基于23S／5SrDNA序列的柑橘黄龙病病原菌亚洲种系统发育研究  

Phylogenetic Analysis of Candidatus Liberibacter Asiaticus of Citrus Huanglongbing Based on 23S/5S rDNA Sequence

作　　者：廖惠红[1,2] 白先进[3] 李杨瑞[1,3] 陈传武 杨丽涛[1] 朱建华[2] 徐宁[2] 

机构地区：[1]广西大学农学院,南宁530004 [2]广西农业科学院园艺研究所,南宁530007 [3]广西农业科学院,南宁530007 [4]广西柑橘研究所,桂林541004

出　　处：《生物技术通报》2009年第5期88-91,100,共5页Biotechnology Bulletin

基　　金：广西壮族自治区人才小高地项目桂发办[2004]47号

摘　　要：根据柑橘黄龙病亚洲种23S/5S的DNA序列设计一对引物对不同地理来源的6个柑橘黄龙病样品DNA进行扩增,扩增片段大小均为1 654 bp包括一个假定细胞壁水解酶假基因(putative cell wall hydrolase pseudogene)和5S rRNA基因。序列同源性分析结果表明;6个柑橘黄龙病病原菌样品与柑橘黄龙病病原菌亚洲种Sihui样品的同源性为99%,然而与土壤杆菌,布鲁氏菌,根瘤菌,中华根瘤菌,巴通体菌和中慢生根瘤菌的同源性只有89%～95%,说明在23S/5S rDNA序列上黄龙病病原菌亚洲种与α变形菌纲根瘤菌目的其他病原菌相差较大。对黄龙病病原菌亚洲种种内的23S/5S rDNA序列进行比较分析,结果发现黄龙病病原菌亚洲种种内之间putative cell wall hydrolase pseudogene和5S rRNA的基因序列非常保守,但不同地理来源的柑橘黄龙病样品碱基序列间确实存在差异,差异的大小与地理的远近无关。利用简约法对黄龙病病原菌亚洲种及α变形菌纲其它病原菌的23S/5S rDNA序列构建的系统发育树显示黄龙病病原菌亚洲种单独聚为一类,其他细菌聚为另一类,该结果与基于rplJ基因及16S rRNA基因的DNA序列构建的分子系统进化树结果一致。A pair of primers based on the 23S/5S rDNA sequences was designed to amplify six Candidatus Liberibacter asiaticus strains collected from different geographical regions in China. The amplicon size is 1 654 bp including putative cell wall hydrolase pseudogene and 5S rRNA gene. BLAST analysis demonstrated that these strains shared 99% of sequence identity with the Ca. L. asiatieus sihui strain, but only 89% -95% of sequence identity with Agrobacterium, BruceUa, Rhizobium, Sinorhizobium, Bartonella and Mesorhizobium. Sequence comparisons were also conducted among Ca. L. asiaticus in 23S/5S rDNA sequence region. The result showed that putative cell wall hydrolase pseudogene and 5S rRNA gene sequences in 23S/5S rDNA region were conserved except a few of nucleotide variations among the multiple strains of Ca. L. asiaticus from different geographical regions. A phylogenetic tree constructed for Ca. L. asiaticus strains and other α-proteobacterias based on the 23S/5S rDNA sequence showed that all Ca. L. asiaticus strains were clustered together, and separated with other α-proteobacterias.

关 键 词：柑橘 黄龙病 亚洲种 23S/5S RDNA 

分 类 号：S436.66[农业科学—农业昆虫与害虫防治]