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Rice hybrid pollen fertility gene and uses thereof

A fertility gene and hybrid technology, applied in the field of genetic engineering, can solve the problems of hybrid pollen sterility or unclear molecular basis of affinity

Inactive Publication Date: 2009-07-29
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The chromosomal location of some rice hybrid fertility genes has been reported, but the molecular basis of hybrid pollen sterility or compatibility (fertility) is still unclear

Method used

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  • Rice hybrid pollen fertility gene and uses thereof
  • Rice hybrid pollen fertility gene and uses thereof
  • Rice hybrid pollen fertility gene and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Example 1: Screening of polymorphic molecular markers and fine mapping of Sa

[0035] It is known that Sa is located in the approximate region on the first chromosome of rice (Zhuang Chuxiong et al. 1994, Acta Genetica Sinica 21: 34-37). The present invention uses as figure 2 The shown map-based cloning method further cloned the Sa gene. First, the japonica rice variety Taichung 65 (abbreviated as T65, with genotype Sa j Sa j ) and its near isogenic line E4 (Sa i Sa i ) crossed to create a F 2 generational segregation groups. The near-isogenic line E4 used in the present invention is except Sa i The allelic locus and its surrounding regions are derived from a line other than indica rice whose nuclear genome background is the same as T65 (Zhuang Chuxiong et al. 1994, Acta Genetica Sinica 21: 34-37). Therefore the genetic segregation of male sterility in their hybrids is limited only by Sa i Sa j Effects of heterozygous gene interaction effects. with 1%I 2 -K...

Embodiment 2

[0041] Example 2: Sequence analysis of genes related to the Sa locus

[0042] According to the genome DNA sequence of rice (japonica) published by GenBank and the RiceGAAS software (http: / / ricegaas.dna.affrc.go.jp) analysis, there are only 2 predicted genes in the positioning region, named SaM respectively and SaF. Genetic analysis showed that in different F 2 Among recombinants, only individuals containing heterozygous SaM and one or two SaF alleles derived from E4, that is, heterozygous SaF or homozygous E4SaF, can produce hybrid pollen semi-sterile ( figure 2 ). It was shown that the Sa locus was composed of SaF and SaM, and the genetic interaction of the three alleles of these two genes produced hybrid sterility.

[0043] Use primer 5'TCTAAG GGATCC AGGCGCGGGTGAAGCAAACTAGAC-3' (underlined as BamHI) and 5'ATTTAA GAATTC GCCGTCTTAACACACATTGGAAAA-3' (underlined as EcoRI) from T65, E4, and multiple indica types (Guangluai 4, 93-11, Minghui 63, etc.) and japonica types (N...

Embodiment 3

[0045] Example 3: Genetic transformation to identify the function of the target gene

[0046] With the SaM described in embodiment 2 + 、SaM - , and SaF + , which include the promoter region, coding region, and downstream sequences, were cloned into the binary transformation vector pCAMBIA1300 (CAMBIA, Australia). The obtained vector transgenes were named tSaM + , tSaM - , and tSaF + . These vectors were introduced into soil Agrobacterium tumefaciens strain EHA105 by electric shock method, and rice with suitable genotype was transformed by Agrobacterium tumefaciens-mediated method (Hiei et al., Plant J.6: 271-282). tSaM - Transform containing genotype to SaM + / SaM + / / SaF + / SaF + E4, obtained from 4 strains of T 0 Transformants showed semi-sterile pollen ( Figure 4 A). tSaM + Transformation of the genotype SaM isolated from the hybrid progeny produced by crossing 65 with E4 - / SaM - / / SaF + / SaF - The pollen fully fertile recombinants obtained 8 T 0 The f...

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Abstract

The invention relates to the technical field of plant genetic engineering, in particular to separation, cloning and an application of two genes named SaM and SaF of a rice hybrid pollen fertility gene locus Sa. The bacterial two-hybrid experiments indicate that proteins coded by specific allele of the SaM and SaF of indica and japonica are subjected to specific interaction. In addition, genetic analyses and transgene functional complementation experiments indicate that the SaM heterozygous in a hybrid and the SaF originated from the indica, namely 3 alleles, interact to lead to infertility of hybrid pollen, while lack of any one of the three or inhibition of expression thereof results in hybrid compatibility, namely pollen fertility. The invention also relates to a method for adopting the genes to nurture rice hybrid fertility compatible line and an application thereof and a molecule marker for appraising rice hybrid fertility gene type and the application thereof.

Description

technical field [0001] The invention relates to the field of genetic engineering, in particular to the isolation and cloning of rice hybrid pollen fertility genes and their function verification, as well as their application in rice hybrid breeding. Background technique [0002] Plants produce hybrids by crossing between different varieties. Hybrids have advantages in growth potential, adaptability, yield, etc., which are called heterosis. The use of heterosis can improve the yield, quality and resistance of crops. Hybrids between indica (Oryza sativa ssp. indica) and japonica (O. sativa ssp. japonica) subspecies of Asian cultivated rice have strong heterosis and great potential for increasing yield. On the other hand, through breeding methods such as hybridization and backcrossing between cultivated rice and wild rice, some useful traits existing in wild rice, such as disease resistance, insect resistance, and high yield, can be introduced into cultivated rice to breed ex...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/63C12N5/10A01H1/00A01H1/02A01H5/00C12N15/11C12Q1/68
Inventor 刘耀光龙云铭赵利锋牛白晓庄楚雄
Owner SOUTH CHINA AGRI UNIV
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