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EP1151119A1 - Verfahren zur hemmung der expression von zielgenen in pflanzen - Google Patents

Verfahren zur hemmung der expression von zielgenen in pflanzen

Info

Publication number
EP1151119A1
EP1151119A1 EP00915151A EP00915151A EP1151119A1 EP 1151119 A1 EP1151119 A1 EP 1151119A1 EP 00915151 A EP00915151 A EP 00915151A EP 00915151 A EP00915151 A EP 00915151A EP 1151119 A1 EP1151119 A1 EP 1151119A1
Authority
EP
European Patent Office
Prior art keywords
plant
dna construct
sequence
chimeric
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.)
Withdrawn
Application number
EP00915151A
Other languages
English (en)
French (fr)
Inventor
Wolfgang Institute für WERR
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.)
Biogemma SAS
Original Assignee
Rhobio SA
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 Rhobio SA filed Critical Rhobio SA
Priority to EP00915151A priority Critical patent/EP1151119A1/de
Publication of EP1151119A1 publication Critical patent/EP1151119A1/de
Withdrawn legal-status Critical Current

Links

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)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • C12N15/8237—Externally regulated expression systems
    • 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)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • 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)
    • C12N15/8216—Methods for controlling, regulating or enhancing expression of transgenes in plant cells
    • C12N15/8218—Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS]

Definitions

  • the term "repression domain of the Drosophila engrailed gene (eng)” means a fragment of the Drosophila engrailed gene or of a derivative sequence thereof, said fragment comprising a nucleotide sequence that encodes a polypeptide interfering with the general transcription machinery and transcriptional activators. Said fragment advantageously comprises a minimal repression sequence coding for a polypeptide of 55 residues (Poole et al., 1985, Han K. et al., 1993). The whole engrailed gene may also be used but most preferably without its homeodomain.
  • Engrailed related domains which share the same repression activity, may also be used, such as those described in Smith et al. (1996).
  • the repressor sequence that is used may be a sequence that codes for at least the Kruppel-associated box-A (KRAB-A) domain of zinc finger proteins (Witzgall et al., 1994) for at least the RE-1- silencing transcription factor (REST) (Thiel et al., 1998), or for at least the BTB (for Broad-complex Tramtrac and Brie) domain, also known as POZ-domain (Ahmad et al., 1998 ; Huynh & Bardwell, 1998).
  • KRAB-A Kruppel-associated box-A domain of zinc finger proteins
  • REST RE-1- silencing transcription factor
  • BTB for Broad-complex Tramtrac and Brie domain
  • said transcription factor may preferably be selected from the group consisting of : STM ("Shootmeristemless"), member of the Knotted class of homeodomain proteins, which is an essential gene for development and function of the shoot apical meristem ;
  • ZmHox - member of the homeobox proteins more particularly ZmHox 1a/1 b and 2a/2b, which are expressed in maize meristems and proliferating cells from the early embryo to late reproductive organs. This expression pattern suggests a contribution to plant growth and morphogenesis;
  • Arabidopsis is a transcription factor belonging the family recently described as ARF1 family for Auxin Response Factor 1 (Ulmasov et al., 1997). Ms41-A analogues may also be used, such as those described in WO 97/23618 or the Arabidopsis gene Monopteros which encodes a transcription factor mediating embryo axis formation and vascular development (Hardtke et al., 1998), and the Arabidopsis gene ETTIN involved in floral development (Sessions, 1997).
  • the plants transformed with a chimeric DNA construct of the invention comprising such Ms-41-A factor are expected to be male sterile.
  • the promoter that is used is a tissue- specific promoter or a developmentally regulated promoter. This allows a conditional loss of functions that may be desired, for example to design new plant varieties.
  • 35S promoter or advantageously the double 35S constitutive promoter of CaMV as described in Kay et al. (1987) ; rice actin promoter followed by the rice actin intron (PAR- IAR) included in plasmid pAct1-F4 as described in Mc Elroy et al. (1991); the constitutive promoter EF-1 ⁇ of the gene encoding for plant elongation factor described in WO 90/02172 or in Axelos et al.
  • HMWG promoter High Molecular Weight Glutenin from barley (Anderson et al., 1989) ; the promoter of maize ⁇ zein (P ⁇ zein) included in p ⁇ 63 plasmid in Reina et al. (1990) allowing the expression in albumen of maize seeds ;
  • PGEA1 and PGEA6 promoters corresponding to the non coding 5' region of the genes GEA1 and GEA6, expressed in the grains in Arabidopsis thaliana (Gaubier et al., 1993) ; and ⁇ -phaseolin promoter (Riggs et al., 1989). • specific promoters which drive expression in particular plant tissues which are involved in the control of fertility :
  • parts of transgenic plants refer in particular to leaves, fruits, seeds, roots or cells that have been genetically transformed.
  • the preferred plants that are used for transformation may be for example selected from the group consisting of Arabidopsis thaliana, rice, tobacco, maize, Brassica, wheat, tomato and flowers (Petunia, rose, carnation).
  • Figure 5 represents a eng-STM construct.
  • Figure 6 represents a eng-AP3 construct.
  • the homeodomain of STM was removed in an independent experiment.
  • the plasmid pRT ⁇ eng-STM was first partially digested with Hindll, secondly to completion with BamHI and ends were ligated after fill in reaction with Klenow enzyme.
  • Plants are regenerated from these callus by modifying the hormonal and osmotic equilibrium of the cells according to the method described by Vain et al, 1989.
  • a method of genetic transformation leading to a stable integration of the modified genes in the genome of the plant involves the use of a gene gun.
  • the target cells are fragments of callus having a surface area of 10 to 20 mm 2 .
  • the fragments are laid in the center of a Petri dish containing a culture medium identical to the initiation medium, further containing 0.2 M of mannitol + 0.2 M of sorbitol (16 fragments per dish). Tissues are then bombarded as previously described.
  • the dishes are then sealed using Scellofrais® and cultivated in dark at 27°C.
  • the first planting is effected 24 hours afterwards, then every two-weeks during three months in a medium identical to the initiation medium but containing a selective agent.
  • the selective agents may be for example active ingredients of herbicidal agents (Basta®, Round up®) or antibiotics (hygromycin, kanamycin).
  • Example 5 Expression of the endogenous STM and AP3 genes
  • transgenic eng-STM or eng-AP3 phenocopy plants were subjected to RNA gel blot and RT-PCR analysis.
  • the chimeric transcripts derived from the CaMV 35S promoter were easily detectable in total or poly(A) + RNA from pooled seedlings or flowers (12 individual progeny) probed with the STM or AP3 coding regions.
  • the high abundance of chimeric transcripts interfered with the detection of the shorter native mRNA in these gel blot experiments.
  • Discrimination was achieved by probing with the natural 3'UTR sequences that are lacking in the transgenes. The result obtained for the STM 3'UTR confirms transcription of the native STM gene although at a significantly reduced level.
  • Example 6 Phenocopies rely on the incorporation of the chimeric eng-STM protein into the nuclear compartment Although the changes in size or identity of the expressing organs might explain the lower transcript levels observed in stm and ap3 phenocopy plants we aimed to elaborate further evidence against homology based gene silencing.
  • the chimeric eng-STM polypeptide therefore was expressed in C- terminal fusion with the hormone-binding domain of the glucocorticoid receptor in transgenic Arabidopsis plants. Due to this addition the resulting eng-STM-GR polypeptide should be cytoplasmatic and incorporated into the nuclear compartment only after hormone application. A linkage between the dexamethasone treatment and alterations in SAM activity thus should depend on the nuclear import of the chimeric eng-STM-GR protein and be incompatible with cosuppression.

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  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Biomedical Technology (AREA)
  • Organic Chemistry (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Physics & Mathematics (AREA)
  • Cell Biology (AREA)
  • Plant Pathology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biophysics (AREA)
  • Virology (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Peptides Or Proteins (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
EP00915151A 1999-02-09 2000-02-09 Verfahren zur hemmung der expression von zielgenen in pflanzen Withdrawn EP1151119A1 (de)

Priority Applications (1)

Application Number Priority Date Filing Date Title
EP00915151A EP1151119A1 (de) 1999-02-09 2000-02-09 Verfahren zur hemmung der expression von zielgenen in pflanzen

Applications Claiming Priority (4)

Application Number Priority Date Filing Date Title
EP99420030 1999-02-09
EP99420030 1999-02-09
EP00915151A EP1151119A1 (de) 1999-02-09 2000-02-09 Verfahren zur hemmung der expression von zielgenen in pflanzen
PCT/EP2000/001524 WO2000047754A1 (en) 1999-02-09 2000-02-09 A method for inhibiting the expression of target genes in plants

Publications (1)

Publication Number Publication Date
EP1151119A1 true EP1151119A1 (de) 2001-11-07

Family

ID=8242281

Family Applications (1)

Application Number Title Priority Date Filing Date
EP00915151A Withdrawn EP1151119A1 (de) 1999-02-09 2000-02-09 Verfahren zur hemmung der expression von zielgenen in pflanzen

Country Status (4)

Country Link
EP (1) EP1151119A1 (de)
AU (1) AU770893B2 (de)
CA (1) CA2359749A1 (de)
WO (1) WO2000047754A1 (de)

Families Citing this family (26)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
AU2002367104A1 (en) 2001-12-26 2003-07-15 National Institute Of Advanced Industrial Science And Technology Transcription regulatory gene and peptide
WO2004018687A2 (en) 2002-08-07 2004-03-04 Basf Plant Science Gmbh Nucleic acid sequences encoding proteins associated with abiotic stress response
EP2184362A3 (de) 2003-04-15 2010-10-06 BASF Plant Science GmbH Nukleinsäuresequenzen, die für Proteine in Zusammenhang mit abiotischer Stressreaktion kodieren, sowie Pflanzenzellen und Pflanzen mit erhöhter Umweltstressverträglichkeit
JP2005295878A (ja) * 2004-04-09 2005-10-27 Japan Science & Technology Agency 花芽形成遅延植物体の生産方法、及びこれを用いて得られる植物体、並びにその利用
JP2005295879A (ja) * 2004-04-09 2005-10-27 Japan Science & Technology Agency 花の形態が改変された植物体の生産方法およびこれを用いて得られる植物体、並びにその利用
JP2006006248A (ja) * 2004-06-28 2006-01-12 Japan Science & Technology Agency 葉の形態形成が制御された植物体の生産方法およびこれを用いて得られる植物体、並びにその利用
JP2006020607A (ja) * 2004-07-09 2006-01-26 Japan Science & Technology Agency 葉の形態が改変された植物体の生産方法およびこれを用いて得られる植物体、並びにその利用
JP2006042730A (ja) * 2004-08-06 2006-02-16 Japan Science & Technology Agency 単子葉植物の雄性不稔体の生産方法およびこれを用いて得られる植物体、並びにその利用
EP1794184A2 (de) 2004-09-24 2007-06-13 BASF Plant Science GmbH Nukleinsäuresequenzkodierende proteine in zusammenhang mit abiotischer stressreaktion und pflanzenzellen und pflanzen mit erhöhter umweltstressverträglichkeit
JP2006101827A (ja) * 2004-10-08 2006-04-20 Japan Science & Technology Agency 雄性不稔形質転換植物体の生産方法およびこれを用いて得られる植物体、並びにその利用
JP2006280242A (ja) * 2005-03-31 2006-10-19 Japan Science & Technology Agency 完全不稔性植物体の生産方法およびこれを用いて得られる植物体、並びにその利用
CA2647115A1 (en) 2006-03-24 2007-10-04 Basf Plant Science Gmbh Proteins associated with abiotic stress response and homologs
MX2009003824A (es) 2006-10-13 2010-04-07 Basf Plant Science Gmbh Plantas con rendimiento aumentado.
EP2064330A2 (de) 2007-05-22 2009-06-03 BASF Plant Science GmbH Pflanzen mit erhöhter toleranz und/oder resistenz gegenüber umweltstress und erhöhter biomasseproduktion
BRPI0817005A2 (pt) 2007-09-18 2019-09-24 Basf Plant Science Gmbh método para produzir uma célula de planta transgênica, uma planta ou uma parte da mesma com rendimento aumentado, molécula de ácido nucleico, construção de ácido nucleico, vetor, célula hospedeira, processo para produzir um polipeptídeo, polipeptídeo, anticorpo, núcleo de célula de planta transgênica, célula de planta, tecido de planta, material de propagação, material colhido, planta ou parte da mesma, semente, processo para identificar um composto, método para produzir uma composição agrícola, composição, e, uso de uma molécula de ácido nucleico
US8809059B2 (en) 2007-09-21 2014-08-19 Basf Plant Science Gmbh Plants with increased yield
US20110098183A1 (en) 2007-12-19 2011-04-28 Basf Plant Science Gmbh Plants with increased yield and/or increased tolerance to environmental stress (iy-bm)
US20110010800A1 (en) 2008-02-27 2011-01-13 Basf Plant Science Gmbh Plants with increased yield
AU2009284172A1 (en) 2008-08-19 2010-02-25 Basf Plant Science Gmbh Plants with increased yield by increasing or generating one or more activities in a plant or a part thereof
CA2736473A1 (en) 2008-09-23 2010-04-01 Basf Plant Science Gmbh Plants with increased yield (lt)
DE112009002342T5 (de) 2008-10-23 2012-06-14 Basf Plant Science Gmbh Verfahren zum Herstellen einer transgenen Zelle mit erhöhtem Gehalt an gamma-Aminobuttersäure (GABA)
CA2740257A1 (en) 2008-10-23 2010-04-29 Basf Plant Science Gmbh Plants with increased yield (nue)
WO2011009801A1 (en) 2009-07-23 2011-01-27 Basf Plant Science Company Gmbh Plants with increased yield
MX2012005719A (es) 2009-11-17 2012-07-30 Basf Plant Science Co Gmbh Plantas con rendimiento aumentado.
EP2961845A4 (de) 2013-03-01 2016-08-10 Univ California Verfahren und zusammensetzungen zum targeting von rna-polymerasen und zur nichtcodierung einer rna-biogenese an spezifischen loci
BR112017016789A2 (pt) 2015-02-11 2018-05-08 Basf Se métodos para produzir uma planta transgênica, para controlar a vegetação indesejada e para o cultivo da planta, molécula de ácido nucleico, construção de ácido nucleico, vetor, polipeptídeo hppd mutado, núcleo de célula vegetal, núcleo de célula vegetal transgênica, planta transgênica, uso do ácido nucleico, combinação útil, processo para a preparação de uma combinação útil e uso de uma combinação útil

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US5866755A (en) * 1993-06-14 1999-02-02 Basf Aktiengellschaft Animals transgenic for a tetracycline-regulated transcriptional inhibitor

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO0047754A1 *

Also Published As

Publication number Publication date
AU770893B2 (en) 2004-03-04
WO2000047754A1 (en) 2000-08-17
CA2359749A1 (en) 2000-08-17
AU3656300A (en) 2000-08-29

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