METHODS FOR ENHANCED ROOT NODULATION IN LEGUMES
Disclosed herein are methods of increasing, enhancing, or accelerating root nodulation in a plant, accelerating growth of nitrogen fixing bacteria in nodules of a plant, increasing protein content in a plant, increasing yield of a plant, improving water retention of a plant, or reducing water use of a plant, the method comprising identifying a plant in need of root nodulation, and applying to the plant a composition comprising a protein component comprising yeast stress proteins resulting from subjecting a mixture obtained from the yeast fermentation to stress.
1 . A method of increasing, enhancing, or accelerating root nodulation in a legume plant, accelerating growth of nitrogen fixing bacteria in nodules of a legume plant, increasing protein content in a legume plant, increasing yield of a legume plant, improving water retention of a legume plant, or reducing water use of a legume plant, the method comprising:
applying to the legume plant a composition comprising:
a yeast preparation component comprising yeast stress proteins resulting from subjecting a mixture obtained from yeast fermentation to stress;
at least one non-ionic surfactant;
at least one anionic surfactant; and
water to make 100% volume;
wherein the yeast preparation component comprising yeast stress proteins is produced by a method comprising:
growing yeast under aerobic fermentation conditions to obtain a fermentation mixture comprising fermented yeast cells, and proteins and peptides secreted therefrom;
subjecting the fermentation mixture to stress in order to stimulate the yeast cells to express heat shock proteins to obtain a stressed fermentation mixture;
separating the stressed fermentation mixture into a solid/precipitate and a supernatant; and
adjusting the pH of the supernatant to produce the yeast preparation component comprising yeast stress proteins.
2 . The method of claim 1 , wherein the composition applied to the legume plant comprises:
23% by volume of the yeast preparation.
3 . The method of claim 1 , wherein the composition applied to the legume plant comprises:
7.5% by volume non-ionic surfactant(s).
4 . The method of claim 1 , wherein the composition applied to the legume plant comprises:
2.5% by volume anionic surfactant(s).
5 . The method of claim 1 , wherein the composition applied to the legume plant comprises:
23% by volume of the yeast preparation;
7.5% by volume non-ionic surfactant(s); and
2.5% by volume anionic surfactant(s).
6 . The method of claim 1 , wherein the at least one non-ionic surfactant comprises at least one of a dodecyl dimethylamine oxide, a coco diethanol-amide alcohol ethoxylate, a linear primary alcohol polyethoxylated, an alkylphenol ethoxylate, an alcohol ethoxylate, an EO/PO polyol block polymer, a polyethylene glycol ester, or a fatty acid alkanolamide.
7 . The method of claim 1 , wherein the at least one anionic surfactant comprises at least one of sodium linear alkylbenzene sulphonate (LABS), sodium lauryl sulphate, sodium lauryl ether sulphate, a petroleum sulphonate, a linosulphonate, a naphthalene sulphonate, a branched alkylbenzene sulphonate, a linear alkylbenzene sulphonate, an alcohol sulphate, or PO and/or PO/EO sulfated alcohol.
8 . The method of claim 1 , wherein the composition applied to the legume plant comprises:
23% by volume of the yeast preparation;
7.5% by volume linear primary C12-C15 alcohol; and
2.5% by volume sodium lauryl ethyl sulfate.
9 . The method of claim 1 , wherein the composition is applied to the soil near the plant.
10 . The method of claim 1 , wherein the composition is applied through irrigation.
11 . The method of claim 10 , wherein the irrigation is spray irrigation or drip irrigation.
12 . The method of claim 1 , wherein the composition is applied with every watering cycle or on an intermittent basis.
13 . The method of claim 1 , wherein the aerobic fermentation conditions comprise continuous aeration and agitation between 30 to 35° C. and at a pH of 4.0 to 6.0 for a period of at least 10 hours.
14 . The method of claim 1 , wherein subjecting the fermentation mixture to stress comprises heating the fermentation mixture to 40-60° C. for at least 2 hours.
15 . The method of claim 14 , wherein subjecting the fermentation mixture to stress further comprises cooling the fermentation mixture to 25° C. after heating it.
16 . The method of claim 1 , further comprising adding sodium benzoate and propylene glycol to the supernatant, to produce the protein component comprising yeast stress proteins.
17 . The method of claim 1 , wherein the stress further comprises physically or chemically disrupting the yeast after fermentation of the yeast.
18 . The method of claim 1 , wherein the stress further comprises lysing the yeast after fermentation of the yeast.
19 . The method of claim 1 , further comprising mixing the protein component with additional nutrients prior to the application to the plant.
20 . The method of claim 1 , wherein the composition further comprises one or more of an anionic surfactant, a non-ionic surfactant, a cationic surfactant, and amphoteric surfactant.
21 . The method of claim 1 , wherein the yeast is selected from the group consisting of Saccharomyces cerevisiae, Kluyveromyces marxianus, Kluyveromyces lactis, Candida utilis (Torula yeast), Zygosaccharomyces, Pichia pastoris , and Hansanula polymorpha.
22 . The method of claim 1 , where separating the stressed fermentation mixture into a solid/precipitate and a supernatant comprises centrifugation.
23 . The method of claim 1 , wherein the legume plant is selected from the group consisting of alfalfa, clover, peas, beans, lentils, lupins, mesquite, carob, soy, peanuts, locust trees ( Gleditsia or Robinia ), wisteria , and the Kentucky coffeetree ( Gymnocladus dioicus ).
24 . A soil mixture comprising soil and a composition comprising:
a yeast preparation component comprising yeast stress proteins resulting from subjecting a mixture obtained from yeast fermentation to stress;
at least one non-ionic surfactant;
at least one anionic surfactant; and
water to make 100% volume;
wherein the yeast preparation component comprising yeast stress proteins is produced by a method comprising:
growing yeast under aerobic fermentation conditions to obtain a fermentation mixture comprising fermented yeast cells, and proteins and peptides secreted therefrom;
subjecting the fermentation mixture to stress in order to stimulate the yeast cells to express heat shock proteins to obtain a stressed fermentation mixture;
separating the stressed fermentation mixture into a solid/precipitate and a supernatant; and
adjusting the pH of the supernatant to produce the yeast preparation component comprising yeast stress proteins.
25 . The method of claim 24 , wherein the aerobic fermentation conditions comprise continuous aeration and agitation between 30 to 35° C. and at a pH of 4.0 to 6.0 for a period of at least 10 hours.
26 . The method of claim 24 , wherein subjecting the fermentation mixture to stress comprises heating the fermentation mixture to 40-60° C. for at least 2 hours.
27 . The method of claim 26 , wherein subjecting the fermentation mixture to stress further comprises cooling the fermentation mixture to 25° C. after heating it.
28 . The method of claim 24 , further comprising adding sodium benzoate and propylene glycol to the supernatant, to produce the protein component comprising yeast stress proteins.
29 . The method of claim 24 , wherein the stress further comprises physically or chemically disrupting the yeast after fermentation of the yeast.
30 . The method of claim 24 , wherein the stress further comprises lysing the yeast after fermentation of the yeast.
31 . The method of claim 24 , further comprising mixing the protein component with additional nutrients prior to the application to the plant.
32 . The method of claim 24 , wherein the composition further comprises one or more of an anionic surfactant, a non-ionic surfactant, a cationic surfactant, and amphoteric surfactant.
33 . The method of claim 24 , wherein the yeast is selected from the group consisting of Saccharomyces cerevisiae, Kluyveromyces marxianus, Kluyveromyces lactis, Candida utilis (Torula yeast), Zygosaccharomyces, Pichia pastoris , and Hansanula polymorpha.
34 . The method of claim 24 , where separating the stressed fermentation mixture into a solid/precipitate and a supernatant comprises centrifugation.
35 . The method of claim 24 , wherein the composition applied to the legume plant comprises:
23% by volume of the yeast preparation.
36 . The method of claim 24 , wherein the composition applied to the legume plant comprises:
7.5% by volume non-ionic surfactant(s).
37 . The method of claim 24 , wherein the composition applied to the legume plant comprises:
2.5% by volume anionic surfactant(s).
38 . The method of claim 1 , wherein the composition applied to the legume plant comprises:
23% by volume of the yeast preparation;
7.5% by volume non-ionic surfactant(s); and
2.5% by volume anionic surfactant(s).
39 . The method of claim 24 , wherein the at least one non-ionic surfactant comprises at least one of a dodecyl dimethylamine oxide, a coco diethanol-amide alcohol ethoxylate, a linear primary alcohol polyethoxylated, an alkylphenol ethoxylate, an alcohol ethoxylate, an EO/PO polyol block polymer, a polyethylene glycol ester, or a fatty acid alkanolamide.
40 . The method of claim 24 , wherein the at least one anionic surfactant comprises at least one of sodium linear alkylbenzene sulphonate (LABS), sodium lauryl sulphate, sodium lauryl ether sulphate, a petroleum sulphonate, a linosulphonate, a naphthalene sulphonate, a branched alkylbenzene sulphonate, a linear alkylbenzene sulphonate, an alcohol sulphate, or PO and/or PO/EO sulfated alcohol.
41 . The method of claim 24 , wherein the composition applied to the legume plant comprises:
23% by volume of the yeast preparation;
7.5% by volume linear primary C12-C15 alcohol; and
2.5% by volume sodium lauryl ethyl sulfate.
42 . A method of improving water retention of a legume plant, reducing water use of a legume plant, accelerating root nodulation in a legume plant, accelerating nitrogen fixation by a legume plant, accelerating growth of nitrogen fixing bacteria in nodules of a legume plant, increasing protein content in a legume plant, or increasing yield of a legume plant, the method comprising: growing the legume plant in the soil mixture of claim 24 .