大豆茎褐腐病病原菌鉴定-论文.pdf

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1、中国油料作物学报2013，35(2)：196—200ChineseJournalofOilCropSciencesdoi：10．7505／j．issn．1007—9084．2013．02．014大豆茎褐腐病病原菌鉴定段灿星，张吉清，丁俊杰，王晓呜，朱振东¨(1．中国农业科学院作物科学研究所／国家农作物基因资源与基因改良重大科学工程，北京，1000812．黑龙江省农业科学院佳木斯分院，黑龙江佳木斯，154007)摘要：对新发生的引起大豆茎维管束和髓褐腐的病原菌进行鉴定。该病原分离物在PDA培养基上的菌落扁平，致密，不规则圆形，白色到浅灰褐色，具放射状褶皱突起，菌落边缘粗糙不整齐，或带有缺刻；

2、在GBA培养基上菌落白色至灰色，菌丝紧贴培养基，气生菌丝稀少，菌落圆形，边缘整齐。在GBA培养基上分离物产生分生孢子，分生孢子卵形至椭圆形，无色；单胞，或形成有分隔的细胞；平均长度为(3．0～5．1)txm×(2．1～3．3)m。致病性测定表明分离物能100％引起合丰25大豆植株出现典型大豆茎褐腐病症状。引物ITSI／ITS4在分离物中扩增的产物与GenBank中Phialophoragregata分离物序列的相似性达99％。利用大豆茎褐腐病菌的特异引物BSR1／BSR2在分离物中扩增出483bp的特异性条带。研究结果表明，上述分离物确为大豆茎褐腐病菌。关键词：大豆；分离物；大豆茎褐腐病菌

3、；大豆茎褐腐病中图分类号：S435．651文献标识码：A文章编号：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