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《Acta Ecologica Sinica》 2004-07
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RAPD analysis of genetic divergence among populations of Oxya japonica, O. chinensis and Pseudoxya diminuta

ZHANG Jian-Zhen~1, GUO Ya-Ping~1, DUAN Yi-Hao~2, MA En-Bo~(1*) (1. College of Life Science and Technology, Shanxi University, Taiyuan 030006, China; 2. College of Environmental Science and Resources, Shanxi University, Taiyuan 030006, China).  
The genetic divergence among 4 populations of Oxya japonica, 2 populations of O. chinensis and 1 population of Pseudoxya diminuta were analyzed using random amplified polymorphic DNA (RAPD) markers. The modified saturated NaCl protocol was developed in this study to extract DNA from the tissue samples fixed in anhydrous ethanol and from dry specimens. Nucleic acids extracted from 60 individuals were then amplified using 10 oligonucleotide primers, each of 10 nucleotides in length. The agarose gel electrophoreses revealed 135 clear and stable bands with molecular weight ranging from 200 to 2500 bps. In average each primer yielded about 8 (3~9) bands per individual. The RAPD patterns demonstrated consistent polymorphisms according to population, species and genus. For P. diminuta 75% bands were commonly shared by the individuals within a population, suggesting the low genetic polymorphic level. Meanwhile, 7.5% bands were shared by the individuals among 4 populations of O. japonica. The percentage of the common bands among populations is much higher than the shared bands between O. chinensis and O. japonica, and no shared bands were observed among P. diminuta, O. japonica and O. chinensis. Shannon 's index showed higher genetic diversity (2.693) within O. chinensis, and the genetic diversity of (O. japonica) (2.319) was close to O. chinensis, while P. diminuta had the lowest level (1.042). The dendrogram based on Nei's genetic distance of RAPD markers was constructed using UPGMA and Neighbor-Joining, indicated high within-population similarities. The clustering followed the order of, first the individuals within a population, then the different populations of the same species, later the species of the same genus, and finally the species among genera. The four populations of O. Japonica fell into 2 branches, two geographically close populations both from Southern China were clustered into one branch, while the other two distant populations were grouped to another branch. The species of O. japonica then was clustered with (O. chinensis,) before clustered with P. diminuta finally. These data suggested that several implications. First RAPD is very useful as an effective molecular marker to distinguish the divergence in Oxya and its closed related genera. Second there seems to be a tendency of correlations between the genetic and geographic distance among the 4 different populations from the south to the north. Third, the RAPD similarity follows the order of individualspopulationsspeciesgenera. Specifically, (O. chinensis) resembles more to O. japonica than to P. diminuta. The high uniformity in both RAPD and chromosomes in (P. diminuta,) along with its narrow distraction range, suggested the species a highly specialized group with close relationship to Oxya. The results of dendrogram based on RAPD markers are consistent with the previous conclusions of morphologic classification and cytotaxonomy.
【Fund】: 国家自然科学基金资助项目 ( 3 0 0 70 112 ;3 0 170 612 ) ;; 山西省自然科学基金资助项目 ( 9910 96)~~
【CateGory Index】: S435.11
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