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1、第40卷第4期植物保護學報Vo1.40No.42013年8月ACTAPHYTOPHYLACICASINICAAug.2013野生大豆內(nèi)生細菌多樣性及其殺線蟲活性分析史鳳玉武云鵬張瑞敬吳楠朱英波(1.河北科技師范學院生命科技學院,秦皇島066600;2.張家口市植物保護植物檢疫站,張家口075000)摘要:為探明河北昌黎野生大豆內(nèi)生細菌的多樣性,尋找新的具有殺線蟲活性的微生物資源,采用平板稀釋法從野生大豆根、莖、葉等組織中分離內(nèi)生細菌,結(jié)合形態(tài)特征以及16SrDNA基因序列進行比對分析,并對大豆孢囊線蟲Heteroderaglycines進
2、行殺線蟲活性比較。從野生大豆分離獲得內(nèi)生細菌180株,分屬于7大類群:變形菌門儀亞群、變形菌門p亞群、變形茵門^y亞群、厚壁菌門、放線茵門、酸桿菌門和疣微茵門,其中變形茵門o【亞群和厚壁菌門為優(yōu)勢類群,分別占分離細菌總株數(shù)的48.9%和18.9%,野生大豆組織內(nèi)生細菌Shannon—Wiener多樣性指數(shù)為2.11~2.91。內(nèi)生細菌Bacillussubtil~WSR93和B.megateriumWSR22菌株能明顯地抑制大豆孢囊線蟲卵的孵化,相對抑制率分別為97.9%和98.2%,對2齡幼蟲也有較強的毒殺作用,校正死亡率分別為88.9
3、%和90.9%。研究表明,野生大豆組織中內(nèi)生細菌具豐富的多樣性,其胞外分泌物中存在天然殺線活性物質(zhì)。關(guān)鍵詞:野生大豆;大豆孢囊線蟲;可培養(yǎng)細菌;多樣性DiversityandnematicidalactivityoftheendophyticbacteriainthewildsoybeanGlycinesoinShiFengyuWuYunpengZhangRuijingWuNanZhuYingbo(1.CollegeofLifeSciences,HebeiNormalUniversityofScienceandTechnology,Qin
4、huangdao066600,HebeiProvince,China;2.ZhangjiakouStationofPlantProtection,Zhangjiakou075000,HebeiProvince,China)Abstract:InordertounderstandthediversityoftheendophyticbacteriafromtissuesofwildsoybeanGlycines咖inChangli,anddiscovernewstrainswithnematicidalactivity,theendophy
5、ticbacteriawereisolatedfromhealthyroot,stem,andleaftissuesusingdilutionplatecountingmethod.Thestrainswereidentifiedaccordingtothemorphologicalcharacterscombinedwith16SrDNAsequencesanalysis,andthenematicidalactivityofthestrainsagainstHeteroderaglycineswasalsotested.Theresu
6、ltsshowedthat180endophyticbacteriastrainsisolatedfromdifferentpartsofwildsoybeanbelongedto7bacterialgroups,Alphaproteobacteria,Betaproteobacteria,Gammaproteobacteria,F(xiàn)irmicutes,Actinobacteria,Acidobacte-riaandVerrucomicrobia.ThestrainsbelongingtogroupAlphaproteobacteriaan
7、dFirmicuteswerethedominantendophytesinwildsoybcan,with48.9%and18.9%oftotalstrains,respectively.TheShan—non—Wienerindexofendophytebacteriainwildsoybeanwasbetween2.11and2.91.TheendophytebacteriaBacillusmegateriumWSR22andB.subtilisWSR93inhibitedegghatchingofsoybeancystnema—r
8、ode,witharelativeinhibitionrateof97.9%and98.2%,andhadstrongtoxiceffectagainst2ndstagejuveniles,w