CN102742606A - Application of gingko endogeny Chaetomium globosum in prevention and control of plant pathogenic fungi - Google Patents
Application of gingko endogeny Chaetomium globosum in prevention and control of plant pathogenic fungi Download PDFInfo
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- CN102742606A CN102742606A CN2012101851030A CN201210185103A CN102742606A CN 102742606 A CN102742606 A CN 102742606A CN 2012101851030 A CN2012101851030 A CN 2012101851030A CN 201210185103 A CN201210185103 A CN 201210185103A CN 102742606 A CN102742606 A CN 102742606A
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Abstract
The invention relates to an application of a strain of gingko endogeny Chaetomium globosum NO.05 in prevention and control of plant pathogenic fungi, and belongs to the technical field of biological prevention and control of plant diseases. A strain of the gingko endogeny Chaetomium globosum NO.04 and fermentation metabolites thereof in the present invention can be applied for prevention and control of northern corn leaf blight, Grapevine white rot, and tomato rhizopus fruit rot caused by plant pathogenic fungi. According to the present invention, a microbe strain is provided for prevention and control of northern corn leaf blight, Grapevine white rot, tomato rhizopus fruit rot, and other plant pathogenic fungi diseases on agriculture; and the provided fermentation metabolites can be applied in prevention and control of plant fungi diseases, and a new approach is provided for development of agricultural fungicides.
Description
Technical field
The invention belongs to the biocontrol of plant disease technical field, be specifically related to the application of the gingko endogenous chaetomium globosum NO.05 of a strain in control of maize leaf blight bacterium (Exserohilum turcicum), fruit white rot of grape bacterium (Coniothyrium diplodiella) and rhizopus stolonifer (Rhizopus stolonifer).
Background technology
The pesticide resistance of phytopathogen is serious problems in the plant disease chemical control, and it brings very big difficulty for the control of various plants (like crops, economic crops and horticultural crop etc.) disease.Because chemical bactericide particularly systemic fungicide uses for a long time and widely, phytopathogen is very outstanding to its pesticide resistance.Need especially combine through integrated control, overcome its drug-fast problem, thereby phytopathogen is control effectively with biological control.
But the biological control agents majority still is in the starting stage of research and development at present, has satisfied not far away and has produced the actual requirement of going up plant pest management.The severe overweight of persticide residue in vegetables, the fruit on the market is unfavorable for the sound development of pollution-free food in addition, and it is imperative therefore to develop the biological control agents product energetically.
The existence of endogenetic fungus has generality and diversity.Plant endogenesis epiphyte is one type of resource microorganism that has a extensive future, and oneself becomes the focus that the biological control microbial inoculum is developed in recent years, and utilizes antagonism property endogenetic fungus to carry out the importance that biological control is ecological agriculture sustainable development.It is to utilize it to carry out the element task of control of plant disease that screening obtains to have the endogenetic fungus that suppresses the pathogen activity, and the endophyte with broad-spectrum disease resistance insect pest effect has great application prospect in biological control damage by disease and insect, promotion agricultural sustainable development.Exploitation to the fungoid bactericide has both at home and abroad given very big concern always, and existing at present more than 20 fungies that belong to are used to the biological and ecological methods to prevent plant disease, pests, and erosion practice of plant disease.
Chaetomium (Chaetomium) is main saprophytic in plant residue, paper, soil, animal wastes be rich in cellulosic old and useless article; Enzyme to produce abundant degraded cellulose is celebrated with having the secondary metabolite of physiologically active, is considered to have the monoid of potential source biomolecule prevention and control capability.About more than 300 of the cupreums of having reported are in the world planted, but the certain micro-organisms in the antagonism soil.Martin and Moore (1954) find; After the seed of each kind oat of Brazil is rolled up cupreum (Chaetomium cochliodes) and is infected by chaetomium globosum (Chaetomium globosum) and spiral shell; Then the effect to the long spore toxin of wriggling of Victoria has produced resistance, thereby makes the seedling of cereal, oat, barley have certain resistance to Fusarium (Fusarium spp.).Walther (1988) Fusarium oxysporum (Fusarium oxysporum schlecht) is the pathogenic bacteria that cause tomato wilt, if use simultaneously with cupreum and chemical bactericide PCNB, the incidence of disease of tomato wilt is reduced to 22% by 43%.Ultimate corruption mould (Pythium ultimum) is to cause that the seed of many plants rots and the pathogen of plant damping off; This pathogen survived in soil with the form of egg spore and sporangiospore in the phase of surviving the winter; When condition is suitable; In 12 hours, just can form bacterium colony, the normal growth of damage to crops then on the beet seed surface.The sugarcane fusarium wilt is because the sugarcane seed is polluted caused by beet stem point mould (Phoma botae) pollution, soil by ultimate corruption mould (Pythium ultimum) or Rhizoctonia solani Kuhn (Rhizoctonia solani); Pietro (1992) handles the sugarcane seed with the ascospore of cupreum, can reduce the incidence of sugarcane fusarium wilt.Equally; Cotton wilt is also owing to cotton seeds is caused by ultimate corruption mould (Pythium ultimum) or Rhizoctonia solani Kuhn (Rhizoctonia solani) pollution; Handle cotton seeds with the ascospore of cupreum, can effectively control this and two grow cotton causing harm that pathogen causes.Handle the Cucurbita seed with ball hair shell, can reduce the seed that the seed fruit body is damaged and pythium spp (Pythium spp.) causes that causes because of Hylemya platura and rot.If seed is not handled with cupreum prior to seeding; Then seed is at duration of germination; Many microorganisms grow at the surface of the seed soon surely very much, and the ovum of fly can promote the growth of the surface of the seed pseudomonad, make pseudomonad become the dominant bacteria of the surface of the seed; Thereby promote the propagation of phytopathogen, cause that then seed rots and the generation of plant damping off.If seed is handled with cupreum before sprouting, to sow again, the ascospore of cupreum is bred very soon and is formed the thick mycelia of one deck at the surface of the seed, has suppressed the logarithmic growth of pseudomonad greatly, and has not influenced the fungus strain of seed surrounding soil.In addition, Christian (1983) and Daniel (1984) foliage spray ball hair shell preparation can be controlled by the caused apple scab of venturia inaequalis (Venturia inaequlis), as with ball hair shell preparation (5 * 10
6Cps/ml) be sprayed on the apple leaf, can reduce the spore count of Venturia inaequlis and reduce the rotten degree of leaf.Play an important role aspect the pathogen that the antimycotic material ball chaetocin (Chaetomin) that is produced by ball hair shell causes the sugarcane damping off in inhibition---the pythium spp (Pythium ultimum).Equally, Soytong (2000) cupreum all has certain inhibitory action to the pathogen of plants such as strawberry, durian and black pepper.
The secondary metabolite that plant endogenesis epiphyte produces has abundant biological function and chemical constitution diversity, in agricultural and pharmaceutical sector, has the important use potentiality, and the some of them compound possibly become the lead compound of medicament research and development.Plant endogenesis epiphyte has become seeks the new resources with novel chemical constitution and bioactivator.
Summary of the invention
The objective of the invention is the dangerous and less deficiency of microbial pesticide kind, a kind of endogenetic fungus that from ginkgo, separates be provided to chemical pesticide---chaetomium globosum (Chaetomium globosum) NO.05; Another object of the present invention provides the application that utilizes this bacterial strain and metabolite antagonism Exserohilum turcicum thereof, fruit white rot of grape bacterium, rhizopus stolonifer.
The gingko endogenous chaetomium globosum of a strain provided by the invention (Chaetomium globosum) NO.05 and fermentating metabolism product thereof can be used in corn northern leaf blight northern corn leaf blight, fruit white rot of grape Grapevine white rot, tomato head mold fruit rot Tomato rhizopus fruit rot that control is caused by plant pathogenic fungi.
The gingko endogenous chaetomium globosum of one strain (Chaetomium globosum) NO.05 and fermentating metabolism product thereof; NO.05 is inoculated in the fermentation medium with the gingko endogenous chaetomium globosum of a strain (Chaetomium globosum); Cultivate 7-8d at 25-30 ℃ of bottom fermentation, the zymotic fluid that obtains is as the microbial inoculum of control plant pathogenic fungi.
The prescription of described fermentation medium is: distilled water 1000mL, oat 30g, maltose 20g, the pH of fermentation medium are 7.0.
Separation screening of chaetomium globosum and evaluation among the present invention:
In in June, 2008 in the ginkgo leaf that gather in Linyi, Shandong, obtain and preserve through steps such as separation, purifying, cultivation, fermentation and determinations of activity; Through morphology and 18SrDNA Protocols in Molecular Biology, be accredited as chaetomium globosum (Chaetomium globosum), tentative chaetomium globosum (C.globosum) NO.05 by name.This bacterial strain is preserved in plant science institute of Jilin University integrated pest control laboratory, preservation date on October 2nd, 2010, preservation registration number NO.05.
Chaetomium globosum NO.05 bacterial strain is cultivated morphological feature basically: bacterium colony light color, and aerial hyphae light color has colourless or light orange secretion; The ascocarp hypergene, sphere, oval or obovate have the aperture, and 7d begins maturation, olive brown or taupe under the reverberation; Appendage is many, Zhousheng, and straight or microbend, brown has verruca; The ascus club-like clusters, and has 8 ascospores, has handle, clears up easily; The ascospore brown, lemon shape, heavy wall, both sides are level and smooth, and the two ends projection has a significantly living germinal aperature in top.
Chaetomium globosum NO.05 of the present invention can obtain the inhibited metabolite of plant pathogenic fungi through fermentation.
Beneficial effect of the present invention:
The one, the present invention separates metabolite the fungal diseases of plants pathogen is had stronger inhibiting chaetomium globosum NO.05 from the plant Ginkgo biloba leaf, and this goes up fungal diseases of plants pathogens such as control of maize leaf blight, fruit white rot of grape, tomato root mould fruit rot a kind of microbial strains is provided for agricultural;
The 2nd, the invention provides the method for utilizing chaetomium globosum NO.05 fermentation to obtain active metabolite; This fermentating metabolism product can be applicable to the control of fungal diseases of plants, for the exploitation of disinfectant use in agriculture has increased new approach.
Description of drawings
Fig. 1 is chaetomium globosum NO.05 4 * photo
Fig. 2 is chaetomium globosum NO.05 10 * photo
Embodiment
Embodiment 1: the separation of chaetomium globosum NO.05 and screening
The present invention gathers ginkgo leaf in Linyi, Shandong in June, 2008; Through surface sterilization, endogenetic fungus separation, cultivation, fermentation, determination of activity and to suppressing the steps such as screening of fungal diseases of plants pathogen active bacterial strain; Therefrom obtain chaetomium globosum NO.05, preserve.
Chaetomium globosum NO.05 bacteriostatic activity is measured: at the dull and stereotyped central side joint kind diameter of PDA be 9cm Exserohilum turcicum, fruit white rot of grape bacterium, rhizopus stolonifer bacterium piece; Apart from bacterium piece 4.5cm place's point cupreum NO.05 that receives, 28 ℃ of constant temperature culture 2-4d observe and have or not antibacterial band; And measure big or small; The contrast that is treated to of the cupreum NO.05 that do not receive is set, and antibacterial band is big more, and the expression fungistatic effect is strong more.
Inhibiting rate is calculated by formula:
Inhibiting rate (%)=(control group pathogen growth radius one processed group pathogen growth radius)/(control group pathogen growth radius one former bacterium cake radius) x 100%
Mensuration result shows: chaetomium globosum NO.05 is respectively 62.69%, 62.01% and 49.36% to the inhibiting rate of Exserohilum turcicum, fruit white rot of grape bacterium, rhizopus stolonifer.
Embodiment 2: the preparation method of chaetomium globosum NO.05 fermentating metabolism product
Comprise the following steps:
(1) activation culture of bacterial classification: will separate the chaetomium globosum NO.05 bacterial classification of preserving and move on the potato glucose PDA solid plate; Under 28 ℃ ± 1 ℃ condition, cultivate; Cover whole culture dish to mycelia, using diameter is that the punching of 9cm card punch obtains the bacterium cake, supplies inoculation to use;
(2) fermented and cultured: fermentation culture is every 1000mL distilled water, oat 30g, maltose 20g, and pH is 6.0-7.0.The bottled fermentation culture 200mL of every 500mL triangle; Prepare back 121 ℃ the sterilization 20min, to be cooled to 40 ℃ under aseptic condition the picking diameter be the bacterium cake of 9cm, under 28 ℃ ± 1 ℃ condition, shaking speed 150rpm, fermented and cultured 7-8d;
(3) fermentation product extracts: centrifugal collection zymotic fluid after fermentation finishes, add the equal-volume ethyl acetate extraction, and get the upper strata, under vacuum condition, be concentrated into medicinal extract, get acetic acid ethyl ester extract, supply bacteriostatic activity to measure.
Embodiment 3: chaetomium globosum NO.05 fermentating metabolism product is to the inhibitory action of Exserohilum turcicum, fruit white rot of grape bacterium, rhizopus stolonifer
Said chaetomium globosum NO.05 fermentating metabolism product is embodiment 2 made zymotic fluids.
Bacteriostatic activity is measured: get above-mentioned zymotic fluid 50 μ l on the aseptic filter paper sheet, and dry up zymotic fluid, the inoculation diameter is Exserohilum turcicum, fruit white rot of grape bacterium, the rhizopus stolonifer bacterium piece of 9cm in the dull and stereotyped central authorities of PDA; Select the zymotic fluid filter paper of the cupreum NO.05 that receives apart from bacterium piece 3cm place; The filter paper that will have only fermentation medium and dry up is set as negative control, 28 ℃ of constant temperature culture 2-4d observe and have or not antibacterial band; And measure size, not put the contrast that is treated to of the cupreum NO.05 that receives.Antibacterial band is big more, and the expression fungistatic effect is strong more.
Inhibiting rate is calculated by formula:
Inhibiting rate (%)=(control group pathogen growth radius one processed group pathogen growth radius)/(control group pathogen growth radius one former bacterium cake radius) x 100%
Mensuration result shows: chaetomium globosum NO.05 zymotic fluid is respectively 60.60%, 59.00% and 50.13% to the inhibiting rate of Exserohilum turcicum, fruit white rot of grape bacterium, rhizopus stolonifer.
Claims (3)
1. the gingko endogenous chaetomium globosum Chaetomium of strain globosum NO.05 and fermentating metabolism product thereof is characterized in that the application in corn northern leaf blight northern corn leaf blight that control is caused by plant pathogenic fungi, fruit white rot of grape Grapevine white rot, tomato head mold fruit rot Tomato rhizopus fruit rot.
2. by a described strain gingko endogenous chaetomium globosum Chaetomium globosumNO.05 of claim 1 and fermentating metabolism product thereof; It is characterized in that the gingko endogenous chaetomium globosum Chaetomium of strain globosumNO.05 is inoculated in the fermentation medium; Cultivate 7-8d at 25-30 ℃ of bottom fermentation, the zymotic fluid that obtains is as the microbial inoculum of control plant pathogenic fungi.
3. by the described strain of claim 2 gingko endogenous chaetomium globosum Chaetomium globosum NO.05 and fermentating metabolism product thereof; The prescription that it is characterized in that described fermentation medium is: distilled water 1000mL, oat 30g, maltose 20g, the pH of fermentation medium are 7.0.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103087923A (en) * | 2012-11-09 | 2013-05-08 | 南京农业大学 | Preparation and application of chaetomium globosum and metabolite flavipin thereof |
| CN107475123A (en) * | 2017-08-16 | 2017-12-15 | 中国人民解放军第二军医大学 | Roxburgh anoectochilus terminal bud endogenetic fungus and its application |
| CN114907992A (en) * | 2022-05-18 | 2022-08-16 | 塔里木大学 | Strain C11 for antagonizing phytopathogen and application thereof |
-
2012
- 2012-06-06 CN CN2012101851030A patent/CN102742606A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103087923A (en) * | 2012-11-09 | 2013-05-08 | 南京农业大学 | Preparation and application of chaetomium globosum and metabolite flavipin thereof |
| CN103087923B (en) * | 2012-11-09 | 2014-10-08 | 南京农业大学 | Preparation and application of chaetomium globosum and metabolite flavipin thereof |
| CN107475123A (en) * | 2017-08-16 | 2017-12-15 | 中国人民解放军第二军医大学 | Roxburgh anoectochilus terminal bud endogenetic fungus and its application |
| CN107475123B (en) * | 2017-08-16 | 2020-02-11 | 中国人民解放军第二军医大学 | Anoectochilus roxburghii endophytic fungus and application thereof |
| CN114907992A (en) * | 2022-05-18 | 2022-08-16 | 塔里木大学 | Strain C11 for antagonizing phytopathogen and application thereof |
| CN114907992B (en) * | 2022-05-18 | 2023-06-16 | 塔里木大学 | Strain C11 antagonizing plant pathogenic bacteria and application thereof |
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Application publication date: 20121024 |