Molecular identification and detection of Eutypa lata in grapevine
Molecular identification and detection of Eutypa lata in grapevine
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DOI：
10.1017/s0953756205002893
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发表时间：
2005-07-01
影响因子：
--
通讯作者：
Scott, ES
中科院分区：
文献类型：
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作者：
Lardner, R;Stummer, BE;Scott, ES
葡萄顶枯病的病原菌Eutypa lata，很难根据菌落形态进行鉴定，并且当从木材中分离时，经常被其他真菌竞争。为了方便病原体的诊断，我们设计了能够扩增大肠杆菌DNA的SCAR引物。lata的基因组DNA文库，并从该文库中筛选出E.鉴定lata并测序。利用SCAR引物对E. lata直接从培养物中分离，而不需要DNA提取或延长孵育期，并且还可以检测从葡萄藤木材中分离的DNA中的病原体。RFLP探针用于狭缝印迹法检测病原体的DNA分离，从1岁的甘蔗以及从成熟葡萄藤树干。本研究中开发的标记物有可能用作研究工具，以收集有关该疾病流行病学的信息，并评估潜在的控制剂对E.拉塔。
Eutypa lata, the causal agent of Eutypa dieback of grapevines, is difficult to identify on the basis of colony morphology and is often out-competed by other fungi when isolated from wood. To facilitate diagnosis of the pathogen, we designed SCAR primers capable of amplifying DNA of E. lata and constructed a genomic DNA library from which DNA sequences specific to E. lata were identified and sequenced. SCAR primers were used to identify E. lata directly from culture without the requirement for DNA extraction or prolonged incubation periods and could also detect the pathogen in DNA isolated from grapevine wood. RFLP probes were used in slot-blot assays to detect the pathogen in DNA isolated from 1 yr old cane as well as from mature grapevine trunks. The markers developed in this study have the potential to be used as a research tool to gather information on the epidemiology of the disease and to assess the efficacy of potential control agents against E. lata.