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Genetic transformation method for agrobacterium tumefaciens mediated fusarium oxysporum watermelon specialization strain

A genetic transformation method, the technology of Fusarium oxysporum, applied in the biological field, can solve the problems of cumbersome transformation steps and no details related to transformation, and achieve the effect of good repeatability and high transformation efficiency

Inactive Publication Date: 2014-04-30
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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Problems solved by technology

[0003] At present, there have been reports at home and abroad on the transformation of fungi using Agrobacterium-mediated genetic transformation methods, but the reported methods either do not involve transformation-related details, or the transformation steps are cumbersome

Method used

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  • Genetic transformation method for agrobacterium tumefaciens mediated fusarium oxysporum watermelon specialization strain
  • Genetic transformation method for agrobacterium tumefaciens mediated fusarium oxysporum watermelon specialization strain
  • Genetic transformation method for agrobacterium tumefaciens mediated fusarium oxysporum watermelon specialization strain

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Embodiment 1

[0038]Agrobacterium-mediated Fusarium oxysporum watermelon specialized strain genetic transformation method, the steps are as follows:

[0039] 1. Preparation of conidia suspension of Fusarium oxysporum watermelon specialized strain

[0040] Use an inoculation loop to pick the watermelon-specific hyphae of the Fusarium wilt fungus Fusarium oxysporum, inoculate it on a potato solid medium, and culture it at 28 °C for 7 days. Use 6 mL of medium A and a sterilized small brush to clean the pathogen spores, and remove the spores. Collect the liquid into a sterilized conidia flask, filter it with sterile double-layer gauze, then dilute the spore liquid with medium A, calculate the concentration of conidia with a hemocytometer, and adjust the concentration of the spore liquid to 1×10 6 cells / mL to obtain the AGL-1 Agrobacterium liquid for future use.

[0041] 2. Preparation of Agrobacterium liquid containing exogenous gene

[0042] Agrobacterium AGL-1 carrying the expression vector...

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Abstract

The invention provides a genetic transformation method for an agrobacterium tumefaciens mediated fusarium oxysporum watermelon specialization strain. The method comprises the following steps: (1) preparing fusarium oxysporum watermelon specialization strain conidiophore suspension; (2) preparing an agrobacterium tumefaciens solution containing exogenous genes; (3) performing exogenous gene transformation on the fusarium oxysporum watermelon specialization strain; and (4) identifying a transformant of the fusarium oxysporum watermelon specialization strain. The exogenous genes are transferred into the genome of the fusarium oxysporum watermelon specialization strain by utilizing an agrobacterium tumefaciens mediated method, and a fusarium oxysporum watermelon specialization transgenic strain is obtained. The method is simple, rapid, high in repeatability and transformation efficiency, and can be used for the construction of a fusarium oxysporum watermelon specialization strain mutant library, host resistance screening, defense response of the host on infection of the fusarium oxysporum watermelon specialization strain and research of the infection and pathogenesis of the fusarium oxysporum watermelon specialization strain.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for genetic transformation of Fusarium oxysporum watermelon-specific strains mediated by Agrobacterium. Background technique [0002] Fusarium wilt of watermelon is a disease of the vascular system caused by Fusarium oxysporum specialized in watermelon. The pathogen is transmitted through soil and seeds, and can survive for 5-6 years without the host, and chlamydospores can survive for more than 10 years. Fusarium wilt of watermelon can occur in various stages of watermelon growth. According to surveys, melon fields with the disease generally reduce production by 30%, and serious plots can reach 80%, or even extinction. It is one of the most serious diseases in watermelon production at home and abroad, and has become a constraint Major obstacle to watermelon production. Breeding new varieties of watermelon that are high-quality, disease-resistant and suitable for facility ...

Claims

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Application Information

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IPC IPC(8): C12N15/80C12N1/15C12R1/77
Inventor 张曼羊杏平徐锦华刘广姚协丰李苹芳
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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