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A method for rapidly and synchronously obtaining male sterile and maintainer transgenic plants of Wucai

A male sterile line and simultaneous acquisition technology, applied in the field of plant genetic engineering, can solve the problems of high cost and complicated operation of Wucai improved varieties

Active Publication Date: 2021-09-14
ANHUI AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The purpose of the present invention is to overcome the deficiencies of the prior art, and provide a method for quickly and synchronously obtaining male sterile and maintainer transgenic plants of Wucai, so as to solve the problems of high cost and complicated operation of Wucai improved varieties in the prior art

Method used

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  • A method for rapidly and synchronously obtaining male sterile and maintainer transgenic plants of Wucai
  • A method for rapidly and synchronously obtaining male sterile and maintainer transgenic plants of Wucai
  • A method for rapidly and synchronously obtaining male sterile and maintainer transgenic plants of Wucai

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Embodiment 1

[0035] 1) At the full flowering stage of Wucai, remove the male sterile line ( figure 1 , there is no pollen on the anthers) and the flowers that have opened on the main inflorescence of the maintainer line, select the flower buds of the maintainer line with a size of 3-5mm, and completely peel off the sepals and petals to expose the anthers ( figure 2 );

[0036] 2) Agrobacterium EHA105 (containing plant expression vector pBI121) was cultured to OD 600 =0.5, centrifuge to collect the cells, and suspend the cells with the infection solution to OD 600 =0.2, stand at room temperature for 30 minutes and set aside;

[0037] 3) Use a syringe and other tools to drop the infection liquid onto the anthers, let it stand for 30 minutes, and then protect it from light for 2 days;

[0038] 4) After 2 days, the flower buds open, use pointed tweezers to remove the anthers of the maintainer line, artificially pollinate the stigma of the male sterile line and its maintainer flower, or use...

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Abstract

The invention discloses a method for synchronously obtaining transgenic plants of male sterile lines and maintainer lines of Chinese cabbage with low cost, short period, good repeatability, simple operation and high transformation efficiency. Its features include the following steps: (1) removing the bloomed flowers of the sterile line and the maintainer plant at the blooming stage, covering the inflorescence with a paper bag and sealing it. (2) Peel off the sepals and petals of the maintainer flower buds with a size of 3-5 mm. (3) Agrobacterium cells were suspended in the infection solution to OD 600 = 0.2. (4) The infection liquid was dropped onto the maintainer anthers and treated in the dark for 2 days. (5) Using pollination tools to artificially pollinate the flower stigmas of the maintainer line and the sterile line. (6) Repeat the above operation 3 times. (7) Harvest the seeds of the sterile line and the maintainer line respectively. (8) The harvested seeds are identified by Kan resistance and GUS staining, and transgenic plants can be obtained. The method can be directly operated in the field without tissue culture to obtain the male sterile line of the transgenic Chinese cabbage.

Description

technical field [0001] The invention belongs to the field of plant genetic engineering, and particularly provides a method for synchronously obtaining transgenic plants of male sterile line and maintainer line of Chinese cabbage with low cost, short period, good repeatability, simple operation and high efficiency. Background technique [0002] Black cabbage, native to my country, is a variety of Brassica cabbage subspecies, and is one of the vegetables with the largest planting area in the Jianghuai region. For a long time, Wucai varieties have been improved mainly through variety hybridization and distant hybridization. There are many disadvantages such as long breeding period, high cost, easy degeneration of parental self-crossing for multiple generations, and difficulty in male sterility transfer. system. At present, the Agrobacterium-mediated method, gene gun method, ultrasonic-assisted transformation method, etc. all require a regeneration system; the injection method ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/82A01H1/02
CPCA01H1/02C12N15/8289
Inventor 陈国户汪承刚曾繁丽朱世东袁凌云侯金锋
Owner ANHUI AGRICULTURAL UNIVERSITY
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