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Brassica napus dominant cell nucleus male sterility gene MS5b and application thereof

A technology of Brassica napus, male sterile line, applied in the application, genetic engineering, plant genetic improvement and other directions, can solve the problems of MS5 sequence, molecular mechanism and function loss that have not been reported in literature.

Active Publication Date: 2019-11-15
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Named as a gene, MS5 b It was reported in the literature very early, but researchers have previously regarded it as a loss-of-function gene, so there is no literature on MS5 b The sequence and molecular mechanism of

Method used

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  • Brassica napus dominant cell nucleus male sterility gene MS5b and application thereof
  • Brassica napus dominant cell nucleus male sterility gene MS5b and application thereof
  • Brassica napus dominant cell nucleus male sterility gene MS5b and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0017] Embodiment 1, MS5 b Segmental transcript analysis

[0018] First, use The Super RNA extraction kit (Promega, Beijing) was used to extract the total RNA from flower buds of the male sterile line FM195A at the meiotic stage, and sent to Nuohezhiyuan (Beijing) for RNA-seq sequencing. Align the sequencing results to MS5 with Hisat2 software b genome segments. MS5 b Genome segment including MS5 a Genomic segment of , and the 8,115 bp MULE transposon insertion sequence located in its second intron. It can be clearly seen from the comparison results that MS5 b Gene segment due to insertion of transposons, resulting in two new transcripts. Through MS5 in Table 1 b Specific Detection Primer MS5 ab -RT-L / MS5 ab -RT-R and MS5 b -RT-L / MS5 b - RT-R, detection of MS5 by RT-PCR and RT-qPCR a MS5 a and MS5 b MS5 b cDNA discovery during meiosis, due to transposon insertion, MS5 b The expression level of the first transcript of the gene decreased ( figure 1 B, C), new ...

Embodiment 2

[0021] Example 2, MS5 b Transgenic verification of functional segments and confirmation of coding regions

[0022] 1. Construction of transgenic vector

[0023] In this example, the plant expression vector pFGC5941 was used as the transgenic vector of Brassica napus. According to the position of the two transcripts, we put the whole MS5 b Segments as well as segments containing only the first transcript were constructed as transgene vectors to verify MS5 b Functional Area. We used Rs1046A genomic DNA as a template. Utilize the primer pFGC5941-MS5 in Table 2 b -L / pFGC5941-MS5 b -2R amplification encompasses a 5,734bp segment of the first transcript, which includes a 1,411bp promoter region, via II One Step Cloning Kit (Vazyme, C112-01) was constructed on the binary vector pFGC5941, and the vector was named MS5 b -2. On this basis, using pFGC5941-MS5 b -2L / pFGC5941-MS5 b-R amplifies the second half of the 6,308bp segment, using II One Step Cloning Kit (Vazyme, C112...

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Abstract

The invention discloses a brassica napus dominant cell nucleus male sterility gene MS5b and application thereof and relates to the technical field of genetic breeding of brassica napus and genetic engineering. An amino acid sequence of function section protein of the brassica napus dominant cell nucleus male sterility gene MS5b is shown in SEQ ID NO:1; a coding sequence of the function section protein of the brassica napus dominant cell nucleus male sterility gene MS5b is shown in SEQ ID NO:3. The gene can be applied to transformation of a dominant cell nucleus male sterility line; through a transgenosis method, the brassica napus dominant cell nucleus male sterility line is directly created. The prepared brassica napus male sterility line has no abnormity in the nutrient growth period, abnormal meiosis only occurs in the reproduction growth period, male sterility is generated, and the gene can be applied to heterosis utilization and population improvement in brassica crops and has important significance on breeding for the brassica crops.

Description

technical field [0001] The invention belongs to the technical field of rapeseed breeding and genetic engineering, in particular, relates to a dominant nuclear male sterility gene MS5 in Brassica napus b The gene can be used to rapidly select and breed excellent sterile lines through genetic transfer and transgenic methods. Background technique [0002] Oil is one of the three main nutrients that human beings depend on for survival, and maintaining the security of edible oil supply is one of the cores of national security strategies. The promotion of hybrids and the utilization of heterosis are one of the effective ways to increase the yield of rapeseed (Fu Tingdong. Current status and prospects of Chinese rapeseed production and variety improvement. Anhui Agricultural Science Bulletin, 2000, 6(1): 2-8). A safe and effective pollination control system is an important prerequisite for the production of rapeseed hybrids. [0003] At present, the commonly used pollination cont...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/415C12N15/29C12N15/82A01H5/00A01H6/20
CPCC07K14/415C12N15/8289
Inventor 杨光圣辛强洪登峰万丽丽董发明
Owner HUAZHONG AGRI UNIV
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