltd. Polynucleotides and polypeptides involved in plant fiber development and methods of using same
BRPI0517294A (pt) 2004-10-22 2008-10-07 Agrinomics Llc geração de plantas com teor de óleo alterado
US20080148432A1 (en) 2005-12-21 2008-06-19 Mark Scott Abad Transgenic plants with enhanced agronomic traits
AU2006204997B2 (en) * 2005-01-12 2011-09-01 Monsanto Technology, Llc Genes and uses for plant improvement
EP1681128A1 (de) 2005-01-14 2006-07-19 Siemens Aktiengesellschaft Verfahren zur Herstellung eines Lochs und Vorrichtung
EP1882392A4 (en) 2005-05-10 2009-07-01 Monsanto Technology Llc GENES AND THEIR USES FOR PLANT IMPROVEMENTS
CN101442903A (zh) 2005-05-26 2009-05-27 孟山都技术有限公司 单子叶植物中油的提高
WO2006138012A1 (en) 2005-06-17 2006-12-28 Ceres Inc. P450 substrates and methods related thereto
US20080301839A1 (en) 2005-08-30 2008-12-04 Ravanello Monica P Transgenic plants with enhanced agronomic traits
RU2487166C2 (ru) 2005-10-24 2013-07-10 Эвоген Лтд Изолированные полипептиды, кодирующие их полинуклеотиды, экспрессирующие их трансгенные растения и способы их использования
CN1840664A (zh) * 2005-10-31 2006-10-04 上海大学 受高盐诱导的盐藻基因片段和钠／磷共转运通道基因
EP2343375A3 (en) 2006-03-24 2012-02-08 BASF Plant Science GmbH Proteins associated with abiotic stress response and homologs
EP2199398A1 (en) 2006-03-31 2010-06-23 BASF Plant Science GmbH Plants having enhanced yield-related traits and a method for making the same
CN100385000C (zh) * 2006-06-08 2008-04-30 江苏省农业科学院 一种真盐生植物细胞质膜水通道蛋白基因的应用
US8362322B2 (en) 2006-10-27 2013-01-29 Ceres, Inc. Modulating lignin in plants
EP2383345B1 (en) 2006-12-20 2017-11-22 Evogene Ltd. Polynucleotides and polypeptides involved in plant fiber development and methods of using same
MX341624B (es) 2007-04-09 2016-08-26 Evogene Ltd Polinucleotidos, polipeptidos y metodos para aumentar el contenido de aceite, la velocidad de crecimiento y biomasa de las plantas.
EP2573178A3 (en) 2007-07-10 2013-07-24 Monsanto Technology LLC Transgenic plants with enhanced agronomic traits
US8362325B2 (en) 2007-10-03 2013-01-29 Ceres, Inc. Nucleotide sequences and corresponding polypeptides conferring modulated plant characteristics
MX357387B (es) 2007-12-27 2018-07-06 Evogene Ltd Polipeptidos, polinucleotidos aislados utiles para modificar la eficacia en el uso de agua, eficacia en el uso del fertilizante, la tolerancia al estres biotico/abiotico, el rendimiento y biomasa en plantas.
CA3148194A1 (en) 2008-05-22 2009-11-26 Evogene Ltd. Isolated polynucleotides and peptides and methods of using same for increasing plant yield, biomass, growth rate, vigor, oil content, abiotic stress tolerance of plants and nitrogen use efficiency
WO2009144311A1 (en) 2008-05-29 2009-12-03 Vib Vzw Minichromosome maintenance complex interacting protein involved in cancer
BR122021014170B1 (pt) 2008-08-18 2022-08-09 Evogene Ltd Método para aumentar a eficiência de uso de nitrogênio, eficiência de uso de fertilizantes, rendimento, taxa de crescimento, vigor, biomassa e/ou tolerância ao estresse abiótico de uma planta
MX345298B (es) 2008-10-30 2017-01-24 Evogene Ltd Polinucleótidos y polipéptidos aislados y métodos para utilizarlos para aumentar el rendimiento de la planta, biomasa, tasa de crecimiento, vigor, contenido de aceite, tolerancia al estrés abiótico de las plantas y eficacia en el uso de nitrógeno.
AR074925A1 (es) 2008-12-29 2011-02-23 Evogene Ltd Polinucleotidos, polipeptidos codificados, y metodos para utilizarlos para modular la tolerancia al estres abiotico, biomasa y/o rendimiento en plantas que los expresan
CA2753616C (en) 2009-03-02 2018-07-17 Evogene Ltd. Isolated polynucleotides and polypeptides, and methods of using same for increasing plant yield and/or agricultural characteristics
EP3238534B1 (en) 2009-06-10 2019-11-13 Evogene Ltd. Isolated polynucleotides and polypeptides, and methods of using same for increasing nitrogen use efficiency, yield, growth rate, vigor, biomass, oil content, and/or abiotic stress tolerance
BR112012002543B1 (pt) 2009-08-04 2021-03-09 Evogene Ltd. método para aumentar a tolerância ao estresse abiótico, o rendimento, a biomassa, taxa de crescimento, e/ou vigor de uma planta, e, construto de ácido nucléico isolado
EP2519097B1 (en) 2009-12-28 2016-03-02 Evogene Ltd. Isolated polynucleotides and polypeptides and methods of using same for increasing plant yield, biomass, growth rate, vigor, oil content, abiotic stress tolerance of plants and nitrogen use efficiency
EP2563112A4 (en) 2010-04-28 2014-03-05 Evogene Ltd INSULATED POLYNUCLEOTIDES AND POLYPEPTIDES AND METHODS OF USING THE SAME FOR INCREASING PLANT YIELD AND / OR AGRICULTURAL CHARACTERISTICS
AR082530A1 (es) 2010-08-30 2012-12-12 Evogene Ltd Polinucleotidos y polipeptidos aislados, y metodos para utilizarlos para aumentar la eficacia en el uso de nitrogeno, rendimiento, tasa de crecimiento, vigor, biomasa, contenido de aceite y/o tolerancia al estres abiotico
MX354193B (es) 2010-12-22 2018-02-16 Evogene Ltd Polinucleótidos y polipéptidos aislados, y métodos para utilizarlos para aumentar la tolerancia al estrés abiótico, rendimiento, tasa de crecimiento, vigor, biomasa, contenido de aceite y/o eficacia en el uso de nitrógeno de las plantas.
CA3139245A1 (en) 2011-05-03 2012-11-08 Evogene Ltd. Isolated polynucleotides and polypeptides and methods of using same for increasing plant yield, biomass, growth rate, vigor, oil content, abiotic stress tolerance of plants and nitrogen use efficiency
CA3177736A1 (en) 2011-08-23 2013-02-28 Evogene Ltd. Isolated polynucleotides and polypeptides, and methods of using same for increasing plant yield and/or agricultural characteristics
CN104254242A (zh) 2011-11-21 2014-12-31 先正达参股股份有限公司 用于增强植物中的线虫抗性的组合物和方法
BR122019023079B1 (pt) 2011-11-28 2021-05-25 Evogene Ltd método para aumento da eficiência no uso de nitrogênio, produção, biomassa, taxa de crescimento, vigor, e/ou tolerância ao estresse abiótico de uma planta, e, construto de ácido nucleico isolado
BR122020022832B1 (pt) 2011-12-28 2021-08-17 Evogene Ltd Método para aumentar a produção, taxa de crescimento, biomassa, energia e/ou produção de semente de uma planta em comparação com uma planta nativa, e, construção de ácido nucleico isolado
MX390593B (es) 2012-02-29 2025-03-20 Evogene Ltd Polinucleótidos y polipéptidos aislados y métodos para utilizarlos para mejorar el rendimiento de la planta, biomasa, tasa de crecimiento, vigor, contenido de aceite, tolerancia al estrés abiótico de las plantas y eficacia en el uso de nitrógeno.
AR092839A1 (es) 2012-05-28 2015-05-06 Evogene Ltd Polinucleotidos y polipeptidos aislados y metodos para utilizarlos para mejorar el rendimiento y/o caracteristicas agricolas de las plantas
BR122019028113B1 (pt) 2012-08-27 2022-08-30 Evogene Ltd Métodos para aumentar o rendimento, taxa de crescimento, biomassa, vigor, capacidade fotossintética, eficiência do uso de nitrogênio, tolerância ao estresse abiótico e/ou redução do tempo de floração ou o tempo de emergência da inflorescência de uma planta, e de cultivar uma colheita, e, construto de ácido nucleico
WO2014102773A1 (en) 2012-12-25 2014-07-03 Evogene Ltd. Isolated polynucleotides and polypeptides, and methods of using same for increasing nitrogen use efficiency of plants
BR122020005657B1 (pt) 2012-12-26 2022-04-05 Evogene Ltd Método para aumentar a eficiência do uso de nitrogênio, rendimento, taxa de crescimento, biomassa, vigor, rendimento de sementes, capacidade fotossintética e/ou tolerância ao estresse abiótico de uma planta, e, construção de ácido nucleico
WO2014188428A1 (en) 2013-05-22 2014-11-27 Evogene Ltd. Isolated polynucleotides and polypeptides, and methods of using same for increasing plant yield and/or agricultural characteristics
CA2916060A1 (en) 2013-08-27 2015-03-05 Evogene Ltd. Isolated polynucleotides and polypeptides, and methods of using same for increasing plant yield and/or agricultural characteristics
WO2015181823A1 (en) 2014-05-28 2015-12-03 Evogene Ltd. Isolated polynucleotides, polypeptides and methods of using same for increasing abiotic stress tolerance, biomass and yield of plants
CN104197792B (zh) 2014-08-22 2015-10-07 哈尔滨工业大学 一类多平衡点饱和切换系统的离散增益调度控制器设计方法及实现btt导弹控制的方法
CA3176207A1 (en) 2014-08-27 2016-03-03 Evogene Ltd. Constructs and methods for increasing plant yield or agricultural characteristics or both

Patent Citations (65)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5504200A (en) * 1983-04-15 1996-04-02 Mycogen Plant Science, Inc. Plant gene expression
US5608152A (en) * 1986-07-31 1997-03-04 Calgene, Inc. Seed-specific transcriptional regulation
US4943674A (en) * 1987-05-26 1990-07-24 Calgene, Inc. Fruit specific transcriptional factors
US5981834A (en) * 1988-10-04 1999-11-09 Monsanto Company Genetically engineering cotton plants for altered fiber
US5495070A (en) * 1988-10-04 1996-02-27 Agracetus, Inc. Genetically engineering cotton plants for altered fiber
US5597718A (en) * 1988-10-04 1997-01-28 Agracetus Genetically engineering cotton plants for altered fiber
US5620882A (en) * 1988-10-04 1997-04-15 Agracetus, Inc. Genetically engineering cotton plants for altered fiber
US5356816A (en) * 1991-11-19 1994-10-18 Board Of Trustees Operating Michigan State University Method and compositions using polypeptides of arabidopsis thaliana
US5296462A (en) * 1992-11-19 1994-03-22 Board Of Trustees Operating Michigan State University Method and compositions using polypeptides of arabidopsis thaliana
US5521708A (en) * 1992-11-25 1996-05-28 Canon Information & Systems, Inc. Correlated color temperature detector
US6094198A (en) * 1994-01-10 2000-07-25 Cognitens, Ltd. System and method for reconstructing surface elements of solid objects in a three-dimensional scene from a plurality of two dimensional images of the scene
US5589330A (en) * 1994-07-28 1996-12-31 Genzyme Corporation High-throughput screening method for sequence or genetic alterations in nucleic acids using elution and sequencing of complementary oligonucleotides
US20060101543A1 (en) * 1994-09-26 2006-05-11 Monsanto Company, Inc. Production of hydroxylated fatty acids in genetically modified plants
US5961466A (en) * 1995-01-03 1999-10-05 Omnicorder Technologies, Inc. Method of detection of cancerous lesions by their effect on the spatial distribution of modulation of temperature and homogeneity of tissue
US5880100A (en) * 1995-08-31 1999-03-09 Research Development Corporation Of Japan Method for inhibiting binding of human lens cells
US6084153A (en) * 1996-02-14 2000-07-04 The Governors Of The University Of Alberta Plants having enhanced nitrogen assimilation/metabolism
US6167151A (en) * 1996-12-15 2000-12-26 Cognitens, Ltd. Apparatus and method for 3-dimensional surface geometry reconstruction
US6080914A (en) * 1997-01-21 2000-06-27 Monsanto Company Strawberry promoters and genes
US20030163839A1 (en) * 1997-08-27 2003-08-28 Pioneer Hi-Bred International, Inc. Genes encoding enzymes for lignin biosynthesis and uses thereof
US6201541B1 (en) * 1997-12-11 2001-03-13 Cognitens, Ltd. System and method for “Stitching” a plurality of reconstructions of three-dimensional surface features of object(s) in a scene defined relative to respective coordinate systems to relate them to a common coordinate system
US20030162294A1 (en) * 1998-08-04 2003-08-28 Cropdesign N.V. Genes involved in tolerance to environmental stress
US20020046419A1 (en) * 1998-08-25 2002-04-18 Yen Choo Regulated gene expression in plants
US6670528B1 (en) * 1998-10-14 2003-12-30 Independent Administrative Institute, Japan International Research Center For Agricultural Sciences Environmental stress-tolerant plants
US6392122B1 (en) * 1999-04-30 2002-05-21 Agritope, Inc. Apple promoters for expression of transgenes in plants
US20030074697A1 (en) * 1999-07-05 2003-04-17 Allen Randy Dale Cotton plants with improved cotton fiber characteristics and method for producing cotton fibers from these cotton plants
US20020049999A1 (en) * 1999-07-05 2002-04-25 Allen Randy Dale Cotton plants with improved cotton fiber characteristics and method for producing cotton fibers from these cotton plants
US6472588B1 (en) * 1999-09-10 2002-10-29 Texas Tech University Transgenic cotton plants with altered fiber characteristics transformed with a sucrose phosphate synthase nucleic acid
US6359196B1 (en) * 1999-09-23 2002-03-19 Finn Lok Germination-specific plant promoters
US6403862B1 (en) * 1999-09-24 2002-06-11 Pioneer Hi-Bred International, Inc. Seed-preferred promoter from maize
US6850862B1 (en) * 1999-10-22 2005-02-01 Meta Instruments Srl Methods and equipment for the measurement of the three-dimensional distribution of the temperatures within dielectric means
US7072504B2 (en) * 2000-02-21 2006-07-04 Sharp Kabushiki Kaisha Image synthesis apparatus
US20010046316A1 (en) * 2000-02-21 2001-11-29 Naoki Miyano Image synthesis apparatus
US6720477B2 (en) * 2000-04-07 2004-04-13 Basf Plant Science Gmbh Signal transduction stress-related proteins and methods of use in plants
US20040123343A1 (en) * 2000-04-19 2004-06-24 La Rosa Thomas J. Rice nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US6701081B1 (en) * 2000-06-06 2004-03-02 Air Controls, Inc. Dual camera mount for stereo imaging
US20060183137A1 (en) * 2000-08-24 2006-08-17 The Scripps Research Institute Stress-regulated genes of plants, transgenic plants containing same, and methods of use
US20020160378A1 (en) * 2000-08-24 2002-10-31 Harper Jeffrey F. Stress-regulated genes of plants, transgenic plants containing same, and methods of use
US6442419B1 (en) * 2000-09-20 2002-08-27 Industrial Technology Research Institute Infrared 3D scanning system
US6965690B2 (en) * 2000-11-22 2005-11-15 Sanyo Electric Co., Ltd. Three-dimensional modeling apparatus, method, and medium, and three-dimensional shape data recording apparatus, method, and medium
US20030005485A1 (en) * 2000-12-01 2003-01-02 Ohlrogge John B. Plant seed specific promoters
US20070044171A1 (en) * 2000-12-14 2007-02-22 Kovalic David K Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US6801257B2 (en) * 2001-01-12 2004-10-05 Cognitens Ltd. Optical three-dimensional digital imaging and mensuration system for industrial applications
US20080076179A1 (en) * 2001-03-16 2008-03-27 Basf Plant Science Gmbh Sugar and lipid metabolism regulators in plants
US6765607B2 (en) * 2001-04-09 2004-07-20 Matsushita Electric Industrial Co., Ltd. Driving aiding system
US20040236225A1 (en) * 2001-04-16 2004-11-25 Murphy John C. Method for imaging and spectroscopy of tumors and determination of the efficacy of anti-tumor drug therapies
US20070061916A1 (en) * 2001-05-07 2007-03-15 Kovalic David K Nucleic acid molecules and other molecules associated with plants and uses thereof for plant improvement
US20030084485A1 (en) * 2001-09-06 2003-05-01 The Arizona Board Of Regents Method for increasing stress tolerance in plants
US20050108791A1 (en) * 2001-12-04 2005-05-19 Edgerton Michael D. Transgenic plants with improved phenotypes
US20030233670A1 (en) * 2001-12-04 2003-12-18 Edgerton Michael D. Gene sequences and uses thereof in plants
US20060168684A1 (en) * 2002-05-08 2006-07-27 Basf Plant Science Gmbh Methods for increasing oil content in plants
US20060123505A1 (en) * 2002-05-30 2006-06-08 National Institute Of Agrobiological Sciences Full-length plant cDNA and uses thereof
US20060174373A1 (en) * 2002-07-10 2006-08-03 Basf Plant Science Gmbh Use of a gene for increasing the oil content in plants
US20060137043A1 (en) * 2003-04-15 2006-06-22 Basf Plant Science Gmbh Nucleic acid sequences encoding proteins associated with abiotic stress response and plant cells and plants with increased tolerance to environmental stress
US20060206961A1 (en) * 2003-04-16 2006-09-14 Basf Plant Science Gmbh Use of genes for increasing the oil content in plants
US20060107345A1 (en) * 2003-09-30 2006-05-18 Nickolai Alexandrov Sequence-determined DNA fragments and corresponding polypeptides encoded thereby
US20050096515A1 (en) * 2003-10-23 2005-05-05 Geng Z. J. Three-dimensional surface image guided adaptive therapy system
US20060048240A1 (en) * 2004-04-01 2006-03-02 Nickolai Alexandrov Sequence-determined DNA fragments and corresponding polypeptides encoded thereby
US20070006346A1 (en) * 2004-06-30 2007-01-04 Ceres, Inc. Nucleotide sequences and corresponding polypeptides conferring modulated plant growth rate and biomass in plants
US20070006345A1 (en) * 2004-06-30 2007-01-04 Ceres, Inc. Nucleotide sequences and corresponding polypeptides conferring modulated plant growth rate and biomass in plants
US7292719B2 (en) * 2004-07-07 2007-11-06 Real Imaging Ltd. System and method for imaging
US20060195943A1 (en) * 2004-12-08 2006-08-31 Ceres, Inc. Nucleotide sequences and corresponding polypeptides conferring modulated plant size and biomass in plants
US20060179511A1 (en) * 2004-12-21 2006-08-10 Chomet Paul S Transgenic plants with enhanced agronomic traits
US20070169219A1 (en) * 2006-01-13 2007-07-19 Greg Nadzan Nucleotide sequences and corresponding polypeptides conferring improved nitrogen use efficiency characteristics in plants
US20100319088A1 (en) * 2007-07-24 2010-12-16 Gil Ronen Polynucleotides, polypeptides encoded thereby, and methods of using same for increasing abiotic stress tolerance and/or biomass and/or yield in plants expressing same
US20110080674A1 (en) * 2009-10-02 2011-04-07 Joel Durand Magnetic azimuth adjustment for tonearm

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
Aharon et al, The Plant Cell. Vol. 15, 439-447, 2003 *
Davletona et al , Plant Physiology, October 2005, Vol 139, pp. 847-856 *
Friedberg (Brief Bioinformatics (2006) 7: 225-242) *

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10953581B2 (en) 2013-05-29 2021-03-23 Synventive Molding Solutions, Inc. Actuator cooling apparatus and method
US9101100B1 (en) 2014-04-30 2015-08-11 Ceres, Inc. Methods and materials for high throughput testing of transgene combinations

Also Published As

Publication number Publication date
US20130219562A1 (en) 2013-08-22
AU2008344935A1 (en) 2009-07-09
WO2009083958A8 (en) 2009-10-15
ZA201004460B (en) 2012-01-25
WO2009083958A2 (en) 2009-07-09
US20170204428A1 (en) 2017-07-20
EP2240510A2 (en) 2010-10-20
EP2240510B1 (en) 2017-05-10
CN101977928A (zh) 2011-02-16
US20120222169A1 (en) 2012-08-30
MX357387B (es) 2018-07-06
CA2709517A1 (en) 2009-07-09
BRPI0819476A2 (pt) 2015-07-14
WO2009083958A3 (en) 2009-12-23
US10407690B2 (en) 2019-09-10
AU2008344935B2 (en) 2013-08-29
MX340504B (es) 2016-07-11
CA2709517C (en) 2019-02-26
AU2008344935C1 (en) 2016-07-14
US9670501B2 (en) 2017-06-06
US8426682B2 (en) 2013-04-23
CN101977928B (zh) 2014-12-10

Similar Documents

Publication Publication Date Title
US20220403404A1 (en) Isolated polypeptides and polynucleotides useful for increasing nitrogen use efficiency, abiotic stress tolerance, yield and biomass in plants
US10407690B2 (en) Isolated polypeptides, polynucleotides useful for modifying water user efficiency, fertilizer use efficiency, biotic/abiotic stress tolerance, yield and biomass in plants
US10961544B2 (en) Polynucleotides, polypeptides encoded thereby, and methods of using same for increasing abiotic stress tolerance and/or biomass and/or yield in plants expressing same
AU2015261690B2 (en) Isolated Polypeptides, Polynucleotides Useful for Modifying Water User Efficiency, Fertilizer Use Efficiency, Biotic/Abiotic Stress Tolerance, Yield and Biomass in Plants
AU2013263801B2 (en) Isolated Polypeptides, Polynucleotides Useful for Modifying Water User Efficiency, Fertilizer Use Efficiency, Biotic/Abiotic Stress Tolerance, Yield and Biomass in Plants
AU2014215945B2 (en) Polynucleotides, Polypeptides Encoded Thereby, and Methods of Using Same for Increasing Abiotic Stress Tolerance and/or Biomass and/or Yield in Plants Expressing Same
AU2015230753A1 (en) Polynucleotides, Polypeptides Encoded Thereby, and Methods of Using Same for Increasing Abiotic Stress Tolerance and/or Biomass and/or Yield in Plants Expressing Same

Legal Events

Date Code Title Description
AS Assignment

Owner name: EVOGENE LTD., ISRAEL

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:RONEN, GIL;VINOCUR, BASIA JUDITH;DIBER, ALEX;AND OTHERS;SIGNING DATES FROM 20100510 TO 20100620;REEL/FRAME:024736/0912

STCB Information on status: application discontinuation

Free format text: ABANDONED -- FAILURE TO RESPOND TO AN OFFICE ACTION