柑橘黄龙病菌亚洲种外膜蛋白基因的遗传变异分析  

Genetic variation analysis of an outer membrane protein gene of Candidatus Liberibacter asiaticus

作　　者：鹿连明 杜丹超 程保平[2] 胡秀荣 张利平 陈国庆 

机构地区：[1]浙江省柑桔研究所,浙江台州318026 [2]广东省农业科学院植物保护研究所,广州510640

出　　处：《浙江大学学报（农业与生命科学版）》2014年第2期125-132,共8页Journal of Zhejiang University:Agriculture and Life Sciences

基　　金：农业部公益性行业科研专项(201003067-08)

摘　　要：为了解柑橘黄龙病菌亚洲种(Candidatus Liberibacter asiaticus)的遗传多样性,对来源于国内外7个不同地区和浙江台州地区11个不同寄主品种的Ca.L.asiaticus的外膜蛋白(outer membrane protein,OMP)基因进行聚合酶链反应限制性片段长度多态性分析(polymerase chain reaction-restriction fragment length polymorphism,PCR-RFLP).PCR产物经克隆测序后,利用NCBI Blast和DNAMAN软件对omp基因序列进行多重比对,并用MEGA软件构建系统发育树.结果显示:在浙江台州地区不同寄主品种上Ca.L.asiaticus各分离物的omp基因的酶切片段长度不具有多态性,其基因序列也完全一致;而不同地理来源的Ca.L.asiaticus的omp基因经限制性内切酶消化后可产生不同的RFLP指纹图谱,其核酸序列和氨基酸序列均存在变异,表明Ca.L.asiaticus存在种内遗传多样性.在系统发育树上,所有的Ca.L.asiaticus分离物聚为1个分支,而Ca.L.africanus和Ca.L.solanacearum分别处于不同的分支上.Summary Huanglongbing （HLB） is a destructive disease that represents a major threat to the world citrus industry.It was first reported from Chaoshan area of Guangdong Province in the early 20th century,and is now known to occur in next to 40 different Asian,African,Oceanian,South and North American countries and areas. Its pathogen is a phloem-limited,non-cultured,Gram-negative bacterium which belongs to the genus Candidatus Liberibacter.Three species,i.e.,Ca. L.asiaticus,Ca. L.americanus,and Ca. L.africanus with Ca. L. africanus subsp.capensis are currently known.So far,the pathogen of HLB disease discovered in China is only Ca.L.asiaticus,which distributes widely in almost all major citrus growing areas such as Guangdong,Guangxi, Fujian,Yunnan,Zhejiang and so on.This pathogen can be transmitted by grafting and Diaphorina citri and infect almost all fruit trees of Citrus Fortunella and Poncirus ,causing different characteristic symptoms including＆amp;nbsp;asymmetric blotchy mottle,vein yellowing or nutrition deficiency on leaves,and ？？red nose”,？？greening” or deformed shape on fruits.While the infected Murraya exotica usually show no symptoms.Based on the above mentioned facts,it is referred that genetic diversity might exist in populations of Ca. L.asiaticus.The gene sequences of 1 6S rDNA,1 6S-23S rDNA ITS and 23S rDNA of Ca.L.asiaticus have been confirmed to have a high degree of conservation and are not suitable for genetic diversity analysis.The omp gene,an outer membrane protein gene of Gram-negative bacterium,has been demonstrated as a potential candidate gene used for research on the genetic diversity. 〈br〉 In this study,genetic variations of omp genes of 18 Ca.L.asiaticus isolates from China and other countries were analyzed aiming to understand the genetic diversity in populations of Ca. L.asiaticus and the effect of geographical source and host species. 〈br〉 First,omp genes of Ca.L.asiaticus isolates from seven different geographical regions and 11 different host species in T

关 键 词：柑橘黄龙病菌亚洲种 外膜蛋白 限制性片段长度多态性分析 遗传变异 

分 类 号：Q78[生物学—分子生物学] S436.661[农业科学—农业昆虫与害虫防治]