CN101897345B - Preparation method of preparation containing hypoxylon bacteriostatic active ingredients for preventing and treating sphaeropsis sapinea - Google Patents
Preparation method of preparation containing hypoxylon bacteriostatic active ingredients for preventing and treating sphaeropsis sapinea Download PDFInfo
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- CN101897345B CN101897345B CN2009100721521A CN200910072152A CN101897345B CN 101897345 B CN101897345 B CN 101897345B CN 2009100721521 A CN2009100721521 A CN 2009100721521A CN 200910072152 A CN200910072152 A CN 200910072152A CN 101897345 B CN101897345 B CN 101897345B
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Abstract
The invention relates to a preparation method of a preparation containing hypoxylon bacteriostatic active ingredients for preventing and treating sphaeropsis sapinea, which comprises the following steps of preparing hypoxylon mycelium fermentation liquid, extracting the bacteriostatic active ingredients from the hypoxylon mycelium fermentation liquid, and preparing the preparation containing the hypoxylon bacteriostatic active ingredients. In the preparation method, reproducible and cheap fungal resources with rich source are taken as raw materials which have obvious inhibition effect on sphaeropsis sapinea; and the preparation method has the characteristics of being efficient, slightly poisonous, nuisanceless and the like, thus having wide application prospect in preventing and treating sphaeropsis sapinea.
Description
Technical field
The present invention relates to a kind of preparation method who prevents and treats the sphaeropsis sapinea biologic product.
Background technology
Sphaeropsis sapinea is a kind of global contagious disease general, that harmfulness is large of falling ill, and more than 20 country successively reported sphaeropsis sapinea generation and harm thereof in the world.In recent years, on the annual loose seedling in the part nursery in Liaoning, the Inner Mongol, Jilin, find die-back, this disease is rapid at the seedling vertical spread, causes that tip buds die, rhizome rot, dead leaf, become current pine grow seedlings produce in urgent problem.Diplodia Shoot-bligt of Pinus Sylvestris Var Mongolica generally occurs in Heilungkiang, Jilin, each province, Liaoning, causes that Pinus Silvestris Var. Mongolica Plantation is withered in flakes, when serious, can cause the death of large tracts of land pine forest.At present, Diplodia Shoot-bligt of Pinus Sylvestris Var Mongolica is paid attention to by people day by day, has classified the important forest disease of China as and has been studied.The control of die-back at present still depends on chemical control largely, although this has brought very high benefit to production of forestry, and, many drawbacks such as destruction of environmental pollution, ecosystem natural equilibrium are also following.Therefore, find efficient biocontrol strains very urgent.
In Answorth 1973 classification of fungi systems, Hypoxylon (Hypoxylon perforatum (Schwein.) Fr.) is subordinate to Ascomycotina (Ascomycotina), gang pyrenomycetes (Pyrenomycetes), Spheeriales (Sphaeriales), Xylariaceae (Xylariaceae), Hypoxylon genus (Hypoxylon).At present, the research that relevant Hypoxylon belongs to is mainly concentrated both ways, and the one, the disease caused by Hypoxylon, the 2nd, the research of several marine fungi metabolites in this is belonged to.Do not find bibliographical information related to the present invention through retrieval.
Summary of the invention
The object of the present invention is to provide a kind of preparation method of preparation containing hypoxylon bacteriostatic active ingredients for preventing and treating sphaeropsis sapinea.
In order to achieve the above object, the technical solution used in the present invention is: the extraction of bacteriostatic active ingredients in the preparation of Hypoxylon mycelium fermentation broth-Hypoxylon mycelium fermentation broth-Hypoxylon bacteriostatic active ingredients preparation preparation.
Advantage of the present invention is:
The raw material that the present invention adopts is reproducible fungal resources, and source is abundant, inexpensive, and Sphaeropsis sapinea is had to significant inhibition, and the present invention has efficient, micro-poison, the characteristics such as nuisanceless, be with a wide range of applications to the control sphaeropsis sapinea.
Embodiment
Below embodiments of the invention are described in further detail.
Embodiment 1,
The preparation of Hypoxylon mycelium fermentation broth: adopt improvement potato glucose (PD) liquid nutrient medium, the formula of medium is: potato 150g, glucose 17g, KH
2pO
42.5g, MgSO
41g, water 1000ml, pH 5, and the Hypoxylon mycelium is carried out to shaker fermentation and cultivate 10 days, inoculum concentration 0.5%, liquid amount 40%, fermentation culture conditions is: 25 ℃ of temperature, shaking table revolution 140r/min,
In the Hypoxylon mycelium fermentation broth, bacteriostatic active ingredients extracts: after the Hypoxylon mycelium fermentation broth that will cultivate 10 days filters with sterile gauze, with ethyl acetate by weight volume ratio mix at 1: 3, under normal temperature, static placement is 5 days, abandon lower floor's filtrate after extraction, the upper strata organic solvent is obtained to bacteriostatic active ingredients with 30 ℃ of rotary evaporation in vacuo except desolventizing
The preparation of Hypoxylon bacteriostatic active ingredients preparation: bacteriostatic active ingredients is dissolved with 10% Tween 80 solution, revert to 1/10 of original fermentation liquor filtered fluid volume.
Embodiment 2,
The preparation of Hypoxylon mycelium fermentation broth: adopt improvement potato glucose (PD) liquid nutrient medium, the formula of medium is: potato 200g, glucose 19g, KH
2pO
43.0g, MgSO
41.5g, water 1000ml, pH 5.5, the Hypoxylon mycelium is carried out to shaker fermentation and cultivates 12 days, inoculum concentration 1.0%, liquid amount 45%, fermentation culture conditions is: 28 ℃ of temperature, shaking table revolution 150r/min,
In the Hypoxylon mycelium fermentation broth, bacteriostatic active ingredients extracts: after the Hypoxylon mycelium fermentation broth that will cultivate 12 days filters with sterile gauze, with ethyl acetate by weight volume ratio mix at 1: 3, under normal temperature, static placement is 8 days, abandon lower floor's filtrate after extraction, the upper strata organic solvent is obtained to bacteriostatic active ingredients with 30 ℃ of rotary evaporation in vacuo except desolventizing
The preparation of Hypoxylon bacteriostatic active ingredients preparation: bacteriostatic active ingredients is dissolved with 10% Tween 80 solution, revert to 1/10 of original fermentation liquor filtered fluid volume.
Embodiment 3,
The preparation of Hypoxylon mycelium fermentation broth: adopt improvement potato glucose (PD) liquid nutrient medium, the formula of medium is: potato 250g, glucose 22g, KH
2pO
43.5g, MgSO
42g, water 1000ml, pH 6, and the Hypoxylon mycelium is carried out to shaker fermentation and cultivate 15 days, inoculum concentration 1.5%, liquid amount 50%, fermentation culture conditions is: 30 ℃ of temperature, shaking table revolution 160r/min,
In the Hypoxylon mycelium fermentation broth, bacteriostatic active ingredients extracts: after the Hypoxylon mycelium fermentation broth that will cultivate 15 days filters with sterile gauze, with ethyl acetate by weight volume ratio mix at 1: 3, under normal temperature, static placement is 10 days, abandon lower floor's filtrate after extraction, the upper strata organic solvent is obtained to bacteriostatic active ingredients with 30 ℃ of rotary evaporation in vacuo except desolventizing
The preparation of Hypoxylon bacteriostatic active ingredients preparation: bacteriostatic active ingredients is dissolved with 10% Tween 80 solution, revert to 1/10 of original fermentation liquor filtered fluid volume.
Hypoxylon bacteriostatic active ingredients preparation is 73.20% to Sphaeropsis sapinea (Sphaeropsis sapinea) mycelial growth inhibition rate, to inhibition of germination, is 91.63%, to the medial lethal dose LD of mouse
50>13200mg/kg, be micro-poison, and the ultraviolet ray under natural environment, temperature are had to very high stability, and have certain anti-oxidant reducing power and good warm tolerance, and long-term storage does not affect bacteriostatic activity.
Claims (1)
1. the preparation method of a preparation containing hypoxylon bacteriostatic active ingredients for preventing and treating sphaeropsis sapinea is characterized in that: the preparation of the extraction of bacteriostatic active ingredients in the preparation of Hypoxylon mycelium fermentation broth-Hypoxylon mycelium fermentation broth-Hypoxylon bacteriostatic active ingredients preparation:
(1) preparation of Hypoxylon mycelium fermentation broth: adopt improvement potato glucose (PD) liquid nutrient medium, the formula of medium is: potato 200g, glucose 20g, KH
2pO
43g, MgSO
41.5g, water 1000ml, pH5-6, carry out shaker fermentation to the Hypoxylon mycelium and cultivate 10 days～15 days, inoculum concentration 0.5%～1.5%, liquid amount 40%～50%, fermentation culture conditions is: 25 ℃～30 ℃ of temperature, shaking table revolution 140r/min～160r/min,
(2) extraction of bacteriostatic active ingredients in the Hypoxylon mycelium fermentation broth: after the Hypoxylon mycelium fermentation broth that will cultivate 10 days-15 days filters with sterile gauze, with ethyl acetate by weight volume ratio mix at 1: 3, under normal temperature, static placement is 5 days-10 days, abandon lower floor's filtrate after extraction, the upper strata organic solvent is obtained to bacteriostatic active ingredients with 30 ℃ of rotary evaporation in vacuo except desolventizing
(3) preparation of Hypoxylon bacteriostatic active ingredients preparation: bacteriostatic active ingredients is dissolved with 10% Tween 80 solution, revert to 1/10 of original fermentation liquor filtered fluid volume, bacteriostatic active ingredients content is 0.2214mg/ml.
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| CN2009100721521A CN101897345B (en) | 2009-06-01 | 2009-06-01 | Preparation method of preparation containing hypoxylon bacteriostatic active ingredients for preventing and treating sphaeropsis sapinea |
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| CN101897345B true CN101897345B (en) | 2013-12-11 |
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Families Citing this family (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102337222B (en) * | 2011-09-30 | 2012-11-07 | 中国科学院南海海洋研究所 | New species of Rhizophora stylosa root cellulose degrading fungus Hypoxylon sp. DPZ-SYz-36 and application thereof |
| CN106434363B (en) * | 2016-08-30 | 2019-08-23 | 中国林业科学研究院森林生态环境与保护研究所 | Produce ring grain Hypoxylon bacterial strain and its application of cineole |
| CN106754399B (en) * | 2016-11-18 | 2020-03-24 | 广西壮族自治区药用植物园 | Method for separating dark ring-shaped Xylaria |
| CN108991020A (en) * | 2018-08-07 | 2018-12-14 | 广西田东县东辰农业开发有限公司 | A kind of extracting method of Hypoxylon fermentation broth extract and its application in mango disease control |
| CN109536390B (en) * | 2018-12-21 | 2022-04-15 | 浙江省柑桔研究所 | Citrus endophytic fungi and application thereof |
| CN110172411B (en) * | 2019-05-29 | 2021-02-26 | 绍兴市曙光科技开发有限公司 | Xylaria cruzi strain ZJ1811 and culture method and application thereof |
| CN112825716B (en) * | 2019-11-22 | 2022-07-22 | 中国林业科学研究院森林生态环境与保护研究所 | Method for regulating and controlling fungal community structure for reducing harm of pine wilt disease pathogenic bacteria to host plants |
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