| 利用基因组步行方法克隆柑橘黄龙病病原菌亚洲种序列 |
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| 引用本文： | 廖惠红,李杨瑞,彭宏祥,白先进,杨丽涛,徐宁.利用基因组步行方法克隆柑橘黄龙病病原菌亚洲种序列[J].园艺学报,2009,36(4):493-500. |
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| 作者姓名： | 廖惠红 李杨瑞 彭宏祥 白先进 杨丽涛 徐宁 |
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| 作者单位： | (广西大学农学院, 南宁530004; 广西农业科学院园艺研究所, 南宁530007; 广西作物遗传改良生物技术重点开放实验室, 南宁530007) |
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| 基金项目： | 广西壮族自治区人才小高地项目 |
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| 摘 要： | 对NCBI基因库黄龙病亚洲种病原菌序列进行拼接, 并根据拼接结果在rplKAJL-rpoBC, 16S/23S rRNA和omp 3个位点上设计引物, 采用基因组步行方法, 在其亚洲种3个位点上共获得了15 168 bp非冗余新序列, 其中, 在rplKAJL-rpoBC位点上5′端上游区和3′端下游区分别获得了3 218和2 653 bp的非冗余新序列; 在16S/23S rRNA 位点上5′端上游区和3′端下游区分别延伸了1 853和2 459 bp的新序列; 在omp基因位点上分别获得了4 014和971 bp的新序列。序列分析表明在rplKAJL-rpoBC位点上的新序列包含2个新基因(suhB和serA); 16S/23S rRNA位点新序列包含4个新基因(膜转运蛋白基因、细胞壁脱氢酶假基因、5S rRNA、tRNAMet ) , omp位点新序列包含6个新基因(rpsB、tsf、pyrH、frr、cdsA2、cds2) 。这些新序列的获得为以PCR为基础的黄龙病相关研究提供了有用的参考依据。
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| 关 键 词： | 柑橘 黄龙病 Candidatus Liberibactor 序列 |
| 收稿时间： | 2008-12-29 |
| 修稿时间： | 2009-3-3 |
Sequences Cloning of Candidatus Liberibactor asiaticus Associated with Huanglongbing Using Genomic Walking Method |
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| LIAO Hui-hong,LI Yang-rui,PENG Hong-xiang,BAI Xian-jin,YANG Li-tao,XU Ning.Sequences Cloning of Candidatus Liberibactor asiaticus Associated with Huanglongbing Using Genomic Walking Method[J].Acta Horticulturae Sinica,2009,36(4):493-500. |
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| Authors: | LIAO Hui-hong LI Yang-rui PENG Hong-xiang BAI Xian-jin YANG Li-tao XU Ning |
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| Institution: | (Agriculture College, Guangxi University, Nanning 530004, China; Horticulture Research Institute, Guangxi Academy of Agricultural Sciences, Nanning 530007, China; Guangxi Crop Genetic Improvement and Biotechnology Lab, Nanning 530007,China) |
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| Abstract: | Huanglongbing (HLB) , previously known as greening disease, is probably the most serious disease in citrus. The presump tive causal agent is believed to be a non-cultured, phloem-restricted α-proteobacterium. Currently, there are three Candidatus L iberibactor spp. associated with citrus Huanglongbing(HLB) disease, Ca. Liberibactor asiaticus, Ca. L iberibactor africanus and Ca. L iberibactor am ericanus. In this study, a genomic walkingmethod was used to obtain new sequences extended from previously known loci,rplKAJL-rpoBC, 16S/23S rRNA and outermembrane protein (omp) of Ca. Liberibactor asia ticus. In rplKA-JL 2rpoBC locus, 3 218 bp of new sequenceswere obtained at the 5′up stream end and 2 653 bp at the 3′downstream. Totally, fragments about 1 853 bp and 2 459 bp were also acquired from 5′up stream and 3′downstream of 16S/23S rRNA region, respectively. Similarly, in om p locus, fragment about 4 014 bp were obtained at 5′up stream and 971 bp at 3′ downstream, respectively. Genomic walking app roach was used in thisstudy, and totally 15 168 bp non-redundancy sequences from asiaticus specieswere obtained. Sequence analy-sis shows that there are two new genes ( suhB and serA) at the rplKAJL-rpoBC gene cluster locus, four newgenes ( transmembrane p rotein gene, cell wall hydrolase p seudogene, 5S ribosomal RNA and tRNAMet ) extended from the 16S/23S rRNA gene locus and six new genes ( rpsB, tsf, pyrH, frr, cdsA2, cds2) at the omplocus. These new sequences obtained in this study will facilitate the PCR-based HLB research. |
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| Keywords: | Cand/datus Liberibactor |
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