Abstract:

【Objective】 To analyze taxonomy of Northeastern Black Bee (Apis mellifera ssp.), the study was carried out by analyzing the genetic relationships among Apis mellifera mellifera, Apis mellifera ligustica, Apis mellifera canica, Apis mellifera carpatica, Apis mellifera anatoliaca, Apis mellifera caucasica and African bees. This is usefull for the protection of genetic resouces of Apis mellifera ssp. 【Method】 Mitochondrial DNA (mtDNA) tRNAleu-COⅡ region of the A.m.ssp. from 9 towns of National Nature Reserve in China’s Heilongjiang province and other Apis mellifera were amplified by PCR. Then PCR products were digested with restriction enzymes DraI, based on the restriction patterns, PCR products were cloned and sequenced, and the dendrogram was constructed by Neighbor-Joining (NJ) method. 【Result】 The length of mtDNA tRNAleu-COⅡ region were 526 base-pair (bp) , and only one Q sequence was found in A.m.ssp. but P sequence was absent. Besides, there were 5 transitions（4T?C、1A?G） and 1transversion（1A?T）in Q sequence. Only one DraⅠ restriction pattern was identified, the length of restriction fragments was 422 bp, 64 bp, 47 bp and 41 bp, respectively, and the restriction sites were validated by DNAMAN. The sequences and restriction pattern of A.m.ssp. were similar with the evolutionary lineages C and O of Apis meliffera, but different from lineages A and M. Based on the NJ tree, A.m.ssp. belongs to lineage O. 【Conclusion】 The sequence of mtDNA tRNAleu-COⅡ region and DraⅠ restriction pattern indicate that A.m.ssp. belongs to O lineage, and has closest relationship with A.m.anatoliaca and A.m. caucasica in the same group, closer with A.m.ligustica, A.m.canica, A.m. carpatica than that with A.m.mellifera and African bees. Based on the NJ tree，A.m.ssp. mixed in A.m.anatoliaca and A.m. caucasica, whether it is a mixed groups of the two geographical races, or it is a new honey bee subspecies need to be further investigated in the future.

Key words: northeastern black bee (Apis mellifera ssp.), genetic relationships, MtDNA tRNAleu-COⅡ, sequence