Development Of Microsatellite Markers From Ramie And Its Application For Preliminary Study Of Heterosis Forecast

This research used three methods to isolate 39 ramie microsatellite sequences, Designed 39 pair of microsatellites sequence by Primer Premier 5.0, 28 cross combinations was established with 26 inbred lines, Investigated the main agronomic characters of 26 inbred lines and 28their hybridized combination , and the relationship was investigated. 30 SSR primers were used to detect genetic polymorphism for 26 inbred lines, DNA molecular genetic distance among these inbred lines were estimated. The relationship between the genetic distances based on SSR markers and heterosis of ramie was investigated.Main result:1) Develops SSR primers from the EST database. Searches NCBI the ramie (Boehmeria nivea (L.) Gaud) EST database, screens to 8 including the SSR sequence from 125 EST sequences, designs 8 pair of SSR primers.2) SSR primers developed using ISSR-suppression PCR and ISSR- reverse PCR technique, Suppression PCR and Reverse PCR primers were designed to conduct genome walking after 10 ISSR primers amplified in genomic DNA. 10 SSR primers was obtained by suppression PCR, 9 SSR primers was obtained by reverse PCR.3) The method is AFLP and magnetic beads enrichment unifies. The fragments of AFLP pre-amplifacation were bond with linkers, those sequences were hybridized with biotin-labeled simple sequence repeat probes (AC)₁₅, Microsatellites were efficiently isolated by using magnetic beads coaled with streptavidin. Genomic library contained the selected microsatellite DNA was constructed, 25 positive clones were isolated with PCR method, we got 25 sequences and successfully designed 20 pairs of SSR primers.4) 30 pair primers developed SSR primer were introduced to determine genetic distance of 26 inbred lines, and SAS 8.0 was used to conduct correlation analysis between genetic distance and yield and quality characters of F₁. The result indicated there was no significant relationship between them.