Study Of Molecular Fingerprint Identification Technology Of Germplasm Resources Of Fritillaria Thunbergii Miq.

RAPD (random amplified polymorphic DNA) technology was built on the basis of PCR, which could analyze the polymorphism of the whole genome that may be unknown. RAPD was started via PCR amplification used genome DNA as template, a single synthetic random polymorphic nucleotide sequence (typically10base pairs) as primer under DNA polymerase (Taq enzyme). ISSR (intersimple sequence repeat) technology was one of molecular markers based on microsatellite. Used anchored microsatellite DNA as primers, that is plus2-4random nucleotides in the3’end or5’end of SSR sequence, which could cause anneal at specific sites, leading to PCR amplification in the PCR reaction.Fritillaria thunbergii Miq was a traditional Chinese herbal medicine, which was one of the famous "Zhe Ba Wei", and had a high medicinal and economic value. Currently, it’s main cultivation base was in Yinzhou, Panan, Jinyun and other regions, while Panan had already become the main origin and the important bases of seed breeding and seed output. However, germplasm mixture phenomenon of Fritillaria thunbergii Miq in Panan was found recently, which had a bad influence on the breeding of Fritillaria thunbergii Miq. In this paper, the genetic diversity of different varieties and the difference within same variety were investigated, which could provide the foundation for the breeding of new varieties.In this paper, RAPD markers and ISSR marker were used to analyze12different germplasm of Fritillaria thunbergii Miq. The results were as follows:(1) RAPD markers had a good percentage of polymorphism and the majority was more than90%. The average of GS of all Fritillaria thunbergii Miq germplasm was0.6006with0.4267～0.7467range. The GS value (0.7467) between multi-seed fritillaria and local big fritillaria in Panan Xinwo was maximum, while the GS value (0.4267) between small fritillaria and multi-seed fritillaria in Panan Xinwo was minimum. The GS value within local big fritillaria in Panan Xinwo was minimum, while the GS value within multi-seed fritillaria Panan Xinwo was maximum. Eight varieties of Fritillaria thunbergii Miq could be divided into two groups. The first group included fraxinus baroniana Fritillaria in panan, local big fritillaria Panan Xinwo, big fritillaria in Panan Xinwo, multi-seed fritillaria in Panan Xinwo, fraxinus baroniana fritillaria in Panan Zhaikou village, middle fritillaria in Panan Xinwo. Meanwhile, it was also found that multi-seed fritillaria in Panan Xinwo and local big fritillariain Panan Xinwo, big fritillaria in Panan Xinwo and middle fritillaria in Panan Xinwo were holded together, respectively. The second group included small fritillaria in Nantong and fraxinus baroniana Fritillaria in Panan Shuangfeng.(2) ISSR marker analysis showed that the average of GS value of all Fritillaria thunbergii Miq germplasm was0.6399with0.4333-0.9333range.The GS value (0.9333) between small fritillaria in Nantong and fraxinus baroniana Fritillaria in Panan Shuangfeng was maximum. The GS value (0.4333) between small fritillaria in Nantong and big fritillariain Panan Xinwo, local big fritillaria in Panan Xinwo were minimum. In the same time, the GS value (0.4333) between local big fritillaria in Panan Xinwo and fraxinus baroniana Fritillaria in Panan Shuangfeng, small fritillaria in Nantong, multi-seed fritillaria in Panan Xinwo was also minimum. Genetic similarity of multi-seed fritillaria in Panan Xinwo was the highest, while that of local big fritillariain Panan Xinwo was the lowest.(3) The results of molecules fingerprint of12Fritillaria tgunbergii Miq germplasm using ISSR markers and RAPD markers were mostly identical. It was that the GS values between three fraxinus baroniana Fritillaria in Panan different regions from two marker methods were all small, while the GS value between small fritillaria in Nantong and fraxinus baroniana Fritillaria in Panan Shuangfeng were both high. The GS value between big fritillaria and local big fritillaria in Panan Xinwo was the highest. As to intraspecific difference, the GS values among multi-seed fritillaria in Panan Xinwo by two methods were both the highest and the lowest was located among local big fritilllaria in Panan Xinwo.(4) There had some different results from ISSR markers and RAPD markers. The GS value between multi-seed fritillaria and local big fritillaria in Panan Xinwo from RAPD markers was the highest, but that from ISSR markers was the lowest. The GS value between multi-seed fritillaria in Panan Xinwo and small fritillaria in Nantong from RAPD markers was very low, but that from ISSR markers was very high.