首页|Detection of DNA Polymorphism in Japanese Rice Cxiltivars Using the Restriction Landmark Genomic Scanning (ELGS) Method
Detection of DNA Polymorphism in Japanese Rice Cxiltivars Using the Restriction Landmark Genomic Scanning (ELGS) Method
扫码查看
点击上方二维码区域，可以放大扫码查看
原文链接
NSTL
Genetic variation among eight Japanese landraces or selections from landrace rice cultivars (cvs. Aikoku, Asahi, Jikkoku, Kameji, Kamenoo, 6ba. Shinriki and Takenari) and seven closely related improved Japanese rice cultivars (cvs. Hatsunishiki, Honenwase, Koshihikari, Koshijiwase, Yamasenishiki, Hitomebore and Sasanishiki) were analyzed using the restriction landmark genomic scanning (RLGS) method followed by image analysis. RLGS was carried out basically according to the protocol of Asakawa (1996)with a slight modification for the analysis of rice DNA. Approximately 250 ng of rice total DNA extracted from young leaves of each cultivar was doubledigested with Not I and EcoR V and the resulting Not I cleaved ends were labeled by a fill-in reactionwith alpha-[~(32)P]dGTP and alpha-[~(32)P]dCTP. About 100 ng of treated DNA fragments were fractionated through agarose gel by the first dimension electrophoresis. After the gel portion containing DNA fragments of 7 kbp to 500 bp was treated with Mbo I,it was fused onto the top of a polyacrylamide gel, and then the second dimension electrophoresis was conducted. The gel was dried and exposed on an X-ray film by autoradiography. The RLGS profile was processed using a two-dimensional electrophoregram analyzer system. A very clearly resolved area with approximately one thousand spots on the profile was chosen to compare the cultivars used. Among the eight old cultivars, the average number of spots that differed between a pair of cultivars was about 166 in the area analyzed, and the calculated value of genetic similarity (GS) varied from 0.902 to 0.928, The level of genetic diversity in Japanese rice revealed by isozymes and RFLP studies was re-appraised by the ELGS analysis. While seven improved cultivars produced very similar but distinguishable RLGS profiles and a higher GS value was obtained, ranging from 0.936 to 0.959. Thus, RLGS is able to reveal a number of informative genetic markers in such closely related cultivars and measure genetic similarity between rice cultivars. The highest GS value was obtained between Koshihikari and Hitomebore, which corresponds with similar pedigree. Consequently, RLGS can effectively detect DNA polymorphism among closely related rice cultivars in Japan.
NSTL
