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Studies on EST-SSR and SRAP Markers in Flax

Author: WangBin
Tutor: DangZhanHai；ZhangJianPing
School: Gansu Agricultural University
Course: Crop Genetics and Breeding
Keywords: Flax EST-SSR SRAP Temperature Sensitive Male Sterile Line Fertility gene
CLC: S563.2
Type: Master's thesis
Year: 2010
Downloads: 75
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Abstract


EST-SSR markers is based on the development of a molecular marker of EST or cDNA data. As a new SSR markers, EST-SSR markers from expressed genes, in addition to with traditional sources of genomic SSR markers all the benefits may be related to the function of gene expression has a direct or indirect relationship, thus reinforcing the SSR markers in genetic research. In this study, flax public database EST sequences developed SSR markers and flax Temperature Sensitive Male sterile line gene screening primers EST-SSR markers. Developed flax the SRAR mark and optimize the reaction system. The main results are as follows: 1 pair of linen EST sequences screening of microsatellite sequences found SSR repeat sequence 222, accounting for 2.73% of the EST database, which trinucleotide repeat unit sequence dominant, accounting for SSR sequences 72.1%; dinucleotide and trinucleotide repeats, respectively, 14.4% and 13.5% of the total sequence of SSR. AGAA advantage in the four-nucleotide repeat type, accounting for 67.67% of the four-nucleotide repeat type. According to the microsatellite sequences obtained by screening EST database design 21 EST-SSR primer pairs. 3 linen EST-SSR reaction system optimization, established the optimal reaction system. A 20uL reaction system: 25mmol/LMg2 the concentration for the 1.9 mmol / L and a 10mmol/LdNTPs the concentration of 0.55 mmol / L and an amount of 1.5U Taq enzyme 25ng/uL primers used in an amount of 75 ng · The amount of 50ng/uLDNA 100ng . 4 EST-SSR primers, 18 pairs of amplification products in 10 flax materials, accounting for 85% of the primers were designed, of which 14 pairs of primers were polymorphic. Cluster analysis based on SSR markers, 10 flax materials can be divided into three groups. By segregating populations packet analysis (BSA), 1S × 81A350 crosses F2 segregating populations the sterility gene pool (BF) and the sterility gene pool (BS) fertility gene SSR markers preliminary study. Repeated experiments, 14 EST-SSR primers screened a the polymorphic primers the SSR757 may linen Temperature Sensitive Male sterile lines fertility gene linkage. 6 flax SRAP markers, the reaction system was optimized. A 20uL reaction system: 25mmol/LMg2 the concentration for the 1.5 mmol / L, 10mmol/LdNTPs concentration of 0.3 mmol / L, 1.5UTaq enzyme, 25ng/uL primers used in an amount of 100ng, 30ng/uLDNA dosage 90ng. 10 flax materials to verify the optimized system, the results show that the clear bands, polymorphism, get good results.

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CLC: > Agricultural Sciences > Crop > Economic crops > Fiber crops > Fiber crops > Flax
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