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Method for improving verticillium wilt resistance of plants by using Verticillium dahlia VdP4-ATPase gene

A Verticillium dahliae, transgenic plant technology, applied in botany equipment and methods, biochemical equipment and methods, plant products, etc., to achieve the effect of improving resistance

Inactive Publication Date: 2016-05-04
SOUTHWEST UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the vast majority of researchers mainly use the former method to achieve the purpose of improving plant resistance, and there is no report on the successful use of foreign gene expression products to detoxify the toxins produced by pathogenic bacteria to improve resistance to Verticillium wilt

Method used

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  • Method for improving verticillium wilt resistance of plants by using Verticillium dahlia VdP4-ATPase gene
  • Method for improving verticillium wilt resistance of plants by using Verticillium dahlia VdP4-ATPase gene
  • Method for improving verticillium wilt resistance of plants by using Verticillium dahlia VdP4-ATPase gene

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preparation example Construction

[0132] 7.3 Preparation of Agrobacterium infusion solution

[0133] Agrobacterium strains containing LGN-35S-VdP4-ATPase carrier stored at -80°C were streak cultured to obtain single colonies, and then picked single colonies and inoculated into YEB supplemented with 50mg / LKm and 125mg / LSm (streptomycin sulfate) Medium (5g / L sucrose, 1g / L bacterial yeast extract, 10g / L bacterial tryptone, 0.5g / LMgSO 4 ·7H 2 0, pH7.0), 28°C, 200rmp shaking culture overnight, when the OD600 value of the bacterial solution reaches 0.8, take the bacterial solution and centrifuge, and resuspend the bacteria with the same volume of MSB liquid medium, and the resuspension is used for transformation Agrobacterium infusion solution.

[0134] 7.4 Genetic transformation and plant regeneration of tobacco

[0135] The leaf disk explants obtained in step 7.2 were immersed in the Agrobacterium infusion solution prepared in step 7.3 for 30 minutes, and then the bacterial solution was poured out, and the toba...

Embodiment 7

[0139] When the transgenic tobacco plants obtained by implementing the genetic transformation and molecular identification of Example 7 grow to 6-10 leaves in the greenhouse, use a scalpel blade to treat the root injury at about 1-2 cm away from the plant, and then water each plant to implement the method prepared in Example 7.1. L2-1 inoculation solution 30mL (injury root irrigation method), placed in 22 ℃ (night) - 26 ℃ (day) artificial climate indoor culture. 30 days after the inoculation, the disease grade of the transgenic plants was counted according to the standard of Example 7.2. The results showed that the average disease grade of 46 independent VdP-ATPase transformants was 1.17, and the disease index was 29.25. While the wild-type non-transgenic control (regenerated wild-type plants) had an average disease grade of 3.03 and a disease index of 75.65 (Table 1), compared with the non-transgenic control, the disease index of the VdP-ATPase transgenic line was reduced by ...

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Abstract

The invention relates to a use of VdP4-ATPase gene for coding Verticillium dahlia P type ATPase in improvement of the verticillium wilt resistance of plants, and a method for improving the verticillium wilt resistance of the plants. The VdP4-ATPase gene for coding the erticillium dahlia P type ATPase is expressed in target plants to improve the verticillium wilt resistance of the plants. Detoxification of pathogen toxins is used to improve the resistance of the plants, and the pathogen race resistance characteristic does not exist. The method can be widely applied in improvement of the resistance of different pathogen physiologic races without the race-specific resistance characteristic.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering. Specifically, it relates to a method for improving plant disease resistance by using genetic engineering technology. technical background [0002] Verticillium wilt seriously affects the yield and quality of crops, and the global annual loss caused by Verticillium wilt reaches billions of dollars (Peggetal., 2002). It is reported that the annual output of potatoes is reduced by more than 50% due to the infection of Verticillium wilt, which can usually cause a 10-15% reduction in production (Powelson et al., 1993; Rowe et al., 1987, 2002); lettuce growing areas, the loss caused by Verticillium wilt It is very easy to reach 100% (Subbarao et al., 1997); Levinet (2003) and other studies have found that the production reduction of olives due to Verticillium wilt can reach more than 70%; Verticillium wilt is also the main disease of cotton in most cotton-growing countries, such as Australia ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/55A01H5/00
CPCC12N15/8271C12N15/82
Inventor 裴炎李玉杰陈杨范艳华侯磊李先碧宋水清
Owner SOUTHWEST UNIV
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