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1、綠僵菌屬4個(gè)生防潛力菌株的多基因鑒nr;王峰劉斌農(nóng)向群魯紅學(xué)王廣君曹廣春劉少?gòu)垵扇A中國(guó)農(nóng)業(yè)科學(xué)院植物保護(hù)研究所/植物病蟲害生物學(xué)國(guó)家重點(diǎn)實(shí)驗(yàn)室長(zhǎng)江大學(xué)農(nóng)學(xué)院摘要:傳統(tǒng)的綠僵菌分類鑒定以形態(tài)特征為主要依據(jù),其局限性使種內(nèi)仍包含著復(fù)雜的類群。DNA分子鑒定使綠僵菌屬、種分類更為細(xì)化和準(zhǔn)確,對(duì)其育種、殺蟲機(jī)理、生態(tài)學(xué)等的研究非常重要。本文對(duì)綠僵菌4個(gè)有生防潛力的菌株IPPMO10202.1PPM2029、1M1330189和M200614的5個(gè)持家基因即核糖體轉(zhuǎn)錄間隔區(qū)序列(ITS1-4)、B微管蛋白(beta-tubulin)、RNA聚合酶II大亞基(RPB1)、RNA聚合酶II第二大亞
2、基(RPB2)和翻譯延伸因子(EF-la)的同源序列進(jìn)行了克隆和測(cè)序,通過構(gòu)建系統(tǒng)發(fā)育樹進(jìn)行了多基因進(jìn)化分析。結(jié)果表明,4個(gè)生防菌株分別位于進(jìn)化樹的3個(gè)分支上,鑒定菌株IPPM010202和IPPM2029同屬于平沙綠僵菌Metarhiziumpingshaense,1M1330189為蝗綠僵菌M.acridum,M200614屬于低溫綠僵菌M.frigidum,它們之間親緣關(guān)系很近,5個(gè)基因的遺傳分化距離只有0.0211036。ITS1-4序列有較高變異頻率和變異幅度,分類效率優(yōu)于其他基因;EF-la序列在鑒別菌株和分析親緣性上有優(yōu)勢(shì)。多基因的系統(tǒng)進(jìn)化分析能夠彌補(bǔ)單基因分析的局限,
3、取得更為準(zhǔn)確的鑒定和更為細(xì)化的分類結(jié)果。關(guān)鍵詞:分了鑒定;多基因標(biāo)識(shí);綠僵菌;系統(tǒng)發(fā)育;親緣性;作者簡(jiǎn)介:王峰,碩士研究生1:wangfeng0925@yeah.net;作者簡(jiǎn)介:農(nóng)向群,研究員,E-mail:xqnong@sina..com0收稿日期:2017-06-22基金:國(guó)家自然科學(xué)基金(61661136004,31471824)MultigeneIdentificationofFourPotentialBiocontrolStrainsinMetarhiziumGenusWANGFengLIUBinNONGXiangqunLUHongxueWANGGuangjunCAOGu
4、angchunLIUShaofangZHANGZehuaStateKeyLaboratoryforBiologyofPlantDiseasesandInsectPests/InstituteofPlantProtection,ChineseAcademyofAgriculturalSciences;CollegeofAgriculture,YangtzeUniversity;Abstract:TheclassicalidentificationofMetarhiziumspp.isbasedonthemorphologicalcharacteristics,whichusuallyr
5、esuItsincomplexgroupswithinaspecies.DNAmolecularidentificationcanofferamorerefinedandaccurateclassifiedtionofthespeciesinMetcirhiziumgenus,whichisveryimportantforstudiesonbreeding,insecticidemechanismandecologyofthefungus.Here,fourpotentialbiocontrolstrainsofMetarhiziumIPPM010202,IPPM2029,IMI33
6、0189andM200614wereidentifiedbyphylogeneticanalysisoffiveident/ifiedgenes,i.e.,ribosomaltranscribedspacersequences(ITS1-4),beta-tubulin,RNApolymeraseIIfirst,secondsubunit(RPB1andRPB2),andtranslationextensionfactor(EF-1a)?Basedonthephylogenetictreeconstructedfromthefourcandidatestrainsandtwenty-t
7、woreferencestrains,thefourstrainswerelocatedonthreebranchesofthephylogenetictree.ThestrainTPPM010202andIPPM2029belongtoM.pingshaense,IMI330189isidentifiedasM.acridumandM200614,asM.frigidum,aspeciesadaptivctolowtemperature-Theyhave