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1、中國油料作物學報2013,35(2):196—200ChineseJournalofOilCropSciencesdoi:10.7505/j.issn.1007—9084.2013.02.014大豆莖褐腐病病原菌鑒定段燦星,張吉清,丁俊杰,王曉嗚,朱振東¨(1.中國農業(yè)科學院作物科學研究所/國家農作物基因資源與基因改良重大科學工程,北京,1000812.黑龍江省農業(yè)科學院佳木斯分院,黑龍江佳木斯,154007)摘要:對新發(fā)生的引起大豆莖維管束和髓褐腐的病原菌進行鑒定。該病原分離物在PDA培養(yǎng)基上的菌落扁平,致密,不規(guī)則圓形,白色到淺灰褐色,具放射狀褶皺突起,菌落邊緣粗糙不整齊,或帶有缺刻;
2、在GBA培養(yǎng)基上菌落白色至灰色,菌絲緊貼培養(yǎng)基,氣生菌絲稀少,菌落圓形,邊緣整齊。在GBA培養(yǎng)基上分離物產生分生孢子,分生孢子卵形至橢圓形,無色;單胞,或形成有分隔的細胞;平均長度為(3.0~5.1)txm×(2.1~3.3)m。致病性測定表明分離物能100%引起合豐25大豆植株出現(xiàn)典型大豆莖褐腐病癥狀。引物ITSI/ITS4在分離物中擴增的產物與GenBank中Phialophoragregata分離物序列的相似性達99%。利用大豆莖褐腐病菌的特異引物BSR1/BSR2在分離物中擴增出483bp的特異性條帶。研究結果表明,上述分離物確為大豆莖褐腐病菌。關鍵詞:大豆;分離物;大豆莖褐腐病菌
3、;大豆莖褐腐病中圖分類號:S435.651文獻標識碼:A文章編號:1007—9084(2013)02—0196—05PathogenidentificationofbrownstemrotofsoybeanDUANCan—xing,ZHANGJi—qing,DINGJun—jie,WANGXiao—ruing,ZHUZhen—dong(1.InstituteofCropSciences,ChineseAcademyofAgriculturalSciences~NationalKeyFacilityforCropGeneResourcesandGeneticImprovement,Beiji
4、ng100081,China;2.JiamusiBranchofHeilongjiangAcademyofAgriculturalSciences,Jiamusi154007,China)Abstract:ThepathogencausingbrowningofvascularsystemandpithofsoybeaninChinawasidentifiedusingmorphologicalandmolecularcharacteristics.ColoniesoftheisolatesonPDAmediumdevelopedintheformofflat,dense,roughlyc
5、ircular,whitetoputty—coloured(orgreymat),withradialfoldsandvariable,usuallyfinelylo—bateornotchedmargin.ColoniesonGBAmediumwerewhitetogrey,circular,withsmoothedgeswhosemyceliaadheredtomediumandtheaerialmyceliaweresparse.OnGBAmedium,theconidiawereproduced,whicharehyaline,aseptate,uniseptateormuhise
6、ptate,ovoidtoelliptical,(3.0—5.1)tzm×(2.1—3.3)p.m.Typicalsymptomsofbrownstemrotoccurredin100%ofseedlingswheninoculatedwiththeisolatesonsoybeancuhivarHefeng25.Toverifythepathogen,rDNA—ITSfragmentsoftheisolatesamplifiedusingprimersITS1andITS4weresequenced,and99%ofsequencesimilarityreachedbetweenthei
7、solateandaPhialophoragregataisolate(GenBankAccessionNo.U66728.1)inGenBank.Thespecific483一bpDNAproductswereamplifiedfromthei—solatesusingprimersBSRI/BSR2specifictoP.gregata.TheaboveresultsindicatedtheisolateswereP