柑橘黄龙病病原的分子检测及其16SrDNA序列分析.doc

柑橘黄龙病原16SrDNA序列异质性分析 孙大光1,2，郑雪芳2，刘波2(，阮传清2，范国成3，Hall D.4，段永平4 （1福建师范大学生命科学学院，福州350108；2福建省农业科学院农业生物资源研究所，福州 350003；3福建省农业科学院果树研究所，福州 350003 ；4美国农业部佛罗里达园艺试验室，弗罗里达34945） 摘要： 关键词：柑橘黄龙病；巢式PCR；16S rDNA；系统进化分析 Analysis about Citrus huanglongbing pathogen 16S rDNA sequence SUN Da-guang1，2，ZHENG Xue-fang2，LIU Bo2*, RUAN Chuan-qing2, FAN Guo-cheng3, Hall D.4, DUAN Yong-ping4 1. College Of Life Science Fujian normal university, Fuzhou 350108, China; 2. Agricultural Bioresource. Research Institute, Fujian Academy of Agricultural Sciences, Fuzhou 350003, China； 3. Research Institute. of Pomology , Fujian Academy of Agricultural Sciences, Fuzhou 350003, China； 4 .USDA-ARS-USHRL, Fort Pierce, FL 34945, U.S.A Abstract：Yellow leaves were counted in different directions of a Citrus grandis Osbeck.Chandler the and Nested-PCR detection of citrus leaf pathogen were carried out and through DNA sequencing phylogenetic tree was built to find out the evolutionary relationship between different parts citrus HLB pathogens. The results showed that positive detection rate was 58.33% in different leaves, and four were dominant character, three were recessive character.The identity for different parts citrus leaf pathogen 16S rDNA was up to 99%. Different parts of the citrus pathogen 16S rDNA phylogenetic analysis showed that Shunchang citrus huanglongbing pathogen is Candidatus Liberibacter asiaticum. And 16S rDNA sequence gap analysis showed that the difference betreen Asian species and African species was 2.75%, African species and American species was 3.90%, Americas species and Asian species was 3.44%. Keywords：Citrus Huanglongbing；Nested-PCR；16S rDNA；Phylogenetic analysis 柑橘黄龙病（Citrus Huanglongbing，简称HLB）又叫柑橘青果病（Citrus greening disease）是柑橘生产中最严重的病害之一[1]。柑橘黄龙病病原菌是一种只限于韧皮部内寄生的革兰氏阴性细菌，属于变形菌门的α亚群，根据病原的热敏性、传播媒介等可以将其分为亚种（Candidatus Liberibacter asiaticum），非种（Candidatus Liberibacter africanus）和美种（Candidatus Liberibacter americanus）3个种。我国的黄龙病病原为亚洲种（Ca.L.asiaticus）[2]。目前柑橘黄龙病病原菌还是一种难以