US20100178303A1 - Stable organic-carrier-based microbial inoculants and method for producing the same - Google Patents

Stable organic-carrier-based microbial inoculants and method for producing the same Download PDF

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Publication number
US20100178303A1
US20100178303A1 US12/663,222 US66322207A US2010178303A1 US 20100178303 A1 US20100178303 A1 US 20100178303A1 US 66322207 A US66322207 A US 66322207A US 2010178303 A1 US2010178303 A1 US 2010178303A1
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Prior art keywords
azospirillum
microbial
biologically pure
pure culture
bacterial microorganism
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Abandoned
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US12/663,222
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English (en)
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Saturnina Halos
Ponciano Halos
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Classifications

    • CCHEMISTRY; METALLURGY
    • C05FERTILISERS; MANUFACTURE THEREOF
    • C05FORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00Other organic fertilisers
    • C05F11/08Organic fertilisers containing added bacterial cultures, mycelia or the like
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/04Antibacterial agents
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00Drugs for immunological or allergic disorders
    • A61P37/02Immunomodulators
    • A61P37/04Immunostimulants
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/04Preserving or maintaining viable microorganisms
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/20Bacteria; Culture media therefor

Definitions

  • the present invention relates to microbial preparations for crop production.
  • the present invention relates to microbial preparations for crop production, specifically microbial inoculants incorporated in carriers that provide protection for the colonizing microorganisms and the method for producing the same.
  • biofertilizers that have an increased colony increased inoculum potential concentration have been developed and commercialized.
  • biofertilizers that have an increased colony increased inoculum potential concentration have been developed and commercialized.
  • Commercial biofertilizers effectively enhance soil fertility because they contain sufficient cells of efficient strains of specific microorganisms, that can fix atmospheric nitrogen, solubilize/mineralize phosphorous and potash or decompose organic wastes and augment plant growth with hormones and other growth promoting substances with their biological activities.
  • the present invention is a carrier-based biofertilizer that promotes growth of plants of any type of terrestrial biome, using special biological inoculants, such as plant growth promoting rhizobacteria.
  • the invention induces extensive root and root hair formation, enable plants to efficiently use soil nutrients, supply plants with nitrogen from the air, increase drought tolerance and protect the plants from pathogens. Further, the inoculant can be mixed with an insecticide or fungicide without any appreciable decrease in effectivity.
  • WO 2007/014974 (“Aseptic Mycorrhization Inoculant and In vitro and Ex vitro Application Methods”), a low-viscosity, light, semi-solid agar-type base culture medium is used to facilitate the penetration of the root in the inoculant, as well as to potentiate and preserve the infectiousness of arbuscular mycorrhizal fungi, such as those of the present invention.
  • WO 2007/014974 sought to obtain the advantage of being as effective in in vitro as well as ex vitro inoculation by using a semi-solid inoculant. However, it is gel-based and has a short shelf-life, which the present invention seeks to address.
  • WO 1994/006732 (“Method for Obtaining a Wettable Powder Inoculant for Use with Leguminous Crops”) reports a method for obtaining a wettable powder inoculant formulation for use with leguminous crops, comprising the industrialization of an inoculant for leguminous from a centrifugated biological mass of Rhizobium spp twice immersed in a cryogenic protective solution rich in sugar and protein which is subjected to a lyophilization method. Like the present invention, it is formulated with substances rich in polysaccharides and associated with a cellular protector which substitutes the previously used peat support. It also has a long shelf life of up to 25 months even when stored at room temperature.
  • WO 1994/006732 can only be used for leguminous crops. Further, the formulation of the carrier uses sugar and protein while the present invention uses milk and glycerin. Also, WO 1994/006732 does not teach that the powder inoculant with a pest protectant such as an insecticide.
  • WO 2006/070061 (“Stable Microbial Inoculants and Methods for Production of Them”) teaches away from the present invention. It describes water containing microbial inoculants in paste form with shelf life of at least 3 months, preferably 12 months. It reports that drying, which is a method utilized in the present invention is complicated, expensive and may even be impossible.
  • WO 2002/015702 (“A Thermo-stable Bio-matrix”) reports a method for producing a thermo-stable biodegradable medium for storage of biological materials wherein the biological material includes: a pesticide; a viricide; a bacteriacide; a fungicide; and a combination of these. While the method is similar in that the biological material can be safely mixed with potentially degradative substances such as pesticides, viricides, bacteriacides, fungicides or a combination of these, the medium is structurally different because it is gel-based and contains a biopolymer selected from the group of xanthan gum, acacia gum, guar gum, gellan, starch or a combination thereof, unlike the present invention which is in powder form. Thus, it is no surprise that WO 2002/015702 discloses a maximum shelf life of only eighteen (18) months, which is considerably shorter than that of the present invention which allows storage of at least three (3) years.
  • WO 2000/034440 (“Microorganisms for Improving Plant Productivity”) is directed to microbes which includes the microorganism Azospirillum brasilense , which belongs to the same genus as some of those in the present invention.
  • two specific strains of microbial inoculant of WO 2000/034440 is effective for increasing the productivity of both nonlegume and legume plants as well as vegetable plants over a wide variety of soil types and climates.
  • the WO 2000/034440 is narrowly directed only to the Azospirillum brasilense , a microorganism not covered by the present application. Further, it does not teach an inventive carrier nor the method of production that prolongs its shelf life.
  • the present invention the microbes, method and product, are absolutely different and improve over other known methods and products and have considerable advantages over the nature and quality of the methods and products, particularly in the storage life of the microbial preparation.
  • the present invention relates to microbial preparations for crop production such as seed, seedling and other planting material inoculants have been the subject of many researches in the past 25 years in the Philippines. They have been shown to be useful in increasing and protecting crop yields. However, they have not been put into commercial production because of their short shelf-lives which limit their effective distribution. This is also true of microbial inoculants for processing foods. Prior art discloses a shelf life of a maximum of only eighteen (18) months. This invention is intended to solve this problem of short shelf-lives and allow for the viable commercialization of microbial inoculants. The invention is expected to produce products with a huge market since biochemical catalysts including microbial preparations have agricultural, food and forestry applications.
  • the invention covers dried, semi-solid and related formulations of microbial inoculants like Azospirillum spp. and Bacillus pumilus . These bacteria are used to inoculate seeds and seedlings of different agriculture and forestry species to promote among others, the extensive root and root hair formation, enable plants to efficiently use soil nutrients, supply plants with nitrogen from the air, increase drought tolerance and protect the plants from root pathogens.
  • microbial inoculants like Azospirillum spp. and Bacillus pumilus .
  • the microbes such as Azospirillum spp. And Bacillus pumilus are grown in fermentors using special media which contain substrates including high protein feed ingredients such as fish meal/bone meal/blood meal/soya meal, table sugar and tablet salt at 30° C.
  • the baterial cells are harvested by methods like filtration or centrifugation, added with powdered milk/glycerol to wet the cells and the Azospirillum mixture lyophilized.
  • the products are formulated under sterile conditions with different proportions of components like the bacterial preparations, minerals, milk, vitamins and flour with or without talc/chalk/diatomaceous earth using standard mechanical devices.
  • the dried and semi-solid products are packed or the tablets are made usually under sterile conditions.
  • the products as invented have extended the shelf-lives of the biochemical catalysts/microbial preparations by at least 3 years way beyond the 3 months observed with the non-lyophilized or other inoculants now available in very limited amounts in the market.
  • the special media used to grow the bacteria improve the tolerance of the bacteria to lyophilization. Milk not only improves the tolerance of the bacteria to lyophilization; it also provides some vitamins, carbohydrates and minerals for a good start of the bacteria with inoculated plants. Bacillus pumilus need not be lyophilized because it is a spore former and can resist adverse conditions for many years.
  • the added vitamins, minerals, flour and talc/chalk/diatomaceous earth also provide additional nutrients to selectively enrich the growth of the desired bacteria and the inoculated plants in problem soils with limiting micronutrients.
  • Azospirillum is made up of curved rod cells, 1 mm in diameter with varying sizes and shapes. It can have polar and peritrichous flagella and undergoes encystment. Its pinkish, whitish orange colonies vary in shape and surface characteristics.
  • Carbon sources include organic acids and sugars.
  • the nitrogen sources are amino acids, N 2 , NH 4 + and NO 3 .
  • Respiration can be aerobic, microaerophilic or anaerobic. It is capable of aerotaxis, chemotaxis and can form clumps.
  • Bacillus pumilus consists of rod-shaped cells, which can form endospores that are very resistant to many adverse conditions. Sporulation is not repressed by exposure to air. It is gram positive, or positive only in the early stages of growth, or negative. The flagellation is peritrichous or degenerately peritrichous. It can be aerobic or facultatively anaerobic. Oxygen is the terminal electron acceptor. Colony morphology and size are very variable on some media. It exhibits a wide diversity of physiological ability; psychrophilic to thermophilic; acidophilic to alkaliphilic. Some strains are salt tolerant while others have a specific requirement for salts. Catalase is formed but may be oxidase positive or negative.
  • Azospirillum halopraeferens UPCC 1370 Azospirillum lipoferum UPCC 1371 Bacillus pumilus UPCC 1372 Azospirillum SC/PH UPCC 1373 Azospirillum AH/EH UPCC 1374
  • a pesticide, a viricide, a bacteriacide, a fungicide, or a combination of these may be added to the microbial inoculant to further prolong its shelf life.
US12/663,222 2007-06-21 2007-06-21 Stable organic-carrier-based microbial inoculants and method for producing the same Abandoned US20100178303A1 (en)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/PH2007/000012 WO2008156380A2 (fr) 2007-06-21 2007-06-21 Inoculants microbiens à base de support organique stable et leur procédé de fabrication

Related Parent Applications (1)

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PCT/PH2007/000012 A-371-Of-International WO2008156380A2 (fr) 2007-06-21 2007-06-21 Inoculants microbiens à base de support organique stable et leur procédé de fabrication

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US13/753,810 Active US9499447B2 (en) 2007-06-21 2013-01-30 Stable organic-carrier-based microbial inoculants and cultures

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US (2) US20100178303A1 (fr)
EP (1) EP2126043A4 (fr)
AU (1) AU2007355202B2 (fr)
WO (1) WO2008156380A2 (fr)

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9499447B2 (en) 2007-06-21 2016-11-22 Saturnina Halos Stable organic-carrier-based microbial inoculants and cultures
CN115927010A (zh) * 2022-12-07 2023-04-07 福建省农业科学院植物保护研究所 一种提升玉米大斑病菌活力和致病力的方法及其应用

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KR20130086122A (ko) * 2010-03-12 2013-07-31 브룩하벤 싸이언스 어쏘씨에이츠 엘엘씨 엔테로박터 에스피. 638 및 이의 사용방법
WO2011154961A1 (fr) * 2010-06-09 2011-12-15 Patel, Babubhai C. Procédé nouveau de préparation de formulation bactérienne utilisant des bactéries mobilisatrices de potasse qui mobilisent la potasse et la rendent disponible pour les plantes de culture
WO2011154960A1 (fr) * 2010-06-09 2011-12-15 Patel, Babubhai C. Matière nouvelle et procédé de préparation de formulation bactérienne utilisant des bactéries fixatrices d'azote qui fixent l'azote atmosphérique et le rendent disponible pour les plantes cultivées
TWI453179B (zh) * 2013-02-19 2014-09-21 Chunghwa Picture Tubes Ltd 顆粒狀肥料及其製造方法
US11136275B2 (en) 2014-06-18 2021-10-05 Kevin Merritt Process for making a composite granule with rock phosphate, sulfur, and humic acid
US11136505B2 (en) 2014-06-18 2021-10-05 Kevin Merritt Potassium Humate sulfur compound granule
US11117840B1 (en) * 2014-06-18 2021-09-14 Kevin Merritt Process for making a semi-soluble granule comprising rock phosphate and humic acid
US11691999B2 (en) 2014-06-18 2023-07-04 Humic Growth Solutions. Llc Homogenous humate powder compositions exhibiting high solubility
US10597337B1 (en) 2014-06-18 2020-03-24 Kevin Merritt Process for making a fully water-soluble granule comprising humic acid and a microbial community composition
CN105624073B (zh) * 2016-03-24 2019-05-03 福建农林大学 一株产脂固氮螺菌及其应用
MX2017008876A (es) 2017-07-04 2019-02-08 Newpek S A De C V Una formulacion inoculante bacteriano a base de un consorcio de microorganismos del genero calothrix sp. para incrementar el rendimiento y calidad de cultivos vegetales, el metodo para la fabricacion de la formulacion, y usos.
RU2678755C1 (ru) * 2018-03-23 2019-01-31 Общество с ограниченной ответственностью "Органик парк" Биологический агент для стимуляции ростовых процессов в растениях

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US9499447B2 (en) 2007-06-21 2016-11-22 Saturnina Halos Stable organic-carrier-based microbial inoculants and cultures
CN115927010A (zh) * 2022-12-07 2023-04-07 福建省农业科学院植物保护研究所 一种提升玉米大斑病菌活力和致病力的方法及其应用

Also Published As

Publication number Publication date
EP2126043A4 (fr) 2010-07-28
WO2008156380A3 (fr) 2009-02-12
WO2008156380A4 (fr) 2009-04-02
WO2008156380A2 (fr) 2008-12-24
AU2007355202A1 (en) 2008-12-24
US9499447B2 (en) 2016-11-22
US20130239633A1 (en) 2013-09-19
EP2126043A2 (fr) 2009-12-02
AU2007355202B2 (en) 2014-10-09

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