Ogura CMS sterility restoring gene RfoB, RfoB plant expression vector and use thereof

A technology for plant expression vectors and gene restoration, which is applied in the field of genetic engineering to achieve the effects of promoting excellent germplasm innovation, improving expression efficiency, and improving expression specificity and expression efficiency

Active Publication Date: 2018-10-12
SOUTHWEST UNIVERSITY
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Problems solved by technology

Therefore, for cabbage vegetables such as cabbage, cauliflower, and kohlrabi that are not easy to bolt and flower, i...

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  • Ogura CMS sterility restoring gene RfoB, RfoB plant expression vector and use thereof
  • Ogura CMS sterility restoring gene RfoB, RfoB plant expression vector and use thereof
  • Ogura CMS sterility restoring gene RfoB, RfoB plant expression vector and use thereof

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Embodiment Construction

[0043] The present invention is further described in conjunction with the following specific implementation examples. It should be understood that these are examples and are not intended to limit the scope of the present invention, and materials not described in this example are commercial products and can be purchased by companies.

[0044] 1. Synthetic Rfo B gene sequence

[0045] According to the published cDNA sequence of the Rfo gene coding region of the radish Ogura CMS cytoplasmic sterility Rfo recovery site (AY285674.1), combined with the codon preference of cabbage (Brassica oleracea var capitata), the codons were optimized, and the optimized The Rfo gene (as shown in SEQ ID No.2). At the same time, a 210bp intron sequence from the potato light-induced tissue-specific expression gene ST-LS1 was added to its coding region to obtain the restored gene sequence Rfo B (as shown in SEQ ID No.4). A NcoI restriction site was added at the 5' end of the sequence, and a BamH...

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Abstract

The invention relates to the field of genetic engineering and particularly to a restoring gene RfoB and an expression vector and application thereof. RfoB utilizes a Brassica oleracea L. Ogura CMS cytoplasmic male sterile line as an object, and according to the restoring gene Rfo sequence, through the codon preference of the Brassica oleracea L. gene, a novel RfoB sequence is artificially synthesized. A PRfo promoter, an RfoB gene and a terminator E9-T are fused to form an RfoB Gene expression kit. After construction of the plant expression vector, through the Agrobacterium tumefaciens-mediated direct transformation method, Brassica oleracea L. Ogura CMS cytoplasmic sterile line is directly transformed so that the transgenic plant of Brassica oleracea L. is obtained. The fertility is restored. The transgenic plants are self-crossed and normally seeded, and they are introduced into other Brassica oleracea L.-like Ogura CMS cytoplasmic sterile line as a male parent so that the individualcarrying the RfoB gene can restore fertility.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to the artificially synthesized Ogura CMS cytoplasmic male sterility restoration gene Rfo B 、Rfo B Plant expression vectors and their application in the creation of Ogura CMS cytoplasmic male sterile restorer lines. Background technique [0002] Currently, Ogura CMS cytoplasmic male sterility, which is widely used in Brassica plants, was discovered in 1968 in radish in Kagoshima, Japan (Ogura H. Studies on the new male-sterility in Japanese radish, with special reference to the utilization of this sterility towards the practical raising of hybrid seeds. Mem Fac Agric Kagoshima Univ, 1968, 6: 39–78), sterility traits are mainly controlled by the orf138 gene on mitochondria (Bonhomme S, Budar F, Lancelin D, Small I, Defrance MC, and Pelletier G. Sequence and transcript analysis of the Nco2.5-specific fragment correlated with cytoplasmic male sterility in Brassica cybrids. Mol Gen ...

Claims

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Application Information

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IPC IPC(8): C12N15/29C12N15/82A01H5/00A01H6/20
CPCC07K14/415C12N15/8289
Inventor 宋洪元杜杨梅徐冰冰李勤菲任雪松司军
Owner SOUTHWEST UNIVERSITY
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