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WO1996034840A1 - Engrais bacterien et son procede d'obtention - Google Patents

Engrais bacterien et son procede d'obtention Download PDF

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Publication number
WO1996034840A1
WO1996034840A1 PCT/ES1996/000097 ES9600097W WO9634840A1 WO 1996034840 A1 WO1996034840 A1 WO 1996034840A1 ES 9600097 W ES9600097 W ES 9600097W WO 9634840 A1 WO9634840 A1 WO 9634840A1
Authority
WO
WIPO (PCT)
Prior art keywords
medium
culture
hours
cect
burk
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/ES1996/000097
Other languages
English (en)
Spanish (es)
Inventor
Antonio Martin Gonzalez
Tilman Sanchez Elsner
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
APPLICACIONES MAGNETICAS SL
Consejo Superior de Investigaciones Cientificas CSIC
Original Assignee
APPLICACIONES MAGNETICAS SL
Consejo Superior de Investigaciones Cientificas CSIC
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by APPLICACIONES MAGNETICAS SL, Consejo Superior de Investigaciones Cientificas CSIC filed Critical APPLICACIONES MAGNETICAS SL
Publication of WO1996034840A1 publication Critical patent/WO1996034840A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • 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
    • C12N1/20—Bacteria; Culture media therefor
    • C12N1/205—Bacterial 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
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • 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
    • C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
    • C12R2001/065—Azotobacter

Definitions

  • the object of the present invention is a bacterial fertilizer consisting of a suspension containing cells of new strains of the microorganisms Rzotobacter and Azo ⁇ plrillum. Said microorganisms and the procedure for obtaining said bacterial fertilizer also constitute another object of the present invention.
  • the main atmospheric nitrogen-fixing microorganisms that have been used as bacterial fertilizers are Rzotobacter and Azo ⁇ pirllum.
  • Rzotobacter has been used as a nitrogenous fertilizer in some plants of agricultural interest such as corn, wheat, potato, beetroot, tobacco, barley, flax, oats, carrots, cabbage and tomato. Inoculation with Rzotobacter as a bacterial fertilizer has been developed for many years (Rubenchic L.I., "Azotobacter and Its Use in Agriculture". Academy of Sciences of the Ukrainian SSR. Microbiological Institute im. D.K. Zabolotnyi, 1963). Of all the strains, the most used as inoculant has been R. chroococcum, as it is the most frequent and abundant in the soil (Krieg N.R, Holt, J.G., Williams and Wilkins Co., Baltimore, 1984).
  • the favorable effect of the inoculation with Azotoiacter and Rzo ⁇ pirillum cells on different agricultural crops is the product of a multiple action because it represents an alteration of the suppressive microbial balance of pathogenic microorganisms, mobilization of phosphates, assimilation of exudates from the roots of the plants (acid p -hydroxybenzoic, ... ) and production of metabolites such as phytohormones that stimulate growth before and after germination, also representing a very considerable saving in nitrogen fertilizers due to the fixation of N 2 in the soil.
  • Rzotobacter and Rzo ⁇ pirillum are related not only to their ability to fix N 2 but also to the ability to produce antibacterial and antifungal compounds, growth regulators, and siderophores (Pandey, A.; "Potential of Azotobacters and Azospirilla as biofertilizers for upland agriculture: a review”; Journal of Scientific and Industrial Research 48.3, 134-144, 1989).
  • the saving of nitrogenous fertilizers supposes a parallel saving of fossil fuels.
  • the object of the present invention is a bacterial fertilizer that contains cells of new strains of the microorganisms Rzotobacter and Rzo ⁇ pirillum specifically developed to improve N 2 fixation and potentiate alginate biosynthesis. .
  • aqueous medium Purk medium
  • the microorganisms suspended in said medium are a strain of Rzotobacter vinelandii deposited in the Spanish Type Culture Collection (CECT) with the number CECT 4534 and a strain of Rzo ⁇ pirillum bra ⁇ ilen ⁇ e deposited in the Spanish Collection of Type Cultures with the number CECT 4533.
  • CECT Type Culture Collection
  • Rzotobacter and Rzo ⁇ pirillum cells By subjecting Rzotobacter and Rzo ⁇ pirillum cells in liquid and solid cultures (with agar-agar) to different substrates, variants of these strains capable of fixing more atmospheric nitrogen, synthesizing more phytohormones (cytokinins, auxins, . . . ) and metabolites such as alginates and essential amino acids, such as usine and methionine, in higher concentration.
  • phytohormones cytokinins, auxins, . . .
  • metabolites such as alginates and essential amino acids, such as usine and methionine
  • strains also present a greater facility for using waste products as substrates, for example, the exudates of the roots of the plants (ortho-, meta- and parahydroxybenzoic acids,%) and agricultural residues such as molasses, vine shoots, stillage, etc
  • waste products for example, the exudates of the roots of the plants (ortho-, meta- and parahydroxybenzoic acids,...) and agricultural residues such as molasses, vine shoots, stillage, etc
  • alginates allows these Rzotobacter and Rzo ⁇ pirillum cells to better adhere to plant roots and at the same time have this capsular substrate available for growth and reproduction in the soil.
  • the contribution of DL glutamic acid is intended to inhibit the nitrogenase enzyme to enhance alginate biosynthesis.
  • cells are obtained, isolated on plates, which in subsequent standardized cultures, with sucrose as a substrate, reach a production of 2.4 g/1 of alginate with a high index of uronic acids (the basis for good gelation). These cells are designated by the initials 382 12 E of R . vinelandii . At the same time that they synthesize a greater amount of alginate, they fix a higher N 2 concentration than the original strains.
  • the strains of Rzo ⁇ pirillum bra ⁇ ilen ⁇ e obtained from the 590 CECT strain are prepared by the same procedure as in the case of Rzotobacter , but with D-Ribose as the only substrate in all steps, obtaining a strain called Rzo ⁇ pirillum with the acronym 40AM.
  • bra ⁇ ilen ⁇ e that has been deposited in the CECT having been assigned the number CECT 4533.
  • the ideal culture medium for the best development of Rzotobacter and Rzo ⁇ pirillum cells is the following:
  • the final pH of the culture medium is adjusted to 7.4.
  • the strains of AzotoJ acter vinelandii and Rzo ⁇ pirillum bra ⁇ ilen ⁇ e are seeded on solid media in Petri dishes and incubated at 30°C for a period of time between 72 and 96 hours. These freshly grown, pure cultures are used to inoculate 300 ml flasks with 100 ml of culture medium, which are incubated at a temperature between 30 and 32°C for 120 hours at 175 revolutions per minute in an orbital incubator.
  • the cell biomass of the mixture of Rzotobacter and Rzo ⁇ pirillum is then available to be inoculated into the seeds and subsequently into the soil.
  • Rzotobacter and Rzo ⁇ pirillum cells will serve the Rzotobacter and Rzo ⁇ pirillum cells as an energy reserve for their subsequent development in the soil, at the same time that they will allow them to adhere more easily to the roots of the plants. 3.- These variants of Rzotobacter and Rzo ⁇ pirillum also have a greater capacity to assimilate the compounds formed by the exudates of the roots of the plants, such as p-hydroxybenzoic acid, which is hardly used by other strains.
  • the speed of reproduction and assimilation of the substrates is also higher in the Rzotobacter and Rzo ⁇ pirillum variants than in the known strains, since the maximum dry weight reached by them is up to 3 times higher after 52 hours of culture. submerged at about 28-30°C. 5.- The performance and effectiveness of these Rzotobacter variants is much higher than the known strains and the initial strain 382 DSM, also synthesizing polysaccharides with a high gelling index (alginates).
  • the cell biomass production of the Rzotobacter strain 24 APV is of the order of 4 g/1 dry weight of lyophilized cells.

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Biotechnology (AREA)
  • Wood Science & Technology (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • Microbiology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Medicinal Chemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Fertilizers (AREA)

Abstract

La présente invention concerne un engrais bactérien (bioengrais) consistant en une suspension qui contient des cellules de nouvelles souches des microorganismes Azotobacter vinelandii et Azospirilum brasilense. Cet engrais bactérien consiste en une suspension de micro-organismes dans un milieu aqueux. Les micro-organismes sont obtenus par un procédé qui comprend: a) la culture en plaques de la souche originale; b) le traitement des colonies précédentes de la culture antérieure avec un autre milieu de culture; et c) la culture des colonies isolées de l'étape antérieure. Les avantages qui découlent de l'utilisation de ces nouvelles souches de micro-organismes sont leur plus grande tolérance à des milieux très alcalins et une meilleure fixation de N2, ainsi qu'une plus grande capacité d'assimilation des composés formés par les exsudats des racines des plantes.
PCT/ES1996/000097 1995-05-04 1996-04-29 Engrais bacterien et son procede d'obtention Ceased WO1996034840A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
ES09500851A ES2093559B1 (es) 1995-05-04 1995-05-04 Fertilizante bacteriano y procedimiento de obtencion.
ESP9500851 1995-05-04

Publications (1)

Publication Number Publication Date
WO1996034840A1 true WO1996034840A1 (fr) 1996-11-07

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ID=8290254

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/ES1996/000097 Ceased WO1996034840A1 (fr) 1995-05-04 1996-04-29 Engrais bacterien et son procede d'obtention

Country Status (2)

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ES (1) ES2093559B1 (fr)
WO (1) WO1996034840A1 (fr)

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998037038A3 (fr) * 1997-02-25 1998-11-12 Arpad Pollak Procede pour extraire l'azote de l'air, pour extraire les composes de phosphore du sol et pour decomposer les sediments d'huile
WO2001032587A1 (fr) * 1999-11-01 2001-05-10 'piksa Inter' Ltd Produit d'addition biologique pour fertilisants organomineraux
WO2001085916A1 (fr) * 2000-05-12 2001-11-15 Biofarmtech Co., Ltd. Materiau de revetement photocatalytique presentant une activite photocatalytique et des proprietes d'adsorption, et procede de preparation associe
WO2003016241A1 (fr) * 2001-08-13 2003-02-27 Agro.Bio Hungary Kft. Micro-organismes pour le traitement du sol et son procede de production
EP1135462A4 (fr) * 1998-12-10 2004-07-14 Tatko Biotech Inc Micro-organismes pour augmenter la productivite vegetale
CN100376666C (zh) * 2006-03-01 2008-03-26 华南农业大学 糖蜜草固氮螺菌及其应用
EP2227936A1 (fr) * 2009-03-09 2010-09-15 Pierre-Philippe Claude Biofertilisation de grandes cultures à l'aide de bactéries azotobacteraceae ayant sur-accumlé du molybdène
CN103159512A (zh) * 2011-12-08 2013-06-19 邵阳市新瑞生物科技有限公司 一种富硒动物蛋白生物有机肥及其制备方法

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US8242053B2 (en) 2007-08-27 2012-08-14 Probelte, S.A. Biological fertilizer, method for obtaining same and use thereof as a plant growth stimulator
ES2378040B1 (es) 2010-03-31 2013-02-18 Probelte, S.A Un preparado biológico bionematicida y estimulador del crecimiento vegetal y cultivos puros de las cepas denominadas n11, sr11 y alo1, contenidas en el mismo.

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1984001686A1 (fr) * 1982-10-28 1984-05-10 Biotech General Corp Nouvelles souches d'azospirillum, procedes de culture de ces souches, compositions les contenant et leur utilisation en tant que bioengrais
EP0241583A2 (fr) * 1986-04-08 1987-10-21 Zhenhua Yang Azotobacter antiammoniac et son utilisation

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES8706826A1 (es) * 1986-05-07 1987-07-01 Consejo Superior Investigacion Procedimiento de obtencion de un mutante de azotobacter vinelandii capaz de fijar n2 en el suelo en concentraciones superiores a las estirpes silvestres conocidas
ES2041219B1 (es) * 1992-04-09 1994-06-01 Consejo Superior Investigacion Procedimiento de activacion microbiologica en la fijacion de nitrogeno.

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1984001686A1 (fr) * 1982-10-28 1984-05-10 Biotech General Corp Nouvelles souches d'azospirillum, procedes de culture de ces souches, compositions les contenant et leur utilisation en tant que bioengrais
EP0241583A2 (fr) * 1986-04-08 1987-10-21 Zhenhua Yang Azotobacter antiammoniac et son utilisation

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
COMMUN. SOIL SCI. PLANT ANAL., Vol. 24 (3 y 4), 1993, MARTIN et al., "Azotobacter and Azospirillum as Potential Nitrogen Fertilizers", pages 255-260. *
FOLIA MICROBIOL., Vol. 35, 1990, FAYEZ M., "Untraditional N2-Fixing Bacteria as Biofertilizers for Wheat and Barley", pages 218-226. *
JOURNAL OF SCIENTIFIC & INDUSTRIAL RESEARCH, Vol. 48, 1989, PANDEY A. & KUMAR S., "Potential of Azotobacters and Azospirilla as Biofertilizers for Upland Agriculture: a Review", pages 134-144. *

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CZ299601B6 (cs) * 1997-02-25 2008-09-17 Zpusob prípravy smesi mikroorganismu na vázání dusíku ze vzduchu, na zvyšování rozpustnosti sloucenin fosforu a na rozklad potravinárského olejového sedimentu
WO1998037038A3 (fr) * 1997-02-25 1998-11-12 Arpad Pollak Procede pour extraire l'azote de l'air, pour extraire les composes de phosphore du sol et pour decomposer les sediments d'huile
EP1135462A4 (fr) * 1998-12-10 2004-07-14 Tatko Biotech Inc Micro-organismes pour augmenter la productivite vegetale
WO2001032587A1 (fr) * 1999-11-01 2001-05-10 'piksa Inter' Ltd Produit d'addition biologique pour fertilisants organomineraux
EP1248754A4 (fr) * 1999-11-01 2003-03-12 Vinarov Alexandr Juryievich Produit d'addition biologique pour fertilisants organomineraux
US6939688B1 (en) 1999-11-01 2005-09-06 Soil Biogenics Ltd Biological addition to organic-mineral fertilizers
WO2001085916A1 (fr) * 2000-05-12 2001-11-15 Biofarmtech Co., Ltd. Materiau de revetement photocatalytique presentant une activite photocatalytique et des proprietes d'adsorption, et procede de preparation associe
US7842494B2 (en) 2001-08-13 2010-11-30 Agro.Bio Hungary Kft. Micro-organisms for the treatment of soil and process for obtaining them
EA009126B1 (ru) * 2001-08-13 2007-10-26 Агро. Био Хангери Кфт. Микроорганизмы для обработки почвы и способ их получения
KR100940615B1 (ko) * 2001-08-13 2010-02-05 아그로.바이오 헝가리 케이에프티. 토양 처리용 미생물 및 이를 얻기 위한 방법
WO2003016241A1 (fr) * 2001-08-13 2003-02-27 Agro.Bio Hungary Kft. Micro-organismes pour le traitement du sol et son procede de production
EP2233458A3 (fr) * 2001-08-13 2011-02-23 Agro.Bio Hungary Kft. Microorganismes pour le traitement du sol et un procédé de leur obtention
HRP20040126B1 (hr) * 2001-08-13 2014-02-14 Agro-Bio Hungary Kft. Mikroorganizmi za tretiranje tla i postupci za njihovo dobivanje
CN100376666C (zh) * 2006-03-01 2008-03-26 华南农业大学 糖蜜草固氮螺菌及其应用
EP2227936A1 (fr) * 2009-03-09 2010-09-15 Pierre-Philippe Claude Biofertilisation de grandes cultures à l'aide de bactéries azotobacteraceae ayant sur-accumlé du molybdène
CN103159512A (zh) * 2011-12-08 2013-06-19 邵阳市新瑞生物科技有限公司 一种富硒动物蛋白生物有机肥及其制备方法

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Publication number Publication date
ES2093559A1 (es) 1996-12-16
ES2093559B1 (es) 1997-07-01

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