中国农业科学 ›› 2009, Vol. 42 ›› Issue (4): 1494-1502 .doi: 10.3864/j.issn.0578-1752.2009.04.047

• 研究简报 • 上一篇    

东北黑蜂(Apis mellifera ssp.)亲缘关系的分析

  

  1. 中国农业科学院蜜蜂研究所
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2009-04-10 发布日期:2009-04-10

Analysis of Genetic Relationships of Northeastern Black Bee (Apis mellifera ssp.) in China

  1. 中国农业科学院蜜蜂研究所
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-04-10 Published:2009-04-10

摘要: 【目的】从分子水平揭示东北黑蜂与欧洲黑蜂、意大利蜜蜂、卡尼鄂拉蜂、喀尔巴阡蜂、安拉托利亚蜂、高加索蜂以及非洲类型的西方蜜蜂的亲缘关系,为东北黑蜂分类地位的正确认识及其合理保护提供参考。【方法】对来自东北黑蜂国家级自然保护区内9个乡镇的东北黑蜂与上述西方蜜蜂的线粒体DNA(mtDNA)tRNAleu~COⅡ序列进行PCR扩增;同时取部分PCR产物进行DraⅠ酶切分析;并根据酶切类型对相应的PCR产物经纯化、克隆和测序后进行序列分析,并构建Neighbor-Joining(NJ)聚类图。【结果】东北黑蜂mtDNA tRNAleu~COⅡ序列长度为526 bp,P序列缺失,Q序列有5个位点出现碱基转换(4T?C、1A?G),1个碱基颠换(1A?T);只有一种DraⅠ酶切类型,其酶切片段长度分别为422、64、47和41 bp,同DNAMAN软件分析的酶切位点一致;其序列特征和酶切类型同西方蜜蜂线粒体C、O分支相似,而与A分支的非洲蜜蜂、M分支的欧洲黑蜂存在较大差异;根据NJ聚类图,东北黑蜂属于线粒体O分支。MtDNA tRNAleu~COⅡ的序列特征和DraⅠ酶切分析表明,东北黑蜂属于线粒体O分支,与同一分支的高加索蜂、安拉托利亚蜂亲缘关系最近,与C分支的意大利蜜蜂、卡尼鄂拉蜂、喀尔巴阡蜂亲缘关系次之,而与M分支的欧洲黑蜂和A分支的非洲蜜蜂亲缘关系较远。【结论】根据NJ聚类图,东北黑蜂处于安拉托利亚蜂和高加索蜂之间,东北黑蜂是否是这两个地理亚种的混合种群,或能否算一个新的地理亚种,还有待进一步的研究。

关键词: 东北黑蜂(A.m.ssp.), 亲缘关系, mtDNA tRNAleu~COⅡ, 序列分析

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