Separation and purification method of beauveria bassiana

Abstract

The invention relates to the technical field of identification, separation and purification application of beauveria bassiana strains, and discloses a separation and purification method of beauveria bassiana. By adopting the separation and purification method, a culture medium is scientifically optimized and improved; simple, quick and effective separation and purification of the beauveria bassiana are achieved by combining improvement of cultivation device coverings on the basis of the culture medium with a scientific formula. The method disclosed by the invention is simple and quick to operate, wide in samples applicable to separation and purification of the beauveria bassiana, low in operation cost and free of pollution on environment; the targets of separation and purification can be well achieved.

Description

technical field [0001] The invention relates to the application technical fields of identification of Beauveria bassiana species, biological control, prevention and control of Beauveria bassiana, etc., and more specifically relates to a separation and purification method of Beauveria bassiana. Background technique [0002] Beauveria bassiana (Beauveria bassiana) is now classified as Cordycipitaceae (Cordycipitaceae), an important fungus of the genus Cordyceps (Cordyceps), as early as 1816, the Italian scientist Bassi (1835) isolated it from the silkworm. [0003] Beauveria bassiana can invade more than 700 kinds of insects and mites belonging to 149 families and 521 genera in 15 orders (Li Zengzhi. Research and Application of Entomogenic Fungi in China (Vol. 1). Beijing: Academic Journal Press, 1988). Because Beauveria bassiana has the advantages of meeting some basic requirements of biological agents, that is, harmless to warm-blooded animals and plants, easy to cultivate, ...

AI Technical Summary

Problems solved by technology

Beauveria bassiana is a filamentous fungus, which can be separated and purified according to traditional fungal streaking, but this method is cumbersome to operate and requires multiple transfers to separate Beauveria bassiana

For improvement, researchers at home and abroad have developed a selective medium for Beauveria bassiana, such as Martin (Martin J P.Use of acid, rose bengal, and streptomycin in the plate method for estimating soil fungi. Soil Science, 1950) in Adding Bengal red (ose bengale) and ox bile to the medium can inhibit the miscellaneous bacteria to a certain extent; Boerema (Boerema G H etl. Some notable fungus infections II. Verslagen van de Plantenziektenkundige Dienst te Wageningen, 1963) made selection with cherry extract (Veen K H, Ferron P.A selective medium for the isolation of Beauveria tenella and of Metarrhizium anisopliae. Journal of invertebrate pathology, 1966) successfully utilized the improved Martin's medium Beauveria bassiana and Metarhizium anisopliae have been isolated accurately, but Beauveria bassiana grows slowly on this medium and is difficult to identify; Doberski et al. (Doberski J W, Tribe H T. Isolation of entomogenous fungi from elm bark and soil with reference to ecology of Beauveria bassiana and Metarhizium anisopliae.Transactions of the British Mycological Society, 1980), the use of crystal violet and cycloheximide can have better selection specificity for Beauveria bassiana; Beilharz et al. (Beilharz V C.etl.Dodine: a selective agent for certain soil fungi.Transactions of the British Mycological Society, 1982) designed a selective medium with dodine as a bacteriostatic agent, which has a good separation effect; Chase et al. (Chase A R .etl.Selective isolation of the entomopathogenic fungi Beauveria bassiana and Metarhizium anisopliae from an artificial potting medium.Florida Entomologist, 1986) optimized medium conditions, adding dodine, benomyl and crystal violet in the oat medium can remove Beauveria bassiana and Metarhizium anisopliae from other fungi Carry out effective separation; Shimazu M, Sato H.Media for selective isolation of an entomogenous fungus, Beauveria bassiana.Applied entomology and zoology, 1996) Utilize the characteristics that Beauveria bassiana can grow in a very wide pH range, made Nutrient-poor alkaline medium obtains better separation effect; domestic Wu Jinwen et al. Leaf extract—pine paste to inhibit the growth of bacteria, actinomycetes and other microorganisms to promote fungal growth, but the specificity is not enough

(Wang Bin, Fan Meizhen, Li Zengzhi. Screening of Beauveria bassiana selective medium. Anhui Agricultural University Journal, 2000) select 25% Sportak emulsifiable oil (containing 25% Prochloraz), 77% Cocide wettable powder (containing 77% copper hydroxide), 5% bacterial poison clear water agent (containing 5% amino acid type systemic fungicide) to replace doditidine and cycloheximide, but the experimental results are not satisfactory; Shi Hongxia etc. (Shi Hongxia, Hu Minglong, Yan Jun . Screening and detection application of Beauveria bassiana selective medium. Journal of Zhejiang Forestry University, 2002) For Beauveria bassiana parasitic in B. bassiana, red bengal, cycloheximide, and chloramphenicol were selected as antibacterial agents. According to the certain host specialization of Beauveria bassiana, Cai Guogui (Cai Guogui. Screening and production application of excellent strains of Beauveria bassiana. Forestry Science, 2003, Highly virulent Beauveria bassiana Screening and application of Bbd3 strains of B. bassiana. Journal of Nanjing Forestry University. 2006) adding dry powder of Phyllostachys basilicum or Caterpillar of cedarwood to the traditional culture medium can provide specific nutrition for Beauveria bassiana invading Phyllostachys basilicum or Caterpillar of Cedarwood; Xu Wenjing et al. (Xu Wenjing, Yin Weihuang, Zhang Zhengkun, etc. Screening of new strains of Beauveria bassiana highly effective in controlling corn borer. Jilin Agricultural Science, 2011) applied PDA medium added with peptone to purify the Beauveria bassiana that invaded corn borer; Wang Taoyuan et al. (Wang Taoyuan, Xu Wenjing, Zhang Zhengkun, et al. Screening and application of isolation medium for Beauveria bassiana. China Agricultural Science Bulletin, 2013) added dodine, cephalosporin, and kanamycin sulfate to PDA medium The element can effectively inhibit miscellaneous bacteria, and screen out a specific medium for detecting the colony content of Beauveria bassiana in the field

[0005] In summary, although a lot of research has been done on selective media in the prior art, only the expensive dodine or cycloheximide can be used as a bacteriostatic agent to separate better, but the actual application cost is too high ; and cycloheximide is carcinogenic and affects human health

The deficiencies in the existing technology limit the practical application of the above technology, and cannot solve the current general demand for the separation and purification of Beauveria bassiana

[0006] At present, there is a lack of a simple, safe, efficient, and low-cost method for isolating and purifying Beauveria bassiana to meet the needs of practical applications

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