乌龟遗传多样性的RAPD分析
GENETIC DIVERSITY ANALYSIS OF CHINESE THREE-KEELED POND TURTLE (CHINEMYS REEVESII) BY RAPD
-
摘要: 运用RAPD技术对乌龟的遗传多样性进行了分析。用20个随机引物对24个个体的基因组DNA进行了PCR扩增,一共扩增出3288条DNA片段,平均每个个体扩增出137条带。在检测到的137个位点中,多态位点数为119个,占869%,标记的分子量在0.2kb-3kb之间。个体间最大的遗传距离为0467,个体间最小的遗传距离为0168。24个个体的平均遗传距离为0324+00631。表明乌龟的遗传多样性水平较高。采用类平均聚类法(NJTREE)构建了24个个体相互关系的分支图。24个个体被分为几个类群,显示种内遗传差异较大,可能存在不同种群。本研究为乌龟的种质保护、合理开发利用以及选择育种提供了新的分析参数。Abstract: Chinese three-keeled pond turtle (Chinemys reevesii Gray) is a member of the family Emydidae and widely ranges in southern China, especially in the regions of the lower and middle Yangtze River. It has high values for food, traditional Chinese medicine and pet trade, and is one of the turtles for aquaculture in China. But the genetic background of Chinese three-keeled pond turtle is still not very clear. In order to realize its population genetic structure and the diversity for the conservation of its germ plasm and prevention of gene loss, genetic diversity analysis was carried out using RAPD-PCR technique. In this paper, genetic diversity of 24 individuals of Chinese three-keeled pond turtle from Hubei province was analyzed by the random amplified polymorphic DNA (RAPD) technique, using 20 10-nucleotide-long primers. Twenty primers produced 3288 well-amplified DNA fragments with reproducible banding patterns and averaged 137 DNA fragments in one individual. 119 of 137 loci detected are polymorphic, amounting to 86.9%. The DNA fragments were from 0.2kb to 3kb. The genetic distance between two individuals was from 0.168 to 0.467. The average genetic distance was 0.324?.0631 among the 24 individuals. The phylogenetic tree of the 24 individuals was obtained by NJTREE analysis in the RAPDistance program and the 24 individuals were divided into several clusters and it indicated the turtle maybe have two populations at least. In conclusion, genetic diversity of Chinese three-keeled pond turtle was high.
-
-
[1] Zhao E M. Studies on taxonomy and distribution of Chinese chelonians[J]. Sichan Journal of Zoology, 1997,15(Suppl): 1-26[赵尔宓. 中国龟鳖动物的分类与分布研究. 四川动物,1997,15(增刊):1-26]
[2] Guo C W. A study on chromosomes of Chinese chelonians[J]. Sichan Journa of Zoology,1997,15(Suppl):93-96[郭超文.中国龟鳖类染色体研究.四川动物,1997,15(增刊):93-96]
[3] Nie L W, Guo C W, Wang M. A comparative study on isozymes of three fresh-water turtle species in China [J]. Sichan Journal of Zoology, 1997, 15(Suppl): 115-120[聂刘旺、郭超文、汪鸣.我国三种龟的同工酶比较研究.四川动物,1997,15(增刊):115-120]
[4] Wu P, Zhou K Y, Yang Q. Phylogeny of Asian freshwater and terrestrial turtles based on sequence analysis of 12SrRNA gene fragment[J]. Acta Zoology Sinica, 1999, 45(3):332-338[吴平、周开亚.亚洲淡水和陆生龟鳖类12SrRNA基因片段的序列分析和系统发生研究. 动物学报,1999,45(3):332-338]
[5] Liu P, Kong J, Shi T, et al. RAPD analysis of wild stock of penaeid shrimp (Penaeus chinensis) in the China's coastal waters of Huanghai and Bohai Seas [J]. Acta Oceanologica Sinica, 2000, 22(5):88-93[刘萍、孔杰、石拓、庄志猛,等.中国对虾黄、渤海沿岸地理群的RAPD分析.海洋学报,2000,22(5):88-93]
[6] Garcia K G.Genetic diversity of cultured Penaeus vannamei shrimp using three molecular genetic techniques [J]. Molecular Marine Biology and Biotechnology,1994, 3(5): 270-280
[7] Zheng G M, Zhu X P, Zhang Y, et al. Genetic differential research on the Mud Carp (Cirrhina molitorella) in different parts of Pearl River Valley[J]. Journal of Agricultural Biotechnology, 2001, 9(2):178-182[郑光明、朱新平、张跃,等.珠江流域不同江段鲮鱼遗传差异研究.农业生物技术学报,2001, 9(2):178-182]
[8] Dinesh K R, Lim T M, Chan W K, et al. Genetic variation inferred from RAPD fingerprinting in three species of Tilapia[J]. Aquaculture, 1996,4(1): 19-30
[9] Trojanowski Jan, Marlene Snyder, and Tom Herman. Assessment of genetic variation of snapping turtle (Chelydra serpentina) populations using RAPD analysis, http:/ /ace. acadiau.Ca/science/boil/honours/biofeedback 99/trojanowsk
[10] Zhou L, Fan L C, Gui J F, RAPD analysis of incorporation of heterologous genetic materials in multiple species of silver crucian carp [J]. Acta Hydrobiol Sinica, 1998,22(4):301-306[周莉、樊连春、桂建芳.银鲫复合种外源遗传物质整入的RAPD分析.水生生物学报,1998,22(4):301-306]
[11] Nei M, Li W H. Mathematical model for studying genetic variation in terms of restriction endonucleases[J]. Pro. Natl.Acad Sci.USA.1985, 76:5269-5273
[12] Apostol B L. Estimation of the number of full sibling families at an oviposition site using RAPD-PCR markers: applications to the mosquito Aedes aegypti [J]. Theor Appl Genet. 1993, 86:991-1000
[13] Galbraraith. D A., White B N., Brooks R J, et al. Multiple paternity in clutcheso of snapping turtles (Chalydra serpensina) detected using DNA fingerprints [J]. Canadian J. Zool., 1993, 71(2): 318-324
[14] Kichler K, Holder M T, Davis S K, Marquez M R, et al.Detection of multiple paternity in the Kemp's ridley sea turtle with limited sampling [J]. Molecular Ecology, 1999, 8:819-830
[15] Rhen T, Lang J W. Phenotypic plasticity for growth in the common snapping turtle: effects of incubation temperature, clutch, and their interaction [J]. American naturalist, 1995,146:726-747
[16] Sugg D W, Chesser R K. Effective population sizes with multiple paternity [J]. Genetics, 1994, 137: 1147-1155
[17] Valenzuela, N. Multiple paternity in side-neck turtles Podocnemis espansa: evidence from microsatellite DNA data [J]. Molecular Ecology, 2000, 9:99-105
计量
- 文章访问数: 1093
- HTML全文浏览量: 1
- PDF下载量: 475