Molecular marker of brassica napus L. grain weight character and preparation method and application
A technology of Brassica napus and molecular markers, applied in the direction of recombinant DNA technology, DNA / RNA fragments, etc., to reduce the breeding workload, speed up the process, and shorten the breeding period
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Embodiment 1
[0048] Example 1: Mapping of grain weight QTLs in Brassica napus
[0049] (1) Construction of DH population of Brassica napus relocation population A254 / A177, field test and thousand-grain weight analysis
[0050] 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 Preservation, its preservation number is CCTCCNO: 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, and ...
Embodiment 2
[0057] Example 2: Acquisition of gene-specific molecular markers for thousand-grain weight traits in Brassica napus
[0058] 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 MIN13F / 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
[0059] Example 3: Verification of the validity of gene-specific molecular markers for thousand-grain weight traits in Brassica napus
[0060] (1) Verification of gene-specific molecular markers for thousand-grain weight traits in Brassica napus
[0061] 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).
[0062] (2) Verification of the combined effects of QTLs for grain weight in Brassica napus
[0063] 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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