中国柑橘黄龙病菌16S rDNA序列研究

园艺学报 ›› 2008, Vol. 35 ›› Issue (5): 649-654.

中国柑橘黄龙病菌16S rDNA序列研究

丁芳^1,2,3;洪霓^1,4;钟云²;易干军^2*;王国平^1,3*

(¹华中农业大学植物科学技术学院, 武汉 430070；²广东省农业科学院果树研究所, 广州 510640；³华中农业大学国家果树脱毒种质资源室内保存中心，武汉 430070；⁴华中农业大学湖北省作物病害监测与安全控制重点实验室，武汉 430070)

收稿日期:2007-10-26 修回日期:2008-04-14 出版日期:2008-05-25 发布日期:2008-05-25
通讯作者: 易干军

Studies on 16S rDNA Sequence of Citrus Huanglongbing Bacteria in China

DING Fang ^{1, 2, 3}, HONG Ni ^{1, 4}, ZHONG Yun ² , YI Gan-jun ^2* and WANG Guo-ping ^{1, 3*}

(¹College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China; ²Institute of Fruit Tree Research, Guangdong Academy of Agricultural Science, Guangzhou 510640, China; ³National Indoor Conservation Center of Virus-free Germplasm of Fruit Crops, Huazhong Agricultural University, Wuhan 430070, China; ⁴The Key Laboratory of Plant Pathology of Hubei Province, Huazhong Agricultural University, Wuhan 430070, China)

Received:2007-10-26 Revised:2008-04-14 Online:2008-05-25 Published:2008-05-25
Contact: YI Gan-jun

摘要/Abstract

摘要：

运用PCR技术对来自中国7省区不同寄主上的黄龙病菌16S rDNA基因区进行了PCR-RFLP-SSCP分析, 采用3种限制性内切酶进行单酶切、双酶切及三酶切反应，对酶切产物进行了单链构象多态性（SSCP）分析，结果表明来自7省区不同寄主上9个黄龙病病菌分离物的16S rDNA无可见变异；同时对9个代表性的分离物16S rDNA进行了克隆测序，序列多重比对结果与PCR-RFLP-SSCP一致，从而在分子水平上证明了中国柑橘黄龙病病菌的16S rDNA序列高度保守, 在不同的地域、寄主内没有发现分子变异，为进一步研究柑橘黄龙病菌系统进化奠定了基础。

关键词: 柑橘, 黄龙病, 病原菌, 16S rDNA, PCR-RFLP-SSCP, 克隆, 序列分析

Abstract:

PCR-RFLP-SSCP analysis was carried out to study the 16S rDNA of the citrus Huanglongbing (HLB) bacterial isolates collected from 7 provinces in China. Three restriction endonucleases were used to digest the target DNA fragment of 16S rDNA. The digested products were subjected to single-strand conformation polymorphism (SSCP) analysis. The results showed that there was no obvious difference among the 9 representative isolates from 7 provinces in 16S rDNA. The 16S rDNAs of the 9 representative isolates were cloned and sequenced. The result of the multiple alignment analysis of the sequences was in agreement with that of PCR-RFLP-SSCP analysis. It revealed that the citrus Huanglongbing bacteria isolated from different areas of China were highly conservative in their 16S rDNA sequence, and no molecular change were found among isolates from different hosts and different geographic areas. The result laid a foundation for further research on systematic evolution of HLB bacteria.

Key words: citrus, Huanglongbing, Candidatus, 16S rDNA, PCR-RFLP-SSCP, cloning, sequence analysis

中图分类号:

S 666

丁芳;;洪霓;钟云;易干军;王国平;. 中国柑橘黄龙病菌16S rDNA序列研究 [J]. 园艺学报, 2008, 35(5): 649-654.

DING Fang;;;HONG Ni;;ZHONG Yun ;YI Gan-jun and WANG Guo-ping; . Studies on 16S rDNA Sequence of Citrus Huanglongbing Bacteria in China[J]. ACTA HORTICULTURAE SINICA, 2008, 35(5): 649-654.

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