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Method for separating endogenetic fungi producing spherosin

A technology of endophytic fungi and swainsonin, applied in the direction of microorganism-based methods, biochemical equipment and methods, fungi, etc., can solve the problems of loss, strain variation, low yield, etc., and achieve a simple and easy separation method, The effect of resolving the source is very difficult

Inactive Publication Date: 2009-08-12
杨凌天力生物技术有限公司
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Problems solved by technology

[0009] The invention provides a method for isolating endophytic fungi producing swainsonine to solve the problems of low yield and loss caused by strain variation in the prior art

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  • Method for separating endogenetic fungi producing spherosin

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Embodiment Construction

[0020] The present invention will be described in detail below in conjunction with examples.

[0021] A method for isolating endophytic fungi producing swainsonine, comprising the steps of:

[0022]1. Separation: wash away the dust on the surface of Oxytropis gansu (collected from the Qinghai-Tibet Plateau) with deionized water, separate the leaves, stems and flowers of the plant with sterilized tweezers, then wrap the leaves, stems and flowers with gauze respectively, Surface disinfection, the surface disinfection procedure is: first soak in 70% ethanol for 30s, then soak in sodium hypochlorite solution with an available chlorine content of 1% for 3min, and finally rinse twice with sterilized deionized water, each time for 1min, put The deionized water is inoculated on the surface of newly prepared PDA or Czapek agar medium, and observed for 30 days to see whether there is fungal growth on the surface of the medium, so as to test the effect of surface disinfection;

[0023] ...

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Abstract

The invention belongs to the technical field of bio-pharmaceuticals, and in particular relates to a method for separating swainsonine-generating endophytic fungi. For solving the problems of low yield and loss caused by strain variation of the prior art, the invention adopts a technical proposal that the method for separating swainsonine-generating endophytic fungi comprises the following steps: 1) separation, which includes removing dust with deionized water, separating leaf, stem and flow, sterilizing surface, absorbing water, cutting, inoculating tissue blocks on the surface of a culture medium respectively, culturing the inoculated tissue blocks in a culture box, selecting edge hyphae when the hyphae grow on the periphery of the surface of the culture medium, inoculating the edge hyphae on the surface of an agar medium, and placing the inoculated edge hyphae in a culture medium for culture, inoculating a bacterial colony growing on a PDA culture medium on the surface of the cultural medium again when the bacterial colony growing on a PDA culture medium is not purification, and repeating the previous operation process till a purified bacterial strain is obtained; and 2) screening by classic thin-layer chromatography. The method has the advantages of high extraction efficiency, simplicity, and easy implementation.

Description

Technical field: [0001] The invention belongs to the technical field of biopharmaceuticals, and in particular relates to a method for isolating endophytic fungi producing swainsonine. Background technique: [0002] Swainsonine is an indolizidine alkaloid, and its anti-tumor activity, immune enhancement effect and anti-radiation effect are very obvious. However, the main problem restricting the development of domestic swainsonine anti-tumor drugs is the The source of swainsonine is very difficult, far from meeting the needs of people's research. [0003] At present, there are three sources of swainsonine. The first one is extracted from plants of the genus Astragalus, Oxytropis, and Swainou in the family Fabaceae, as well as plants in the genus Ipomoea genus of the family Convolvulaceae and plants of the genus Astragalus in the Malvaceae family. Because the amount of swainsonine contained in the plant itself is very low, the extraction rate is low, the output is limited, and...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12Q1/04C12R1/645
Inventor 赵宝玉来航线马尧贺武利刘彬慧
Owner 杨凌天力生物技术有限公司
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