2009年3月2日 星期一

尾突也可抽DNA- 保育類蛾種的非致死取樣方法之效果


Effect of non-lethal sampling on life-history traits of the protected moth Graellsia isabelae (Lepidoptera: Saturniidae)
Ecological Entomology (Accepted 4 December 2008, 將於2009近期出刊,尚未上線)

M . Vila 1, M . A . Auger-Rozenberg2 , F. Goussard 2and C . Lopez-Vaamonde2
1 Department of Molecular and Cell Biology, Faculty of Sciences, University of A Coruna, A Coruna, Spain and 2 INRA, UR 633 Zoologie Forestière, Orléans, France

在昆蟲方面使用非致死的取樣方法通常是由一隻腳或翅膀的一部分中萃取DNA,雖然比殺死動物以取得樣本更好,但關於缺少一隻腳或一部分翅膀對昆蟲個體的生存或繁衍有何影響並不清楚。Graellsia isabelae是一種受到生境指令與伯恩公約保護的歐洲蛾類,在保育遺傳學上的取樣必須採用非致死方式,作者的研究目標是(1)量化從腳和後翅尾突取得DNA樣本對於雌雄成蟲的生存和繁殖的影響,以及(2)評估由這些組織中萃取DNA的量與品質。結果顯示後翅尾突和中足都是品質良好的核DNA與粒線體DNA來源,且取自大於130mm2的後翅尾突的DNA濃度顯著高於來自大約半截中足的DNA濃度。對成蟲進行捉放法的結果顯示無論是後翅尾突或是中足被取樣,對於雄性的存活率或交配總次數都沒有顯著的減少;然而雖然少了中足對於母蛾的生存沒有顯著影響,卻對其能否成功交配有負面的影響。修剪雄蛾後翅對G. isabelae而言似乎是最好的非致死取樣方式,既快速又與自然造成的翅膀損傷類似,對其生存或交配行為沒有顯著影響

Abstract
1. Non-lethal genetic surveys in insects usually extract DNA from a leg or a piece of wing. Although preferable to lethal sampling, little is known about the effect of leg/wing non-lethal sampling on fitness-related traits. 2. Graellsia isabelae (Graells, 1849) is a European moth protected by the Habitats Directive and the Bern Convention. Conservation genetics surveys on this species should therefore use non-lethal sampling. 3. The present study aimed to (1) quantify the effects of both leg and hind-wing tail sampling on survivorship and reproductive behaviour of adult males and females, and (2) assess the quality and quantity of DNA obtained from those tissues. 4. Both hind-wing tails and mid-legs proved to be good sources of high quality nuclear and mitochondrial DNA. DNA concentration was significantly higher when extracted from a large (130 mm 2 ) piece of the hind-wing tails than from about half of the mid-leg. Using mark – release – recapture experiments with adults, it was found that neither midleg nor hind-wing tail sampling significantly reduced male survivorship or total number of matings. However, although mid-leg sampling did not significantly affect female survivorship, it had a negative effect on female mating success. 5. Wing-tail clipping on males appeared to be the best non-lethal sampling procedure for G. isabelae . It is a fast procedure, similar to natural wing impairment, and did not significantly affect survival or mating behaviour.

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