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Brassica napus nucleic male sterility gene Bnms4 and preparation method and application

A technology of Brassica napus, recessive genital sterility, application of in situ locus, application of Brassica napus genic male sterile gene Bnms4b, Brassica napus genic male sterile gene Bnms4b, preparation of Brassica napus genic male sterile gene Bnms4b It can solve problems such as complex genetic basis, unfavorable breeding and strong advantage combination, and difficulty in breeding of temporary insurance lines

Inactive Publication Date: 2016-03-30
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the disadvantage of this system is that both the sterile line and the restorer line must be transgenic, and the system is currently protected by a patent
However, the genetic basis of this system is complicated, and the selection of temporary insurance lines is difficult, which is not conducive to the selection of strong dominant combinations.

Method used

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  • Brassica napus nucleic male sterility gene Bnms4<b> and preparation method and application
  • Brassica napus nucleic male sterility gene Bnms4<b> and preparation method and application
  • Brassica napus nucleic male sterility gene Bnms4<b> and preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] Example 1: Fine mapping and chromosome landing of the sterile gene Bnms4

[0037] A Genic Sterile Gene Bnms4 in Brassica napus b The preparation method, its steps are:

[0038] 1. Experimental materials:

[0039] for Bnms4 b The near-isogenic line population of gene map site cloning is 7365AC, consisting of 7365A (Huang et al., Theor Appl Genet., 2007, 115:113-118) and 7365C (Xiao et al., 2008, Euphytica, 164(2):377-384), Then it was backcrossed with 7365C, and the offspring showed segregation between sterile and fertile. After the high-generation backcross, siblings were preserved. The segregation ratio of fertile and sterile in this population was 1:1, and the sterile single plant contained Bnms4 b Gene. The DNA of 5050 fertile and sterile individual plants in the near-isogenic line 7365AC extracted in Wuhan in the spring of 2009 (see: Stewart and Via, 1993, 14(5): 748-749 for the method of extracting DNA) for gene identification Fine positioning.

[0040] 2. Fi...

Embodiment 2

[0053] Example 2: The sterility gene Bnms4 b Isolation and functional verification of

[0054] A Genic Sterile Gene Bnms4 in Brassica napus b Application in the genetic improvement of nuclear male sterility of Brassica napus and other cruciferous plants (taking Arabidopsis as an example), the steps are:

[0055] 1. Bnms4 b Determination of Candidate Genes and Transformation Experiment of Arabidopsis Rapeseed

[0056] The sequence comparison program adopted in this example comes from the Blast (http: / / brassicadb.org / brad / blastPage.php) software provided by the genome website of Chinese cabbage Chiifu-401, and includes Bnms4 b The sequence of BAC clone B107 was analyzed, and a candidate gene CC2 was predicted from it. Analyze the CDS sequences of other homologous genes by BAC sequence alignment to preliminarily determine the start and end sites of the ORF of the candidate gene CC2, develop primers to amplify the CDS sequence of the target gene, and use RACE (rapid amplificat...

Embodiment 3

[0065] Example 3: Utilization of the restorer gene BnMs3 and the sterility gene Bnms4 b Construction of sterile material and corresponding maintenance material of cruciferous plants (taking Col-0 wild-type Arabidopsis as an example)

[0066] Using Bnms4 b The nuclear male sterility system of other cruciferous plants can be created: Chinese cabbage, cabbage, and Arabidopsis, using the same method as Brassica napus, all use this gene to lead to stable and complete male sterility.

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Abstract

The invention discloses a brassica napus nucleic male sterility gene Bnms4 and a preparation method and application. The method comprises the steps that 1, a CDS nucleotide sequence or promoter nucleotide sequence or coded aminoacid sequence of the gene is utilized for designing and developing a primer capable of amplifying the gene, and the primer is synthesized; 2, by means of the synthesized primer, the gene or any section of interested nucleotide or one section of polynucleotide homologous with the gene is amplified from a genome, mRNA and cDNA. The gene or a homologous gene is screened and obtained from the cDNA and a genome library by adopting the cloned polynucleotide as a probe. The method is easy to carry out and easy and convenient to operate, the primer is used for amplifying the gene, the gene is cloned to various carriers and converted into brassica napus, stable and thorough male sterility can be caused, and breeding application is conducted; a sequence of the gene itself is used for designing a specific primer for molecular marker-assisted selection, and the advantages of being rapid, economical, effective and accurate are achieved.

Description

technical field [0001] The invention belongs to the technical field of rapeseed molecular breeding. More specifically relates to a Brassica napus nuclear sterile gene Bnms4 b , also involves a Brassica napus genic male sterile gene Bnms4 b The preparation method also relates to a Brassica napus nuclear sterile gene Bnms4 b Applications. A multiple allelic locus related to male sterility involved in the present invention: Bnms4 a To restore the gene, Bnms4 b For the sterile gene, Bnms4 c It is a null allele, and its dominant-recessive relationship is: Bnms4 a >Bnms4 b >Bnms4 c . sterile gene Bnms4 b The cloning, verification and application of the method can not only speed up the selection of recessive nuclear male sterile lines and temporary insurance lines of Brassica napus, but also create the nuclear sterile systems of other Brassicaceae plants based on this, for example, will not Bnms4 b Stable sterile material can be obtained by transformation into Arabi...

Claims

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Application Information

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IPC IPC(8): C12N15/29C07K14/415C12N15/84A01H5/00
Inventor 涂金星夏胜前文静易斌马朝芝沈金雄傅廷栋
Owner HUAZHONG AGRI UNIV
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