CN104642018A - Biological control disease-resistant promoting method for Zanthoxylum bungeanum - Google Patents
Biological control disease-resistant promoting method for Zanthoxylum bungeanum Download PDFInfo
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- CN104642018A CN104642018A CN201510058447.9A CN201510058447A CN104642018A CN 104642018 A CN104642018 A CN 104642018A CN 201510058447 A CN201510058447 A CN 201510058447A CN 104642018 A CN104642018 A CN 104642018A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G13/00—Protecting plants
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- 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
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
- Y02W30/00—Technologies for solid waste management
- Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse
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- Pest Control & Pesticides (AREA)
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Abstract
The invention discloses a biological control disease-resistant promoting method for Zanthoxylum bungeanum. The method comprises the following steps of making bacillus subtilis into high-density bacterium suspension of which the density is 109-1010cfu (colony-forming unit)/mL; cutting healthy branches of the Zanthoxylum bungeanum into small sections for composting; spraying the high-density bacterium suspension of the bacillus subtilis in the composting process; fermenting to obtain biological fertilizer capable of effectively suppressing and killing pathological fungi of the Zanthoxylum bungeanum, and applying the biological fertilizer to the biological control disease-resistant promoting of the Zanthoxylum bungeanum. According to the biological control disease-resistant promoting method disclosed by the invention, as the bacillus subtilis producing beta-glucanase is plant probiotics of playing a role of disease-resistant promoting on the Zanthoxylum bungeanum, by adding the bacillus subtilis to compost, the biological control action on pathogenic bacteria of phytophthora capsici of the Zanthoxylum bungeanum is remarkably enhanced, and the bacillus subtilis has a promoting effect; the composting process treatment is simple; no waste water is drained; the investment is small; the energy consumption is low; the operation is easy; the biological control disease-resistant promoting method is easy for industrialized mass production, and has a good economic benefit and a broad application prospect.
Description
Technical field
The present invention relates to the preparation field of biological control fertilizer, be specifically related to the method for the disease-resistant growth-promoting of a kind of Chinese prickly ash biological control.
Background technology
Chinese prickly ash (Zanthoxylum bungeanum Maxim.) Bie Ming Luen, large green pepper, long and thin hot pepper, another name for Sichuan Province green pepper, pericarpium zanthoxyli or Chinese pepper, be Rutaceae, Zanthoxylum machaka or dungarunga, can isolated planting can do again to protect thorn hedge.Its pericarp can be used as flavoring, and can extract aromatic oil, can be used as medicine again, seed edible, also can processing and fabricating soap.Chinese prickly ash except the stink smell of various meat, can promote salivary secretion, and increase appetite, distend the blood vessels, thus play hypotensive activity, population all can eat, pregnant woman, fire excess from yin deficiency person's diet.Pericarpium Zanthoxyli fruit is rounded, mung bean size, and its crust is a kind of conventional perfumes.Red or aubergine pericarp is red green pepper during fruit maturation, seed is in green pepper order, and be all traditional Chinese medicine, family commonly uses flavouring, and Chinese prickly ash plant is drought-enduring, like sunlight, and the many plantations in various places, are distributed in the provinces such as Shanxi, Shaanxi, Sichuan, Shandong, Hebei, Henan.
The Chinese prickly ash phytophthora root rot (also known as balck shank) caused by phytophthora (Phytophthora spp.) is a kind of important trunk disease, and the winter raw epidemic disease mould (Phytophthora hibernalis) is one of the main pathogenic fungi.Plant pathology fungi is mainly present in soil, infection plant in plant growth, causes plant disease, causes production loss.Therefore, fungal diseases of plants to be eliminated from the angle of biological control, kill or suppress the pathology fungi in soil most important, and use the bio-fertilizer with biological control effect to be the effective way realizing this purpose.In various bio-fertilizer, what China applied for a long time is all without gemma mushroom.But low without Bacillus resistance, the resistivity for dry high temperature is poor, so stability is not high when making liquid or adsorbent.The new trend of bio-fertilizer development should be greatly develop resistance high, can the bacillus of existence steady in a long-term.Particularly bacillus can produce multiple antifungal substance, has remarkable inhibitory action to phytopathogen, and to person poultry harmless, does not also pollute the environment, and will play an important role in control fungal diseases of plants.
In straight or closely straight shaft-like in bacillus form, aerobic or amphimicrobian, Gram-positive, can produce resistance endospore under certain environmental conditions.Bacillus extensively surely grows in plant, is considered to generally acknowledged grade-safe microorganism (generally regarded as safety, GRAS).Bacillus subtilis is a kind of typical bacillus, produce the antibacterial materials such as antibacterial protein, polypeptides matter, lipid material and beta dextranase, particularly hydrolase plant pathology fungi anti-with it ability such as beta dextranase has substantial connection, because the composition of fungal cell wall is glucan.
Summary of the invention
The invention provides the method for the disease-resistant growth-promoting of a kind of Chinese prickly ash biological control, by being incorporated in Chinese prickly ash natural flora by the bacillus subtilis (CICC 10339) producing beta dextranase, make the bio-fertilizer that effectively can suppress to kill by biological control approach Chinese prickly ash pathology fungi.
A method for the disease-resistant growth-promoting of Chinese prickly ash biological control, comprises the following steps:
1) bacillus subtilis (CICC 10339) is cultivated, make density 10
9~ 10
10the high density bacteria suspension of cfu/mL;
2) healthy for Chinese prickly ash branch is cut into segment and carries out compost, spray step 1 in composting process) in high density bacteria suspension, compost ferments after terminating, after fermentation, obtain the bio-fertilizer that can effectively suppress to kill Chinese prickly ash pathology fungi, this bio-fertilizer is used in the disease-resistant growth-promoting of Chinese prickly ash biological control.
In the present invention, healthy for Chinese prickly ash branch is cut into trifle compost, and make the flora in prepared bio-fertilizer be healthy Chinese prickly ash plant naturally flora, this flora and Chinese prickly ash plant coevolution grow, define mutual beneficial syntaxial system, have certain biological control effect to disease fungi.Further, the bacillus subtilis (CICC 10339) that can produce beta dextranase adds in compost, significantly strengthens the biological control effect to Chinese prickly ash disease fungi.Bacillus subtilis (CICC 10339), owing to producing beta dextranase, has good decomposition to Chinese prickly ash plant fiber, by composting fermentation process is necessary, therefore fully can be dissolved in compost flora and breeds stably and lastingly.As the prebiotic effect of lactic acid bacteria for animal, the microorganism killing plant pathology fungi is suppressed to be referred to as plants probiotics.In bio-fertilizer prepared by the present invention, bacillus subtilis (CICC 10339) is namely Chinese prickly ash probio, has remarkable biological control effect to Chinese prickly ash pathology fungi.
Step 1) in, bacillus subtilis (CICC 10339) adopts prior art, from Chinese industrial Microbiological Culture Collection administrative center (CICC), and address: No. 24, Road, Jiuxianqiao, Chaoyang District, Beijing City institute No. 6 building; Preservation date is 2007, and bacterial classification keeps being numbered 10339.Bacillus subtilis (CICC 10339) can be bought to Chinese industrial Microbiological Culture Collection administrative center.
Bacillus subtilis (CICC 10339) is cultivated, comprising: the density of setting out of bacillus subtilis (CICC10339) is 10
6~ 10
7individual/mL, being seeded to inoculum concentration in liquid nutrient medium is 1% ~ 5%, 33 DEG C ~ 39 DEG C shaken cultivation 20 ~ 28h, is 10 to the liquid-tight degree of bacterium
9~ 10
10the high density bacteria suspension of cfu/mL.
Further preferably, bacillus subtilis (CICC 10339) is cultivated, comprising: the density of setting out of bacillus subtilis (CICC 10339) is 10
6~ 10
7individual/mL, being seeded to inoculum concentration in liquid nutrient medium is 2% ~ 3%, 36 DEG C ~ 37 DEG C shaken cultivation 22 ~ 26h, is 10 to the liquid-tight degree of bacterium
9~ 10
10the high density bacteria suspension of cfu/mL.
Described liquid nutrient medium, in 1L, comprises following component:
Regulate pH=6.5 ~ 7.5.
Further preferably, described liquid nutrient medium, in 1L, comprises following component:
Regulate pH=7.0.
Liquid nutrient medium is at 121 DEG C of sterilizing 20min.
Step 2) in, spray step 1 in composting process) in high density bacteria suspension, comprising: when compost internal temperature reaches 30 DEG C ~ 34 DEG C, spray step 1) in high density bacteria suspension, continue compost, when compost internal temperature reaches 41 DEG C ~ 45 DEG C, compost terminates.
Spray step 1 in composting process) in high density bacteria suspension, comprising: when compost internal temperature reaches 32 DEG C, spray step 1) in high density bacteria suspension, continue compost, when compost internal temperature reaches 43 DEG C, compost terminates.
Step 2) in, after compost terminates, compost is stirred spreading for cooling.Described fermentation adopts nature bottom fermentation.
Compared with prior art, tool of the present invention has the following advantages:
In the present invention, healthy for Chinese prickly ash branch is cut into trifle compost, and make the flora in prepared bio-fertilizer be healthy Chinese prickly ash plant naturally flora, this flora and Chinese prickly ash plant coevolution grow, define mutual beneficial syntaxial system, have certain biological control effect to disease fungi.
Bacillus subtilis (CICC 10339) produces beta dextranase, and strong destruction pathology fungal cell wall, has pathology fungi and significantly suppress killing action.As the prebiotic effect of lactic acid bacteria for animal, the microorganism killing plant pathology fungi is suppressed to be referred to as plants probiotics.Namely the bacillus subtilis (CICC 10339) of being rich in bio-fertilizer prepared by the present invention is Chinese prickly ash probio, has remarkable biological control effect to Chinese prickly ash pathology fungi.
Composting process carries out in its natural state, the stable Chinese prickly ash plant natural flora of Tiny ecosystem is relied on to ferment, and bacillus subtilis (CICC 10339) is owing to producing beta dextranase, good decomposition is had to Chinese prickly ash plant fiber, by composting fermentation process is necessary, therefore, bacillus subtilis (CICC10339) fully can be dissolved in compost flora and breed stably and lastingly.Composting process process is easy, non-wastewater discharge, and small investment, energy consumption are low, easy to operate, is easy to industrialization large-scale production, has good economic benefit and wide application prospect.
Embodiment
Embodiment 1
1) produce the High Density Cultivation of the bacillus subtilis (CICC 10339) of beta dextranase, the density of setting out of bacillus subtilis (CICC 10339) is 1.11 × 10
6individual/mL, is seeded to liquid nutrient medium (glucose 14g/L, beef extract 25g/L, potassium dihydrogen phosphate 2.45g/L, groundnut meal 10g/L, pH=7.0,121 DEG C of sterilizing 20min) in, inoculum concentration is 2%, 37 DEG C of shaken cultivation 24h, and the liquid-tight degree of bacterium is 2.50 × 10
9cfu/mL, obtains high density bacteria suspension.
2) the common 100kg of healthy for Chinese prickly ash branch is cut into trifle and carries out Natural compost, spray 1 when compost internal temperature reaches 32 DEG C) in the high density bacteria suspension 2L for preparing, continue compost, when compost internal temperature is more than 43 DEG C, compost terminates, stir spreading for cooling, after fermentation under nature, becomes the bio-fertilizer being rich in the bacillus subtilis producing beta dextranase, effectively can suppress the bio-fertilizer killing Chinese prickly ash pathology fungi.
3) step 1) in the high density bacteria suspension for preparing and step 2) in the bio-fertilizer for preparing significantly there is Beta dextranase vigor, have the mould Phytophthorahibernalis of Chinese prickly ash pathology fungi winter raw epidemic disease and significantly suppress killing action, specifically see the following form 1.
Mould Phytophthora hibernalis is purchased from Chinese industrial Microbiological Culture Collection administrative center (CICC) for winter raw epidemic disease, bacterium numbering 3.986.
Beta dextranase vitality test: adopt reducing sugar test method, Beta glucan can be degraded into oligosaccharides and monose by Beta dextranase, the reduction group that it has under boiling water bath condition can with DNS reagent generation chromogenic reaction, the colour developing depth is directly proportional to reducing sugar growing amount, and reducing sugar growing amount is directly proportional to reactant liquor Beta glucanase vigor, thus utilize the light absorption value of colorimetric estimation reactant liquor to calculate reducing sugar growing amount, obtain Beta dextranase vigor.Beta glucanase unit of activity defines: under condition determination, and hydrolysis Beta glucan per minute produces the enzyme amount needed for 1 μm of ol reducing sugar (with glucose meter), is defined as a Ge Meihuo unit (IU).
The antibacterial broadband of dull and stereotyped suppression: adopt dull and stereotyped spray-on process, test strains point is connected on NA flat board, often dull and stereotyped 2 strains, each 3 repetitions, cultivates 24h for 28 DEG C, the filter paper being moistened with chloroform is put into stifling 20min bottom inverted culture dish, repeats once, kill bacterium to be measured; Disease fungus is made into 10
8bacteria suspension, is sprayed on rapidly with aseptic laryngeal spray (0.1ml/ ware) on the flat board being connected to bacterium to be measured, and after 48-60h is cultivated in 28 DEG C of continuation, record forms inhibition zone size.
Dull and stereotyped antagonism: pour NA medium into flat board, becomes triangle to place Oxford cup, adds zymotic fluid 100 μ L in the cup of Oxford on medium after solidifying, 28 DEG C of incubators are cultivated, after disease fungus growth, record inhibition zone size.
Table 1
Note: antibacterial broadband refers to the difference of antibacterial radius and colony radius to be measured ,-: indicate without antibacterial band; +: represent antibacterial broadband <5mm; ++ represent antibacterial broadband >5mm.
Embodiment 2
1) produce the High Density Cultivation of the bacillus subtilis (CICC 10339) of beta dextranase, the density of setting out of bacillus subtilis (CICC 10339) is 1.65 × 10
6individual/mL, is seeded to liquid nutrient medium (glucose 14g/L, beef extract 25g/L, potassium dihydrogen phosphate 2.45g/L, groundnut meal 10g/L, pH=7.0,121 DEG C of sterilizing 20min) in, inoculum concentration is 3%, 37 DEG C of shaken cultivation 24h, and the liquid-tight degree of bacterium is 2.92 × 10
9cfu/mL, obtains high density bacteria suspension.
2) the common 100kg of healthy for Chinese prickly ash branch is cut into trifle and carries out Natural compost, spray 1 when compost internal temperature reaches 32 DEG C) in the high density bacteria suspension 3L for preparing, continue compost, when compost internal temperature is more than 43 DEG C, compost terminates, stir spreading for cooling, after fermentation under nature, becomes the bio-fertilizer being rich in the bacillus subtilis producing beta dextranase.
3) step 1) in the high density bacteria suspension for preparing and step 2) in the bio-fertilizer for preparing significantly there is Beta dextranase vigor, have Chinese prickly ash pathology fungi and significantly suppress killing action, specifically see the following form 2.
Table 2
Note: antibacterial broadband refers to the difference of antibacterial radius and colony radius to be measured ,-: indicate without antibacterial band; +: represent antibacterial broadband <5mm; ++ represent antibacterial broadband >5mm.
Embodiment 3
1) produce the High Density Cultivation of the bacillus subtilis (CICC 10339) of beta dextranase, the density of setting out of bacillus subtilis (CICC 10339) is 1.57 × 10
6individual/mL, is seeded to liquid nutrient medium (glucose 14g/L, beef extract 25g/L, potassium dihydrogen phosphate 2.45g/L, groundnut meal 10g/L, pH=7.0,121 DEG C of sterilizing 20min) in, inoculum concentration is 3%, 37 DEG C of shaken cultivation 24h, and the liquid-tight degree of bacterium is 2.80 × 10
9cfu/mL, obtains high density bacteria suspension.
2) the common 100kg of healthy for Chinese prickly ash branch is cut into trifle and carries out Natural compost, spray 1 when compost internal temperature reaches 32 DEG C) in the high density bacteria suspension 3L for preparing, continue compost, when compost internal temperature is more than 43 DEG C, compost terminates, stir spreading for cooling, after fermentation under nature, becomes the bio-fertilizer being rich in the bacillus subtilis producing beta dextranase.
3) step 2) in the bio-fertilizer for preparing to Chinese prickly ash, there is obvious growth-promoting prebiotic effect, improve individual plant fresh pod yield 6.14% (p<0.05), specifically see the following form 3 and table 4.Chinese prickly ash plant is the wild type picking up from area, Qaidam.
Table 3
Table 4
Claims (9)
1. a method for the disease-resistant growth-promoting of Chinese prickly ash biological control, is characterized in that, comprise the following steps:
1) bacillus subtilis CICC 10339 is cultivated, make density 10
9~ 10
10the high density bacteria suspension of cfu/mL;
2) healthy for Chinese prickly ash branch is cut into segment and carries out compost, spray step 1 in composting process) in high density bacteria suspension, compost ferments after terminating, after fermentation, obtain the bio-fertilizer that can effectively suppress to kill Chinese prickly ash pathology fungi, this bio-fertilizer is used in the disease-resistant growth-promoting of Chinese prickly ash biological control.
2. the method for the disease-resistant growth-promoting of Chinese prickly ash biological control according to claim 1, is characterized in that, step 1) in, bacillus subtilis CICC 10339 is cultivated, comprising: the density of setting out of bacillus subtilis CICC 10339 is 10
6~ 10
7individual/mL, being seeded to inoculum concentration in liquid nutrient medium is 1% ~ 5%, 33 DEG C ~ 39 DEG C shaken cultivation 20 ~ 28h, is 10 to the liquid-tight degree of bacterium
9~ 10
10the high density bacteria suspension of cfu/mL.
3. the method for the disease-resistant growth-promoting of Chinese prickly ash biological control according to claim 2, is characterized in that, step 1) in, bacillus subtilis CICC 10339 is cultivated, comprising: the density of setting out of bacillus subtilis CICC 10339 is 10
6~ 10
7individual/mL, being seeded to inoculum concentration in liquid nutrient medium is 2% ~ 3%, 36 DEG C ~ 37 DEG C shaken cultivation 22 ~ 26h, is 10 to the liquid-tight degree of bacterium
9~ 10
10the high density bacteria suspension of cfu/mL.
4. the method for the disease-resistant growth-promoting of Chinese prickly ash biological control according to Claims 2 or 3, is characterized in that, described liquid nutrient medium, in 1L, comprises following component:
5. the method for the disease-resistant growth-promoting of Chinese prickly ash biological control according to claim 4, is characterized in that, described liquid nutrient medium, in 1L, comprises following component:
6. the method for the disease-resistant growth-promoting of Chinese prickly ash biological control according to claim 1, it is characterized in that, step 2) in, spray step 1 in composting process) in high density bacteria suspension, comprise: when compost internal temperature reaches 30 DEG C ~ 34 DEG C, spray step 1) in high density bacteria suspension, continue compost, when compost internal temperature reaches 41 DEG C ~ 45 DEG C, compost terminates.
7. the method for the disease-resistant growth-promoting of Chinese prickly ash biological control according to claim 6, it is characterized in that, step 2) in, spray step 1 in composting process) in high density bacteria suspension, comprise: when compost internal temperature reaches 32 DEG C, spray step 1) in high density bacteria suspension, continue compost, when compost internal temperature reaches 43 DEG C, compost terminates.
8. the method for the disease-resistant growth-promoting of Chinese prickly ash biological control according to claim 1, is characterized in that, step 2) in, after compost terminates, compost is stirred spreading for cooling.
9. the method for the disease-resistant growth-promoting of Chinese prickly ash biological control according to claim 1, is characterized in that, step 2) in, described fermentation adopts nature bottom fermentation.
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|---|---|---|---|
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105165537A (en) * | 2015-09-23 | 2015-12-23 | 柳小明 | Method for preventing disease of zanthoxylum bungeanum maxim |
| CN106489629A (en) * | 2016-11-15 | 2017-03-15 | 筠连县强生农副产品综合开发有限公司 | A kind of pest and disease damage processing method in rattan green pepper growth course |
-
2015
- 2015-02-04 CN CN201510058447.9A patent/CN104642018B/en not_active Expired - Fee Related
Non-Patent Citations (2)
| Title |
|---|
| 李建嫄等: "《花椒枯穂病生防菌的筛选及鉴定》", 《华北农学报》 * |
| 沈平等: "《枯草芽孢杆菌可湿性粉剂对花椒黑胫病田间防效评价》", 《林业实用技术》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105165537A (en) * | 2015-09-23 | 2015-12-23 | 柳小明 | Method for preventing disease of zanthoxylum bungeanum maxim |
| CN106489629A (en) * | 2016-11-15 | 2017-03-15 | 筠连县强生农副产品综合开发有限公司 | A kind of pest and disease damage processing method in rattan green pepper growth course |
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