New botrytis cinerea gene related to pathogenicity and application of new botrytis cinerea gene

A botrytis cinerea and gene technology, applied in application, genetic engineering, plant genetic improvement and other directions, can solve the problems of botrytis cinerea that are prone to drug resistance, environmental pollution, pesticide residues, etc.

Inactive Publication Date: 2013-03-13
HEBEI AGRICULTURAL UNIV.
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the prevention and control of Botrytis cinerea is still dominated by chemical control, but because Botrytis cinerea is easy to produce drug resistance, and many problems such as pesticide residues and environmental p

Method used

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  • New botrytis cinerea gene related to pathogenicity and application of new botrytis cinerea gene
  • New botrytis cinerea gene related to pathogenicity and application of new botrytis cinerea gene
  • New botrytis cinerea gene related to pathogenicity and application of new botrytis cinerea gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Bcdp1 Obtaining of Gene T-DNA Insertion Mutants

[0034] The mutant involved in the present invention is obtained by Agrobacterium tumefaciens mediated transformation method (ATMT).

[0035] 1.1 Cultivation of Agrobacterium. Pick a single colony of Agrobacterium and place it in LB liquid medium (containing 50 μg / mL kanamycin and 50 μg / mL rifampicin), shake it at 120 rpm at 28°C for 48 h, take 1 mL of the bacterial liquid at 4°C Centrifuge at 10000 rpm for 1 min and discard the supernatant. Wash twice with induction medium (IM), dilute to OD600≈0.15 with IM medium, add AS at a final concentration of 200-400 μM, and activate at 28°C for about 7 h at 180 rpm.

[0036] 1.2 Collection of conidia of Botrytis cinerea. Wash the conidia of Botrytis cinerea from the PDA plate cultured for about 7 days with 10 mL of sterilized distilled water, filter through two layers of gauze, count with a hemocytometer, and dilute the spore concentration with sterile water to 10...

Embodiment 2

[0039] Example 2 Screening and Stability Detection of Pathogenicity Loss Mutants

[0040] The T-DNA insertion mutant library of Botrytis cinerea was screened by in vitro fruit inoculation. First select fresh tomatoes of the same size and scrub the fruit with 75% alcohol for surface disinfection. At the same time, the rejuvenated Botrytis cinerea wild-type BC22 and transformants were cultured for 3 days, and the colonies were punched out with a puncher to get a 5 mm-diameter plate from the edge of the colony, and they were symmetrically inoculated on tomato fruits (without wounds). The tomato fruit was put into a moisturizing tank, and the condition was investigated after 6 days. The difference in pathogenicity of the strains was evaluated according to the size of the lesion area. The mutant Bct89 that lost the pathogenicity of Botrytis cinerea was subcultured in PDA medium (containing hygromycin 100 μg / mL) for 15 generations, and the pathogenicity of each generation was te...

Embodiment 3

[0041] Example 3 Biological studies of mutants

[0042] Botrytis cinerea wild-type strain BC22 and mutant strain Bct89 were rejuvenated. After rejuvenation, they were respectively inoculated on PDA medium. After 3 days of cultivation, the wild-type and mutant strains with a diameter of 9 mm were punched on the edge of the colony. , cultured at 20°C for 10 days, observed the colony shape, color, measured growth rate, spore production, etc.

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Abstract

The invention relates to a new botrytis cinerea gene which is named as Bcdp1. The gene contains three exons and two introns, the total length of DNA (Deoxyribonucleic Acid) is 1608bp, the total length of cDNA (complementary deoxyribonucleic acid) is 597bp, and 198 amino acids are encoded. The invention also relates to an expression vector and a host containing the gene, and application of the gene in the prevention and control of plant diseases (especially botrytis) and application of the gene used as a target of a pesticide for preventing and controlling the plant diseases.

Description

technical field [0001] The present invention relates to the field of plant disease prevention and control, in particular, the present invention relates to a method related to the pathogenicity of Botrytis cinerea Bcdp1 Gene. The present invention also relates to the application of the gene in plant disease control and as the target of medicine for plant disease control. Background technique [0002] Botrytis cinerea ( Botrytis cinerea ) belongs to Deuteromycotina, Hyphospora, Hyphospora, Botrytis, and it is one of the earliest studied fungi. The hyphae of the fungus have septa, and the sexual reproduction is mainly heterothallic, and almost no ascospores are produced under natural conditions. Conidiophores are mostly brown, erect, with septa, the size is 960~1 200 μm×16~22 μm, the top has irregular tree-like branches, the top is slightly enlarged, and it is club-headed, and a large number of Conidia, conidia aggregated into grape spikes. Conidia are round or ovoid, lig...

Claims

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Application Information

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IPC IPC(8): C12N15/31C07K14/37A01N61/00A01P3/00C12R1/645
Inventor 邢继红董金皋王凤茹赵斌司贺龙郑会欣赵福鑫
Owner HEBEI AGRICULTURAL UNIV.
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