Development Of EST-SSR Maker And Application In Sunflower

This article adopts the method of bioinformatics to search, analysis sunflower EST-SSR and the design of primers, EST-SSR molecular marker is discoverd, with a significant difference was found in drought in sunflower oil are optional for the parent inbred lines K55, K58 187 F7 recombinant inbred lines (RILs) groups for mapping, EST-SSR molecular marker detection will detect polymorphic EST-SSR markers encryption to built a SSR markers and SRAP markers on the high density genetic map in laboratory, in order to the sunflower map-based gene cloning, molecular marker assisted breeding and so on work to lay the important research foundation. The main results were as follows:1. Through the download from the NCBI 134468 sunflower EST sequence, is obtained by pretreatment, clustering, joining together the 56998 sequences. The MISA script search from 56998 sunflower EST-SSR loci, including contigs8029, singletons48969,search 3602 SSR loci, accounted for 2.68% of the whole EST database, distributed in 3169 without redundant EST, its frequency is 5.6% (contains SSR EST and not the ratio of the redundant EST). Among 370 of the EST containing more than one SSR.2. In search of SSR, two to six nucleotide repeat all exist, but the freque ncy difference is bigger. The highest trinucleotide repeats, a total of 1947, acc ounting for 54.05%,followed by the dinucleotide repeat,1348,and accounted for 34.42%, four, five, six repeated nucleotide repeat is lower than the two, three nucleotide repeat, accounted for only 11.53%.From the point of the frequency of occurrence, dinucleotide and trinucleotide repeat types dominate, the freque ncy of close, accounted for 88.47% of total SSR.3.The use of any material in the recombinant inbred lines 187 and canopy K55 parents and K58 combination as a template to design for 130 EST-SSR primers screened, selected 40 primer combinations with good results in polymorphism, accounting for 30.77% of the total design primers.40 EST-SSR primer combination testing 187 recombinant inbred lines group, amplify the 137 sites, including polymorphism loci86, and the average of EST-SSR primer combination amplified out 2.15 polymorphism loci, total polymorphism rate of about 62.77%.4. This study use EST-SSR molecular marker encrypt sunflower genetic linkage map, the map will be 78 EST-SSR polymorphism markers loci locate on 17 linkage group, the encrypted map increased from 3846.04 cM to 3863.93 cM in length, average distance is shortened by 3.48 cM to 3.27 cM. As you can see, mark in the new encryption map site of the communist party of China has 370 partial separation of site, accounting for 31.28% of the total site.27 the SDR, respectively at 1,2,4,5,6,7,8,9, 11,12,15,16 chain group, showed that partial separation of loci may exist in these chromosomal regions. EST-SSR molecular markers for genetic map of sunflower encryption, prove the advantage of rich EST-SSR molecular marker to encrypt the feasibility of the genetic map.