西方蜜蜂的線粒體DNA細(xì)胞色素氧化酶亞基-Ⅰ～細(xì)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(xué)研究所，吉林132108）

西方蜜蜂的線粒體DNA細(xì)胞色素氧化酶亞基-Ⅰ～細(xì)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(xué)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(xué)分析），將分布于那里的歐洲黑蜂等A.m.地理亞種（及其相關(guān)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(fù)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(xué)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(guān)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(xué)分析），將分布于那里的高加索蜂等A.m.地理亞種（及其相關(guān)生態(tài)類型）概括為A.m.亞洲世系[25]。基于DraI 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(fù)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(jié)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(xué)研究提供了新思路。

在同領(lǐng)域的相關(guān)研究中，相關(guān)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(shù)限制了相關(guān)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(shù)和低通量第一代DNA序列測定技術(shù)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(xì)管DNA電泳技術(shù)和高通量第二代DNA序列測定技術(shù)，能夠克服上述研究的技術(shù)瓶頸[9]。鑒于高分辨率、高通量技術(shù)具有方法學(xué)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(lǐng)域的相關(guān)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(jì)初期引入的歐洲黑蜂等A.m.西部歐洲世系后代，19世紀(jì)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(jì)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(lǐng)域的相關(guān)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(guān)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(shù)，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(shù)手段。因此，在蜜蜂屬范圍內(nèi)，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(shù)體系（CARS-45-KXJ2；CA2S-45-SYZ4）

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