Constructionand Analysis Of BAC Contigs Around The Yellow-Seeded Locus Of The Chromosome B03 In Brassica Juncea

Oilseed brassica can be divided into yellow-seededrape,black-seeded rape,brown-seeded rape.In comparison, in the same genetic background, Yellow-Seeded than Black-Seeded more advantages in oil and cellulose content, protein content and shavings feeding and so on. So yellow-seeded oil occupies an important position in the modern improvements in breeding and improvement.In previous studies, we have been positionedBrassica juncea yellow seed genes in A09 and B03 two linkage groups. In this study, Through the constructed Brassica juncea genomic library,using the B03 chromosome yellow seed region of the gene markers closely linked B15₃3151, B15₃884, Ni4F11, B15₃9736, ST8, SY15₅263 by PCR walking screening build Brassica juncea B03 chain BAC contigs group of yellow seed gene regions.Using the purpose gene region of BAC end sequence designed 219 pairs of STS primers; In addition, the use of Brassica juncea unigene （through the mustard RNA_seq technology of rape seed non redundant genes） sequence design for 36 primers;use Brassica juncea genome Survey sequences designed 41 pairs of primers; Also synthesized 15 IP marker primers. With nearly isogenic line on the part of the tag polymorphism analysis, whether the inspection tag and objective traits chainThrough the screen ZBjuH libraries available from a total of 640 BAC clones, of which 435 are at the end of sequencing, get 866 BAC end sequence. And utilization Brassica juncea B03 chromosome markers closely linked B15₃3151, B15₃884, B15₃9736, SY15-5263 and BAC as a fulcrum, constructed sixth chromosome yellow Brassica juncea seeds B03 gene region totaled approximately 3.3Mb of BAC contigs.Selected the five BAC for full-length sequencing, return sequence is determined two full-length sequences （ZBjuH036F18（138109bp） and ZBjuH090N23 （125151bp））located on Brassica junceaB03 chromosome,Analysis by the laboratory notes, found that the two BAC consisting of overlapping groups （long 254376bp） encoding 12 genes, and contains the transcription factor gene TT8. Through Markers and BAC end sequencing,it was found that highly repetitive sequences are present in the area with yellow seed B03, A03, A09.Get a large number of linked markers and sequence of Brassica juncea, to reveal the molecular mechanism of yellow seed Brassica juncea formed a foundation to provide reference for yellow seed-assisted breeding.