| 周汶静,蒲雪莲,张丽娜,岑伊静,邓晓玲.柚木虱体内“Candidatus Liberibacter asiaticus”多基因位点的分子鉴定[J].中国南方果树,2012,41(3): |
| 柚木虱体内“Candidatus Liberibacter asiaticus”多基因位点的分子鉴定 |
| Molecular characterization of multilocus of "Candidatus Liberibacter asiaticus" in Psylla citrisuga |
| 投稿时间：2012-02-13 修订日期：2012-03-08 |
| DOI： |
| 中文关键词: 柑橘黄龙病 柚木虱 柠檬 “Candidatus Liberibacter asiaticus” |
| 英文关键词:Citrus Huanglongbing Psylla citrisuga Lemon "Candidatus Liberibacter asiaticus" |
| 基金项目:现代农业（柑橘）产业技术体系项目（CARS-27）；农业部公益性行业项目（201003067） |
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| 中文摘要: |
| 利用与柑橘黄龙病相关的韧皮部杆菌（“Candidatus Liberibacter sp.”，简称“Ca. L. sp.”）16S rRNA基因、β-操纵子核糖体蛋白基因、外膜蛋白基因以及16S-23S rRNA间隔转录区基因位点的4对特异性引物，对采自云南瑞丽柠檬园的柚木虱（Psylla citrisuga）DNA样品进行PCR扩增，将扩增产物进行纯化、回收，连接至PEASY-T3载体并转化大肠杆菌感受态细胞Trans1-T1，筛选阳性克隆进行测序，利用NCBI Blastn软件以及MEGA5.05软件对所克隆的序列进行同源性分析与聚类分析。结果表明：柚木虱体内韧皮部杆菌亚洲种在这4个基因位点与已报道的“Ca. L. asiaticus” psy62株系全基因组相应基因位点的同源率均为99%。本研究证实柚木虱能够携带韧皮部杆菌亚洲种，是该菌新的昆虫宿主。 |
| 英文摘要: |
| In the study, four pair of primers based on the 16S rRNA gene, β-operon ribosomal protein gene, outer membrane protein gene, 16S-23S rRNA intergenic transcribed spacer gene sequence were designed to amplify \"Candidatus Liberibacter asiaticus\" strains from Psylla citrisuga, Diaphorina citri and Lemon which were collected from Yunnan province. Amplicons were collected, purified, ligated to PEASY-T3 and transformed into E.coli Trans1-T1. The cloned DNAs were sequenced and compared to published DNA sequences in GenBank through BLAST for phylogenetic analyses. A phylogenetic tree constructed for \"Ca. L. sp.\" of Psylla citrisuga and other proteobacterias based on the 16S rDNA sequence showed that the \"Ca. L.sp.\" strains from Psylla citrisuga were clustered with \"Ca. L. asiaticus\". The result showed that the similarity of the four sequences amplified from Psylla citrisuga were respectively 99% compared with the corresponding DNA sequences of \"Ca. L. asiaticus\" str. psy62 in GenBank, suggesting that Psylla citrisuga is a new host of \"Ca. L. asiaticus\". |
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