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Gene for controlling male reproduction and development of rice and application of gene

A male reproductive and genetic technology, applied in the fields of agriculture and plant biology, can solve problems such as difficult to use and difficult to reproduce efficiently, and achieve the effect of increasing yield and expanding the use of heterosis

Inactive Publication Date: 2017-12-08
JIANGXI ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

GMS lines are difficult to utilize in crop hybrid production because they are difficult to reproduce efficiently

Method used

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  • Gene for controlling male reproduction and development of rice and application of gene
  • Gene for controlling male reproduction and development of rice and application of gene
  • Gene for controlling male reproduction and development of rice and application of gene

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Design of targets; design 2 targets for genes with unknown functions. One target is designed in the 5' region and the functional domain of the ORF, so that mutation of any one target can result in loss of function, or the sequence between the two targets can be knocked out. A high GC% of the target sequence can improve the targeting efficiency, so the target preferably contains 11-14 C / G (including U6 transcription initiation point G). Blast the target genome with the candidate target sequence + NGG (add tens of bases upstream and downstream) (choose Somewhat similar sequences (blastn) to avoid similarity between the 3' end of the target sequence + NGG and other functional genes and genome sequences.

Embodiment 2

[0022] Detection of mutation sites; detection of targeting effects of transgenic T0 plants: Transformants with successful targeting often have single base insertions or deletions of several bases at the target site. With the target as the center, primers were synthesized for PCR amplification at about 200-300 bp on both sides. Key points: Design internal primers at about 150-250bp upstream of the target as sequencing primers (the sequencing quality at this distance is the best, but not less than 100bp and not greater than 400bp), and directly sequence the PCR products (do not use PCR amplification) If the modified primers are used again for sequencing, a high level of heterogeneous signal will be generated). If there are all overlapping peaks from a certain base, it indicates a heterozygous mutation or a biallelic mutation.

[0023] Table 1

[0024]

[0025]

[0026]

[0027]

[0028]

[0029]

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Abstract

The invention discloses a gene for controlling male reproduction and development of rice and application of the gene. The gene has a nucleotide sequence represented by SEQ ID NO.1, and a protein encoded by the gene has an amino acid sequence represented by SEQ ID NO.2. By targeting a LOC_Os08g20730 gene design target by virtue of a CRISPR / Cas9 gene editing system, a receptor is a middle flower 11, and a targeted T0 generation phenotype are shown in the figures 1, 2, 3 and 4 in the description; and sequencing analysis shows that anther development abnormality occurs in a homozygously mutated plant or a biallelically mutated plant and pollen does not exist. The analysis of a homozygously mutated T1 generation shows that the separation proportion of fertile plants to sterile plants is 3 to 1. The integration shows that the gene is capable of regulating and controlling the reproduction and development of rice, once the gene in the rice is damaged, the anther development abnormality occurs, and non-pollen sterility occurs. According to the obtained cell nucleus gene for controlling the reproduction and development of the rice, an intelligent sterile line can be created by virtue of a molecular measure, and excellent parents in the production are improved into excellent sterile lines, so that the heterosis utilization is expanded.

Description

technical field [0001] The present invention relates to fields such as agriculture and plant biotechnology, in particular to basic research on plant genetics and breeding, plant physiology, and gene functions at the molecular level, and specifically relates to a gene for controlling male reproductive development in rice and its application. Background technique [0002] The male reproductive development of rice is an important process of reproductive growth, and it is also a key process for the reproduction of offspring and the formation of yield. The discovery of male sterile lines enabled the application of heterosis in rice. With the birth of the "three-line method" and "two-line method" seed production techniques, hybrid rice began to be planted on a large scale in China and gradually expanded to the world, playing a pivotal role in food production and food security. Male sterility (Male Sterility, MS) refers to the abnormal development or degeneration of male organs, t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415
CPCC07K14/415
Inventor 杨平陈春莲熊运华黄永萍尹建华刘耀光
Owner JIANGXI ACAD OF AGRI SCI
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