GAMYB gene based cultivation method for transgenic rice sterile line

A technology of genetically modified rice and breeding methods, applied in genetic engineering, botany equipment and methods, biochemical equipment and methods, etc., capable of solving problems such as delayed programmed death, unfree mating, and sterility

Inactive Publication Date: 2018-11-20
QINGDAO YUANCE GRP CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the three-line method, the available parents with good shape are limited, the combination is not free, and the frequency of wild-lost maintenance lines is low
However, the rice GAMYB gene mutation will cause the tapetum cells to fail to degrade in time, and the programmed death will be delayed, so the callose enzyme secreted by the tapetum will affect the development and separation of microspores, and finally show sterility

Method used

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  • GAMYB gene based cultivation method for transgenic rice sterile line
  • GAMYB gene based cultivation method for transgenic rice sterile line
  • GAMYB gene based cultivation method for transgenic rice sterile line

Examples

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Embodiment 1

[0039] A method for cultivating a transgenic rice sterile line based on the GAMYB gene, characterized in that it comprises the following steps:

[0040] A, the acquisition of the rice gene expression cassette;

[0041] B, amplification of the corn lethal gene ZMAA1;

[0042] C, amplification of the corn pollen-specific promoter Pg47;

[0043] D, the acquisition of the ECFP gene expression cassette;

[0044] E. Connection of each gene expression element.

[0045] 1. Obtaining the rice gene expression cassette

[0046] Extract the genomic DNA of rice plants, use the genomic DNA as a template, and use F2 / R2 as upstream and downstream primers to amplify the complete GAMYB gene expression cassette. During the amplification process, SmaI and DmaI enzymes were introduced into the upstream primer and downstream primer site, the amplified rice GAMYB gene contains its upstream promoter and downstream terminator, as well as all exons and introns.

[0047] 2. Amplification of maize l...

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Abstract

The application discloses a GAMYB gene based cultivation method for a transgenic rice sterile line. The GAMYB gene based cultivation method comprises the steps as follows: A, acquisition of a rice gene cassette; B, acquisition of a maize lethal gene ZMAA1; C, amplification of a maize pollen specific promoter Pg47; D, acquisition of an EGFP gene cassette; and E, connection of each gene expression element. The step E of connection of each gene expression element comprises: a first step of introducing a complete rice GAMYB gene into a pCAMBIA1300 vector, and then connecting the maize lethal geneZMAA1, the maize pollen specific promoter pPg47 and the EGFP gene expression element onto the pCAMBIA1300 vector. According to the invention, a F1-generation heterozygote follows the Mendel segregation law in the self fruitfulness process, and generated progenies not only contain heterozygotes which can keep a triple linked gene, but also contain the sterile line without fertility.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering genetics and breeding, and in particular relates to a method for cultivating a transgenic rice sterile line based on GAMYB gene. Background technique [0002] Rice is native to tropical Asia, and after being widely planted in China, it gradually spread to all parts of the world. Rice is the staple food of nearly half of the world's population. In addition to being edible, rice can also be used for wine making, sugar refining and as an industrial raw material. Years of production practice have shown that hybrid rice can generally increase yield by more than 20% compared with conventional rice, so hybrid rice shows great potential for increasing yield. [0003] The development of hybrid rice depends on the cultivation of male sterile lines of hybrid rice. Research on hybrid rice in my country began in the 1960s, and it began to be planted on a large scale in the 1970s. The first-gener...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N15/65C12N15/66A01H5/00A01H6/46
CPCC12N15/65C12N15/66C12N15/8289
Inventor 刘佳音米铁柱张国栋修旺珊万吉丽徐春莹
Owner QINGDAO YUANCE GRP CO LTD
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