Method for directional separation and authentication of swainsonine fungal endophyte Undifilum oxytropis produced through oxytropis kansuensis

A swainsonine, endophytic fungi technology, applied in the directions of microorganism-based methods, microbial determination/inspection, biochemical equipment and methods, etc., can solve the problems of low separation rate, loss of other endophytic fungi, etc. The effect of reducing interference, simple and easy separation method, and expanding scale production

Inactive Publication Date: 2014-09-10
NORTHWEST A & F UNIV
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Problems solved by technology

[0007] The purpose of the present invention is to provide a kind of endophytic fungus producing swainsonine from Oxytropis chinensis Undifilum oxytropis The directio

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  • Method for directional separation and authentication of swainsonine fungal endophyte Undifilum oxytropis produced through oxytropis kansuensis
  • Method for directional separation and authentication of swainsonine fungal endophyte Undifilum oxytropis produced through oxytropis kansuensis
  • Method for directional separation and authentication of swainsonine fungal endophyte Undifilum oxytropis produced through oxytropis kansuensis

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Embodiment Construction

[0035] The present invention will be described in detail below in combination with specific embodiments.

[0036] Gansu Oxytropis producing swainsonine endophytic fungus related to the present invention Undifilum oxytropis The directional separation and identification method is realized by the following steps:

[0037] Step 1: Separation:

[0038] The stems and mature seeds of Oxytropis gansu were rinsed with distilled water, rinsed with 75% ethanol solution for 30 s, sterile water for 3 times, and sodium hypochlorite solution with 2% available chlorine for 3 times. min. Rinse with sterile water for 3-5 times to complete surface cleaning and disinfection;

[0039] Then cut the tissue into small pieces of 5mm×5mm and place them on PDA plates filled with CO 2 And sealed with plastic wrap, cultivated at room temperature under light conditions, after the mycelia grow out of the incision, pick the top mycelium and transfer it to PDA medium for purification 2-3 times, scrape the ...

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Abstract

The invention relates to a method for directional separation and authentication of swainsonine fungal endophyte Undifilum oxytropis produced through oxytropis kansuensis. When the fungal endophyte Undifilum oxytropis is separated from oxytropis kansuensis, bacterial strains of other species can be inevitably separated, and therefore separation efficiency is affected. Stems and mature seeds of oxytropis kansuensis are sequentially cleaned through distilled water, ethanol, sterile water and sodium hypochlorite, tissue is sheared into small blocks and placed on a PDA flat plate, the flat plate is filled with CO2 and sealed by a fresh-keeping film, the tissue is cultured at indoor temperature under the illumination condition, after mycelia grow on incisions, the mycelia at the top ends are picked and transferred to a culture medium for purification, the mycelia are scraped, and then the bacterial strains are inoculated to a test tube slant for refrigeration; genome DNA of the separated bacterial strains is extracted, and sequence alignment is conducted in a Gene Bank. Only one fungus is separated from locoweed, interference of fungal endophyte of other species is reduced, the method is easy and convenient to implement and feasible, and in the storage process, the bacterial strains are not prone to mutation or activity of the bacterial strains is not reduced.

Description

technical field [0001] The invention belongs to the technical field of biopharmaceuticals, and in particular relates to an endophytic fungus producing swainsonine from Oxytropis chinensis in Gansu Undifilum oxytropis directional separation and identification methods. Background technique [0002] Endophytic fungi (fungal endophyte) refer to fungi that live in living plant tissues for a certain period of their life history or do not cause obvious disease symptoms to plant tissues, including saprophytic fungi, latent pathogenic fungi and mycorrhizal fungi. The current study found that locotoxin is closely related to locoweed endophytic fungi. On the one hand, endophytic fungi assist locoweeds to produce toxins, thereby enhancing the toxicity of locoweeds and reducing animals’ consumption of locoweeds; on the other hand, endophytic fungi can improve the stress resistance of locoweeds, which makes locoweeds Grass is extremely tenacious in the arid and semi-arid grasslands, wh...

Claims

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Application Information

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IPC IPC(8): C12N1/14C12Q1/68C12Q1/04C12R1/645
Inventor 路浩杨晓雯赵宝玉曹丹丹薛瑞旭权海云
Owner NORTHWEST A & F UNIV
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