CN104630107A - Method for biologically preventing and controlling stem rot of nitraria tangutorum bobr and promoting growth of nitraria tangutorum bobr - Google Patents
Method for biologically preventing and controlling stem rot of nitraria tangutorum bobr and promoting growth of nitraria tangutorum bobr Download PDFInfo
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- CN104630107A CN104630107A CN201510058590.8A CN201510058590A CN104630107A CN 104630107 A CN104630107 A CN 104630107A CN 201510058590 A CN201510058590 A CN 201510058590A CN 104630107 A CN104630107 A CN 104630107A
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- 239000008103 glucose Substances 0.000 description 4
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Classifications
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
-
- 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
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- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Pest Control & Pesticides (AREA)
- Biochemistry (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Fertilizers (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a method for biologically preventing and controlling the stem rot of nitraria tangutorum bobr and promoting the growth of the nitraria tangutorum bobr. The method comprises the steps of preparing a 109-1010cfu/mL high-density bacterial suspension from bacillus subtilis; shearing healthy branches of the nitraria tangutorum bobr into small segments to compost, spraying the high-density bacterial suspension of the bacillus subtilis in the composting process, and fermenting to obtain a biological fertilizer capable of effectively inhibiting and killing pathological fungi of the nitraria tangutorum bobr; and applying the biological fertilizer to biological prevention and control of the stem rot of the nitraria tangutorum bobr and growth promotion of the nitraria tangutorum bobr. According to the method, the bacillus subtilis capable of generating beta-glucanase is a plant probiotic capable of taking disease resisting and growth promoting effects, and if the bacillus subtilis is added into compost, the effects for biologically preventing and controlling pathological fungi of the nitraria tangutorum bobr and promoting the growth of the nitraria tangutorum bobr are remarkably improved. The method is simple and convenient in composting process treatment, free of wastewater discharge, low in investment and energy consumption, easy to operate, capable of easily realizing large-scale industrial production, favorable in economic benefit and wide in application prospect.
Description
Technical field
The present invention relates to the preparation field of biological control fertilizer, be specifically related to a kind of method that biological control resists white thorn stem rot and growth-promoting.
Background technology
White thorn is the shrub of zygophyllaceae Nitraria; branch is many and intensive; in shape of growing thickly; there is the husky ability of very strong sand resistance; be distributed on the lake rims in area such as China Qaidam, Tarim Basin, Alxa, Hexi Corridor and the clay of stream terrace and sand; the characteristic such as there is drought resisting, weather-proof sand buries, Salt And Alkali Tolerance, sprouting ability are strong; be arid-desert area important build mass-planting thing; the widest in the distribution of lower reaches of river arid-desert area, area is maximum, in Revegetation, check winds and fix drifting sand, safeguard the important role such as desert ecosystem balance, protection oasis.Bai thorn is the fruit that zygophyllaceae Nitraria plant stings Nitraria tangutorum Bor. in vain, has strengthening the spleen and stomach, strengthening by means of tonics, regulating menstruation and activating blood, effect of lactagogue.
White thorn stem rot endangers comparatively serious one in white thorn disease, and plant mobile jib of being injured is dry rot shape, causes the whole strain of white thorn withered, cause Bai Ci group serious degradation, and the final thorn sand dune in vain that formed activates.Fusarium oxysporum Fusarium oxysporum F.Sp. is one of white thorn stem rot pathogenic bacteria.
Plant pathology fungi is mainly present in soil, infection plant in plant-growth, causes Plant diseases, causes production loss.Therefore, fungal diseases of plants to be eliminated from the angle of biological control, except killing the pathology fungi on plant, killing or suppressing the pathology fungi in soil also 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 sporeformer resistance, the resistivity for dry high temperature is poor, so stability is not high when making liquid or sorbent material.The new trend of bio-fertilizer development should be greatly develop resistance high, can the genus bacillus of existence steady in a long-term.Particularly genus bacillus can produce multiple antifungal substance, has remarkable restraining effect 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 genus bacillus form, aerobic or amphimicrobian, Gram-positive, can produce resistance statospore under certain environmental conditions.Genus 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 genus bacillus, produce the antimicrobial substances 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 dextran.
Summary of the invention
The invention provides a kind of method that biological control resists white thorn stem rot and growth-promoting, by being incorporated in white thorn natural flora by the subtilis (CICC 10339) producing beta dextranase, make the bio-fertilizer that effectively can suppress to kill by biological control approach white thorn pathology fungi.
Biological control resists a method for white thorn stem rot and growth-promoting, comprises the following steps:
1) subtilis (CICC 10339) is cultivated, make density 10
9~ 10
10the high-density bacteria suspension of cfu/mL;
2) be cut into segment carry out compost by stinging healthy branch in vain, 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 white thorn pathology fungi, this bio-fertilizer is used in the anti-white thorn stem rot of biological control and growth-promoting.
In the present invention, being cut into trifle compost by stinging healthy branch in vain, making the flora in prepared bio-fertilizer be healthy white thorn plant naturally flora, this flora with sting plant coevolution in vain and grow, define mutual beneficial syntaxial system, have certain biological control effect to disease fungi.Further, the subtilis (CICC 10339) that can produce beta dextranase adds in compost, significantly strengthens the biological control effect of dialogue thorn disease fungi.Subtilis (CICC 10339) is owing to producing beta dextranase, and dialogue thorn plant fiber has good Decomposition, by composting fermentation process is necessary, therefore fully can be dissolved in compost flora and breeds stably and lastingly.As the prebiotic effect of milk-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, subtilis (CICC 10339) is namely white thorn probiotic bacterium, and dialogue thorn pathology fungi has remarkable biological control effect.
Step 1) in, 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.Subtilis (CICC 10339) can be bought to Chinese industrial Microbiological Culture Collection administrative center.
Subtilis (CICC 10339) is cultivated, comprising: the density of setting out of subtilis (CICC 10339) is 10
6~ 10
7individual/mL, being seeded to inoculum size in liquid nutrient medium is 1% ~ 5%, 33 DEG C ~ 39 DEG C shaking culture 20 ~ 28h, is 10 to the liquid-tight degree of bacterium
9~ 10
10the high-density bacteria suspension of cfu/mL.
Further preferably, subtilis (CICC 10339) is cultivated, comprising: the density of setting out of subtilis (CICC 10339) is 10
6~ 10
7individual/mL, being seeded to inoculum size in liquid nutrient medium is 2% ~ 3%, 36 DEG C ~ 37 DEG C shaking culture 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:
Further preferably, described liquid nutrient medium, in 1L, comprises following component:
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 state of nature bottom fermentation.
Compared with prior art, tool of the present invention has the following advantages:
In the present invention, being cut into trifle compost by stinging healthy branch in vain, making the flora in prepared bio-fertilizer be healthy white thorn plant naturally flora, this flora with sting plant coevolution in vain and grow, define mutual beneficial syntaxial system, have certain biological control effect to disease fungi.
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 milk-acid bacteria for animal, the microorganism killing plant pathology fungi is suppressed to be referred to as plants probiotics.Namely the subtilis (CICC 10339) of being rich in bio-fertilizer prepared by the present invention is white thorn probiotic bacterium, and dialogue thorn pathology fungi has remarkable biological control effect.
Composting process carries out in its natural state, the stable white thorn plant natural flora of Tiny ecosystem is relied on to ferment, and subtilis (CICC 10339) is owing to producing beta dextranase, dialogue thorn plant fiber has good Decomposition, by composting fermentation process is necessary, therefore, subtilis (CICC 10339) fully can be dissolved in compost flora and breed stably and lastingly.Composting process process is easy, non-wastewater discharge, and less investment, energy consumption are low, easy to operate, is easy to industrialization scale operation, has good economic benefit and wide application prospect.
Embodiment
Embodiment 1
1) produce the high-density culture of the subtilis (CICC 10339) of beta dextranase, the density of setting out of subtilis (CICC 10339) is 1.18 × 10
6individual/mL, is seeded to liquid nutrient medium (glucose 14g/L, extractum carnis 25g/L, potassium primary phosphate 2.45g/L, groundnut meal 10g/L, pH=7.0,121 DEG C of sterilizing 20min) in, inoculum size is 2%, 37 DEG C of shaking culture 24h, and the liquid-tight degree of bacterium is 2.65 × 10
9cfu/mL, obtains high-density bacteria suspension.
2) by sting in vain healthy branch altogether 100kg be cut into trifle and carry 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, secondary fermentation under state of nature, becomes the bio-fertilizer being rich in the subtilis producing beta dextranase, effectively can suppress the bio-fertilizer killing white thorn 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, dialogue thorn pathology fungi has and significantly suppresses killing action, specifically sees the following form 1.
White thorn stem rot pathogenic bacteria Fusarium oxysporum Fusarium oxysporum F.Sp. is purchased from Chinese industrial Microbiological Culture Collection administrative center (CICC), and bacterium numbering is 2532.
Beta dextranase vitality test: adopt reducing sugar test method, Beta dextran 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 color reaction, the colour developing depth is directly proportional to reducing sugar growing amount, and reducing sugar growing amount is directly proportional to reaction solution Beta glucanase vigor, thus utilize the light absorption value of colorimetric estimation reaction solution to calculate reducing sugar growing amount, obtain Beta dextranase vigor.Beta glucanase unit of activity defines: under condition determination, and per minute hydrolysis Beta dextran 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 method, 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; Pathogenic fungi is made into 10
8bacteria suspension, is sprayed on rapidly with aseptic throat 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 substratum into flat board, becomes triangle to place Oxford cup, adds fermented liquid 100 μ L in the cup of Oxford on substratum after solidifying, 28 DEG C of incubators are cultivated, after pathogenic fungi 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 culture of the subtilis (CICC 10339) of beta dextranase, the density of setting out of subtilis (CICC 10339) is 1.70 × 10
6individual/mL, is seeded to liquid nutrient medium (glucose 14g/L, extractum carnis 25g/L, potassium primary phosphate 2.45g/L, groundnut meal 10g/L, pH=7.0,121 DEG C of sterilizing 20min) in, inoculum size is 3%, 37 DEG C of shaking culture 24h, and the liquid-tight degree of bacterium is 3.03 × 10
9cfu/mL, obtains high-density bacteria suspension.
2) by sting in vain healthy branch altogether 100kg be cut into trifle and carry 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, secondary fermentation under state of nature, becomes the bio-fertilizer being rich in the 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, dialogue thorn pathology fungi has and significantly suppresses killing action, specifically sees 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 culture of the subtilis (CICC 10339) of beta dextranase, the density of setting out of subtilis (CICC 10339) is 1.68 × 10
6individual/mL, is seeded to liquid nutrient medium (glucose 14g/L, extractum carnis 25g/L, potassium primary phosphate 2.45g/L, groundnut meal 10g/L, pH=7.0,121 DEG C of sterilizing 20min) in, inoculum size is 3%, 37 DEG C of shaking culture 24h, and the liquid-tight degree of bacterium is 2.79 × 10
9cfu/mL, obtains high-density bacteria suspension.
2) by sting in vain healthy branch altogether 100kg be cut into trifle and carry 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, secondary fermentation under state of nature, becomes the bio-fertilizer being rich in the subtilis producing beta dextranase.
3) step 2) in prepare bio-fertilizer dialogue thorn there is obvious growth-promoting prebiotic effect, improve individual plant fruit yield 13.0% (p<0.05), specifically see the following form 3 and table 4.White thorn plant is the wild-type picking up from area, Qaidam.
Table 3
Table 4
Claims (9)
1. biological control resists a method for white thorn stem rot and growth-promoting, it is characterized in that, comprises the following steps:
1) subtilis CICC 10339 is cultivated, make density 10
9~ 10
10the high-density bacteria suspension of cfu/mL;
2) be cut into segment carry out compost by stinging healthy branch in vain, 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 white thorn pathology fungi, this bio-fertilizer is used in the anti-white thorn stem rot of biological control and growth-promoting.
2. biological control according to claim 1 resists the method for white thorn stem rot and growth-promoting, it is characterized in that, step 1) in, subtilis CICC 10339 is cultivated, comprising: the density of setting out of subtilis CICC 10339 is 10
6~ 10
7individual/mL, being seeded to inoculum size in liquid nutrient medium is 1% ~ 5%, 33 DEG C ~ 39 DEG C shaking culture 20 ~ 28h, is 10 to the liquid-tight degree of bacterium
9~ 10
10the high-density bacteria suspension of cfu/mL.
3. biological control according to claim 2 resists the method for white thorn stem rot and growth-promoting, it is characterized in that, step 1) in, subtilis CICC 10339 is cultivated, comprising: the density of setting out of subtilis CICC 10339 is 10
6~ 10
7individual/mL, being seeded to inoculum size in liquid nutrient medium is 2% ~ 3%, 36 DEG C ~ 37 DEG C shaking culture 22 ~ 26h, is 10 to the liquid-tight degree of bacterium
9~ 10
10the high-density bacteria suspension of cfu/mL.
4. the biological control according to Claims 2 or 3 resists the method for white thorn stem rot and growth-promoting, it is characterized in that, described liquid nutrient medium, in 1L, comprises following component:
5. biological control according to claim 4 resists the method for white thorn stem rot and growth-promoting, it is characterized in that, described liquid nutrient medium, in 1L, comprises following component:
6. biological control according to claim 1 resists the method for white thorn stem rot and growth-promoting, 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. biological control according to claim 6 resists the method for white thorn stem rot and growth-promoting, 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. biological control according to claim 1 resists the method for white thorn stem rot and growth-promoting, it is characterized in that, step 2) in, after compost terminates, compost is stirred spreading for cooling.
9. biological control according to claim 1 resists the method for white thorn stem rot and growth-promoting, it is characterized in that, step 2) in, described fermentation adopts state of nature bottom fermentation.
Priority Applications (1)
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| CN102965320A (en) * | 2012-12-04 | 2013-03-13 | 赵斌 | Bacillus subtilis for preventing and controlling plant fungal disease and application of bacillus subtilis |
| CN104016745A (en) * | 2014-06-12 | 2014-09-03 | 河海大学 | Microorganism organic fertilizer with functions of preventing and eliminating successive cropping obstacle of artemisia selengensis and preparation method thereof |
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| CN102965320A (en) * | 2012-12-04 | 2013-03-13 | 赵斌 | Bacillus subtilis for preventing and controlling plant fungal disease and application of bacillus subtilis |
| CN104016745A (en) * | 2014-06-12 | 2014-09-03 | 河海大学 | Microorganism organic fertilizer with functions of preventing and eliminating successive cropping obstacle of artemisia selengensis and preparation method thereof |
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| Title |
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| 林福呈: "枯草芽抱杆菌(Bacillus subtilis)S9对植物病原真菌的溶菌作用", 《植物病理学报》 * |
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