华侨大学学报（自然科学版）2018，Vol.39Issue(2)：221-226,6.DOI:10.11830/ISSN.1000-5013.201709055
快速简单组织定位甘蔗RSD致病菌Lxx的原位PCR方法
Fast and Simple In Situ PCR Method for Localizing RSD Pathogen Lxx in Sugarcane Tissue
摘要
Abstract
An in situ polymerase chain reaction (in situ PCR) method was developed for visualizing the localization of the pathogen Lxx (Leisonia xyli subsp.xyli) in sugarcane frozen section with ratoon stunting disease (RSD),providing a tool for the studyon RSD in sugarcane,especiallyon the mechanism of Lxx and sugarcane interaction.Frozen sections were obtained from both RSD-infected and-uninfectedsugarcane plants (cultivar Badila).After the digestion with lysozyme,in situ PCR was performed directly on the sections by amplif ying Lxx 16S～23S rDNA internal transcriptional spacer (ITS) using the specific primers,the resulting products,which contained the random Digoxigenin-11-dUTP in the amplified fragments,were visualized based on immunohistochemistry,enabling a direct in situ PCR detectionfor Lxx on sugarcane frozen sections.The results showed that the positive signalsin the stem of RSD-infected samples were detected and localized on sugarcane stem xylem and phloem,while no signals was found in the healthy samples,suggesting this method was excellent in specificity.关键词
甘蔗宿根矮化病/革兰氏阳性苛养致病菌/冰冻切片/原位多聚酶链式反应Key words
ratoon stunting disease/Leisonia xyli subsp.xyli/frozen sections/in situ polymerase chain reaction分类
农业科技引用本文复制引用
胡敏,汪文华,何恩铭,孟红岩,郭莺..快速简单组织定位甘蔗RSD致病菌Lxx的原位PCR方法[J].华侨大学学报（自然科学版）,2018,39(2):221-226,6.基金项目
国家自然科学基金青年科学基金资助项目(31301381) （31301381）
