| Castanopsis fissa and Camellia semiserrata are most local characteristics of native trees. C. fissa also has characteristics of natural seeding update and strong sprout regeneration, meanwhile, it is the better species of secondary forest, such as water supply conservation, energy, water and soil conservation forest. C. semiserrata is important mywood edible oil trees in southern of China. Studying on the genetic of C. fissa and C. semiserrata can contribute to understanding the genetic background and geographical distribution, which lay a foundation for germplasm resources protection and excellent provenance collection. Based on C. fissa and C. semiserrata as the research object by sequence-related amplisied polymorlpism(SRAP) can find its DNA molecular polymorphisms, at the same time, important economic characters of the evaluation system can provide powerful science basis. The mean results are as follows:1. Establishment and optimization of SRAP-PCR system in C. fissa and C. semiserrataOrthogonal experiment designing method was used to get the stable, reliable and repeatability system. The SRAP- PCR reaction system of 25 ?L was followed : 10 x PCR Buffer(Mg2+ free)2.5 ?L, DNA template 40 ng, Mg2+ 2.0 mmol·L-1, d NTPS 0.175 mmol·L-1, each primer 0.5 ?mol·L-1, Taq polymerase 0.2 U?2. Genetic Diversity Analysis of C. fissa13 pairs of SRAP primers were selected from 594 combinations and applied to analyze the genetic diversity. The results showed that a total of 236 fragments was amplified, and the percentage of polymorphic bands was 82.63%. The Nei's gene diversity(h), Shannon's information index(I) and percentage of polymorphic loci(PPL) of different populations were 0.0992-0.3066, 0.1488-0.4555 and 28.81%-85.59%, with the average 0.1956, 0.2923 and 55.24%, respectively, showing medium genetic diversity at population of C. fissa. The dendrogram indicated that when the threshold was 0.88, 28 populations of C. fissa was divided into six major categories that the first group consisted of the 5 populations from Yingde, Gaotian, Xingning, Yangdong and Xinxing of Guangdong Provinces. The second group had 6 accession, Huaiji, Wengyuan, Longchuan and Wuhua, which originated from Guangdong Province, Nanping and Yongan of Fujian province. The 5 populations from Fujian(excluding Fuzhou), Guangdong(including Wujiang, Yangchun, Dabu and Pingyuan) were grouped in the third cluster. Group four was composed of populations from Zengcheng(Guangdong), Shunchang(Fujian) and Zhenlong(Guangxi). Group five contained mostly accessions from Guangdong(Jiedong, Dinghu and Fengkai). The rest of the accessions assigned to group six. Genetic distance of Jiedong and Dinghu was minimum(0.0134), showing a closer relationship for these two populations. The analysis of molecular variance(AMOVA) showed the 60% of the genetic variation existed within populations and 40% among them.3. Genetic Diversity Analysis of C. semiserrata13 pairs of SRAP primers were selected from 594 combinations and applied to analyze the genetic diversity. The results showed that a total of 206 fragments was amplified, and the percentage of polymorphic bands was 90.78%. The Nei's gene diversity(h), Shannon's information index(I) and percentage of polymorphic loci(PPL) of different populations were 0.2179-0.3563, 0.3217-0.5248 and 58.25%-92.72%, with the average 0.3078, 0.4563 and 83.62%, respectively, showing rich genetic diversity at population of C. semiserrata. The dendrogram indicated that when the threshold was 0.93, 4 populations of C. semiserrata was divided into two major categories that Qingyuan population was one group and the second group included populations of Guangning County, Zengcheng and Yunfu. Genetic distance of Yunfu and Zengcheng was minimum(0.0193), showing a closer relationship for these two populations. The analysis of molecular variance(AMOVA) showed the 95% of the genetic variation existed within populations and 5% among them. |
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