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Extraction method for citrus brown blotch germ crude toxin and toxin production condition

A technology for citrus brown spot and crude toxin, which is applied in the biological field, can solve the problem that the crude toxin of citrus brown spot has not been extracted, and achieves the effects of easy large-scale preparation, large output and simple operation.

Inactive Publication Date: 2015-03-04
CITRUS RES INST OF CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the prior art, there is no relevant report on extracting the crude toxin of citrus brown spot bacteria

Method used

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  • Extraction method for citrus brown blotch germ crude toxin and toxin production condition
  • Extraction method for citrus brown blotch germ crude toxin and toxin production condition
  • Extraction method for citrus brown blotch germ crude toxin and toxin production condition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] The screening of embodiment 1 toxin-producing bacterial strains

[0046] 1.1. Strains and plants

[0047] Tested strains: CJBHJF-2, TJCHJ35B, CJBHJL-2, SP-36 and other 29 strains were isolated from leaves, branches and fruits of typical brown spot disease in Chongqing, Spain, Guangdong, Guangxi, Zhejiang and other places.

[0048] Bioassay materials: collected from 1-year-old tangerine leaves with a length of 3 to 4 cm in the net room.

[0049] Media formulations involved in the experiment:

[0050] PDA medium (PDA Medium): 200 g of potatoes, 20 g of glucose, 18 g of agar, and distilled water to make up to 1 L.

[0051] PSK medium (PSK Medium): potato 200g, sucrose 30g, K 2 HPO 4 1g, add distilled water to make up to 1L.

[0052] Richard’s Medium: 50g of sucrose, KNO 3 10g, KH 2 PO 4 5g, MgSO4 2.5g, FeCl 2 0.02g, add distilled water to 1L.

[0053] Improved Richard Medium (Improved-Richard Medium): Glucose 50g, KNO 3 10g, KH 2 PO 4 5g, MgSO4 2.5g, FeCl 2 0...

Embodiment 2

[0064] The optimization of embodiment 2 toxin production conditions

[0065] 2.1 Selection of extractant

[0066] After the high-toxin-producing strains are cultured according to the determined toxin-producing conditions, the culture filtrate is obtained. Take 15ml of the culture filtrate and add an equal volume of acetone, methanol, ethyl acetate, chloroform, carbon tetrachloride, and extract at 300r / min for 30min. The organic solvent was evaporated and removed by a rotary evaporator, and the obtained crude toxin was bioassayed by the needle-pricking method of detached leaves.

[0067] 2.2 Optimization of toxin-producing culture conditions

[0068] 2.2.1 The strains screened by the method in Example 1 were used for the optimization experiment of toxin-producing culture conditions.

[0069] 2.2.2 Time: According to the method 1.2 in Example 1, samples were taken after culturing in liquid medium for 5, 10, 15, 20, 25, and 30 days to prepare crude toxin.

[0070] 2.2.3 Tempe...

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Abstract

The invention relates to the field of biotechnology, particularly to an extraction method for citrus brown blotch germ crude toxin and a toxin production condition. According to the invention, the preservation number of citrus brown blotch germ is CCTCC M2014484, the method is used for extracting crude toxin, and the obtained citrus brown blotch germ crude toxin is high in content and purity and big in output, short in period and easy to realize large-scale preparation, and besides, the toxin replacing the pathogenic bacteria as a studying object avoids mutual interference and cross contamination among organisms, and the like. According to the invention, a simple, convenient, economic and efficient extraction method for citrus brown blotch germ crude toxin is established, and an excellent foundation is laid for the study of pathogenesis of germ and toxin thereof.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for extracting crude toxin from brown spot fungus citrus and toxin-producing conditions. Background technique [0002] In the process of interaction with host plants, pathogenic fungi can produce metabolites that cause physiological and biochemical reactions of the host, mainly including enzymes, hormones and toxins. Among them, toxins are sensitive inducing factors for host plant-induced diseases, which are research hotspots in recent years. Toxins produced by pathogenic fungi can be divided into host-selective toxins (HST) and non-host-selective toxins (NHST) according to their pathogenicity to the host. At present, about 20 kinds of pathogenic fungi are known to produce HST, most of which are secondary metabolites of pathogenic bacteria. In 1933, Tanaka first isolated the HST-AK toxin (Tanaka) produced by Alternaria kikuchiana from pear black spot fungus - Alternaria ki...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/14C07K14/37C07K1/14C12R1/645
CPCC07K1/14C07K14/37C12N1/14C12N1/145C12R2001/645
Inventor 唐科志王玲杰李中安周常勇
Owner CITRUS RES INST OF CHINESE ACAD OF AGRI SCI
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