文章摘要
链格孢菌Alternaria alternata和细极链格孢A. Tenuissima是浙贝母黑斑病的致病菌
Alternaria alternata and A. tenuissima are the Pathogenic Fungi of Black Spot Disease of Fritillaria thunbergii
投稿时间:2019-07-29  修订日期:2019-07-29
DOI:
中文关键词: 浙贝母  黑斑病  DNA条形码  链格孢菌  细极链格孢菌
英文关键词: Fritillaria thunbergii  black spot  DNA barcode  Alternaria alternata  Alternaria tenuissima
基金项目:浙江省科技计划项目(2015C32076)
作者单位E-mail
徐云飞 浙江中医药大学生命科学学院 13735592593@163.com 
祁依佳 浙江中医药大学生命科学学院  
温思思 浙江中医药大学生命科学学院  
王丹依 浙江中医药大学生命科学学院  
赵伟春 浙江中医药大学生命科学学院 weichunzhao@zcmu.edu.cn 
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中文摘要:
      摘要: 目的 分离、纯化和鉴定浙贝母黑斑病的病原真菌,为浙贝母黑斑病的监测及防治提供基础。方法 采用病组织分离法在PDA培养基上分离纯化病原真菌,PCR扩增基因组DNA的核糖体内转录间隔区(Internal Transcribed Spacer,ITS)、翻译延伸因子-1α(Translation Elongation Factor 1 Alpha,TEF-1α)、甘油醛-3-磷酸脱氢酶(glyceraldehyde 3-phosphate dehydrogenase,G3PDH)和RNA聚合酶II亚基(the second largest subunit of the nuclear RNA polymerase enzyme II,RPB2)序列并双向测序、拼接,经与GenBank中序列同源性比对后构建系统发育邻接树,确定真菌种类。摩擦接种法检测致病性。结果 从浙贝母黑斑病病株中分离纯化获得2种链格孢属Alternaria真菌。分离物1的ITS、TEF-1α、G3PDH、RPB2扩增片段与GenBank中链格孢菌Alternaria alternata(登录号:MG182428、KY175227、XM_018523704、MG250634)的同源性分别均为100%。分离物2则与细极链格孢菌Alternaria tenuissima(登录号:MK967997、LC136862、KY290574、LT707523)的同源性分别均为100%。经摩擦接种确定两种链格孢属真菌均为浙贝母黑斑病致病菌。结论 浙贝母黑斑病的病原真菌有链格孢菌和细极链格孢菌,其中细极链格孢菌首次被证明可引起浙贝母黑斑病。
英文摘要:
      Abstract: Objective To isolate, purify and identify the pathogenic fungi of black spot disease of Fritillaria thunbergii, and provide experimental basis for the monitoring and control of black spot disease of F. thunbergii. Methods Pathogenic tissue isolation method was used to isolate and purify pathogenic fungi from F. thunbergii. Partial fragments of ribosomal internal transcription spacer (ITS), translation elongation factor-1α (TEF-1α), glyceraldehyde-3-phosphate dehydrogenase (G3PDH) and the second largest subunit of the nuclear RNA polymerase enzyme II (RPB2) of the fungi were obtained by PCR amplification with specific primers. After two-way sequencing and splicing, the amplified fragments were homologous with the corresponding sequences in GenBank, and the phylogenetic adjacent trees were constructed to identify fungal species. The pathogenicity of fungi to F. thunbergii was detected by friction inoculation method. Results Two fungi belonging to Alternaria genus were isolated and purified from F. thunbergii plants infected with black spot disease. The ITS, TEF-1α, RPB2, and G3PDH amplified fragments of the isolate 1 were 100% homologous to Alternaria alternata (accession numbers: MG182428, KY175227, XM_018523704, MG250634, respectively) in GenBank, while those of the isolate 2 was 100% homologous to A. tenuissima (accession numbers: MK967997, LC136862, KY290574, LT707523, respectively). Both of the fungi were identified as pathogenic fungi of F. thunbergii black spot by friction inoculation. Conclusion A. alternata and A. tenuissima are the pathogenic fungi of black spot disease of F. thunbergii. A. tenuissima was first reported to cause black spot of F. thunbergii.
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