水稻细菌性条斑病菌的实时荧光PCR检测技术研究  

Real-time Quantitative PCR for Detection of Xanthomonas oryzae pv. oryzicola

作　　者：韩阳[1] 张丽辉[1] 王永吉[1] 林琳[1] 周丽洪[1] 姬广海[1] 

机构地区：[1]云南农业大学植物保护学院,云南昆明650201

出　　处：《云南农业大学学报（自然科学版）》2012年第3期315-320,共6页Journal of Yunnan Agricultural University:Natural Science

基　　金：农业部公益性(农业)行业科研专项(3-23);公益性行业(农业)科研专项(201003029)

摘　　要：水稻细菌性条斑病是我国水稻上的检疫性病害之一。设计并筛选出特异性引物对Xoc2071F/Xoc2071R(Xoc2071),其扩增片段大小为329 bp,利用SYBR GreenI荧光定量PCR方法进行水稻细菌性条斑菌的实时荧光PCR检测。结果表明,引物对Xoc2071能特异性检测到目标病原菌产生荧光信号而其他菌种不产生荧光信号。检测的灵敏度是2.25 fg/μL质粒DNA和1.0×102cfu/mL的菌悬液,相当于1个细菌的基因,比常规PCR电泳检测的灵敏度高约100倍。此实时荧光PCR可以检测到仅需10 g的带菌种子上目标菌的存在。Xanthomonas oryzae pv. oryzicola was listed as a plant quarantine disease of China. Primers pair of Xoc2071F/Xoc2071R （Xoc2071） is specific primers based on cysteine synthase gene to amplify fragment 329 bp. Using SYBR Green I fluorescence quantitative PCR method and primers Xoc2071, the detection method of Xooc by qPCR was established. The results showed that the primers Xoc2071 could specifically detect the target bacteria, but the other pathogenic bacteria can not have fluorescence signals. The level of detection was 2.25 fg/μL plasmid DNA and 1.0 ×10^2 cfu/mL bacterial suspension, roughly equaling to one cell and 100 times sensitive than PCR gel electrophoresis detection. Xooc was detected from seed washes of infected seeds as small as 10 g.

关 键 词：水稻细菌性条斑病菌 检测 荧光定量PCR 

分 类 号：S435.111.49[农业科学—农业昆虫与害虫防治]