Genetic Diversity Earthworms Family Megascolecidae in Northeastern Thailand using PCR-RFLP Technique

บทคัดย่อ

The objectives of this study were to identify the taxonomic identifi cation of earthworm family Megascolecidae Samples were collected from 7 provinces (Kalasin, Nakhon Phanom, Buriram Roi Et, Sisaket, Sakon Nakhon and Nongbua Lamphu) in northeastern Thailand. The external and internal morphology of 275 specimens of family Megascolecidae were classifi ed as 12 species including Metaphire peguana, M. bahli, M. houlleti, M. bipora, M. anomala, M. planata, M. posthuma, M. sp 1, M. sp 2, M. sp 3, Amynthas seiboldi and unknown species. The genetic diversity was determined using PCR-RFLP technique restriction analysis of Cytochrome oxidase. Total DNA was extracted from the muscular tissue of each individual earthworm, and then amplifi ed with COI primers as 710 bp PCR products. After that the PCR products were digested with 3 restriction enzyme (Dde I, Alu I, Hinf I). The results of these samples showed 13 composite haplotypes as AAA, BBB, CCC, DDD, EEE, FJF, GFG, HGH, IHI, JII, KJI, LJI and MJI. The UPGMA dendrogram of restriction pattern was analyzed using NTSYSpc version 2.10p. The earthworms were divided into 2 groups as group 1 consisting of Dichogaster bolaui and group 2 consisted of Metaphire peguana, M. bahli, M. houlleti, M. bipora, M. anomala, M. planata, M. posthuma, M. sp. 1, M. sp. 2, M. sp. 3, Amynthas seiboldi and unknown species. The similarity coeffi cient between 2 groups was 0.66.