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Specific molecular markers of related genes of brassica napus grain weight and application thereof

A molecular marker technology for Brassica napus, applied in the fields of application, genetic engineering, plant gene improvement, etc., to achieve the effects of reducing the breeding workload, speeding up the process, and shortening the breeding period

Inactive Publication Date: 2011-02-02
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] There is no report on the cloning and analysis of genes related to grain weight in rapeseed

Method used

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  • Specific molecular markers of related genes of brassica napus grain weight and application thereof
  • Specific molecular markers of related genes of brassica napus grain weight and application thereof
  • Specific molecular markers of related genes of brassica napus grain weight and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1: Mapping of grain weight QTLs in Brassica napus

[0047] (1) Construction of DH population of Brassica napus relocation population A254 / A177, field test and thousand-grain weight analysis

[0048] Adopt Brassica napus A A254 (large grain pure line, the seed of this material has been delivered to the Chinese Type Culture Collection Center (CCTCC) in Wuhan University, Wuhan City, Hubei Province on November 20, 2009 for preservation, and its preservation number is CCTCC NO: P200909) as the female parent, Brassica napus A A177 (small grain pure line, the seeds of this material have been sent to the Chinese Type Culture Collection Center (CCTCC) in Wuhan University, Wuhan, Hubei Province on November 20, 2009 Preserved, and its preservation number is CCTCC NO: P200908) is used as the male parent to cross to obtain F1, and the flower buds of F1 are subjected to microspore culture to obtain DH segregation population. A total of 238 lines of DH lines were obtained, an...

Embodiment 2

[0055] Example 2: Acquisition of gene-specific molecular markers for thousand-grain weight traits in Brassica napus

[0056] In Arabidopsis, the MINI3 and TTG2 genes have been shown to be important genes controlling seed size and grain weight. In order to explore the possibility of using gene-specific markers of grain weight-related genes MINI3 and TTG2 in Brassica napus, an experiment was designed to isolate homologous sequences in Brassica napus, by searching the NCBI nucleotide database (http: / / www .ncbi.nlm.nih.gov / nucleotide / ), two cabbage BAC clones with high homology to Arabidopsis MINI3 and TTG2 gene sequences were found: AC189531 and AC232555, and these two BAC clones were located on chromosome A5 superior. According to the gene sequence information, two pairs of primers MINI3F / R and TTG2F / R were designed to amplify the full length of the gene. The genome fragments of these two genes were amplified from the two parents A254 and A177 of the DH population, cloned and se...

Embodiment 3

[0057] Example 3: Verification of the validity of gene-specific molecular markers for thousand-grain weight traits in Brassica napus

[0058] (1) Verification of gene-specific molecular markers for thousand-grain weight traits in Brassica napus

[0059] Compared with the two loci on A7, the contribution of TTG2a and MINI3a loci to phenotypic variation is small. In order to detect the genetic effect of TTG2a and MINI3a loci on phenotypic variation, the genotype pair The lines of the DH population were grouped and the average value of the thousand-grain weight was calculated, and the values ​​of the thousand-grain weight of each of the two genotypes of the TTG2a and MINI3a loci in the DH population were significantly different (Table 3).

[0060] (2) Verification of the combined effects of QTLs for grain weight in Brassica napus

[0061] The combined effects of QTLs loci for grain weight were detected in the DH population. The lines of the DH population were grouped by the geno...

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Abstract

The invention belongs to the field of rape molecular breeding, and relates to preparation of specific molecular markers of related genes MINI3 and TTG2 of the brassica napus grain weight. Double haploid colony (DH) is constructed with brassica napus I A 254 as a female parent and a brassica napus I A 177 as a male parent through hybridization, and the DH colony genotype and the thousand seed weight data are analyzed to obtain a QTLs locus of grain weight character. The MINI3 and the TTG2 genes of the IA254 and the IA177 are cloned by using a homology based candidate gene method, specific molecular markers MINI3a and TTG2a of the MINI3 and the TTG2 genes are designed according to sequence different locuses, and the molecular markers MINI3a and TTG2a are located on two grain weight QTLs locus of an A5 linkage colony for related verification and application, which proves that the molecular marker prepared by the invention is a novel genetic marker. The gene sequence is obtained firstly. The invention provides a novel marker for the molecular breeding of the brassica napus grain weight, and also provides useful information for candidate gene clone and marker auxiliary selection of thethousand seed weight character locuses of the brassica napus.

Description

technical field [0001] The invention belongs to the field of rapeseed molecular breeding and biotechnology, and specifically relates to the discovery, cloning and identification of genes related to grain weight of Brassica napus, and the development and application of specific molecular markers of related genes. Background technique [0002] Brassica napus L. (hereinafter referred to as rapeseed) is one of the most important oil crops in the world. The seeds of rapeseed are not only the storage organs of oil and protein, but also the organs of plant life cycle continuation. Seed size or weight are very important economical traits. First of all, grain weight is one of the three major factors constituting plant yield per plant (effective silique number per plant, number of silique grains per plant, and grain weight), and therefore also determines yield (Clarke and Simpson, 1978; Butruille et al., 1999; Shi et al., 2009); secondly, seed size is also related to oil and protein...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/29C12N15/11C12N15/10
Inventor 周永明傅廷栋范楚川蔡光勤
Owner HUAZHONG AGRI UNIV
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