CN120456819A - Endogenous microbial biostimulants - Google Patents
Endogenous microbial biostimulantsInfo
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- CN120456819A CN120456819A CN202380076652.0A CN202380076652A CN120456819A CN 120456819 A CN120456819 A CN 120456819A CN 202380076652 A CN202380076652 A CN 202380076652A CN 120456819 A CN120456819 A CN 120456819A
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The present invention relates to a composition comprising-an isolated endophyte, wherein the endophyte is a strain of Paenibacillus xylan, and-an adjuvant, wherein the strain of Paenibacillus xylan has accession number DSM 34353.
Description
Technical Field
The present invention relates to a novel isolated plant microbiome strain, in particular an endophyte, plants infected with such strain and related methods. In particular, the isolated endophytes may be used in compositions for use as biofertilizers and/or biostimulants or for bioprotection in plants. The composition may further comprise adjuvants.
Background
In many cases, the growth and/or yield of crops is limited by the amount of nitrogen that can be used or absorbed by the plants. To overcome this limitation, exogenous nitrogen is added to the soil either before or after sowing. Nitrogen fertilizers may be of organic (e.g., urea, amino acids, manure, corner pieces) or mineral (e.g., ammonium nitrate, ammonium sulfate, potassium nitrate) origin in nature. All these artificial nitrogenous fertilizers have in common that their production, transportation and/or application are energy intensive and thus result in a forward CO 2 balance. In particular, ammonium nitrate and urea, which are the nitrogen fertilizers most widely used worldwide so far, have a large CO 2 footprint (footprint) because their common precursor ammonia (NH 3) is derived from the Haber-Bosch process (Haber-Bosch process). The process uses atmospheric nitrogen (N 2) and hydrogen (H 2) to synthesize ammonia (NH 3) at high temperature and pressure. This approach thus exacerbates climate change and is environmentally unfriendly.
A more sustainable method of providing plants with at least some portion of their required nitrogen substitution is the use of biofertilizers. Biofertilizers consist of living microorganisms that are able to reduce atmospheric nitrogen and thus make them available to plants in the form of ammonium/ammonia or organically bound nitrogen. These organisms must be applied to the seed, root or, in rare cases, to the leaves in a metabolically active form.
Previous and current uses of these biofertilizers apply nitrogen-fixing microorganisms to the surface of seeds, foliage or soil. Due to this fact, organisms are exposed to high stresses from abiotic factors such as temperature, drought, pH, flushing from rain water and high humidity. The effectiveness and efficacy of biofertilizer is reduced. Thus, there is a need for a large amount of biofertilizer that is effective in providing biological protection for plants and/or increasing crop yield. This increases agricultural costs.
Further, biological factors such as high competition with already prevailing plants or soil microbiome or the presence of antibiotic actives also reduce the survival rate of exogenously added nitrogen fixing microorganisms on the respective surfaces. This also reduces the efficacy of currently available biofertilizers.
Accordingly, there is a need in the art for means to better utilize these endophytes as biofertilizers and/or biostimulants to improve sustainable agriculture and environment.
Brief Description of Drawings
Fig. 1 is a picture of maize growth (no treatment) after 6 to 8 weeks at different nitrogen fertilization (0 kg n/ha, 21kg n/ha, 42kg n/ha). Oxisol soil, which is poor in nutrients, was used as test soil (substrate).
FIG. 2 shows the next time at 21kg N/ha the application of Paenibacillus xylan (Paenibacillus xylanexedens) DSM 34353 (left) and the one time of Paenibacillus xylan DSM 34353 (which is in contact withApplied to leaves only) maize growth pictures.
FIG. 3 is a graph showing the presence and absence of Paenibacillus xylanisolvens DSM 34353 at different nitrogen fertilizer levels (which is shown in conjunction withApplied in a mixture of (c) applied corns.
FIG. 4 is a graph showing the presence and absence of Paenibacillus xylanisolvens DSM 34353 at different nitrogen fertilizer levels (which is shown in conjunction withApplied in a mixture of (c) applied corns.
Description of the invention
The present invention attempts to solve the above problems by providing a novel isolated endophyte from Paenibacillus xylan (Paenibacillus xylanexedens) that can be an effective and efficient biostimulant and/or biofertilizer. Due to their unique genetic equipment and/or in the presence of specific adjuvants, these newly isolated endophytes are able to allow and in particular to enhance the penetration of these endophytes into the plants or parts thereof with which they come into contact. In particular, the newly isolated Paenibacillus xylanisolvens strain is capable of penetrating and proliferating in plant tissue or seed in the presence of suitable adjuvants. In this way, the (non) biotic stress can be reduced and thus the viability or effectiveness of the organism in the plant is significantly increased. Within the plant or seed or any part thereof, the active Paenibacillus xylanisolvens strain is in a regulated steady state which increases both the efficacy and viability of the nitrogen fixing endophyte.
According to one aspect of the present invention there is provided a substantially purified or isolated endophyte, wherein said endophyte is a strain of paenibacillus xylan, wherein said strain of paenibacillus xylan has accession No. DSM 34353 and which provides a bio-protective and/or bio-stimulant phenotype to the plant into which it is introduced.
According to any aspect of the invention, the newly isolated endophyte, paenibacillus xylan with accession number DSM 34353, solves the problem of low stability of microbial nitrogen fixation agents, as it is an endophyte with both unprecedented high nitrogen fixation capacity and capacity to penetrate into the plant interior (endosphere), which is not available for most other existing endophytes used in agriculture.
Endophytes according to any aspect of the invention are isolated from the internal plant tissue of copper flowers (Minuartia verna subsp. Hercynica) and typed and sequenced as a xylan paenibacillus species. Greenhouse experiments have demonstrated that paenibacillus xylan is able to provide significant amounts of enzymatically immobilized nitrogen to plants in both seed and foliar applications for maize (maize), variety LG 31.224. Thus, maize inoculated or sprayed with paenibacillus xylan can show the same or even improved growth as fully fertilized maize (42 kg/ha) when fertilized with only half of the synthetic nitrogen fertilizer (21 kg/ha). In contrast, significantly reduced growth was observed in maize that was not inoculated with Paenibacillus xylan at 21kg n/ha.
As used herein, the term "endophyte" is an endosymbiont (endosymbiont) that refers to a bacterial or fungal strain that lives within a plant for at least a portion of the plant's life cycle without causing significant disease. In particular, bacteria or fungi are closely related to plants, wherein the term 'closely related' refers to the living of bacteria or fungi on, within or near the plant. For example, it may be endogenous and live within the internal tissues of the plant, or may be epiphytic and grow externally on the plant. There are many different endophytes that have been found. However, only a few have been used commercially as endophyte inoculants for agriculture, such as arbuscular mycorrhiza (arbuscular mycorrhizae), rhizobia (rhizobia) and azoospira (Azospirillium). The ergot (Clavicipitaceous) fungus is also an endophyte used in agriculture. The endophyte according to any aspect of the invention is a strain of Paenibacillus xylan having accession number DSM 34353.
As used herein, the term "substantially purified" refers to endophytes that are free of other organisms. The term includes, for example, endophytes in sterile culture. In particular, the endophyte is at least about 90% pure, more particularly at least about 95% pure, even more particularly at least about 98%, 99% or 99.5% pure.
As used herein, the term 'isolated' refers to an endophyte according to any aspect of the invention that is removed from its original environment (e.g., the natural environment if it is naturally occurring). For example, naturally occurring endophytes present in living plants are not isolated, but the same endophytes isolated from some or all of the coexisting materials in the natural system are isolated. In particular, the endophyte isolated according to any aspect of the invention may be a pure culture of a single strain and the single strain was submitted to the German collection of microorganisms and cell cultures (German Collection of Microorganisms and Cell Cultures) (DSMZ) at Inhoffenstra βe7B, 38124Braunschweig, germany on day 11 of 2022 and has accession number DSM 34353. Particles for preserving and modifying cells are available from the prior art, for example, sambrook/Fritsch/Maniatis (1989).
As used herein, the term "bioprotectant and/or biostimulant" may refer to endophytes according to any aspect of the present invention that have genetic and/or metabolic characteristics that result in a beneficial phenotype in a plant containing or otherwise associated with the endophyte. Such beneficial properties or phenotypes arising from endophytes present in plants include increased resistance to pests and/or diseases, improved tolerance to water and/or nutrient stress, increased biotic stress tolerance, increased drought tolerance, increased water use efficiency, reduced toxicity and increased vigor in plants to which the endophytes are related, as compared to plants not related to endophytes according to any aspect of the invention or to endophytes such as standard virulence (ST) endophytes. In particular, the bioprotective and/or biostimulant phenotype according to any aspect of the present invention includes nitrogen fixation in plants into which endophytes are introduced.
Pests and/or diseases may include, but are not limited to, fungal and/or bacterial pathogens, particularly fungi. In one example, an endophyte may lead to the production of a biological agent protecting compound in a plant with which it is associated.
As used herein, the term 'bioprotectant compound' refers to a compound that provides or helps provide biological protection against pests and/or diseases, such as bacterial and/or fungal pathogens, for plants with which it is associated. The bioprotectant compound may also be referred to as a 'biocidal compound'.
As used herein, the term 'biostimulant' refers to any substance or microorganism applied to a plant for the purpose of enhancing nutritional efficiency, abiotic stress tolerance, and/or crop quality traits, regardless of its nutrient content. Endophytes according to any aspect of the invention act as biostimulants for the plants and/or parts thereof with which it is in contact. A more detailed definition of biostimulants is provided at least in Ricci,M.,General Principles to Justify Plant Biostimulant Claims,Frontiers in Plant Science(2019),10.
As used herein, the term 'introduced' refers to contacting and/or treating a plant or part thereof with an endophyte, wherein the endophyte is delivered to the plant. In particular, endophytes are introduced into plants or parts thereof to promote the growth of endophytes therein. Any method of introducing an endophyte according to any aspect of the invention into a plant or part thereof may be used. For example, endophytes may be sprayed onto or inoculated into plants or parts thereof. In particular, endophytes may be inoculated for at least 1,2, 3, 4,5, 6, 7, 8, 9, 10, 11, 12, 13 or 14 days before they begin growing in plants. In one example, endophytes may be sprayed onto the foliage of plants (i.e., foliar application). In this example, the spraying process is performed only at an early developmental stage of the plant (about six-leaf stage), or multiple times (about six-leaf stage and several stages of the new/adult plant). The skilled person is able to identify the best introduction procedure to use on plants. In particular, the plant or part thereof may be infected with endophytes by methods known in the art. More particularly, the plant or portion thereof may be infected with endophytes using a method selected from the group consisting of inoculation, spraying, breeding, crossing (cross), crossing (hybridisation), transduction, transfection, transformation, and/or gene targeting, and combinations thereof.
In particular, the endophyte according to any aspect of the invention is a paenibacillus xylan strain having accession No. DSM 34353. In particular, the sequence of the strain has been divided into loci and comprises the nucleotide sequences of SEQ ID NOS 1-70 and variants thereof. In particular, the sequence of the 16S ribosomal RNA is SEQ ID NO. 78 and/or SEQ ID NO. 41.
As used herein, the term "variant" comprises an amino acid or nucleic acid sequence having at least 70, 75, 80, 85, 90, 92, 94, 95, 96, 97, 98 or 99% identity to a reference amino acid or nucleic acid sequence, respectively, wherein preferably amino acids other than those essential for the function (e.g. catalytic activity of a protein) or folding or structure of the molecule are deleted, substituted or replaced by insertion, or essential amino acids are replaced in a conservative manner in which the biological activity of the reference sequence or the molecule derived therefrom is substantially preserved. The prior art contains algorithms that can be used to align two given nucleic acid or amino acid sequences and calculate the degree of identity, see Arthur Lesk (2008), thompson et al, 1994 and Katoh et al, 2005. The term "variant" is synonymous and is used interchangeably with the term "homolog". Such variants may be prepared by introducing deletions, insertions or substitutions in the amino acid or nucleic acid sequence, as well as fusions comprising such macromolecules or variants thereof. In one example, the term "variant" with respect to an amino acid sequence comprises, in addition to the sequence identity described above, an amino acid sequence comprising one or more conservative amino acid changes with respect to the reference or wild-type sequence, respectively, or a nucleic acid sequence encoding an amino acid sequence comprising one or more conservative amino acid changes. In one example, the term "variant" of an amino acid sequence or nucleic acid sequence comprises, in addition to the degree of sequence identity described above, any active part and/or fragment of an amino acid sequence or nucleic acid sequence, respectively, or any nucleic acid sequence encoding an active part and/or fragment of an amino acid sequence. As used herein, the term "active moiety" refers to an amino acid sequence or nucleic acid sequence that is less than the full length amino acid sequence or encodes less than the full length amino acid sequence, respectively, wherein the amino acid sequence or encoded amino acid sequence retains at least some of its requisite biological activity, respectively.
According to another aspect of the invention there is provided a biostimulant comprising an isolated endophyte, wherein the endophyte is a strain of Paenibacillus xylane having accession number DSM 34353. Endophytes are in accordance with any aspect of the invention.
According to a further aspect of the present invention there is provided a composition comprising:
-isolated endophyte, wherein the endophyte is a strain of paenibacillus xylan having accession No. DSM 34353, and
-An auxiliary agent.
The composition according to any aspect of the invention solves the problems of low uptake rate and uptake kinetics of the endophyte into the plant interior. The adjuvants enable efficient uptake through stomata on the upper and most predominantly lower surfaces of the plant, small damaged wounds and growth cracks in the cuticle. The use of adjuvants to support the process of ingrowth (penetration of microorganisms, in particular paenibacillus xylan, into plant cells) allows for lower drug concentrations (CFU/mL or CFU/g) to be used, since uptake occurs more effectively than without the addition of adjuvants.
The endophyte is a Paenibacillus xylan strain with accession number DSM 34353.
The isolated Paenibacillus xylanisolvens according to any of the aspects of the invention have suitable genetic equipment to invade plant tissue and survive and proliferate within the tissue. This, in combination with the appropriate penetration sites on the surface of the plant and/or seed to which the Paenibacillus xylan strain is inoculated, enables successful penetration of the endophyte into the plant tissue via stomata of leaves or via small wounds or growing cracks. Active penetration of the non-uniformly shaped stratum corneum has also been reported.
However, since both the speed and kinetics of the endophyte process are limited in nature and therefore in most cases very slow and host-specific occurs, the presence of adjuvants in the composition according to any aspect of the invention allows for efficient and effective successful uptake of the endophyte according to any aspect of the invention into plant tissue (inside).
In general, when paenibacillus xylan is applied alone to soil, leaves or seeds, slow endogenous processes result in the organism being exposed to biotic and abiotic stresses, respectively, for a long period of time. This may result in a significant reduction in the titer of the Paenibacillus xylan and thus reduced product efficacy. The presence of adjuvants according to any aspect of the invention eliminates this negative effect and in fact increases the efficacy of the composition by accelerating the uptake of the organism into the target plant or allowing its complete uptake (to enableit at all).
As used herein, 'adjunct' refers to an ingredient or substance in a composition according to any aspect of the invention that increases or modifies the activity of other ingredients (i.e., isolated paenibacillus xylan strains). In particular, the adjuvant according to any aspect of the present invention is a biocompatible adjuvant (active ingredient medium) which is added to a suspension of microorganisms to form a composition according to any aspect of the present invention. Any adjuvant known in the art may be used in the composition according to any aspect of the present invention. In particular, the auxiliary agent according to any aspect of the present invention may be selected from (a), (B) or (C) and mixtures thereof, wherein (a), (B) or (C) is:
(A) Polyglycerol esters of the general formula (I),
M j Dk Tl (I)
Wherein M is [ C 3H5(OR")2O1/2 ],
D is [ C 3H5(OR")1O2/2 ],
T is [ C 3H5O3/2 ],
J=1 to 10, preferably 2 to 3, more preferably 2;
k=0 to 10, preferably greater than 0 to 5, more preferably 1 to 3;
l=0 to 3, preferably 0 to 1, more preferably 0;
wherein the sum of j+k+l is 1 to 20, preferably 2 to 4, more preferably 3
Wherein the groups R "are each independently selected from acyl groups R' -C (=0) -and H, provided that at least one group R" is not equal to H;
Wherein each group R' is independently selected from monovalent aliphatic, saturated or unsaturated hydrocarbon groups having 3 to 39, preferably 7 to 21, more preferably 9 to 17 carbon atoms;
(B) Polyether modified siloxanes of formula (II)
M 1 o D1 p D′q (II)
Wherein M 1 is P 1 3SiO1/2,
D 1 is P 1 2SiO2/2, D' is P 1P2SiO2/2,
O is 2, p is 0 to 0.1, q is 1.0 to 1.15,
P 1 is independently a hydrocarbon group having 1 to 8 carbon atoms,
P 2 is independently a polyether radical of the formula (III),
-P 3O[CH2CH2O]m[CH2CH(CH3)O]nP5 formula (III)
Wherein the method comprises the steps of
M is 3.4 to 11.0, n is 2.5 to 8.0,
And provided that
M/n is 1.9 to 2.8,
P 3 is independently a divalent hydrocarbon group having 2 to 8 carbon atoms,
P 5 is hydrogen, and/or
(C) An organomodified polysiloxane of formula (IV)
(IV)
Wherein a+b+c+d +2=20 to 210,
A is 15 to 205, b is 1 to 12, c is 1 to 12, d is 1 to 12,
Each R group is independently the same or different aliphatic or aromatic hydrocarbyl group having 1 to 10 carbon atoms,
R4 radicals are each independently identical or different R, R, R2 or R3 radicals,
R1, R2 and R3 are each independently a different polyether radical of the formula (V)
(V)
Wherein the method comprises the steps of
E is 3 to 11, f is 6 to 30, g is 0 to 15, h is 0 to 5,i, 0 to 5, and
R5 are independently the same or different and are each a methyl, acetyl or hydrogen group.
More particularly, the auxiliary agent may be selected from BREAK-S 301、BREAK-SP 133、BREAK-S255. The adjuvants used according to any aspect of the invention result in a reduction of the surface tension at the stomata or where the damage is present and thus in a lower rejection or improved flow of particles (microorganisms) through the orifice into the plant or part thereof into which the microorganisms, in particular the paenibacillus xylan, are inoculated. In particular, the use of adjuvants allows/accelerates the endogenous processes of bacterial uptake. In addition, enhanced uptake also allows translocation of bacteria from leaf to root across the phloem, from where nitrogen fixation can also be enhanced. This results in the locally applied biostimulant acquiring systemic properties. Based on its systemic properties, there are advantages in that in the plants forming shoots, the shoots have also been inoculated with biostimulants and thus deliver the active ingredient to the next generation.
The use of adjuvants also allows for uniform distribution of the Paenibacillus xylan from the upper surface of the leaf to the lower surface of the leaf where most (open) pores are present. Thus, a faster and more extensive penetration of endophytes into plant tissue can be achieved. If no biocompatible auxiliary is used, particularly in the case of foliar application, it would be particularly difficult, if not impossible, to reach the air holes on the underside of the leaf. The use of adjuvants with "anti-rinse" properties also increases the residence time on the upper surface of the leaf and thus promotes uptake of endophytes into the plant tissue. The presence of the auxiliary agent reduces early wash-off of endogenous nitrogen fixing organisms.
In one example, the adjuvant is (a):
(A) Polyglycerol esters of the general formula (I),
M j Dk Tl (I)
Wherein M is [ C 3H5(OR")2O1/2 ],
D is [ C 3H5(OR")1O2/2 ],
T is [ C 3H5O3/2 ],
J=1 to 10, preferably 2 to 3, more preferably 2;
k=0 to 10, preferably greater than 0 to 5, more preferably 1 to 3;
l=0 to 3, preferably 0 to 1, more preferably 0;
wherein the sum of j+k+l is 1 to 20, preferably 2 to 4, more preferably 3
Wherein the groups R "are each independently selected from acyl groups R' -C (=0) -and H, provided that at least one group R" is not equal to H;
Wherein each group R' is independently selected from monovalent aliphatic, saturated or unsaturated hydrocarbon groups having 3 to 39, preferably 7 to 21, more preferably 9 to 17 carbon atoms.
In particular, at least one group R 'corresponds to a group of formula R' -C (O) -.
More particularly, M, D and T may be:
even more particularly, the polyglycerol ester of the mixture according to any aspect of the invention has the formula (I (a)):
Wherein the method comprises the steps of
A=1 to 10, preferably 2 to 3, more particularly 2;
b=0 to 10, preferably greater than 0 to 5, more particularly 1 to 3;
The conditions are as follows:
a+b=2 to 20, preferably 2 to 4, in particular 3;
Wherein the radicals R ' are each independently selected from the group consisting of acyl radicals R ' -C (=0) -and H, with the proviso that at least one radical R is not equal to H, wherein the radicals R ' are each independently selected from the group consisting of monovalent aliphatic, saturated or unsaturated hydrocarbon radicals having from 3 to 39, preferably from 7 to 21, in particular from 9 to 17, carbon atoms.
The polyglycerol ester of the composition according to any aspect of the invention may have more than one, in particular at least 2, more particularly at least 3 groups R of the form R' -C (=0) -.
The radicals R 'of the formulae R' -C (O) -may be, independently of one another, acyl radicals of saturated or unsaturated fatty acids which are identical or different and comprise 4 up to 40 carbon atoms, in particular fatty acids selected from butyric acid (butyric acid) (butyric acid (butanoic acid)), caproic acid (caproic acid) (caproic acid (hexanoic acid)), caprylic acid (CAPRYLIC ACID) (caprylic acid), capric acid (CAPRIC ACID) (capric acid (decylic acid)), lauric acid (lauric acid), myristic acid (myristic acid), palmitic acid (palmitic acid), stearic acid (stearic acid), arachic acid (eicosanoic acid), behenic acid (behenic acid), lignoceric acid (lignoceric acid), palmitoleic acid ((Z) -9-hexadecenoic acid), oleic acid ((Z) -9-hexadecenoic acid), elaidic acid ((E) -9-octadecenoic acid), cis-iso-oleic acid ((Z) -11-octadecenoic acid), linoleic acid ((9Z, Z) -9, 12-octadecadienoic acid), alpha-linolenic acid ((9Z, 15Z) -9,12, 15-octadecenoic acid), tric-9, 6-linolenic acid ((6Z) -6-tricyclooleic acid, 8-6-tricyclooleic acid Arachidonic acid ((5Z, 8Z,11Z, 14Z) -5,8,11, 14-eicosatetraenoic acid), erucic acid ((Z) -13-docosenoic acid), nervonic acid ((Z) -15-tetracosenoic acid), ricinoleic acid, hydroxystearic acid, undecylenic acid, and mixtures thereof. In one example, the fatty acid may be a mixture of rapeseed oleic acid, soybean fatty acid, sunflower fatty acid, peanut fatty acid, and tall oil fatty acid. In particular, for this context, the fatty acid may be a group of oleic acid. When calculating the HLB value, the molar mass of the lipophilic molecule fraction is the arithmetic mean of the sum of the molar masses of all the radicals R' present in the molecule.
Suitable sources of fatty acids or fatty acid esters, especially glycerides, may be vegetable or animal fats, oils or waxes. For example, the number of the cells to be processed, lard, tallow, goose fat, duck fat, chicken fat, horse fat, whale oil, fish oil, palm oil, olive oil, avocado oil, kernel oil, coconut oil, palm kernel oil, cocoa butter, cottonseed oil, pumpkin seed oil, corn seed oil, sunflower oil, wheat germ oil, grape seed oil, soybean oil, peanut oil, lupin oil, rapeseed oil, mustard oil, castor oil, jatropha oil (jatropa oil), walnut oil, jojoba oil, lecithin (e.g., based on soybean, rapeseed or sunflower), bone oil, claw oil, borage oil, lanolin, emu oil, deer fat, woodchuck oil, mink oil, safflower oil, hemp oil, pumpkin oil, evening primrose oil, tall oil, carnauba wax, beeswax, candelilla wax, ouricuri wax, retamow wax, caranday wax, raffia wax, spanish wax, alfalfa wax, bamboo wax, hemp wax, douglas fir wax, sisal wax, cotton wax, daar, cotton wax, coffee, sugar cane wax, and/or a fatty acid ester of a source such as may be a sugar cane wax, a fatty acid or a fatty acid ester.
In particular, the polyglycerol ester compound has formula (I), or (I (a)) having an arithmetic average of 2.9 to 3.1 groups of the form R' -C (=0) -and an HLB value of 4 to 6.5.
More particularly, the polyglycerol ester compound has the formula (I (a)) having an arithmetic average of 2.9 to 3.1 groups with a sum a+b of 3, and a form R' -C (=0) -and an HLB value of 4 to 6.5.
Even more particularly, the polyglyceryl ester compound may have formula (I (a)) having an arithmetic average of 2.9 to 3.1 groups of the form R' -C (=0) -and an HLB value of 4 to 6.5, wherein the acyl residues of the fatty acid mixture of oleic acid, stearic acid, palmitic acid and gamma-linolenic acid are contained, and the fatty acids comprise in particular at least 85% by weight of the fatty acid mixture.
In one example, the polyglyceryl ester compound may have formula (I (a)) having an arithmetic average of 2.9 to 3.1 groups of the form R' -C (=0) -and an HLB value of 4 to 6.5, the acyl residues being derived from a fatty acid mixture containing oleic acid, stearic acid, palmitic acid and gamma-linolenic acid, and the fatty acids comprising in particular at least 85% by weight of the fatty acid mixture.
In another example, the polyglyceryl ester compound used according to any aspect of the present invention may have formula (I (a)) having an arithmetic average of 2.9 to 3.1 groups of form R' -C (=0) -and an HLB value of 4 to 6.5, the mass fraction of oleic acid acyl residues being at least 75%, particularly 85%, more particularly 95% based on the mass of all acyl residues. Even more particularly, the polyglycerol ester is triolein (triglyceroltrioleate).
A more complete disclosure of the adjuvant (B) is provided at least in US10390530B 2.
In one example, the adjuvant is (B):
(B) Polyether modified siloxanes of formula (II)
M 1 o D1 p D′q (II)
Wherein M 1 is P 1 3SiO1/2,
D 1 is P 1 2SiO2/2, D' is P 1P2SiO2/2,
O is a number of times, where o is 2,
P is 0 to 0.1, especially 0
Q is from 1.0 to 1.15, in particular from 1.0 to 1.10, in particular from 1.00 to 1.05,
P 1 is independently a hydrocarbon radical having from 1 to 8 carbon atoms, in particular a methyl, ethyl, propyl or phenyl radical, in particular a methyl radical,
P 2 is independently a polyether radical of the formula (III),
-P 3O[CH2CH2O]m[CH2CH(CH3)O]nP5 formula (III)
Wherein the method comprises the steps of
M is 3.4 to 11.0, especially 3.6 to 9.9, more especially 4.5 to 8.5,
N is 2.5 to 8.0, especially 2.7 to 7.5, more especially 3.0 to 6.0,
And provided that
M/n is 1.9 to 2.8,
P 3 is independently a divalent hydrocarbon radical having 2 to 8 carbon atoms, in particular ethylene, propylene, 1-methylpropene, 1-dimethylpropenyl radical, in particular-CH 2CH2CH 2-,
P 5 is hydrogen.
The polyether modified siloxanes of formula (II) have a biodegradability of greater than 60%, more particularly greater than or equal to 63% and especially greater than or equal to 65%, with a maximum of 100%.
In particular, the polyether groups calculated without P 3 O and calculated without P 5 have a molar mass M (PE) calculated by 44g/mol×m+58g/mol×n, where the indices M and n relate to formula (III). More particularly, the value of M (PE) is such that the lower limit M (PE) is greater than 520g/mol, in particular greater than 530g/mol, more particularly greater than 535g/mol, and the upper limit M (PE) is less than 660g/mol, in particular less than 630g/mol, more particularly less than 600g/mol. Even more particularly, the value of M (PE) is greater than 520g/mol and less than 660g/mol, in particular greater than 535g/mol and less than 600g/mol. In particular, the sum of m+n is greater than 9 up to 19, more particularly greater than 9.5 up to 15, and even more particularly greater than 10 up to 12.
In one example, the polyether modified siloxane used in the composition according to any aspect of the present invention is a polyether modified siloxane of formula (II) wherein the index c is from 1 to 1.05, wherein the index of the polyether groups of formula (III) is from m from 3.4 to 11.0 and n is from 2.5 to 8.0. In particular, the polyether modified siloxane used in the composition according to any aspect of the present invention is a polyether modified siloxane of formula (II), wherein the index c is from 1 to 1.05, wherein the ratio m/n is from 1.9 to 2.8. More particularly, the polyether modified siloxane used in the composition according to any aspect of the present invention is a polyether modified siloxane of formula (II), wherein the index c is from 1 to 1.05, wherein the molar mass of the polyether residues M (PE) is greater than 520g/mol and less than 660g/mol. Even more particularly, the polyether modified siloxane used in the composition according to any aspect of the present invention is a polyether modified siloxane of formula (II), wherein the index c is from 1 to 1.05, wherein the P 5 groups are hydrogen, or wherein the index c is 1 and 1.05, wherein the molar mass of the polyether residue M (PE) is greater than 520g/mol and less than 660g/mol and the P 5 groups are hydrogen. In particular, the polyether modified siloxane used in the composition according to any aspect of the present invention is a polyether modified siloxane of formula (II) and does not include any further polyether modified siloxane other than those of formula (II).
A more complete disclosure of the adjuvant (B) is provided at least in US10299471B 2.
In one example, the adjuvant is (C), an organomodified polysiloxane of formula (IV)
(IV)
Wherein a+b+c+d+2=20 to 210, preferably 30 to 100, especially 40-60,
A is 15 to 205, preferably 35 to 45,
B is 1 to 12, preferably 1 to 8, in particular 2 to 6,
C is 1 to 12, preferably 1 to 8, in particular 2 to 6,
D is 1 to 12, preferably 1 to 8, in particular 2 to 6,
Each R group is independently the same or different aliphatic or aromatic hydrocarbyl group having 1 to 10 carbon atoms, preferably a methyl group,
R4 radicals are each independently identical or different R, R, R2 or R3 radicals,
R1, R2 and R3 are each independently a different polyether radical of the formula (V)
(V)
Wherein the method comprises the steps of
E is 3 to 11, preferably 3,
F is 6 to 30, preferably 10 to 30,
G is 0 to 15, preferably 0 to 10,
H is a number from 0 to 5,
I is 0 to 5, and
R5 are independently identical or different and are each a methyl, acetyl or hydrogen radical, preferably provided that the polyether radical of the formula (V) has a molecular weight of more than 200g/mol, preferably from more than 400 to 2000g/mol, and the proportion of ethylene oxide is more than 45% by mass of the polyether and the proportion of ethylene oxide in the polyether radical R2 is at least 9% by mass greater than the proportion of ethylene oxide in the polyether radical R1, in each case based on the polyether radical of the formula (V), where the radicals of the formula (V) can each be formed randomly, in a gradient or in blocks.
In formula (V), the units specified by the index 'g' are those units already derived from propylene oxide, the units specified by the index 'h' are those units already derived from butylene oxide, and the units specified by the index 'i' are those units already derived from styrene oxide.
The indices 'a to d' and 'e to i' may be natural integers or weighted averages (WEIGHT AVERAGES). The index is preferably a weighted average.
A more complete disclosure of the adjuvant (C) is provided at least in US8580225B 2.
The composition according to any aspect of the present invention may comprise any one of the adjuvants (a), (B) or (C). In one example, the composition may comprise a mixture of adjuvants, such as (a) and (B), (a) and (C), (B) and (C) or (a), (B) and (C). In another example, the composition according to any aspect of the invention may comprise more than one adjuvant (a) or more than one adjuvant (B) or more than one adjuvant (C).
The composition according to any aspect of the invention may further comprise an emulsifier. The emulsifier in the mixture according to any aspect of the invention may be different from the adjuvant. The emulsifier may be selected from the group consisting of fatty acid esters of polyols and polyalkylene glycol derivatives thereof, polyethylene glycol derivatives of fatty acids and fatty alcohols, sorbitan fatty acid esters, ethoxylated and/or propoxylated sorbitan fatty acid esters, alkylphenol ethoxylates, propoxylates, alkylphenol ethoxylates, amino-oxo (aminoxylated) oxides, amine oxides, propoxylated amine oxides, amino-oxo propylene oxide, acetylene glycol surfactants, ethoxylated and/or propoxylated acetylene glycol, silicon surfactants (silicone surfactants), and mixtures thereof. In particular, the emulsifier is selected from sorbitan fatty acid esters and ethoxylated sorbitan fatty acid esters. More particularly, the emulsifier is an ethoxylated sorbitan fatty acid ester or a mixture thereof.
The acyloxy groups of the sorbitan fatty acid esters or ethoxylated sorbitan fatty acid esters have from 4 to 40, in particular from 8 to 22, more in particular from 10 to 18, carbon atoms and/or the sorbitan fatty acid esters or ethoxylated sorbitan fatty acid esters have from 0 to 40, in particular from 10 to 30, more in particular from 15 to 25, oxyethylene groups. Fatty acids or fatty acid residues of sorbitan fatty acid esters are defined in particular as fatty acids or fatty acid residues of polyglycerol esters. The acyl groups (also referred to as alkanoyl groups) are derived in particular from a fatty acid mixture comprising oleic acid, stearic acid, palmitic acid and gamma-linolenic acid, said fatty acids representing in particular at least 85% by weight of the fatty acid mixture. In particular ethoxylated sorbitan fatty acid esters are used, the mass fraction of oleic acid acyl residues being at least 75%, in particular 85%, more in particular 95% based on the mass of all acyl residues.
The emulsifier according to any aspect of the invention has an HLB value of greater than or equal to 9, specifically greater than or equal to 10, more specifically greater than or equal to 11. The HLB value may be a maximum of 16, more particularly a maximum of 15, even more particularly a maximum of 13. In particular, the emulsifier has an HLB value of 9 to 16, particularly 10 to 15, more particularly 11 to 13. The HLB value is determined as described above. The HLB value of the sorbitan fatty acid ester and/or the ethoxylated sorbitan fatty acid ester is specifically as determined for the polyglycerol ester. The molar mass of the lipophilic part of the molecule results from the arithmetic average of the sum of the molar masses of all the groups R '"present in the molecule as part of the acyl group R'" - (CO) -. The radical R' "is preferably as defined for polyglycerol esters. The group R '"as part of the acyl group R'" - (CO) -of the sorbitan fatty acid ester or of the ethoxylated sorbitan fatty acid ester is in particular selected from monovalent aliphatic, saturated or unsaturated hydrocarbon groups having from 3 to 39, preferably from 7 to 21, in particular from 9 to 17, carbon atoms. The calculation of the molar mass of the whole molecule is carried out as defined above. In particular, the emulsifier is polyethylene glycol-20-sorbitan trioleate. Number 20 indicates the average number of ethylene oxide units in the polyethylene glycol residue.
The HLB values of the polyglycerol ester and the emulsifier are matched with each other. The polyglycerol ester has an HLB value of less than or equal to 8, specifically less than or equal to 7, more specifically less than or equal to 6.5, and the at least one emulsifier has an HLB value of greater than or equal to 9, specifically greater than or equal to 10, specifically greater than or equal to 11. At least one polyglycerol ester has an HLB value of from 0.5 to 8, particularly from 1 to 7, more particularly from 2 to 6.5, and at least one emulsifier has an HLB value of from 9 to 16, particularly from 10 to 15, more particularly from 11 to 13. In particular, the emulsifier is polyethylene glycol 20 sorbitan trioleate.
According to a further aspect of the invention there is provided a composition comprising a medium of isolated endophyte and an endophyte, wherein the endophyte is a strain of paenibacillus xylan having accession No. DSM 34353.
Strains and compositions according to any aspect of the invention may be obtained by culturing a strain of Paenibacillus xylanisolvens according to methods well known in the art, including by using an appropriate medium. Conventional large scale microbial culture methods include submerged fermentation, solid state fermentation, or liquid surface culture. Endophytes may be cultivated under aerobic or anaerobic conditions and may be cultivated in a bioreactor. Endophytes and metabolites resulting from the culture medium may be used directly or concentrated by conventional industrial methods such as centrifugation, tangential flow filtration, depth filtration and evaporation. The concentrated fermentation broth may be washed, for example, via a diafiltration process, to remove residual fermentation broth and metabolites.
The fermentation broth or fermentation broth concentrate may be dried, with or without the addition of a carrier, using conventional drying processes or methods such as spray drying, freeze drying, tray drying, fluid bed drying, roller drying or evaporation. The resulting dried product may be further processed, for example, by grinding or granulating, to achieve a particular particle size or physical form. The carrier may also be added after drying. In particular, the strain is prepared as a supernatant of a fermentation broth. In one example, a composition according to any aspect of the invention may be prepared according to the method provided in EP21198571 or EP21202623, wherein endophytes according to any aspect of the invention are first spray dried and then contacted with at least one adjunct. In particular, the auxiliary agent may be (a), (B) or (C) according to any aspect of the invention. More particularly, the auxiliary is (A), a polyglycerol ester of general formula (I). Even more particularly, the polyglycerol ester is combined with at least one emulsifier according to any aspect of the invention.
According to a further aspect of the present invention there is provided a plant or part thereof infected with one or more endophytes according to any aspect of the present invention. In particular, plants or parts thereof infected with endophytes may produce bioprotectant compounds. In particular, the plant or part thereof comprises an endophyte-free host plant or part thereof stably infected with said endophyte.
Any plant or part thereof may be inoculated with an endophyte according to any aspect of the invention. In particular, the plants inoculated with endophytes may be herbaceous plants or non-herbaceous plants suitable for agriculture, in particular forage grass, turf grass or bioenergy grass, or cereal crops or industrial crops. More particularly, the cereal crop or industrial crop species may be selected from wheat, barley, oat, maize/maize, any cereal legume such as chickpea, triticale, fava bean, lupin, purple pea, canola (canola), cereal rye, field pea, lentil, chestnut/millet, safflower, linseed, sorghum, sunflower, maize, canola, mung bean, soybean, oilseed crops, tomato and cotton.
Endophytes according to any aspect of the invention may be transferred from one generation of plants to the next by seed. Endophytes may then spread or localize to other tissues, i.e., roots, as the plant grows. Alternatively or additionally, endophytes may be recruited to plant roots, for example from the soil, and spread or localized to other tissues. According to a further aspect of the present invention there is provided a plant, plant seed or other plant part derived from a plant or part thereof according to any aspect of the present invention. Plants, plant seeds, or other plant parts may produce the bioprotectant compound.
Endophyte-infected plants or parts thereof may be cultivated by known techniques. The person skilled in the art can easily determine the appropriate conditions depending on the plant to be cultivated or parts thereof.
According to another aspect of the present invention there is provided a method of producing a composition for bioprotection and/or biostimulation, the method comprising combining:
-an isolated endophyte, wherein the endophyte is a strain of paenibacillus xylan, and
-An auxiliary agent.
According to a further aspect of the present invention there is provided a method of providing biological protection to a plant or part thereof, the method comprising contacting a composition according to any aspect of the present invention with a plant or part thereof.
The part of the plant may be, for example, a seed. In one example, a composition according to any aspect of the invention is introduced into the leaves, roots or seeds of a plant. In particular, the composition according to any aspect of the invention is introduced into the foliage of plants. Application of endophytes to the leaves allows the user (if required) to treat the growing or adult plant at several different times after sowing. In contrast, during soil application (prior to sowing) or seed application (during seed treatment), only a single application is possible. Further, foliar application of the paenibacillus xylan allows for reduced use of microbial agents, since higher CFU losses (several log levels) over a short period of time (lower stability of gram negative organisms on seeds or in soil) can be expected when inoculating seeds or soil.
According to a further aspect of the present invention there is provided the use of a composition according to any aspect of the present invention to provide biological protection to plants or parts thereof.
Examples
The foregoing describes preferred embodiments that may be subject to variations or modifications in design, construction, or operation without departing from the scope of the claims, as will be understood by those skilled in the art. For example, such variations are intended to be covered by the scope of the claims.
Example 1
Strain collection
For the collection of the Paenibacillus xylanisolvens strain with accession number DSM 34353, different plants were collected from different sites. Sites are selected based on their unique habitat. A list of locations and plant types for collection of paenibacillus xylanisolvens DSM 34353 is provided in table 1.
TABLE 1 sites and plant lists for collection of Paenibacillus xylanisolvens DSM 34353
A trowel or small shovel is used to gently dig the soil around each plant and lift the root with minimal disturbance. The removal of the root portion is performed gently and without damaging the entire root system. Similarly, leaf material is harvested from individual plants without damage to the plants. The plant material is placed in separate plastic bags and the bags are then carefully packed into insulated and refrigerated transport bags. After on-site collection, the plant material is refrigerated in a refrigerator at 5 ℃ until further use.
As part of the pretreatment, the leaves and roots of each plant were washed in sterile distilled water under slow-flowing tap water for 15 minutes, respectively, to remove adherent soil particles and most microbial surface probiotics (epiphytes). The samples were then rinsed three times, one minute each, with sterile distilled water in a laminar flow cabinet.
The roots and leaves are cut into sections. Plant material was placed in a petri dish, soaked in distilled water and drained. It was rinsed in 70% ethanol for 30 seconds, then sterilized with 3% sodium hypochlorite for 3 minutes for roots and 5 minutes for leaves. The tissue was then washed ten times with sterile water. Verification of the surface sterilization procedure was accomplished by culturing aliquots of water from the last rinse on nutrient media.
After the surface sterilized plant material has been properly dried in a laminar flow cabinet, the roots and leaves are cut into 2-3cm long pieces. The outer portion of the leaf approximately 0.5cm from the edge was removed with a sterile blade. Each of the roots and leaves was placed on nutrient agar medium and supplemented with 100mg L-1 cycloheximide to inhibit possible fungal development. Plates with plant tissue were sealed with sealing film tape and incubated at 28±2 ℃ in order to recover as many bacterial endophyte colonies as possible. After 48 hours, morphologically distinct bacterial colonies were selected from root tips and leaf nodes, and streaked cultures were repeated after four-fold serial dilutions to obtain enriched bacterial isolates. To obtain pure cultures, the enriched cultures were streaked on individual 50% TS agar plates and incubated for 2-3 days at 30 ℃.
The colony morphology was studied visually to check the purity of the cultures. Colonies that appeared different were isolated and streaked again on separate 50% TS agar plates for new culture. Thus, the mixed culture is separated into single-cell (monoseptic) isolates. Purity was confirmed by microscopic analysis and subsequent alignment of the 16S sequence with the corresponding sequence from the NCBIrRNA/ITS database using "blastn". The latter procedure is provided as a service by VERMICON (Hallbergmoos, germany).
The cellular material from these re-plated cultures was resuspended in 50% glycerol solution, as done with primary cellular material, and stored at-80 ℃. To test reactivation (re-vitalizing), new cultures were inoculated from these frozen stocks. The growing cultures were labeled "pure" and "cryopreserved". For genome sequencing, strains of particular interest were grown in liquid culture, centrifuged and resuspended in 70% ethanol. Cells of about 1x 10 8 of these strains were sent to LGC Biomics (Berlin, germany) for whole genome sequencing. It was confirmed as a novel strain. The strain is Paenibacillus xylan DSM 34353.
Example 2
Efficacy analysis
To determine the nitrogen fixation efficacy of Paenibacillus xylan DSM 34353 as an endophyte stimulator, untreated maize (maize) seeds (Zea mays, variety LG 31.224) were planted into poor sandy soil (Oxisol) with little organic matter (importantly low nitrogen content, such that the effect of independently surviving nitrogen fixing bacteria became apparent).
Five seeds were planted per pot, which had been further thinned to three plants after germination, in order to avoid a small number of replicates. Fertilizer dosage rate was calculated on the basis of 1.000.000kg soil/ha. The fertilization was performed with phosphate, potassium and magnesium sulfate (25 kg P/ha, 50kg K/ha and 15kg Mg/ha, respectively). With water-soluble compositions containing 21% N and 24% S (24 and 48kg N/ha)45 (Domo, germany) fertilization with nitrogen was performed. A 2L basin was used and filled with 2kg of sandy soil.
In the sixth foliar stage (about 2 weeks after planting), paenibacillus xylan DSM34353 was applied to the leaves in combination with the adjuvant. In the process, a spray volume of 1mL was applied to the leaves of young maize plants. The mixture contained 300.000CFU of Paenibacillus xylan (Paenibacillus xylanexedens) DSM34353 and 0.1% adjuvant(EvonikIndustries, germany) for surface wetting and pore flooding.Having the formula of the auxiliary (B) according to any aspect of the invention. In particular the number of the elements to be processed,Has the following structure
The 1mL spray for the leaves of the pot corresponds to about 400L spray/ha, which is similar to the standard application rate for field application. Maize plants are grown at temperatures of 20 to 25 ℃ with regular irrigation. Pictures (fig. 1 and 2) at eight weeks after treatment (8 WAT) were recorded and the dry mass of roots (fig. 4) and shoots (fig. 3) were determined.
Shows the combination with an auxiliary agentThe efficacy of the combined Paenibacillus xylan DSM 34353 was excellent.
Further, mixtures with other microorganisms such as biofertilizer Blue-N with Methylobacterium symbioticum (Corteva, inc, USA), kreotec (Biofa) with Bacillus belicus (Bacillus velezensis), azospirillum brasilense (Azospirillum brasilense) and Herbaspirillum seropedicae, biofertilizer Utrisha TM -N with Methylobacterium symbioticum SB23 (Corteva, inc, USA), blue-N with Methylobacterium symbioticum (Corteva, inc, USA), bacillus atrophicus (Bacillus atrophaeus)(AbiTep, germany) RhizoFos (Rizobacter, USA) with Pseudomonas fluorescens (Pseudomonas fluorescens) was used as a control for comparison studies. These comparative products also contain microorganisms capable of fixing atmospheric nitrogen and are also applied to the leaves. However, using reduced artificial nitrogen fertilization (21 kgN/ha), the comparative products showed no or lower growth promoting effect.
These comparative experiments showed that the use of Paenibacillus xylan in combination with adjuvants resulted in better plant growth under nitrogen limitation compared to when Utrisha TM -N, blue-N or Kreotec were used.
Example 3
Research on the effect of different mineral nitrogen fertilization doses on the efficacy of endophyte xylan paenibacillus of nitrogen fixing bacteria
The general purpose of example 3 was to find out how much nitrogen the bacterial endophyte xylan paenibacillus could fix under field conditions, i.e. at different mineral fertilisation doses for crop selectivity and yield (grain and shoots).
When treating seeds, bacterial endophytes are carried out by wet seed treatment. When foliar application, bacterial endophytes are applied in the 5-6 leaf stage.
Corn seed treatment
100,000 Seeds were planted and 1,000,000CFU (bacterial units) were applied per seed. 1X 10 11 CFU/ha at 100,000 grains/ha
Each bacterial suspension had 2.5X10 8 CFU/mL, thus requiring 400mL of the test product per 100 000 pellets. This resulted in 4mL of experimental product (with 2.5x10 8 CFU) per 1000 pellets. A mixture of 4mL of the test product with 1mL PREMAX and a marinade (pickle) was prepared for 1000 kernels, and then left to dry in the dark (e.g., open paper bag). Nothing was added to the control.
Foliar treatment
50,000CFU (units) of experimental bacteria were added to each plant. Plants are in the 5-6 leaf stage, where only about 10% or less of the soil area is covered by plants. 100,000 plants/ha and 50,000CFU were used, which resulted in a total of 10% area of 5x10 9 CFU/1 ha-i.e. at 5x10 10 CFU/ha on 1 ha. 200mL/ha of the experimental bacterial product with 2.5X10 8 CFU/mL was sprayed-this resulted in 5X10 10 CFU/ha.
Dilution of 10mL of the experimental product (2.5X10 8 CFU/mL) with 90mL of demineralised water gives 100mL of 2.5X10 7 CFU/mL. In this 100mL (with 2.5X10 7 CFU/mL), 20mL/100m 2 -or 2000mL/ha was applied. The water application rate (spray liquid) was 300L/ha, and BREAK was addedSP 133:300mL/ha (=0.1%). All experiments were performed in germany.
The following experiments were performed:
The results are provided in table 2 below and fig. 5.
TABLE 2 results for example 3
The results show that the overall yield increases when a biostimulant (i.e., paenibacillus xylan) is applied in addition to the synthetic fertilizer.
Claims (13)
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
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| EP22193373.2 | 2022-09-01 | ||
| EP22193373 | 2022-09-01 | ||
| PCT/EP2023/073235 WO2024046864A1 (en) | 2022-09-01 | 2023-08-24 | Endophytic microbial biostimulants |
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| EP (1) | EP4580408A1 (en) |
| CN (1) | CN120456819A (en) |
| AR (1) | AR130354A1 (en) |
| AU (1) | AU2023331560A1 (en) |
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| BR112013030521B1 (en) * | 2011-05-27 | 2022-03-29 | Bayer Cropscience Biologics Gmbh | Liquid preparation for biological plant protection, method for producing a liquid preparation, and use of the liquid preparation |
| DE102011078624A1 (en) | 2011-07-05 | 2013-01-10 | Evonik Goldschmidt Gmbh | Use of hydrophilic organo-modified siloxanes as process aids for melt granulation |
| PL3203838T3 (en) | 2014-10-08 | 2020-09-07 | Evonik Operations Gmbh | Use of hydrophobic, self-emulsifying polyglycerol esters as adjuvants and anti-spray drift agents |
| PL3106033T3 (en) | 2015-06-16 | 2017-09-29 | Evonik Degussa Gmbh | Biodegradable super dispersing organomodified trisiloxane |
| KR101621732B1 (en) * | 2016-02-03 | 2016-05-17 | 충북대학교 산학협력단 | Halotolerant Paenibacillus xylanexedens strain promoting plant growth condition and uses thereof |
| CA3097236A1 (en) * | 2018-04-19 | 2019-10-24 | Evonik Operations Gmbh | Biocompatible siloxanes for formulation of microorganisms |
| PL3972412T3 (en) * | 2019-05-20 | 2026-01-26 | Evonik Operations Gmbh | APPLICATION OF GLYCEROL POLYESTERS AS CARRIERS FOR MICROBIOLOGICAL ACTIVE SUBSTANCES |
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