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WO2010037167A1 - Utilisation de penicillium spp pour stimuler la croissance des plantes - Google Patents

Utilisation de penicillium spp pour stimuler la croissance des plantes Download PDF

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Publication number
WO2010037167A1
WO2010037167A1 PCT/AU2009/001294 AU2009001294W WO2010037167A1 WO 2010037167 A1 WO2010037167 A1 WO 2010037167A1 AU 2009001294 W AU2009001294 W AU 2009001294W WO 2010037167 A1 WO2010037167 A1 WO 2010037167A1
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Prior art keywords
strain
penicillium
plant
composition
soil
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PCT/AU2009/001294
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English (en)
Inventor
Paul Robert Harvey
Steven Alan Wakelin
Maarten Harm Ryder
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Commonwealth Scientific And Industrial Research Organisation
Grains Research And Development Corporation
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Publication of WO2010037167A1 publication Critical patent/WO2010037167A1/fr

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Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • 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
    • C12N1/14—Fungi; 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
    • A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
    • A01N63/36—Penicillium
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05B—PHOSPHATIC FERTILISERS
    • C05B17/00—Other phosphatic fertilisers, e.g. soft rock phosphates, bone meal
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00—Other organic fertilisers
    • C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
    • 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
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/145—Fungal isolates
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
    • C12R2001/00—Microorganisms ; Processes using microorganisms
    • C12R2001/645—Fungi ; Processes using fungi
    • C12R2001/80—Penicillium

Definitions

  • the isolated strain of Penicillium bilaiae is as deposited with the National Measurement Institute on 12 August 2008 under deposit reference V08/021001.
  • composition of the invention may be suitable for coating plant seeds and/or for application to a field.
  • the present invention further provides a method for increasing the phosphorus uptake of a plant, the method comprising applying an effective amount of an inoculum of the Penicillium bilaiae strain of the invention and/or the composition of the invention comprising the Penicillium bilaiae strain to soil in which the plant is grown.
  • the fungus of the invention may also be used to coat a seed of the plant.
  • the method of the invention comprises coating a seed of the plant with the Penicillium bilaiae strain of the invention.
  • the plant seed is further coated with a soil compatible carrier.
  • the soil-capable carrier may be capable of acting as a carbon source for the fungus.
  • the isolated strain of Penicillium gaestrivorus of the invention is as deposited with the National Institute of Measurement on 25 August 2008 under deposit reference V08/021260.
  • the present invention further provides a composition comprising the Penicillium gaestrivorus strain of the invention.
  • composition of the invention comprising the Penicillium gaestrivorus strain further comprises a solid particulate source of phosphorus.
  • the solid particulate source of phosphorus may comprise rock phosphate, mono-ammonium phosphate, superphosphate, triple superphosphate, diammonium phosphate, and/or ammonium polyphosphate.
  • Figure 3 Genetic differentiation among PeniciUium strains using Interspersed Simple Sequence Repeat (ISSR) primer (GAA) 6 .
  • MW Molecular weight marker (from top) 2000 bp, 1200 bp, 800 bp, 400 bp, 200bp; 1: PeniciUium bilaiae P-208 (2002 stock); 2: PeniciUium bilaiae P-208 (2006 stock); 3: PeniciUium bilaiae P-201 (2007 stock); 4: PeniciUium bilaiae 3391 (2007 stock); 5: PeniciUium bilaiae P-208 (2007 stock); 6: PeniciUium radicum Pr70 (2007 stock); 7: PeniciUium gaestrivorous P-209 (2002 stock); 8: PeniciUium gaestrivorous P-209 (2006 stock).
  • ISSR Interspersed Simple Sequence Repeat
  • Figure 4 Dendrogram showing phenetic relationships among strains of PeniciUium bilaiae (3391, P-201, P-208)) and P. gaestrivorus (P-209) based on 43 ISSR and 122 AFLP loci.
  • SEQ ID NO:1 Nucleotide sequence of Australian P. bilaiae strain (P-208) ⁇ -tubulin gene (BenA).
  • SEQ ID NO:2 Nucleotide sequence of Canadian P. bilaiae strain (P-201) ⁇ -tubulin gene (BenA).
  • SEQ ID NO:6 Oligonucleotide primer for ISSR analysis.
  • Penicillium gaestrivorus was deposited on 25 August 2008 with the National Measurement Institute under Deposit Reference V08/021260.
  • the phrase "which comprises” means that a particular sequence of nucleotides is contained within the genome of the Penicillium.
  • the term "inoculum” as used in this specification is intended to mean any form of fungus cells for example mycelium, or spores, which are capable of propagating on or in the soil when the conditions of moisture, temperature, etc. are favourable for fungal growth.
  • the terms "coated” or “coating” when used in relation to a plant seed means that a fungus or composition of the invention is applied to the seed.
  • the seed need not be completely covered with the fungus or composition of the invention, but need only be at least partially covered.
  • the fungi may also be routinely propagated on a suitable carbon source such as autoclaved moist ground wheat straw with added glucose, bran, or a peat or vermiculite-based substrate, or grains including, but not limited to oats, wheat, barley or rice. Propagation normally takes place for a period of approximately one week or more before the fungal inoculum is ready to use.
  • the resulting fungus propagated on a solid support may be used as such for incorporation into soil, preferably at root level, but may alternatively be coated onto seeds.
  • a liquid culture of the fungus may be prepared using a conventional nutrient solution and then used as such or dried and applied to the soil either with or without a suitable carrier and/or nutrient source.
  • the fungal culture and/or spores may be stored using any suitable means know in the art. For example, for longer term storage, cultured fungus may be lyophilised or alternatively stored frozen in an appropriate solution, such as a glycerol solution.
  • compositions comprising a Penicillium strain of the invention.
  • a composition of the invention is used to inoculate a seed of a plant with Penicillium.
  • a composition of the invention may be for application to a field.
  • the field may contain existing plants, for example crop plants, or it may be intended that the field be sown with seed of a crop plant.
  • the source of phosphorus is organic.
  • An organic fertilizer refers to a soil amendment derived from natural sources that guarantees, at least, the minimum percentages of nitrogen, phosphate, and potash. Examples include plant and animal by-products, rock powders, seaweed, inoculants, and conditioners. These are often available at garden centres and through horticultural supply companies.
  • said organic source of phosphorus is from bone meal, meat meal, animal manure, compost, sewage sludge, or guano.
  • the skilled person will be able to determine the amount of inoculum to be applied to soil using routine methods. For example, if an insufficient inoculum of fungus is used, there will be no noticeable effect, for example there will be no reduction in the required amount of phosphate containing fertilizer, whereas the addition of large amounts of inoculum will see no further improvement in phosphate solubilzation and/or reduction in fertilizer usage.
  • the suitable application rates vary according to the type of soil, the type of crop plants, the amount of insoluble phosphate, etc. and a suitable rate can be found without difficulty by routine trial for each particular case. Normally, the application rate falls into the range of 0.001 - 1.0 kg fungal spores and mycelium (fresh weight) per hectare.
  • the present inventors compared the 5.8 S rRNA, cytochrome c oxidase and ⁇ - tubulin gene sequences of the P. bilaiae and the P. gaestrivorus strains of the invention to the gene sequences from Canadian P. bilaiae strain P-201 (American Type Culture Collection No. ATCC 20851) and P. bilaiae strain 3391. It was found that there was a difference in the ⁇ -tubulin gene (BenA) sequence which can be used to differentiate the P. bilaiae strain P-208 of the invention from P. bilaiae strain P-201.
  • the query sequence is at least 45 nucleotides in length, and the GAP analysis aligns the two sequences over a region of at least 45 nucleotides.
  • the query sequence is at least 150 nucleotides in length, and the GAP analysis aligns the two sequences over a region of at least 150 nucleotides.
  • the query sequence is at least 300 nucleotides in length and the GAP analysis aligns the two sequences over a region of at least 300 nucleotides.
  • the query sequence is at least 400 nucleotides in length and the GAP analysis aligns the two sequences over a region of at least 400 nucleotides Alternatively, the two sequences are aligned over their entire length.
  • DNA marker technology particularly PCR based markers, such as randomly amplified polymorphic DNA markers (RAPD; Welsh et al., 1990), amplified fragment length polymorphisms (AFLP; Bleeker et al., 1995) and microsatellite markers (Tautz, 1989) are useful resources for genetic analyses of a wide variety of genomes.
  • PCR technology finds increased use in various genetic analyses, additional variations of this technique are emerging in order to augment the high-resolution genotyping and genetic mapping of various complex animal, plant and microbial genomes.
  • the PCR analysis using anchored simple sequence repeat primers, referred to as ISSR-PCR (ISSR), or anchored SSR-PCR provides a means of characterizing complex genomes (Williams et al., 1990).
  • ISSR interleukin-semiconductor-semiconductor-semiconductor-semiconductor-semiconductor-semiconductor-semiconductor-semiconductor-semiconductor-semiconductor-semiconductor-semiconductor-semiconductor-semiconductor-semiconductor-derived cloning and sequencing inherent in the original microsatellite-based approach.
  • ISSR has been used for genetic linkage analysis of various plant species (Godwin et al., 1997; Tsumura et al., 1996; Ratnaparkhe et al., 1998) and the silkworm, Bombyx mori (Reddy et al., 1999).
  • AFLP employs basically three steps. Initially, a sample of genomic DNA is cut with restriction enzymes and oligonucleotide adapters are ligated to the restriction fragments of the DNA. The restriction fragments are then amplified using PCR by using the adapter and restriction sequence as target sites for primer annealing. The selective amplification is achieved by the use of primers that extend into the restriction fragments, amplifying only those fragments in which the primer extensions match the nucleotide flanking the restriction sites. These amplified fragments are then visualized on a denaturing polyacrylamide gel. In a preferred embodiment, AFLP analysis is performed as described herein in Example 5.
  • AFLP analysis may comprise digesting DNA isolated from a Penicillium bilaiae strain with Hael ⁇ l and Pstl, amplifying the digested DNA fragments with the pre-selective primers H (5'-GATGAGTCC TGAGCC) (SEQ ID NO:8) and P (5 '-GACTGCGTACATGCAG) (SEQ ID NO:9), and the selective primer combinations P-AAG (SEQ ID NO: 13) + H-AG (SEQ ID NO14), P-ACC (SEQ ID NO: 15) + H-G (SEQ ID NO: 16), P-AGG (SEQ ID NO: 17) + H-CG (SEQ ID NO: 18), P-ACC (SEQ ID NO: 15) + H-GT SEQ ID NO: 19) and P-AGG (SEQ ID NO: 17) + H- TA (SEQ ID NO:20), and analysing the DNA fragments produced by electrophoresis in 2% TAE agarose gels stained with ethi
  • Penicillium strains were grown on either Malt Extract Agar (MEA) or in 100 ml flasks containing 25 ml of Czapek-Dox medium without shaking for 7-14 days at 25 °C.
  • MEA Malt Extract Agar
  • the Penicillium strains used in this study are listed in Table 1. Table 1. Penicillium strains used in this study.
  • figures in parenthesis refer to the % decrease (-) or increase (+) in crop biomass resulting from treatment with P-201 & P-208 relative to the untreated control.
  • DNA-based identifications were made using nucleotide sequence data from the conserved regions of the nuclear 5.8S ribosomal RNA (rDNA), mitochondrial cytochrome c oxidase gene (COl) and the ⁇ -tubulin (BenA) genes. Isolation of fungal genomic DNA from pure cultures (Harvey et al., 2000) and PCR amplification of 5.8S rDNA (White et al., 1990; Wakelin et al., 2004b), COl (Seifert et al., 2007) and BenA (Sampson et al., 2004) sequences were performed in an Eppendorf (Australia) thermocycler, as described previously.
  • rDNA nuclear 5.8S ribosomal RNA
  • COl mitochondrial cytochrome c oxidase gene
  • BenA BenA
  • PCR primers used for internal-simple sequence repeat (ISSR) analyses were Rl, (5'-TCGTGGCTGACTTACCTG (SEQ ID NO:6)) (Weining and Langridge, 1991), the M13 minisatellite core sequence M13-MS (5'-GAGGGTGGCGGTTCC (SEQ ID NO:7)) (Stenlid et al, 1994) and the microsatellite sequences (GA) 8 (SEQ ID NO: 10), (GAA) 6 (SEQ ID NO: 11), and (GACA) 4 (SEQ ID NO: 12).
  • DNA amplifications were performed in a Eppendorf (Australia) thermocycler and each 25 ⁇ l reaction contained 10-20 ng Penicillium DNA, 25 pmol of primer(s), 5.0 ⁇ mol each of dATP, dCTP, dGTP and dTTP, 2.5 ⁇ l 10x reaction buffer and 0.5 units Taq polymerase (Promega).
  • a two-step program was used for all primers and consisted of an initial denaturing step of 94 °C for 4 minutes followed by 6 cycles of 1 min. denaturation at 94 °C, 1 min. annealing at 40 °C and 1 min. extension at 72 °C. This was followed by a further 28 cycles of 94 °C for 1 min., 58 °C for 1 min. and 72 °C for 1 min. A final extension for 7 min. at 72 °C was performed before cooling to ambient temperature.
  • AFLP amplified fragment length polymorphism
  • Multilocus genotype analysis with ISSR and AFLP markers clearly differentiated all 4 Penicillium strains, with each isolate having a unique genotype (DNA fingerprint).
  • DNA fingerprint DNA fingerprint
  • 41% of loci were unique to P. gaestrivorus, whereas within northern hemisphere P. bilaiae only 3% and 7% of loci were unique to the phosphorous solubilising inoculant P-201 and type culture 3391 respectively (Table 9). In contrast 16% of loci were unique to the Australian phosphorous solubilising strain P- 208 (Table 9). Overall only 10% of all ISSR and AFLP loci were common to all 3 P.
  • Table 9 Summary statistics of genetic differentiation among strains of Penicillium bilaiae and P. gaestrivorus based on 43 ISSR and 122 AFLP loci.

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Abstract

Cette invention concerne des souches isolées de Penicillium qui sont capables de stimuler la croissance des plantes et des compositions contenant lesdites souches de Penicillium. Les souches de Penicillium isolées et les compositions contenant lesdites souches peuvent être utilisées dans des procédés visant à stimuler la croissance des plantes.
PCT/AU2009/001294 2008-10-01 2009-09-30 Utilisation de penicillium spp pour stimuler la croissance des plantes WO2010037167A1 (fr)

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Cited By (14)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102503691A (zh) * 2011-11-04 2012-06-20 安徽省神农农业技术开发有限公司 一种新拌种肥料制备方法
WO2014078647A1 (fr) * 2012-11-16 2014-05-22 Novozymes Bioag A/S Souches microbiennes, compositions et procédés pour améliorer le phosphate disponible pour des plantes
US20140342904A1 (en) * 2011-09-14 2014-11-20 Novozymes Bioag A/S Use of lipochito oligosaccharides and/or chito oligosaccharides in combination with phosphate solubilizing microorganisms to enhance plant growth
US20150166420A1 (en) * 2011-09-08 2015-06-18 Novozymes Bioag A/S Seed treatment methods and compositions
US9101088B2 (en) 2008-10-01 2015-08-11 Novozymes Biologicals Limited Methods and compositions for increasing the amounts of phosphorus available for plant uptake from soils
CN106747712A (zh) * 2016-11-18 2017-05-31 青岛蔚蓝生物集团有限公司 一种新型、多功能生物肥及其应用
US20180055047A1 (en) * 2010-04-19 2018-03-01 Iden Biotechnology S. L. Method for changing the development pattern, increasing the growth and the accumulation of starch, changing the structure of starch and increasing the resistance to water stress in plants
CN107771860A (zh) * 2017-10-24 2018-03-09 慕恩（北京）生物科技有限公司 一种微生物复合型种衣剂及其生产方法
CN108697098A (zh) * 2015-11-20 2018-10-23 孟山都技术公司 用于减少玉米连续种植产量损失的组合物和方法
US10206396B2 (en) 2011-09-23 2019-02-19 Novozymes Bioag A/S Agricultural compositions comprising chitin oligomers
CN112375691A (zh) * 2020-11-20 2021-02-19 青岛农业大学 一株产紫青霉及其作为解磷真菌的应用以及在促进玉米生长中的应用
RU2794356C1 (ru) * 2022-08-19 2023-04-17 Федеральное государственное бюджетное научное учреждение Уфимский федеральный исследовательский центр Российской академии наук Композиция с фунгицидной и росторегулирующей активностью для предпосевной обработки семян зерновых культур
WO2023126461A1 (fr) * 2021-12-30 2023-07-06 Aphea.Bio Nv Produits et procédés pour améliorer des caractéristiques de croissance de plantes
CN118667669A (zh) * 2024-08-26 2024-09-20 南京农业大学三亚研究院 一株青霉属真菌q50及其应用

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US11059759B2 (en) 2008-10-01 2021-07-13 Novozymes Biologicals Limited Methods and compositions for increasing the amounts of phosphorus available for plant uptake from soils
US12172936B2 (en) 2008-10-01 2024-12-24 Novozymes Biologicals Limited Methods and compositions for increasing the amounts of phosphorus available for plant uptake from soils
US10035735B2 (en) 2008-10-01 2018-07-31 Novozymes Biologicals Limited Methods and compositions for increasing the amounts of phosphorus available for plant uptake from soils
US10308561B2 (en) 2008-10-01 2019-06-04 Novozymes Biologicals Limited Methods and compositions for increasing the amounts of phosphorus available for plant uptake from soils
US9101088B2 (en) 2008-10-01 2015-08-11 Novozymes Biologicals Limited Methods and compositions for increasing the amounts of phosphorus available for plant uptake from soils
US9732007B2 (en) 2008-10-01 2017-08-15 Novozymes Biologicals Limited Methods and compositions for increasing the amounts of phosphorus available for plant uptake from soils
US20180055047A1 (en) * 2010-04-19 2018-03-01 Iden Biotechnology S. L. Method for changing the development pattern, increasing the growth and the accumulation of starch, changing the structure of starch and increasing the resistance to water stress in plants
US20150166420A1 (en) * 2011-09-08 2015-06-18 Novozymes Bioag A/S Seed treatment methods and compositions
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US10239798B2 (en) * 2011-09-08 2019-03-26 Novozymes Bioag A/S Seed treatment methods and compositions
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US11999666B2 (en) 2011-09-14 2024-06-04 Novozymes Bioag A/S Use of lipo-chitooligosaccharides and/or chitooligosaccharides in combination with phosphate-solubilizing microorganisms to enhance plant growth
US20140342904A1 (en) * 2011-09-14 2014-11-20 Novozymes Bioag A/S Use of lipochito oligosaccharides and/or chito oligosaccharides in combination with phosphate solubilizing microorganisms to enhance plant growth
US10206396B2 (en) 2011-09-23 2019-02-19 Novozymes Bioag A/S Agricultural compositions comprising chitin oligomers
US11134683B2 (en) 2011-09-23 2021-10-05 Novozymes Bioag A/S Chitooligosaccharides and methods for use in enhancing plant growth
US12185720B2 (en) 2011-09-23 2025-01-07 Novozymes Bioag A/S Chitooligosaccharides and methods for use in enhancing plant growth
CN102503691A (zh) * 2011-11-04 2012-06-20 安徽省神农农业技术开发有限公司 一种新拌种肥料制备方法
US9758438B2 (en) 2012-11-16 2017-09-12 Novozymes Bioag A/S Microbial strains, compositions, and methods for increasing available phosphate for plants
CN104797701A (zh) * 2012-11-16 2015-07-22 诺维信生物农业公司 用于增加植物的有效磷酸盐的微生物菌株、组合物、和方法
US10450237B2 (en) 2012-11-16 2019-10-22 Novozymes Bioag A/S Microbial strains, compositions, and methods for increasing available phosphate for plants
WO2014078647A1 (fr) * 2012-11-16 2014-05-22 Novozymes Bioag A/S Souches microbiennes, compositions et procédés pour améliorer le phosphate disponible pour des plantes
RU2668832C2 (ru) * 2012-11-16 2018-10-02 Новозимс Биоаг А/С Микробные штаммы, композиции и способы увеличения доступного фосфата для растений
US9365464B2 (en) 2012-11-16 2016-06-14 Novozymes Bioag A/S Microbial strains, compositions, and methods for increasing available phosphate for plants
CN108697098A (zh) * 2015-11-20 2018-10-23 孟山都技术公司 用于减少玉米连续种植产量损失的组合物和方法
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CN107771860A (zh) * 2017-10-24 2018-03-09 慕恩（北京）生物科技有限公司 一种微生物复合型种衣剂及其生产方法
CN112375691B (zh) * 2020-11-20 2022-11-01 青岛农业大学 一株产紫青霉及其作为解磷真菌的应用以及在促进玉米生长中的应用
CN112375691A (zh) * 2020-11-20 2021-02-19 青岛农业大学 一株产紫青霉及其作为解磷真菌的应用以及在促进玉米生长中的应用
WO2023126461A1 (fr) * 2021-12-30 2023-07-06 Aphea.Bio Nv Produits et procédés pour améliorer des caractéristiques de croissance de plantes
RU2794356C1 (ru) * 2022-08-19 2023-04-17 Федеральное государственное бюджетное научное учреждение Уфимский федеральный исследовательский центр Российской академии наук Композиция с фунгицидной и росторегулирующей активностью для предпосевной обработки семян зерновых культур
CN118667669A (zh) * 2024-08-26 2024-09-20 南京农业大学三亚研究院 一株青霉属真菌q50及其应用
CN118667669B (zh) * 2024-08-26 2024-10-18 南京农业大学三亚研究院 一株青霉属真菌q50及其应用

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