李興安 牛慶生 薛運波

（吉林省養(yǎng)蜂科學研究所，吉林132108）

西方蜜蜂的線粒體DNA細胞色素氧化酶亞基-Ⅰ～細胞色素氧化酶亞基-ⅠⅠ富含A+T非編碼區(qū)單倍型類群（續(xù)）

李興安 牛慶生 薛運波

（吉林省養(yǎng)蜂科學研究所，吉林132108）

續(xù)《中國蜂業(yè)》2016年第10期

因此，根據(jù)A.m.的COI-COII NC單倍型類群分析的原始數(shù)據(jù)，A.m.非洲世系為DraI RFLP譜圖表征的3個COI-COII NC單倍型類群（表1）。

在A.m.進化過程中，阿爾卑斯山以北直至烏拉爾山的寬闊地域及其比鄰的突尼斯、摩洛哥、撒哈拉沙漠近地中海地帶等地是A.m.的一個自然分布區(qū)[5，8]。昆蟲形態(tài)變異分析（和昆蟲系統(tǒng)地理學分析），將分布于那里的歐洲黑蜂等A.m.地理亞種（及其相關生態(tài)類型）概括為A.m.西部歐洲世系[25]。基于DraI RFLP分析，本文將A.m.西部歐洲世系樣本的90種DraI RFLP譜圖歸納為1種COI-COII NC單倍型類群，即P、Q元件組成類群[19，21-24，26，27，30，32-35]。這個單倍型類群具有1個P元件和5個不同串聯(lián)重復頻率Q元件，至少包括PQ、PQQ、PQQQ、PQQQQ以及PQQQQQ 5種mtDNA單倍型類型[19，22-24，26，27，30，32-34，36]。因此，根據(jù)A.m.的COI-COII NC單倍型類群分析原始數(shù)據(jù)，A.m.西部歐洲世系為RFLP譜圖表征的1個COI-COII NC單倍型類群（表1）。

在A.m.進化過程中，阿爾卑斯山南麓、地中海東北部沿岸以及喀爾巴阡山之間的廣袤地域是A.m.的另外一個自然分布區(qū)[5，8]。昆蟲形態(tài)變異分析（和昆蟲系統(tǒng)地理學分析），將分布于那里的意大利蜂、卡呢鄂拉蜂等地理亞種（及其相關生態(tài)類型）概括為A.m.東部歐洲世系[25]?；赟NP分析，本文將A.m.東部歐洲世系樣本的25種SNP譜圖歸納為1種COI-COII NC單倍型類群,即Q元件組成類群[23，24，37-39]。這個單倍型類群僅為Q元件1種mtDNA單倍型類型。因此，根據(jù)A.m.的COICOII NC單倍型類群分析的原始數(shù)據(jù)，A.m.東部歐洲世系為SNP譜圖表征的1個COI-COII NC單倍型類群（表1）。

西部亞洲或（和）中部亞洲是A.m.的第四個自然分布區(qū)[5，8]。昆蟲形態(tài)變異分析（和昆蟲系統(tǒng)地理學分析），將分布于那里的高加索蜂等A.m.地理亞種（及其相關生態(tài)類型）概括為A.m.亞洲世系[25]?；贒raI RFLP分析，本文將A.m.亞洲世系樣本的29種DraI RFLP譜圖歸納為3種COI-COII NC單倍型類群，即P0、Q元件組成類群，P0'、Q元件組成類群以及P1'、Q元件組成類群[22，35，40，41]。其中，P0、Q元件組成類群具有1個P0元件和3個不同串聯(lián)重復頻率Q元件，至少包括P0Q、P0QQ以及P0QQQ 3種mtDNA單倍型類型；P0'、Q元件組成類群具有1個P0'元件和1個Q元件，僅為P0'Q 1種mtDNA單倍型類型[22]；P1'、Q元件組成類群具有1個P1'元件和1個Q元件，僅為P1'Q 1種mtDNA單倍型類型[42]。因此，根據(jù)A.m.的COI-COII NC單倍型類群分析的數(shù)據(jù)，A.m.亞洲世系為DraI RFLP譜圖表征的3個COI-COII NC單倍型類群（表1）。

3 總結與展望

通過上述分析，本文依據(jù)A.m.地理亞種的COICOII NC多態(tài)性規(guī)律，將A.m.歸納為8個COI-NCCOII單倍型類群。A.m.地理亞種的COI-COII NC單倍型類群為西方蜜蜂分類學研究提供了新思路。

在同領域的相關研究中，相關文獻既缺乏A.m.非洲世系、A.m.西部歐洲世系以及A.m.東部歐洲世系的SNP分析數(shù)據(jù)，又缺乏A.m.亞洲世系的DraI RFLP分析數(shù)據(jù)（表1）。其原因可能是低通量分析技術限制了相關研究在研究期間獲得大量COI-COII NC數(shù)據(jù)，即低分辨率瓊脂糖（和聚丙烯酰胺）凝膠電泳技術和低通量第一代DNA序列測定技術不適合進行大量A.m.地理亞種樣品的COI-COII NC多態(tài)性分析。然而，在一些其它生物樣品的mtDNA單倍型分析中，高分辨率毛細管DNA電泳技術和高通量第二代DNA序列測定技術，能夠克服上述研究的技術瓶頸[9]。鑒于高分辨率、高通量技術具有方法學優(yōu)勢，A.m.的mtDNA多態(tài)性分析最終將在分子水平建立A.m.地理亞種的COI-COII NC單倍型數(shù)據(jù)庫，COI-COII NC單倍型數(shù)據(jù)庫將更透徹地反映A.m.的COI-NC-COII多態(tài)性規(guī)律。

表1 A.m.的COI-COII NC單倍型類群分析統(tǒng)計表

表1 A.m.的COI-COII NC單倍型類群分析統(tǒng)計表(續(xù))

表1 A.m.的COI-COII NC單倍型類群分析統(tǒng)計表(續(xù))

在同領域的相關研究中，除了美洲A.m.的COICOII NC多態(tài)性分析之外，較少文獻系統(tǒng)地報道其它非自然分布區(qū)A.m.的研究進展。美洲A.m.包括17世紀初期引入的歐洲黑蜂等A.m.西部歐洲世系后代，19世紀引入的意大利蜂、卡呢鄂拉蜂等A.m.東部歐洲世系后代和高加索蜂等A.m.亞洲世系后代，以及20世紀中期引入的非洲蜂等A.m.非洲世系后代。本文將美洲A.m.非洲世系的20種DraI RFLP譜圖歸納為P0、Q元件組成類群的3種mtDNA單倍型類型（即P0Q、P0QQ和P0QQQ），將美洲A.m.西部歐洲世系的7種DraI RFLP譜圖歸納為P、Q元件組成類群的3種mtDNA單倍型類型（即PQ、PQQ和PQQQ），美洲A.m.東部歐洲世系的2種SNP譜圖歸納為Q元件組成類群的1種mtDNA單倍型類型（即Q），將美洲A.m.亞洲世系的4種DraI RFLP譜圖歸納為P0、Q元件組成類群的3種mtDNA單倍型類型（即P0Q、P0QQ和P0QQQ）[29]。美洲A. m.的COI-COII NC多態(tài)性分析將為其它非自然分布區(qū)A.m.的同類型研究提供可靠的文獻依據(jù)。

在同領域的相關研究中，除了東方蜜蜂、沙巴蜂、綠努蜂以及蘇拉威西蜂等A.m.近緣物種的COI-COII NC多態(tài)性分析外，較少文獻報道大蜜蜂等蜜蜂屬其它物種的同類型研究。盡管如此，這些A.m.近緣物種的COI-COII NC多態(tài)性分析還是揭示了這樣一個事實：相對于A.m.，它們具有短的COI-COII NC，其長度范圍是30～100堿基。這或許為我們透露出這樣一點信息：蜜蜂屬生物的COI-NC-COII多態(tài)性與蜜蜂屬生物的物種多樣性之間是否存在某種關系。而且，鑒于A.m.的COI-COII NC多態(tài)性分析已積累了豐富的知識和成熟的技術，這些蜜蜂屬其它物種的同類型研究有了可借鑒的知識儲備和技術手段。因此，在蜜蜂屬范圍內，有必要系統(tǒng)研究每個物種的COI-COII NC單倍型類群。

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吉林省科技發(fā)展計劃項目（20130101094JC；20140101132JC）、國家蜂產(chǎn)業(yè)技術體系（CARS-45-KXJ2；CA2S-45-SYZ4）

李興安（1965-），男，研究員，E-mail∶Lxingan@sina.com