Different Provenances of Vernicia Montana’s Diversity Analysis in Genetic Structure
|School||Fujian Agriculture and Forestry University|
|Keywords||Aleurites montana RAPD Genetic structure Variance Analysis|
Tung tree species Aleurites montana (Vernicia Montana), industrial oilseed species endemic to China, has a long history of cultivation, high oil content, a wide range of uses, our nation's most ideal biodiesel feedstock species one, at home and abroad Aleurites montana mainly focused on the physiology and biochemistry, seedling cultivation and resource development and utilization, very few of its research in molecular markers and genetic breeding, the paper by RAPD Aleurites montana 33 individuals of 11 kinds of sources The molecular marker experiment, Aleurites montana differentiation of differences in the genetic structure and phylogenetic relationship Variation, provides a theoretical basis for the genetic structure of different provenances Aleurites montana Variation kinship division germplasm resources protection and breeding. Compare CTAB method and plant genomic DNA extraction kit, selected DNA purity and impurity content is less and less protein and phenol residual RNA contamination less moderate concentrations of DNA amplified bands relatively complete, the total more clear and bright, tail-less plant genomic DNA extraction kit as the best method for total DNA extraction Aleurites montana. Content and annealing temperature on the the dNTP, Tag enzymes primer, template DNA, and Mg 2 sup> five reactants do gradient optimization experiments, to finalize the Millennium Tong RAPD-PCR amplification procedure was as follows: 94 ℃ degeneration 3min denaturation at 94 ° C for 1 min, 36.4 ° C annealing 1min extend an extension at 72 ° C for 1.5min, cycle 44 times, then 72 ° C 5min. PCR reaction system for a total volume of 25μl dNTP (2.5mmol / l), 3.0 μl Tag enzyme (2.5U/μl) 0.4 μl primer (5 μmol / l) 1.5μl DNA (10ng/μl) 2.0μl mg 2 sup> (25 mmol / l) 3.0μl, 10 × PCR buffer2.5μl, sterilized ultrapure water 12.6μl. 105 random primers screened and Aleurites montana sample of 33 of the 11 kinds of sources, filter out 14 specific primers for RAPD amplification, a total of 130 DNA bands, of which 119 polymorphic bands polymorphic percentage of 91.54%, on average, each primer 8.5 polymorphic bands, indicating a large genetic differences between the Millennium Tong samples of different provenances. 11 kinds of source Nei's observed number of alleles Na 1.8923 and 1.8769, Nei's effective allele number Ne = 1.5248 and 1.4896, Nei's genetic diversity H = 0.3088 and .2949, Shannon's information index I = 0.4639 and 0.4472, these four important index to measure the genetic diversity of the various populations shows a high level of genetic diversity among 11 kinds of sources. Governance (H = .2631, I = 0.3892), Lake City (H = 0.2669, I = 0.3887), Bookstore (H = 0.2617, I = 0.3819), organ (H = 0.2533, I = .3751) and a small lake (H = 0.2218, I = 0.3316) five kinds of sources Nei's genetic diversity index and Shannon's information index is relatively high and close to the five kinds of sources constitute the first level of genetic diversity. Shaxian (H = 0.1679, I = 0.2452), Youxi (H = 0.1287, I = 0.1886) and Putian (H = 0.1226, I = 0.1797) three kinds of sources Nei's gene diversity and Shannon's information index were lower, The three kinds of sources geographically dispersed long distances, hybridization and interspecific exchanges less, better preservation of germplasm resources, homozygous high. Huangkeng Xu City (H = 0.0898, I = 0.1289) and Zhangzhou pier (H = 0.0748, I = 0.1079), three kinds of sources of Nei's genetic diversity index and Shannon's information index (H = 0.1045, I = 0.1530), compared with low and close to the high genetic diversity. Aleurites montana 11 kinds of sources, kinship farthest provenance authorities, provenance Zhanglong pier between genetic distance 0.3693, distantly related the Provenance authorities and the provenance of the pit, as well as the kinds of sources Zhangzhou pier governance with the provenance and provenance small lake, the genetic distances were 0.3271,0.3286 and 0.3174. Between kinship recent provenance Youxi, provenances Xu City, the genetic distance was 0.0510, closer relationship is the provenance Huangkeng provenances Xu, and genetic distance 0.0603 11 provenances Millennium Tong similarity coefficient range of 0.9503 to 0.6912 between the highest similarity source Youxi provenances Xu City. similarity lowest provenance authorities and provenances the Zhangzhou pier, the frequent exchanges between various provenance polymorphism rich, wide range of genetic and geographic complexity of the environment will also affect the exchange variation between provenances. Similarity coefficients between the various provenances basically 1, but the difference is not the greatest similarity coefficient is 0.9503, the minimum is 0.6912, the difference is only 0.2591, each between individuals coming together, most of the individual gathered at a distance of 0.27 about similar genetic background, most of the samples between the effects of the geographical environment of. The different provenances Millennium Tong kinship clustering popgen software, 11 kinds of sources Aleurites montana divided into two major clustering and cluster of four small, obvious Putian provenance become a separate branch Putian provenance and other kinds of source distance far fewer exchanges, homozygosity higher; another two sanming source the distance them closer Xu provenance coming together, indicating their due The close geographical distance leaving more exchanges between them, the similarity is relatively high, close to kinship; the Jianyang Huangkeng Zhangzhou pier two kinds of source terrain is relatively complex, causing them to communicate with the outside world other provenances fewer, larger differences between Jianyang other provenances, Lake City, governance, Bookstore, small lakes and bodies of the five kinds of source aggregation into a large cluster, which showed that between five kinds of source Geographic proximity, relatively close genetic relationship. Genetic clustering using the the Dps software on Aleurites montana 33 individuals, 33 of the 11 kinds of sources the individual generally divided into four parts, basically, and the results of the provenances clustering map, showing the cross the provenances internal provenances phase polymerization, the high degree of polymerization between geographical distance closer provenance, contrary lower provenances degree of polymerization of the geographical border area is also high, the discrimination more carefully.