CN108300670A - One plant can antagonism anthrax-bacilus yeast strain and its application - Google Patents

One plant can antagonism anthrax-bacilus yeast strain and its application Download PDF

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CN108300670A
CN108300670A CN201711430109.9A CN201711430109A CN108300670A CN 108300670 A CN108300670 A CN 108300670A CN 201711430109 A CN201711430109 A CN 201711430109A CN 108300670 A CN108300670 A CN 108300670A
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bacilus
anthrax
culture
strain
plant
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乔敏
余泽芬
贾东晨
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Yunnan University YNU
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Yunnan University YNU
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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
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • C12N1/18—Baker's yeast; Brewer's yeast
    • C12N1/185—Saccharomyces 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
    • C12R2001/85—Saccharomyces
    • C12R2001/865—Saccharomyces cerevisiae
    • 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

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  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Mycology (AREA)
  • Zoology (AREA)
  • Chemical & Material Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Genetics & Genomics (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Biochemistry (AREA)
  • Virology (AREA)
  • Environmental Sciences (AREA)
  • Dentistry (AREA)
  • Plant Pathology (AREA)
  • Pest Control & Pesticides (AREA)
  • Agronomy & Crop Science (AREA)
  • Botany (AREA)
  • Medicinal Chemistry (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Biomedical Technology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

The present invention relates to the yeast strain of one plant of energy antagonism anthrax-bacilus, microorganism belonging to genus applied technical fields.The present invention carries out strain isolation, culture by conventional method from nature, obtain one plant of energy antagonism anthrax-bacilus saccharomyces cerevisiae (Saccharomyces cerevisiae)-MLB1, the bacterial strain are deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number on January 6th, 2014：CGMCC No.8689.The yeast MLB1 bacterial strains of the present invention, the culture in malt extract medium have antagonism to anthrax-bacilus, and the inhibiting rate to fruit anthracnose is more than 75%.

Description

One plant can antagonism anthrax-bacilus yeast strain and its application
Technical field
The present invention relates to the yeast strain of one plant of energy antagonism anthrax-bacilus, microorganism belonging to genus applied technical fields.
Background technology
Plant disease is the formidable enemy of agricultural production, is counted according to FAO (Food and Agriculture Organization of the United Nation) (FAO), the whole world is because of harmful organism institute Caused by economic loss be up to 120,000,000,000 dollars, be equivalent to the half of the Chinese agriculture gross output value.The generation of plant disease is not only straight The income for influencing China peasant is connect, the development of other related industries, especially farming industry are also affected.1880 Year, Dogue de Bordeaux area wine-growing causes wine-making industry to be on the verge of bankruptcy to stop doing business due to the harm by downy mildew.It is the tightest The long-range and sustainable development for being detrimental to agricultural and its related industry of weight.
Currently, the main means of controlling plant diseases are chemical pesticide.It is anti-due to carrying out plant disease using chemical pesticide Quick, wide sterilization spectrum is controlled, it is at low cost, the advantages that using simplicity, it is made to be rapidly developed in cultural control, becomes diseases prevention high yield One of effective means.But it is long-term, using chemical pesticide, there is potentially hazardous repeatedly and largely.Not only bring ring Border pollution and the potential hazard to human health, but also bring the influence to non-target organism and phytopathogen drug resistance Development the problems such as so that the development of chemical pesticide receives the limitation from variant aspect.Find wide spectrum, efficient, low toxicity Biological pesticide have become the common recognition of scientific research personnel and Pesticide use person.
The secondary metabolite generated using antagonistic microbe or antagonistic microbe is plant disease come controlling plant diseases One of highly important effective measures in prevention.Antagonistic microbe, which has, is not easy to make pathogen to generate drug resistance, to safety of human and livestock Nontoxic, free from environmental pollution, noresidue does not injure beneficial microbe, can keep ecological balance, and can avoid due to the use of chemistry A series of problems caused by pesticide control plant disease, and safely, effectively and persistently, many antagonistic microbes also have simultaneously There is the features such as growth-promoting functions to crop and gets growing concern for and pay attention to.
In biocontrol of plant disease Antagonistic Fungus study it is more be trichoderma (TrichodermaSp.) fungi, And it is considered as a kind of most promising bio-control factors that can replace a variety of chemical bactericides.Fungus Trichoderma is in nature point Cloth is very extensive, can be mushroomed out on many plants, has antagonism to a variety of disease fungus and certain pathogenetic bacterias, is one The ideal Biocontrol microorganism of kind.It is mainly used for the prevention of plant silborne fungal diseases, has cotton spoting verticillium wilt, samping off, stands Blight, wheat rust, banded sclerotial blight, various vegetables sprout term disease etc..A kind of trichoderma wettable powder of oneself registration of China is used It is prevented in the downy mildew of cucumber and Chinese cabbage, another trichoderma wettable powder (special Garrick, trichoderma virideT.viride) It is then used for the prevention of the crops fungoid root diseases such as booth vegetable gray mold and wheat, and field practical application shows it also to Huang The downy mildew and powdery mildew of melon have good effect.Trichoderma is also used for the fruits and vegetables storage phase disease such as green mold of citrus Prevention.
Anthracnose is the general designation of a kind of plant disease caused by anthrax-bacilus, and the main harm of the disease is can to cause leaf Withered, fruit rot even dead seedling of spot, branch etc..Germ is imbedded in after infecting plant with conidium in epidermis or cuticula, can be into Suspend mode of the row up to the several months.Anthracnose is one of the Major Diseases of fruit especially tropical fruit (tree), wide in each fruit belt in the world General generation.In recent years, anthracnose grows in intensity, area especially damp and hot in some high temperature, such as Perenniporia martius The generation in area, disease causes extremely serious loss to many vegetables, fruit tree, ornamental plant and crops etc., As one of crop disease the most serious.Colletotrichum (Colletorichum) there are about 590 kinds, numerous species can be endangered Host leads to the generation of anthrax disease, and prodigious threat is brought to the agricultural production of countries in the world.
Anthrax bacteria, in field latent infection fruit, is difficult control after adopting before fruit is adopted, thus during easily leading to storing The commodity shelf phase of fruit is shortened in fruit rot.Varietal resistance, fruit before the method for control fruit anthracnose is mainly adopted at present Cultivation management condition, rainfall, the temperature other factors in garden, air conditioned storage, heat treatment after adopting, chemical bactericide processing and life Object prevention etc., but all neither one stabilization and effective methods.
To the biological control of anthracnose, there is presently no molding products to appear on the market.Huang Siliang etc. passes through micro- to antagonism in air The investigation of biology is found, there are a large amount of anthracnose antagonistic microbe in air, is followed successively by the antagonism of anthracnose：Bacterium >Actinomyces>Fungi (Huang think good etc., 1999).Have been reported that Penicillium (Penicillium) many bacterial strains have to mango anthracnose Very strong antagonism, wherein act on it is strongest be a watt gram mould (Penicillium waksmanZaleski) (Zhang Yixian, 2006).Zhang Lubin etc. reports the bacillus M35 that can effectively inhibit anthrax bacteria, and bacteria liquid suspension a concentration of 1 × 108When cfu/ml, incidence is 45% (Zhang Lubin etc., 2010).Patino-Vera M (2005) etc. are before the harvest season with small RhodotorulaRhodotorula minutaSuspension sprays mango, adopts the incidence of rear anthracnose and using chemical bactericide Maintain an equal level or lower.
The emphasis of biological pesticide research and development at present and it is with fastest developing speed be farm antibiotics, such as jinggangmeisu, kasugarnycin, more It is true that antimycin, new Polyoxin, Zhongshengmycin, pesticide corrosion 120, Wuyiencin, qingfengmeisu have been widely used in prevention plant Fungus diseases.China develop some possess and obtain only more than the ten of agriculture chemical registration in the farm antibiotics of independent intellectual property right and plant, And be mainly used for preventing the disease of rice and field crops, and be used to prevent veterinary antibiotics etc. important fungal disease then very It is few.In addition, due to the difficulty of fungal disease prevention and the deficiency of biological control preventive effect, the preventive effect of anthracnose biological control rarely has Chemical pesticide is reached and exceeded, the inhibiting rate of anthrax-bacilus is difficult to stablize 70% or more.
MLB1 bacterial strains of the present invention are isolated from natural environment, which is the energy antagonism anthrax found for the first time The Wine brewing yeast strain of bacterium, it is different from the other yeast strains for the energy antagonism anthrax-bacilus reported before, be on category significance not Same yeast monoid, while the strain culture energy antagonism anthrax-bacilus, the inhibiting rate to fruit anthracnose are more than 75%.
Invention content
The purpose of the present invention is to provide the bacterial strain of one plant of energy antagonism anthrax-bacilus, the energy which belongs to inventor's discovery is short of money The Saccharomyces strain of anti-anthrax-bacilus, the culture energy antagonism anthrax-bacilus of the bacterial strain.
The present invention bacterial strain be saccharomyces cerevisiae (Saccharomycescerevisiae)-MLB1, the bacterial strain in January, 2014 It is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number within 6th：CGMCC No.8689；Preservation Address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.
The bacterium source of the present invention is in natural environment, and culture has antagonism to anthrax-bacilus, to the inhibiting rate of anthrax-bacilus More than 75%.
The bacterial strain of the present invention is different from other energy Saccharomyces strains of antagonism anthrax-bacilus, belongs to the inhomogeneity on taxology Group, and be the Wine brewing yeast strain for the energy antagonism anthrax-bacilus that inventor has found for the first time.
The present invention is accomplished by the following way：
One, the acquisition of bacterial strain：
The bacterial strain of the present invention is detached by the conventional commonsense method that air microbe detaches, purifies culture acquisition, the training of use Support base be malt extract medium, through ITS be accredited as saccharomyces cerevisiae (Saccharomyces cerevisiae), it names as MLB1；
Form：25 DEG C of cultures, cell circle, oval or foreign pyriform.In childhood bacterium colony, cell is 3 ~ 7 × 4 ~ 10 microns, long It is 1 with wide ratio:1~2:1；20 DEG C of cultures precipitate for 21 days in brewer's wort.Ascospore is round, smoothly；For wild-type strain, Rather than engineering strain.
Two, the culture of MLB1 bacterial strains：
First, malt is prepared into malt broth by microorganism routine malt processing method spare, 115 DEG C of sterilising conditions 20 minutes.
Secondly, MLB1 bacterial strains are carried out by microbial fermentation conventional liq seed culture method, with sterilizing malt medium Culture, after shaking flask, a series of cultures of seeding tank, when every milliliter of culture solution is more than 1 × 10 containing yeast count8When a, by fermentation The amount of tank culture solution 2%, which is linked into fermentation tank, is cultivated, and temperature is controlled at 27 DEG C, is continually fed into filtrated air, is ventilated Than controlling 1:1.1, dissolved oxygen is maintained at 40%.Culture 48-72 hours.
Three, MLB1 strain cultures measure the antagonistic ability of anthrax-bacilus：Using conventional pathogen as indicator bacteria External plate antagonism measuring method carries out (Zheng Aiping, 2003).
Specific implementation mode：
Embodiment one：
First, malt is prepared into malt broth by microorganism routine malt processing method spare, 115 DEG C of sterilising conditions 20 minutes.
Secondly, by MLB1 bacterial strains, by microbial fermentation conventional liq seed culture method, to sterilize, malt medium carries out Culture, the sugar a concentration of 3% of seed culture fluid, the sugar a concentration of 5% of fermentation tank culture liquid.Through shaking flask, a series of cultures of seeding tank Afterwards, when every milliliter of culture solution contains yeast count 1 × 108When a, it is linked into fermentation tank and carries out by the amount of fermentation tank culture liquid 2% Culture, temperature are controlled at 27 DEG C, are continually fed into filtrated air, and ventilation ratio is controlled 1:1.1, dissolved oxygen is maintained at 40%.Culture 48 hours.
Finally, collect zymotic fluid, i.e., it is spare for purpose culture, antagonistic ability is measured, the inhibiting rate to anthrax-bacilus is 75%。
Embodiment two：
With one difference of embodiment：Seed culture fluid yeast count reaches 1.8 × 108Fermentation tank, fermentation tank training are accessed when a The sugar a concentration of 7% of nutrient solution, incubation time 72 hours.Measure antagonistic ability.Inhibiting rate to anthrax-bacilus is 80%.
Embodiment three：
With one difference of embodiment：Seed culture fluid yeast count reaches 2.5 × 108Fermentation tank, fermentation tank training are accessed when a The sugar a concentration of 7% of nutrient solution, incubation time 60 hours.Measure antagonistic ability.Inhibiting rate to anthrax-bacilus is 85%.
Example IV：
With one difference of embodiment：Seed culture fluid yeast count reaches 2.0 × 108Fermentation tank, fermentation tank training are accessed when a The sugar a concentration of 9% of nutrient solution, incubation time 72 hours.Measure antagonistic ability.Inhibiting rate to anthrax-bacilus is 90%.
Embodiment five：
Cultured effective culture is diluted with water the solution after 100 times, fruit tree blade face and fruit are uniformly sprayed, not spray The fruit tree of culture solution is control, and the fruit tree of investigation control and application culture after 30 days compares the morbidity of Fruit Anthracnose Rate, culture are 75% to the inhibiting rate of fruit anthracnose.
Embodiment six：
Cultured effective culture is diluted with water the solution after 100 times, is uniformly sprayed in the fruit surface harvested, with not The fruit for spraying culture solution is control, investigates control after 60 days and sprays the fruit of culture, compares fruit anthracnose Incidence, culture are 79% to the inhibiting rate of fruit anthracnose.

Claims (1)

1. the yeast strain of one plant of energy antagonism anthrax-bacilus, it is characterised in that：
1.1 strain isolations from soil environment, it is categorized be accredited as saccharomyces cerevisiae (Saccharomyces cerevisiae)— MLB1, the bacterial strain are deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation on January 6th, 2014 Number：CGMCC No.8689；
The 1.2 bacterial strain energy antagonism anthrax-bacilus, the culture in malt extract medium can effectively inhibit anthrax-bacilus；
1.3 bacterial strains effectively inhibit the acquisition of anthrax-bacilus culture that MLB1 bacterial strains are pressed microbial fermentation conventional liq seed culture Method is cultivated with sterilizing malt medium, after shaking flask, a series of cultures of seeding tank, when every milliliter of culture solution contains yeast Bacterium number is more than 1 × 108When a, it is linked into fermentation tank and is cultivated by the amount of fermentation tank culture liquid 2%, temperature is controlled at 27 DEG C, Holding is continually fed into filtrated air, and ventilation ratio is controlled 1:1.1, dissolved oxygen is maintained at 40%, cultivates 48-72 hours；
1.4 strain culturings are more than 75% to the inhibiting rate of fruit anthracnose；
Saccharomyces cerevisiae described in right 1 (Saccharomyces cerevisiae)-MLB1 bacterial strains answering in antagonism anthrax-bacilus With.
CN201711430109.9A 2017-12-26 2017-12-26 One plant can antagonism anthrax-bacilus yeast strain and its application Pending CN108300670A (en)

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114829321A (en) * 2019-12-17 2022-07-29 植物响应公司 Methods and systems for reducing pathogens in organic materials

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104946548A (en) * 2015-06-26 2015-09-30 云南大学 Saccharomyces cerevisiae strain and application thereof

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN104946548A (en) * 2015-06-26 2015-09-30 云南大学 Saccharomyces cerevisiae strain and application thereof

Non-Patent Citations (3)

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王润菡等: "采后芒果炭疽病拮抗酵母菌的分离和筛选", 《食品研究与开发》 *
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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN114829321A (en) * 2019-12-17 2022-07-29 植物响应公司 Methods and systems for reducing pathogens in organic materials
EP4077243A4 (en) * 2019-12-17 2024-01-17 Plant Response, Inc. Methods and systems for pathogen mitigation in organic materials

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