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Separating clone of rice wide compatibility gene S5 and uses thereof

A gene and rice technology, which is applied in the field of isolation and cloning of the rice broad-compatibility gene S5, can solve the problems of heterosis restricting the utilization of heterosis and the like

Active Publication Date: 2008-06-18
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Rice indica and japonica subspecies have stronger heterosis than rice cultivars, but the sterility of indica and japonica hybrids limits the use of heterosis

Method used

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  • Separating clone of rice wide compatibility gene S5 and uses thereof
  • Separating clone of rice wide compatibility gene S5 and uses thereof
  • Separating clone of rice wide compatibility gene S5 and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Isolation and cloning of DNA fragments comprising S5 gene segments

[0028] 1. Identification of rice wide-compatibility gene S5 by map-based cloning

[0029] In the present invention, three parents were used in constructing the targeting population: 02428, Nanjing 11 and Balilla. 02428 is a wide-compatibility japonica variety, which is selected from two japonica rice varieties Yunnan Xiegu and Shanghai Jibangdao, treated with radiation mutagenesis respectively, and crossed with materials selected by high light efficiency, and bred from the offspring ( Zou Jiangshi et al., Preliminary utilization of the wide-compatibility line 02428 in hybridization between indica and japonica subspecies. Chinese Agricultural Sciences, 1989, 22:6-14). Nanjing 11 is a medium indica variety bred by Jiangsu Academy of Agricultural Sciences, which is a derivative of Shengli indica. Balilla is a japonica variety imported from Italy.

[0030]In the preliminary work, the present ...

Embodiment 2

[0278] Example 2: Structural analysis of S5 gene in different subspecies of rice

[0279] The sequences obtained by sequencing in Example 1 were analyzed: the full-length cDNAs of rice varieties 02428 (wide affinity), Ballila (japonica rice) and Nanjing 11 (indica rice) were 1778bp, 1912bp and 1911bp respectively, and their structures consisted of three exogenous sub and two introns (Figure 9). Indica rice and japonica rice have one and only three base differences, of which Nanjing 11 has a single base deletion in the 3' untranslated region, Nanjing 11 and Ballila have two single base polymorphisms (single nucleotide polymorphism) in the coding region , SNP), these two SNPs form two missense mutations in the encoded protein (Figure 10). The S5 gene encodes a protein of 472 amino acids, and BLASTp analysis (http: / / www.ncbi.nlm.nih.gov / ) shows that the protein is highly homologous to aspartyl protease. PROSITE (http: / / www.expasy.ch / cgi-bin / prosite / ) analysis found that the pro...

Embodiment 3

[0280]Example 3: Using chip data to detect the expression level of S5 gene in the whole growth period of rice

[0281] The present invention utilizes the chip data (Database of the National Center of Plant Gene Research, available at http: / / crep.ncpgr.cn / .) to detect the expression level of the S5 gene in different tissues during the whole growth period of rice, and the chip data used include indica rice Zhenshan The expression profiles of 25 different tissues (Table 3) under different growth stages of 97 and Minghui 63 (Fig. 11). The results showed that the S5 gene was expressed at a very low level during the whole growth period.

[0282] Table 3 Microarray data corresponding to rice tissue name

[0283] Sample Tissue

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Abstract

The present invention relates to the technical field of plant gene engineering and concretely relates to the separation clone of paddy wide compatibility gene S5, a function validation and an application thereof for improving the paddy. The segment of the gene contains paddy wide compatibility gene coding aspartic proteinase. Subspecific Indica-Japonica paddy has strong hybrid advantage than varietal paddy, but the sterility of the subspecific Indica-Japonica hybrid restricts using the hybrid advantage thereof. The wide compatibility gene cloned by the present invention can conquer the sterility of the subspecific Indica-Japonica hybrid of the paddy and accordingly uses the strong hybrid advantage of the subspecific Indica-Japonica to further improve the output of the paddy.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering. In particular, it relates to the isolation and cloning, functional verification and application of a rice wide-affinity gene S5 in rice improvement. Background technique [0002] Oryza sativa belongs to Gramineae, Oryzeae of Pooideae, and is divided into two cultivated rice species: Asian cultivated rice (Oryza sativa L.) and African cultivated rice (Oryza glaberrima). Asian cultivated rice is divided into indica subspecies (ssp.indica) and japonica subspecies (ssp.japonica). There are obvious differences in morphology and genetics between the two subspecies, and there are also reproductive disorders. The hybrids between indica and japonica subspecies are often sterile or semi-sterile (Kato S, Kosaka H, ​​Hara S(1928) On the affinity of rice varieties as shown by fertility of hybrid plants. Bull Sci Fac Agric Kyushu Univ, 3: 132-147; Oka HI (1988) Origin of cultivated rice. Sc...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82
Inventor 张启发欧阳亦聃陈炯炯丁寄花刘克德
Owner HUAZHONG AGRI UNIV
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