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Comparative Analysis of the Genomic Region Containing Recessive Genic Male Sterility Gene in Rapeseed and Development of Sequence-specific Markers

Author: MaLei
Tutor: YangGuangSheng
School: Huazhong Agricultural University
Course: Crop Genetics and Breeding
Keywords: Brassica napus Recessive genic male sterility gene Comparative genomics Micro - collinearity Fine mapping
CLC: S565.4
Type: Master's thesis
Year: 2009
Downloads: 14
Quote: 1
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Abstract

Brassica napus is formed by the cabbage and cabbage hybridization of natural double A, C two genomic information contained in the genome. A, C genome is about 400 million years ago by a common Brassica genus ancestors differentiation from two highly conserved genome structure and micro-collinearity (Rana et al, 2004). Brassica napus genetic mapping studies are likely to be homologous genome. Study the variation between the A, C genome from the DNA sequence level, and micro collinearity between homologous genomic help distinguish between the two, will Brassica napus. 9012AB is a new the Brassica napus recessive genic male sterility material has completely Abortion, sterility and stability, restore the characteristics of a wide range of sources, and therefore has an important supporting breeding of the three-line use value. Chen Fengxiang of the material: its fertility by two duplicate recessive male sterile gene (ms3ms3, ms4ms4) to inhibit gene (rfrf) and a pair of recessive epistasis interaction control. He Junping has been one of the infertility gene MS3 fine mapping in 0.034cM genetic interval. The physical mapping showed that the interval of at least 6 BAC clone consisting of a complete contig covering, and wherein a BAC clone containing the target gene subcloned sequencing method. In order to further the development of closely linked markers Ms3 and verify that the BAC clone screening results and compare the interval between cabbage and cabbage genome micro-collinear MS4 possible convenience, this study from the BAC clone sequencing sequence randomly selected 20kb region in different materials to compare sequencing and analysis, for the following main results: 1. designed according to 20kb region of 18 pairs of primers amplified products was theoretically able to form a complete contigs and completely covered the interval. 18 pairs of primers in the 9012A, 9012B, cabbage and kale amplification, amplification product stitching three materials 52 segment sequence information. To compare and analyze section sequence of the resulting material found there is a good micro-collinearity the Brassica napus Tapidor, 9012A, 9012B, cabbage and kale, and the similarity between Brassica napus 9012A and cabbage high 9012B, Tapidor and cabbage similarity between the high. Reference model plant Arabidopsis further gene prediction: section 3 the corresponding gene in the same direction, the exon regions consistent and more conservative; the intron segment sequence a base mutations, insertions or deletions. A larger area based on the sequencing of the resulting sequence differences developed between the 9012A and 9012B 6 polymorphic markers, cabbage and kale the polymorphic markers DM1 and HWY. After recombinant monoclonal verify 9012A and 9012B sequence differences between the development of new markers in a segregating population of 20,391 individuals ms3 gene were isolated. And compared to the original positioning results, developed a new mark, but not narrowed ms3 gene physical range. According to the sequence divergence of cabbage and kale to develop a stable polymorphism between 49 cabbage and 8 cabbage codominant marker 2. Brassica napus BAC clone library, previously identified using the marker screening markers in the cloning, amplification product with cabbage genome size and sequence is not found with the Brassica genome size consistent with clone. According to previous studies, the MS3 gene positioning N19 linkage groups, corresponding cabbage chromosome 9. Presumably, the screening of the BAC clone may be ms3 homologous region of the gene located in the genome of the cabbage. Developed SSR and the experimental use of BAC clones developed A, C genome specific markers DM1 MS4 mapping population polymorphism, all markers in the mapping population MS4 gene chain, suggesting that ms4 gene may simply not Ms3 of a homologous gene, but two different genetic origin. Therefore, MS3 and MS4 this statement overlap recessive gene that may require further argument.

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