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1、大豆蚜捕食性天敌捕食行为的COI基因标记检测宋新元,丛斌,钱海涛,董辉(沈阳农业大学植物保护学院,沈阳110161)摘要:【目的】探索大豆田捕食性天敌对大豆蚜的捕食作用。【方法】根据基因库NCBI中的一段大豆蚜细胞色素氧化酶I(COI)的基因序列(登录号为AY842503),设计了2对大豆蚜特异引物A和B,其扩增片段大小分别约为197和253bp,应用DNA标记方法检测天敌对大豆蚜的捕食行为。【结果】种特异性检验表明,所设计引物只对大豆蚜DNA具有扩增效果,对与其同域发生的其它种类不具扩增作用;室内以引物对A与B分别检测喂食单头大豆蚜的异色瓢
2、虫成虫和大草蛉成虫腹内靶标食物的衰变情况,结果表明,异色瓢虫成虫阳性比率的半衰期分别为喂食后3.371h和2.814h,大草蛉成虫分别为喂食后1.312h和1.032h;以A作为引物对田间捕食性天敌进行检测,结果表明,在所检测的7类群14种捕食性天敌中,5种不同种类或虫态的天敌对大豆蚜的捕食作用检出率高于50%,从高到低依次为异色瓢虫幼虫、草蛉成虫、东亚小花蝽若虫、异色瓢虫成虫、东亚小花蝽成虫;此外,田间检测表明,捕食性天敌的阳性比率与大豆蚜的种群密度呈显著正相关。【结论】DNA标记技术是探索捕食行为的有效方法之一。关键词:大豆蚜;细胞色素氧
3、化酶I基因;捕食性天敌;消化道内容物检测;DNA标记IdentificationoftheKeyPredatorsofAphisglycinesMatsumura(Homoptera:Aphididae)UsingCOIGeneMarkersSONGXin-yuan,CONGBin,QIANHai-tao,DONGHui(CollegeofPlantProtection,ShenyangAgriculturalUniversity,Shenyang110161)Abstract:【Objective】Inordertounderstandth
4、ekeypredatorsofAphisglycinesMatsumura(Homoptera:Aphididae)insoybeanfields,theCOIgenewasintroducedtomarktheaphiscapturedbythepredator.【Method】COImarkersofA.glycinesweredevelopedtodetecttheremainsofA.glycinesinthegutofHarmoniaaxyridis(Pallas)(Coleoptera:Coccinellidae)andChrys
5、opapallens(Rambur)(Neuroptera:Chrysopidae)rearedinthelabandvariouspredatorscollectedinthesoybeanfields.Twopairsofsequence-characterizedprimers(AandB)weredesigned,primerAamplifiedsinglebandwasabout197bpandprimerBabout253bp.【Result】Thespecificitytestsperformedwiththeprimerssh
6、owedthebandpresentedonlyinA.glycines;UsingprimersAandB,thehalf-livesofdetectabilityfortheDNAfromasingleA.glycinesconsumedbyadultH.axyridiswas3.371hand2.814h,thehalf-lifeofdetectabilityfortheDNAfromasingleA.glycinesconsumedbyadultC.pallenswas1.312hand1.032h.UsingprimerA,thep
7、ercentageofpositiveresponsesofA.glycinesDNAinthepredatorscollectedinfields,suchasthelarvaeofH.axyridis,thenymphofOriussauteri(Poppius)(Hemiptera:Anthocoridae),theadultofH.axyridis,O.sauteriandChrysopas,wereover50%.Moreover,thetestsofsamplescollectedfromthefieldshowedthatthe
8、detectionrateofpredatorsbyusingprimerAsignificantlycorrespondedtothedensityofA.gly