Identification Of Purity Of Oil Sunflower (Helianthus Annuus L.) Hybrids In Xinjiang With Fingerprinting Technology

In this thesis, we conducted research work in identification of purity of oil sunflower (H. annuus) hybrids grown in Xinjiang with protein electrophoresis technique and RAPD molecular markers. We used several oil sunflower varieties --- Kangdi No.4, No.5, No.6, No.7 and G101 (including hybrid and its parents) as experimental materials, results were shown as following. Results of protein electrophoresis showed: (1) proper concentration and crosslinking degree of gel could improve the resolution of the electrophoresis bands; (2) comparing results of SDS-PAGE of seed protein with various protein extraction solutions, we chose the one which included 1mol/L urea in it as the extraction solution, its protein was rich and electrophoresis bands appeared better, but the special and characteristic band was few; (3) results of electrophoresis of the esterase and peroxidase isozymes in cotyledon stage showed: in Kangdi No.6 hybrid, there were some special bands of esterase isozyme coming from its parents separately, that is, the hybrid had parental complementary characteristic bands. With this result, production hybrids were tested and the results were identical with that of field planting identification; in G101 hybrid, there was one new band of peroxidase isozyme in cotyledon stage which didn't appear in result of parents electrophoresis bands. Result of purity testing to G101 production hybrid with this special band also matched well with that of field planting identification.Results of RAPD-PCR showed: (1) comparing different methods (CTAB, SDS) of total DNA extraction from different materials (young leaves, seed), we considered that SDS method using seed as material was much simpler and quicker than others; (2) In optimizing of the reaction components (concentration of template DNA, Taq DNA polymerase, primer, dNTPs etc) and procedure (annealing temperature, extention time, etc.) of RAPD-PCR, we developed the optimization reaction system of RAPD-PCR for oil sunflower in our lab; (3) using 100 random 10-mer oligonucleotide primers screened varieties mentioned above with the optimization reaction system of RAPD-PCR, results showed that primer OPB05 generated the different parental complementary characteristic bands in Kangdi No.6, No.7 hybrids separately; Kangdi No.4 hybrids with primer OPB03, Kangdi No.6 hybrid with primer OPC14 could also generate parental complementary special bands; (4) with above results, three hybrids (Kangdi No.4, No.6, No.7) of their production seeds were tested, and the results all matched well with those of field planting identification.