WO2015137928A1 - Compositions de traitement des semences contenant des métabolites dérivés de rhizobium - Google Patents

Compositions de traitement des semences contenant des métabolites dérivés de rhizobium Download PDF

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Publication number
WO2015137928A1
WO2015137928A1 PCT/US2014/023537 US2014023537W WO2015137928A1 WO 2015137928 A1 WO2015137928 A1 WO 2015137928A1 US 2014023537 W US2014023537 W US 2014023537W WO 2015137928 A1 WO2015137928 A1 WO 2015137928A1
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Prior art keywords
rhizobium
product
seed
composition
combinations
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PCT/US2014/023537
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English (en)
Inventor
Gary Michael NIJAK JR.
Mark O'donnell
Samuel Jacob VIGUE
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Green & Grow, Inc.
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Priority to PCT/US2014/023537 priority Critical patent/WO2015137928A1/fr
Publication of WO2015137928A1 publication Critical patent/WO2015137928A1/fr

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    • 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
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
    • 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
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change

Definitions

  • the invention relates generally to a composition of materials containing a rhizobium derived composition that when combined with conventional and novel seed treatment technologies promotes growth of all types of plants.
  • Seed treatments are a significant and growing industry with an estimated $3.2 billion value as of 2013. Over the next 5 years they're expected to have an annual compounded growth rate of more than 8 percent reaching $4.8 billion by 2018 (PRWeb, 2013).
  • the quick growth rates in the seed treatment industry have been catalyzed through the increased role of the seed in agriculture as a high-value input. This has occurred as biotechnological advances have increased the expectations of producers for maximum performance of input and output traits (Munkvold, 2009).
  • Seed treatment therefore, provides a highly economical method for maximizing food production world-wide, and will play a significant global role in reducing food insecurity and feeding a growing population.
  • seed treatments allows for the close proximity of active materials to the seed and, thus, can induce an effective and rapid response.
  • the intended responses include pest control, plant health through nutrition, and disease control.
  • systemic fungicides and insecticides that provide residual pest control after the emergence of the seedlings has greatly expanded the use and effectiveness of seed treatments as well as the number of seed treatment products on the market today (Hairston, 2013).
  • Traditional seed treatments include fungicides and insecticides in varying combinations to promote early growth of the plant.
  • Chemical stimulants such as gibberellic acids and other stimulants promote early growth and germination of the seed.
  • Macronutrients are sometimes used to provide starter nutrients at germination.
  • micronutrient deficiencies in the soil can be corrected by applying micronutrients as a seed treatment due to the very small quantity required by plants (Farooq, 2012).
  • the present invention meets the above-identified needs by providing a composition containing a metabolite mixture derived from a Rhizobium spp. bacteria that can be applied with various seed treatment packages and that can provide significant and unexpected benefits across a range of crops.
  • a composition containing a metabolite mixture derived from a Rhizobium spp. bacteria that can be applied with various seed treatment packages and that can provide significant and unexpected benefits across a range of crops.
  • the ability of these types of derived materials to provide additional benefits to a seed treatment package on top of an existing optimized, extensively
  • One embodiment relates to a product including at least one Rhizobium spp. derived metabolite and a seed treatment material.
  • the seed treatment material can include a fungicide, an insecticide, a plant growth regulator, a plant growth promoter, a micronutrient, and combinations thereof.
  • the Rhizobium spp. derived metabolite is not derived from a lipochitooligosaccharide (LCO).
  • the fungicide can be, but is not limited to, metalaxyl, carboxin, menfenoxam, triticonazole, fludioxonil and combinations thereof.
  • the insecticide can be, but is not limited to, imidacloprid, thiamethoxam, clothianidin, cyromazine, permethrin and combinations thereof.
  • the plant growth regulator or promoter can be, but is not limited to, gibberellic acid, cytokinin, indolebutyric acid, kelp, indoleacetic acid, amino acid and combinations thereof.
  • the micronutrient can be, but is not limited to, zinc, copper, iron, molybdenum, manganese, and combinations thereof.
  • the nematicide can be, but is not limited to abamectin, harpin, spinosad, chitin and combinations thereof.
  • the product can be produced by a process including, but not limited to the steps of adding a broth to a biological reactor, culturing the broth to produce a culture containing Rhizobium derived metabolites; terminating viability of the culture; and adding the seed treatment material to the culture to make a seed treatment composition.
  • the broth can include a carbon source, the carbon source including at least one
  • the step of terminating viability of the culture can be performed by lysis, stabilization, thermal treatment, combinations thereof, and other suitable methods.
  • the seed treatment composition can be concentrated 2 to 10 times to produce a final composition that has an increased concentration of Rhizobium derived metabolites.
  • the Rhizobium spp. can be a Rhizobiaceae.
  • the Rhizobiaceae can be, but is not limited to, Rhizobium etli, Rhizobium leguminosarum, Rhizobium phaseoli, Rhizobium tropici, Rhizobium fredii, Rhizobium meliloti, and combinations thereof.
  • Various embodiments relate to a method comprising applying a product to one or more seeds.
  • the product can include at least one Rhizobium spp. derived metabolite and a seed treatment material.
  • the seed treatment material can include, but is not limited to, a fungicide, an insecticide, a plant growth regulator, a plant growth promoter, and combinations thereof.
  • the product can be applied by a method including, but not limited to, film coating, encrusting, pelleting, priming, film coating, drenching, and combinations thereof.
  • the product can be applied at a rate of from 0.00025 to 10 mg per seed.
  • Applying the product can increase the overall yield of the plants grown from the treated seed as compared to plants grown from seeds not treated with the product. Applying the product can increase the harvestable fruit by at least 0.5 bushels per acre as compared to plants grown from seeds not treated with the product. Applying the product can increase the harvestable foliage by at least 5% as compared to plants grown from seeds not treated with the product. Applying the product can promote the overall growth of the plant for the treated seed as compared to plants not treated with the product. Applying the product can increase the overall growth of the plant for the treated seed as compared to the plants not treated with the product by at least 5% of the maximum above ground length.
  • Various embodiments relate to a method for producing a product that includes at least one Rhizobium spp.
  • the seed treatment material can include, but is not limited to a fungicide, an insecticide, a plant growth regulator, a plant growth promoter, and combinations thereof.
  • the method can include, but is not limited to the steps of adding a broth to a biological reactor, culturing the broth to produce a culture; terminating viability of the culture; and adding the seed treatment material to the culture to produce the product.
  • the broth can include a carbon source, the carbon source including at least one monosaccharide and at least one higher order saccharide; a source of nutrients, the source of nutrients comprising at least one phosphate, a nitrogen source, at least one source of magnesium; and a Rhizobium spp.
  • the step of terminating viability of the culture can be performed by lysis, stabilization, thermal treatment, and combinations thereof.
  • Various embodiments relate to a method of manufacturing a seed treatment composition, the seed treatment composition including at least an exudates produced by Rhizobium spp. and one other seed treatment material. More specifically, the method can include, but is not limited to the steps of cultivating a composition; adding a broth to a biological reactor, allowing a culture of bacteria to grow in an aerobic biological reactor for 2-6 weeks;
  • the broth can include a carbon source consisting of at least one monosaccharide and at least one higher order saccharide; a source of nutrients including one or more phosphates, a nitrogen source, at least one source of magnesium; and a Rhizobium spp.
  • Figure 1 is an HPLC spectra demonstrating a lack of LCOs in the
  • Rhizobium spp. derived composition Rhizobium spp. derived composition
  • Figure 2 is a chart plotting romaine lettuce shoot length against
  • Figure 3 is a chart showing Corn seed treatment comparison
  • Rhizobium spp. derived composition Rhizobium spp. derived composition
  • Figure 4 is a chart showing Soybean seed treatment comparison between Rhizobium spp. derived composition plus chemical combination versus composition without
  • Rhizobium spp. derived composition Rhizobium spp. derived composition
  • Figure 5 is a chart showing Soybean seed treatment comparison between Rhizobium spp. derived composition plus chemical combination versus composition without
  • Rhizobium spp. derived composition Rhizobium spp. derived composition
  • Rhizobium derived, Rhizobium exudate, derived by Rhizobium, Rhizobium produced, produced by Rhizobium means a composition that is derived from maintaining a culture of Rhizobium in stationary phase for an extended period of time (at least 20 days after achieving stationary phase), in a highly enriched carbon environment as compared to the nitrogen content of the culture (in excess of 4 to 1 and preferred to be above 10 to 1 ) during the entire stationary phase to induce the production of exudates of which the bulk exudate will be an
  • exopolysaccharide At no point are flavonoids introduced which are typically considered a means to produce lipochitooligosaccharides, a known exudate that has beneficial plant and bacteria interactions. After at least 20 days in stationary phase, the culture is then thermally lysed and concentrated denaturing any unstable exudates in the mixture and terminating the viability of the culture.
  • the then produced exudates exhibit a response and interaction with living biologicals which is not solely due to the production of vitamins such as B-12, hormones such as auxin, cytokin, gibberrilic acid, indole acetic acid, ACC deaminase (1 -aminocyclopropane-1 -carboxylate) or plant signaling compounds such as lipochitooligosaccharides or due to the
  • exopolysaccharide contained within the metabolite mixture All of these chemicals are measurable by assays known in the literature.
  • the biological reactor can be aerated to an aerobic state and fed the raw materials necessary for the culture.
  • the batch can be feed every other day with a sugar source.
  • the suitable period of time can be within a range having a lower limit and/or an upper limit.
  • the range can include or exclude the lower limit and/or the upper limit.
  • the lower limit and/or upper limit can be selected from 1 , 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 1 10, 120, 130, 140, 150, 160, 170, 180, 190, 200, 300, 400, 500, 600, 700, 800, 900, and 1000 days.
  • the suitable period of time for allowing the materials to remain in the biological reactor can be from 20 to 49 days.
  • the reactor can be brought to full volume by expanding the culture and maintaining a sugar concentration below a Brix of 1 .5 such that the free or unutilized sugar concentration is reduced to nearly zero once the bacteria reach steady state. These steps can be repeated until the bacteria culture reaches full volume. Once the full reactor volume is achieved, the sugar can be spiked.
  • the suitable level of carbon to nitrogen within the culture can be within a range having a lower limit and/or an upper limit. The range can include or exclude the lower limit and/or the upper limit.
  • the lower limit and/or upper limit can be selected from 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 , 32, 33, 34, 35, 36, 37, 38, 39, 40, 41 , 42, 43, 44, 45, 46, 47, 48, 49, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 1 10, 120, 130, 140, and 150 C:N ratio.
  • the suitable C:N for allowing the materials to produce the Rhizobium spp. derived metabolites can be from 10-120 C:N ratio and maintained at this level +/- 10 C:N.
  • the bacteria culture can be maintained in a decay rather than growth stage, i.e. the number of viable cells is decreasing.
  • the C:N ratio can be increased once the tank has reached steady state.
  • the tank after the appropriate period can be treated to lyse the cells and evaporate the material into a condensed form.
  • the suitable level of concentration of the culture composition can be within a range having a lower limit and/or an upper limit.
  • the lower limit and/or upper limit can be selected from 1 .5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, and 15.
  • the suitable concentration factor to produce the concentrated composition can be from 2-10 times. In doing so, unstable metabolites are degraded in the final composition.
  • No flavonoids are introduced into the process which results in a mixture that does not contain flavonoid induced products such as lipochitooligosaccharides (LCOs).
  • LCOs lipochitooligosaccharides
  • This material does not contain unstable materials such as LCOs and the
  • concentration process eliminates and would degrade any of these materials if they were to be produced in the culturing process.
  • a microbial growth inhibitor such as an alkaline material like sodium hydroxide can be added. Additional amendments and combinations can be added to this Rhizobium spp. derived composition.
  • the broth in which the Rhizobium spp. materials are cultured can include at least a nitrogen source, sugar source, and magnesium sulfate.
  • the nitrogen source suitable level of concentration of the culture composition can be within a range having a lower limit and/or an upper limit.
  • the lower limit and/or upper limit can be selected from 0.005, 0.006, 0.007, 0.008, 0.009, 0.01 , 0.01 1 , 0.012, 0.013, 0.014, 0.015, 0.016, 0.017, 0.018, 0.019, 0.02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0.1 , 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1 , 2, 3, 4, 5, 6, 7, 8, 9, and 10% of the culture by weight.
  • the suitable nitrogen source concentration to produce composition can be from 0.005 to 0.5% by weight of the culture composition.
  • the sugar source suitable level of concentration of the culture composition can be within a range having a lower limit and/or an upper limit.
  • the lower limit and/or upper limit can be selected from 0.05, 0.10, 0.20, 0.30, 0.40, 0.50, 0.60, 0.70, 0.80, 0.90, 1 .0, 1 .1 , 1 .2, 1 .3, 1 .4, 1 .5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 1 1 , 12, 13, 14, 15, 16, 17, 18, 19, 20, 21 , 22, 23, 24, 25, 26, 27, 28, 29, and 30% by weight of the culture.
  • the suitable sugar source concentration to produce composition can be from 0.05 to 15% by weight of the culture composition.
  • magnesium sulfate suitable level of concentration of the culture composition can be within a range having a lower limit and/or an upper limit.
  • the lower limit and/or upper limit can be selected from 0.0005, 0.0010, 0.0020, 0.0030, 0.0040, 0.0050, 0.0060, 0.0070, 0.0080, 0.0090, 0.010, 0.01 1 , 0.012, 0.013, 0.014, 0.015, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, and 0.10% by weight of the culture.
  • the magnesium sulfate concentration to produce composition can be from 0.005 to 0.10% by weight of the culture composition.
  • the remaining portions of the culture can be water.
  • Rhizobium bacteria can be selected from Rhizobium alamii, Rhizobium alkalisoli, Rhizobium cellulosilyticum, Rhizobium daejeonense , Rhizobium endophyticum, Rhizobium etli, Rhizobium galegae, Rhizobium gallicum, Rhizobium giardinii, Rhizobium hainanense, Rhizobium herbae, Rhizobium huautlense, Rhizobium indigoferae, Rhizobium
  • leguminosarum including biovars viciae, phaseoli, and trifolii, Rhizobium loessense formerly Rhizobium huanglingense, Rhizobium lusitanum,
  • Mesorhizobium metallidurans Mesorhizobium opportunistum, Mesorhizobium plurifarium, Mesorhizobium robiniae, Mesorhizobium shangrilense,
  • Sinorhizobium morelense Ensifer adhaerens, Ensifer numidicus, Ensifer saheli, Ensifer sojae, Ensifer terangae, Bradyrhizobium canariense,
  • Bradyrhizobium pachyrhizi and Bradyrhizobium yuanmingense.
  • triazolopyrimidines sulfonylureas, imidazolinones, and sulfonylamino carbonyl triazolinones
  • carbamates triazines
  • glycerol ethers EPSPS inhibitors (glyphosate)
  • Synthetic auxins (2,4-D, 2,4-DB, 2,4-DP, MCPA, MCPB, MCPP)
  • acetamides dinitroanilines
  • bipirydiums diquat and
  • diphenilethers nitrofen , nitrofluorfen, and acifluoren
  • pyridazinones uracils, phenoxys, ureas, and benzoic acids growth regulators, pigment inhibitors, and photosystem inhibitors
  • Seed treatment herbicides which can include clopyrallid, dicamba, imazapyr, picloram, prosulfuron, and Phomopsis amaranthicola; Herbicide safeners such as fluxofenim, benzoxazine, benzhydryl derivitives, ⁇ , ⁇ -diallyl dichloroacetamide, dihaloacyls, oxazolidinyls, thiazolidinyls, ethanone, naphthalic anhydrides, and oxime derivatives; Algicides such as copper sulfate, hydrogen peroxide, and peracetic acid; Avicides; Bactericides (acibenzolar -S-methyl, bacteriophages, chlorine, copper sulfate, high valency silver, hot water, oxidized copper, sodium hypochlorite
  • mandipropamid maneb, mefenoxam, metalaxyl, metalaxyl-m, metconazole, myclobutanil, penconazole, penflufen, pentachloronitrobenzene,
  • penthiopyrad phosphorus acid, plant extracts, plant oils, propiconazole, propineb, prothioconazole, pyraclostrobin, Reynoutria sachalinensis, sedaxane, tebuconazole, thiabendazole, thiophanate methyl, thiram, triadimefon, triadimenol, trifloxystrobin, trisodium phosphate, triticonazole, and valifenalate; Insecticides such as neonicotinoids (acetamiprid, clothianidin, dinotefuran, imidacloprid, nitenpyram, nithiazine, sulfoxaflor, thiacloprid, and thiamethoxam etc.), carbamates (carbofuran etc.), insect growth regulators (cryomazine etc.), organochlorides, organophosphates (chlorpyrifos etc.),
  • Rodenticides such as anthraquinone and methiocarb
  • Bird repellents such as anthraquinone and methiocarb
  • Macronutrients such as nitrogen (ammonium sulfate
  • brassinosteroids morphactin, aminotriazole, chloroethylphosphonic acid, jasmonic acids, salicylic acids, mepiquat salts, 1 -methylcyclopropane, aviglycine, 1 -naphthnaleneacetic acid, chlormequat chloride, mepiquat, mepiquat pentaborate, chlorofluren, chloroflurenol, dichloroflurenol, flurenol, daminozide, flurprimidol, mefluidide, paclobutrazole, cyproconazole, tetcyclacis, uniconazole, karrikins, clofibric acid, 2,3,5-tri-iodobenzoic acid, 2,4-dichlorophenoxy acetic acid, dichloroprop, fenoprop, naphthalene acetamide, 1 -naphthole, naphthoxy acetic acid, potassium
  • polyethylenes polychloroprene, alginic acid, pectic acid, celluloses, natural and synthetic waxes, kaolin clay, charcoal, and silicates
  • Seed treatment components such as colorants (dyes), solvents, surfactants, emulsion stabilizers, antifreeze compounds, adjuvants, inert fillers (vermiculte, talc, woodflours, clays, activated carbon, sugars, diatomaceous earth, cereal flours, fine-grain inorganic soilds, calcium carbonate, calcium bentonite, kaolin, china clay, perlite, mica, silicas, quartz powders, montmorillonite etc.), and dessicants; Viroid control agents; RNA interface technology; Seed Disinfectants such as sodium hypochlorite, hydrogen peroxide, fatty acid monoesters, calcium hypochlorite, potassium permanganate, steam, hot humid air.
  • seed treatment includes the finished product of the present invention through its multiple forms and is applicable for all forms of seed coatings and enhancements. These include but are not limited to priming, solid matrix priming, pelleting, coating, dusting, encrustation, drenching, dressing, soaking, and any combination of these methods.
  • methods used pertain to all forms and mixtures in single and additive products during and following seed coating and enhancement processes.
  • priming additive products within a prescribed environment are used to initialize the physiological processes of the seed, while preventing germination until planting.
  • the pelleting process involves adding substantial inert materials to seeds for ease of planting and emergence benefits.
  • Coating involves material application with a binder for chemical adherence to the seed.
  • Dusting involves adding a powdered substance to the seed for increased flow-ability and/or active enhancement compounds.
  • Encrustation involves increasing the seed size with inert and active compounds beyond a film coating, while without greatly altering the seed's natural shape.
  • Drenching involves multiple methods of the applying liquids to seeds, typically during planting. Using farm implements to apply seed enhancement products to the soil surrounding the seed is a method of use applicable for the present invention.
  • seed includes plant propagules of all types such as but not limited to true seeds, somatic embryos, synthetic seeds, seed pieces, suckers, corms, bulbs, fruit, tubers (eg. potatoes etc.), grains, cuttings, and shoots.
  • composition was transferred into a Spectra Por Dialysis Membranes (MWCO 1000). Dialysis was performed against 4 L of running Dl water at ambient temperature for one week to remove salt, monomers and other contaminants. After dialysis, the sample was lyophilized.
  • MWCO 1000 Spectra Por Dialysis Membranes
  • MALDI matrix-assisted laser desorption/ionization
  • Flavonoids when introduced into a culture of Rhizobium bacteria nodulate and induce the production of LCOs.
  • the most commonly utilized flavonoid is Naringenin, which is a known nodulator and inducer of LCO production. No flavonoids are used in the production of the Rhizobium derived metabolite mixture and thus, no LCOs are expected and found.
  • Romaine lettuce seeds were tested in a sterile agar media that simulates a readily available soil nutrient package that includes a calcium chloride, a magnesium sulfate, a monopotassium phosphate, a potassium nitrate, and a ammonium nitrate in a nutrient agar.
  • a Rhizobium spp. derived composition previously described is incorporated into the agar medium plate at several concentrations from 0.25% of the volume per plate to 1 % of the agar media volume of the poured plate, ensuring that the overall amount of nutrients provided on each agar plate remains constant with the addition of the Rhizobium spp. derived composition.
  • the nutrient agar is prepared by adding 20 g of agar to 1 L of water, heating the solution to activate the agar, then pouring the material into plates.
  • Romaine lettuce seeds are sterilized by quick immersion in a dilute (less than 0.1 M) sodium hypochlorite washing solution to eliminate any potential bacteria contamination in the lettuce seed during the length of the assay.
  • a dilute (less than 0.1 M) sodium hypochlorite washing solution to eliminate any potential bacteria contamination in the lettuce seed during the length of the assay.
  • Nine sterilized romaine lettuce seeds are then placed onto each agar medium plate test. Four replications for each test or rate are measured.
  • the plates are placed in an incubator chamber for a period of 7 days in the dark and maintained at a constant 27°C chamber. At the end of the 7 day assay, the seeds are removed and the shoot is measured. The resulting
  • Table 1 shows Rhizobium spp. derived composition concentration and the corresponding and resulting shoot length average for each of the concentrations showing a significant increase in the average length of the shoot of the Romaine lettuce seeds when in proximity to the Rhizobium spp. derived composition, even when composed on an optimum growth medium with an optimal availability of micronutrients. Results are shown in Figure 2.
  • Field corn trial plots were planted in a partially randomized block design with four rows per block and one block for each treatment replication. Four replications are planted for each treatment at each location of the
  • Rhizobium spp. derived composition plus a combination of consisting of fludioxonil, mefenoxam, azoxystrobin, thiabendazole, thiamethoxam and a control treated consisting only of fludioxonil, mefenoxam, azoxystrobin, thiabendazole, thiamethoxam (all components except Rhizobium spp. derived composition).
  • the active rate of each of the components is in the range of 0.0001 mg of ai per seed to 1 .25 mg of active ingredient (ai) per seed where the optimum rate is 0.0025 mg of azostrobin per seed, 0.0065 mg of fludioxonil per seed, 0.005 mg of mefenoxam per seed, 0.05 of thiabendazole per seed, 0.05 mg of thiamethoxam per seed, and 0.2 mg of Rhizobium spp. derived composition per seed.
  • the individual blocks had a length of 20 feet. Portions at the end of the plot not used to plant blocks are planted with a filler. Edges of the field around the trials are planted with a filler variety of the same crop type so that all test areas are at least 20 feet from field edges.
  • Each block is planted with four rows of commercially treated corn seed per treatment.
  • two varieties are planted at each location. All treatments and one control are planted with four blocks (reps) per each treatment.
  • Trials may be either center pivot irrigated or dry land (non-irrigated natural rainfall dependent). Standard production practices are followed uniformly for the field throughout the season.
  • the two center rows harvested for the field yield results. Each harvested block is moisture tested and corrected for actual yield to 15 percent moisture. The yield data for each replication and block is converted to a per acre value. Results are shown in Figure 3.
  • Soy bean trial plots were planted in a partially randomized block design with four rows per block and one block for each treatment replication. Four replications are planted for each treatment at each location of the Rhizobium spp. derived composition plus a combination of consisting of ipconazole (0.005 mg/seed), imidacloprid (0.1 mg/seed), metalaxyl (0.006 - 0.02 mg/seed), and thiram (0.1 mg/seed), one control consisting only of
  • ipconazole imidacloprid, metalaxyl, and thiram (all components except Rhizobium spp. derived composition), and another control consisted of no treatment onto the seed.
  • Portions at the end of the plot not used to plant blocks are planted with a filler. Edges of the field around the trials are planted with a filler variety of the same crop type so that all test areas are at least 20 feet from field edges.
  • Each block is planted with four rows of treated soybean seed per treatment. For all the trials, two varieties are planted at each location. All test are planted with four blocks (reps) per each treatment. Trials may be either center pivot irrigated or dry land. Standard production practices are followed uniformly for the field throughout the season. At harvest, the two center rows harvested for the field yield results. Each harvested block is moisture tested and corrected for actual yield to 13 percent moisture. The yield data for each replication and block is converted to a per acre value. Results are shown in Figures 4 and 5.
  • Example 1 the increased shoot length in the romaine lettuce seeds when in the proximity of the Rhizobium spp. derived composition is surprising.
  • the increase on average from the un-augmented romaine lettuce agar plates to the Rhizobium spp. derived composition augmented plates from 4.88 ⁇ 0.99 cm to 6.39 ⁇ 1 .1 1 cm, a 30.9% increase in the shoot length, at a concentration of 0.1 % of the nutrient agar plate is surprising and unexpected.
  • Rhizobium spp. derived composition is involved in promoting early growth and vigor in a variety of treated plants.
  • the use of a Rhizobium spp. derived composition does not involve the use of a living biological organism except that of the plant which is unique. Benefits can be seen with the use of a living biological, not the derived materials from a biological composition of a beneficial biological.
  • Figure 2 is a chart plotting romaine lettuce shoot length against
  • Rhizobium spp. derived composition percentage Figure 1 shows Agar plates containing an optimal mixture of nutrients showed significantly increased growth patterns when combined with a Rhizobium spp. derived composition at a wide range of composition percentages. All levels tested with the Rhizobium spp. derived composition resulted in a statistically significant response in the Romaine lettuce shoot length which is unexpected and novel.
  • Example 2 harvested corn yields were compared between the treatment compositions containing the Rhizobium spp. derived composition vs without the Rhizobium spp. derived composition and reported as a yield change (i.e. difference between the two treatments) in bushels per acre (bu/ac).
  • a yield change i.e. difference between the two treatments
  • bo/ac bushels per acre
  • Figure 3 is a chart showing Corn seed treatment comparison between Rhizobium spp. derived composition plus chemical combination versus chemical composition without Rhizobium spp. derived composition indicating a 2.03 bu/ac increase when using the Rhizobium spp. composition over the non-Rhizobium spp. derived composition that represents a standard and widely utilized seed treatment composition.
  • the hypothesis for the mechanism involves early season (early stage plant life) growth promotion and vigor of the plant which is induced by the Rhizobium spp. derived composition, which in turn results in improved yield.
  • results are unexpected from a composition derived from Rhizobium to show a response in corn.
  • the amplitude of the results seen are also unexpected.
  • a significant positive response by the addition of a Rhizobium spp. derived composition in corn is surprising.
  • the low levels (0.2 mg / seed) at which the Rhizobium spp. derived composition is applied to the seed that results in a significant response is also surprising and novel.
  • Example 3 harvested soybean yields were compared between the treatment composition containing the Rhizobium spp. derived composition vs without the Rhizobium spp. derived composition vs without any treatment and reported as a yield change (i.e. difference between the two treatments) in bushels per acre (bu/ac).
  • a yield change i.e. difference between the two treatments
  • bo/ac bushels per acre
  • Figure 4 is a chart showing soybean seed treatment comparison between Rhizobium spp. derived composition plus chemical combination versus composition without Rhizobium spp. derived composition indicating a 2.1 bu/ac increase when using the Rhizobium spp. composition over the non- Rhizobium spp. derived composition that is would represent a standard and often utilized seed treatment composition.
  • the Rhizobium spp. derived composition provided a positive response, i.e. a response above the untreated control
  • Figure 5 is a chart showing soybean seed treatment comparison between Rhizobium spp. derived composition plus chemical combination versus composition without Rhizobium spp. derived composition indicating a 0.6 bu/ac increase when using the Rhizobium spp. composition over the non- Rhizobium spp. derived composition that is would represent a standard and widely utilized seed treatment composition.
  • the Rhizobium spp. composition provided a positive response that is a yield that greater than the control for that individual test.
  • the hypothesis for the mechanism involves early season growth promotion and vigor of the plant which is induced by the Rhizobium spp. derived composition, which in turn results in improved yield.
  • the consistency (i.e. the percent positives) and the amplitude of the response is unexpected over the untreated seed and the base chemical seed treatment composition at the very low levels
  • Rhizobium The amplitude of the results seen are also unexpected.
  • the levels at which the Rhizobium spp. composition is applied to the seed that results in a response is very low and a surprising result.
  • the other non-Rhizobium spp. derived chemicals are expected to produce a certain response over untreated seed but the response seen on top of those is unexpected.
  • Living Rhizobium are known to produce an increase in nodulation but their performance is highly variable and inconsistent and limited to legume crops (Deaker, 2004). The consistency and amplitude of the response is uncharacteristic and unique.

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Abstract

La présente invention porte sur un produit comprenant au moins un métabolite dérivé de Rhizobium spp. et un matériau de traitement des semences. Le matériau de traitement des semences peut comprendre un fongicide, un insecticide, un régulateur de la croissance des plantes, un promoteur de croissance végétale, un micronutriment et des combinaisons de ceux-ci. L'invention concerne des procédés de fabrication du produit. Des procédés pour utiliser le produit sont divulgués, comprenant un procédé pour appliquer un produit à une ou à plusieurs semences. L'application du produit peut augmenter le rendement global des plantes provenant des semences traitées par rapport à des plantes provenant de semences non traitées avec le produit.
PCT/US2014/023537 2014-03-11 2014-03-11 Compositions de traitement des semences contenant des métabolites dérivés de rhizobium WO2015137928A1 (fr)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105950520A (zh) * 2016-07-18 2016-09-21 武汉市农业科学技术研究院作物科学研究所 一种高效解磷的根瘤菌及其应用
CN107382483A (zh) * 2017-08-01 2017-11-24 天津有茂食用菌开发有限公司 一种金针菇培养基
CN110810433A (zh) * 2019-12-04 2020-02-21 河南农贝得农业科技有限公司 一种针对农作物粗缩病的拌种剂及其制备方法

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1987006796A1 (fr) * 1986-05-13 1987-11-19 The Australian National University Modification de la reaction de plantes a des micro-organismes
US20130047505A1 (en) * 2011-08-25 2013-02-28 Loveland Products, Inc. Aqueous composition for accelerating secretion of alpha-amylase in plant seed germination
US20130338003A1 (en) * 2010-12-06 2013-12-19 United States As Represented By The Secretary Of The Army Rhizobium tropici produced biopolymer salt
US20130345058A1 (en) * 2011-03-10 2013-12-26 Wolfram Andersch Use of lipochito-oligosaccharide compounds for safeguarding seed safety of treated seeds

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1987006796A1 (fr) * 1986-05-13 1987-11-19 The Australian National University Modification de la reaction de plantes a des micro-organismes
US20130338003A1 (en) * 2010-12-06 2013-12-19 United States As Represented By The Secretary Of The Army Rhizobium tropici produced biopolymer salt
US20130345058A1 (en) * 2011-03-10 2013-12-26 Wolfram Andersch Use of lipochito-oligosaccharide compounds for safeguarding seed safety of treated seeds
US20130047505A1 (en) * 2011-08-25 2013-02-28 Loveland Products, Inc. Aqueous composition for accelerating secretion of alpha-amylase in plant seed germination

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105950520A (zh) * 2016-07-18 2016-09-21 武汉市农业科学技术研究院作物科学研究所 一种高效解磷的根瘤菌及其应用
CN105950520B (zh) * 2016-07-18 2019-05-10 武汉市农业科学技术研究院作物科学研究所 一种高效解磷的根瘤菌及其应用
CN107382483A (zh) * 2017-08-01 2017-11-24 天津有茂食用菌开发有限公司 一种金针菇培养基
CN110810433A (zh) * 2019-12-04 2020-02-21 河南农贝得农业科技有限公司 一种针对农作物粗缩病的拌种剂及其制备方法

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