Study Of The Population Structure Of Apis Cerana In China

The purpose for this paper is taking a step forward to clear the taxonomy status of Apis cerana subspecies distributed in china. The mitochondrial DNA tRNAIleND2and16S rRNA genes as well as nuclear gene EFl-α of A. cerana collected from19locations in china were amplified using the reported pairs of primers and sequenced. Sequences of these genes were compared with the corresponding sequences from other locations. Accordind to the results we obtained in this study:1. Sequence alignment showed that the length of tRNAIle～ND2gene partial sequence was471bp-474bp. There are13variation sites in these sequences, which contains14transition sites,2transvertion sites,3insertions or deletions sites. The sequence similarity analysis showed that The lowest similarity of tRNAIle-ND2gene sequences was92.7%, The highest similarity of tRNAIle-ND2gene sequences was97.9%. The results of ND2gene protein Sequence alignment indicated that there was one amino acid mutation in Qinyuan sample,by Q→K. The results of Phylogenetic analysis indicated that Phylogenetic tree based on tRNAIle-ND2gene partial sequence has4major branches. Apis koschevnikovi, A.c.in Thailand and A.c. in hainana was a big branch respectively.The other branch is A. c. in Viet Nam, A.c. in Nepal and A.c. in other18locations.2. The length of16S rRNA gene partial sequence was581bp-585bp. There are6variation sites in these sequences, which contains1transition sites,5insertions or deletions sites. The lowest similarity of16S rRNA gene sequences was97.9%, The highest similarity of16S rRNA gene sequences was100%. Phylogenetic tree based on16S rRNA gene partial sequence has2major branches. A.c. in hainana and A.c.in other18locations was a branch respectively.3. The length of EF1-α gene partial sequence was306bp. There are3variation sites in these sequences, which contains1transition sites,2transvertion sites. The lowest similarity of EF1-α gene sequences was99.3%, The highest similarity of EF1-α gene sequences was100%. Phylogenetic tree based on EF1-α gene sequence has4major branches. Apis andreninformis and Apis florae, Apis dorsata, Apis mellifera are three branches.The other brance was A.c. in Chinese19locations, Apis nulunsis and Apis koschevnikovi. 4. The sequencing results of this study was storaged in National Center for Biotechnology Information (NCBI),The aceession number was KF663574～KF663592,KF572594～KF572612and KF663570～KF663573.5. Phylogenetic tree based on tRNAUe～ND2and16S rRNA gene partial sequence indicated that A. cerana in hainana was significantly different from A. cerana in the other18locations. A. cerana in Aba may belong to a high altitude ecotype. The result didn’t support the conclusion that A. cerana in Aba was a separate subspecies. A. cerana in3Jilin locations have closer relationship from each other, which may belong to the same taxon with those in South korea and Japan. The variation of A. cerana in Yunnan was rich.6. The EF1-α gene partial sequences of A. cerana in19chinese locations have a higher similarity with A.c.in Indonesia, A.c. in Sri Lanka and A.c.in Malaysia. EF1-α gene can not be good for the subspecies taxonomy of Apis cerana. EF1-α gene is a good molecular marker in the classification of high level of bees.