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Paddy rice interspecific hybrid pollen fertility gene and application thereof

A fertility gene, rice hybrid technology, applied in the application field of rice hybrid breeding, can solve the problem that the positioning interval is not precise enough and the like

Active Publication Date: 2018-10-23
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These loci are called different names in different hybrid combinations. Subsequent studies have found that some loci may widely exist in indica and japonica subspecies or rice species, or there may be epistasis interactions between them, which together determine hybrid Infertility; on the other hand, due to the limitation of the research conditions at that time, the positioning interval was not fine enough, and it is possible that a previously reported locus was later decomposed into several loci

Method used

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  • Paddy rice interspecific hybrid pollen fertility gene and application thereof
  • Paddy rice interspecific hybrid pollen fertility gene and application thereof
  • Paddy rice interspecific hybrid pollen fertility gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] Example 1: Screening of polymorphic molecular markers and fine mapping of qHMS7

[0048] In the present invention, firstly, the japonica rice variety DJY1 is used as the recurrent parent, and the southern wild rice (O. meridionalis, Mer) with a relatively distant relationship is used as the donor parent to construct the BC of DJY1 / Mer / / DJY1 1 f 1 QTL detection was carried out in the population, and it was found that qHMS7 had a large effect, and it was located near the site found in different combinations by the predecessors. It may be a hybrid sterility site commonly found among rice species, so it is a good choice for the study of rice hybrid sterility. Ideal sites (Lin and Ikehashi, 1993; DOI et al., 1998; Sobrial et al., 2000; Yan et al., 2003a; Chen et al., 2006; Miyazaki et al., 2006a). A backcross approach via molecular marker-assisted selection, ultimately in BC 6 f 2 A single plant with a small Mer insert, a clean genetic background, and semi-sterile pollen ...

Embodiment 2

[0063] Example 2: Sequence analysis of genes associated with the qHMS7 locus

[0064] According to the prediction and analysis of NCBI and RGAP (Rice genome annotation project) websites, within the interval: there are three expressed proteins in the japonica rice line DJY1, named ORF1 respectively D , ORF2D , ORF3; while only ORF1 exists in southern wild rice (Mer) M and ORF2 M Two genes, the DNA sequences of ORF1, ORF2 and ORF3 were finally clarified through the first-generation sequencing technology, and the ORF2 of DJY1 D , the nucleotide sequences corresponding to ORF3 are SEQ ID NO: 1, SEQ ID NO: 7; their corresponding protein sequences are SEQ ID NO: 2, SEQ ID NO: 8; the ORF2 of Mer M The corresponding nucleotide sequence is SEQ ID NO:3; the corresponding protein sequence is SEQ ID NO:4. Gene and protein structures of rice qHMS7 loci ORF2 and ORF3 image 3 shown, where image 3 A shows ORF2 D , ORF2 N , ORF2 M A schematic diagram of the protein structure, image...

Embodiment 3

[0065] Example 3: Genetic transformation to clarify the function of the target gene

[0066] ORF2 in Example 2 D , the complete genome segment of ORF3 (including the promoter region and the 3' non-coding region) was cloned into the binary transformation vector pCAMBIA1305 (CAMBIA, Australia). The vector confirmed by sequencing was transformed into Agrobacterium EH105 by chemical transformation method to infect the callus. When NIL-qHMS7 was used as the transgenic recipient, the recipient genotype was D / M, and the ORF3 from DJY1 would partially restore its pollen fertility to about 75% ( Figure 4 , Figure 4 A shows that the pollen of the heterozygous strain (D / M type) used as the transformation recipient is semisterile, Figure 4 B shows that the pollen fertility of the heterozygous line (D / M type) is partially restored (about 75%) after the transgene is introduced into ORF3, Figure 4 C shows that M / M type plants carrying the transgenic ORF3 fragment appeared after ORF3 ...

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Abstract

The invention discloses a paddy rice interspecific hybrid pollen fertility gene and an application thereof. The gene concretely relates to a paddy rice interspecific hybrid pollen fertility related locus qHMS7, is positioned at a 7th chromosome long arm, and accurately positioned in a 31.6 kb area between molecule marks D17 and YU-33, and a DNA fragment of the related gene of the paddy rice interspecific sterility site qHMS7 is provided, a protein sequence coded by the hybrid pollen fertility gene is provided, a concrete application method of the hybrid pollen fertility gene in a seed selection affinity system during a paddy rice heredity improvement process is provided, and the tight linkage molecule mark generated according to the hybrid pollen fertility gene sequence and an applicationof the tight linkage molecule mark in the paddy rice cross breeding are provided.

Description

technical field [0001] The invention relates to the technical field of plant genetic engineering, in particular to the isolation and cloning of two genes of the rice interspecific hybrid pollen fertility locus qHMS7, functional verification and application in rice hybrid breeding. Background technique [0002] There is strong heterosis in hybrids between rice subspecies or between Asian cultivars and wild relatives, and there is great potential for yield increase. Wild relatives also provide excellent germplasm resources for genetic improvement of Asian cultivated rice. Rice heterosis tends to increase with the increase of genetic distance between parents, but the accompanying reproductive isolation will also become more serious, which adds natural obstacles to the exploration of rice yield potential (Peng et al., 1999). Therefore, in order to make full use of these rich genetic resources, improve existing rice varieties, and utilize heterosis among species, the first prob...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/29C07K14/415C12N15/82C12N15/11
CPCC07K14/415C12N15/8213C12N15/8287C12Q1/6895C12Q2600/13C12Q2600/156
Inventor 万建民余晓文赵志刚张欣江玲朱杉杉刘喜刘世家田云录陈亮明刘裕强王益华
Owner NANJING AGRICULTURAL UNIVERSITY
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