期刊文献+

沙田柚黄龙病病原β-操纵子DNA片段的克隆与序列分析 被引量:4

Cloning and sequencing of huanglongbing pathogen in shatianyou pomelo
下载PDF
导出
摘要 采集表现斑驳症状的沙田柚叶片,根据柑桔黄龙病特异保守区域β-操纵子序列设计引物,通过对其病叶脉总DNA进行PCR扩增、扩增产物纯化、与pMD18-T Vector连接、转化大肠杆菌(Escherichia coli)DH 5a菌株、筛选阳性克隆重组子、重组子测序,并将测序结果与柑桔黄龙病病原β-操纵子部分DNA相应序列进行比较.结果表明,引起沙田柚斑驳症状的病原与亚洲柑桔黄龙病原(Liberobacter asiaticum)相应序列的同源率为99%,而与非洲柑桔黄龙病病原的同源率仅为79.9%.因此,从分子水平上证明沙田柚斑驳症状是由黄龙病病原侵染所致,该病原属于亚洲种中的一个成员. The amplification of leaf samples on shatianyou pomelo with yellow symptom was carried out using HLB - specific primers. The products were ligated into pMD18 - T vector after purified, and then were transferred into DH5a strain of Escherichia coli respectively. The positive recombinant plasmids were sequenced and compared with β- operon of the citrus Huanglongbing pathogen. The results showed that the cloned DNA fragment shared 99% and 79.9% homogeneity of sequence with those of Liberobacter asiaticum and Liberobacter africanum respectively. This indicated on level of molecule that the shatianyou pomelo disease was the same as citrus Huanglongbing and that the pathogen belonged to Liberobacter asiaticum.
出处 《仲恺农业技术学院学报》 CAS 2005年第4期45-48,共4页 Journal of Zhongkai Agrotechnical College
基金 广东省自然科学基金(032262) 广东省重点科技计划(2004B20901010)资助项目
关键词 沙田柚 黄龙病 多聚酶链式反应 克隆 序列分析 shatianyou pomelo Huanglongbing PCR clone sequence analysis
  • 相关文献

参考文献11

  • 1GRACA J V.Citrus greening disease[J]. Phytopathology, 1991, 29(1): 109- 136.
  • 2PLANET P, JAGOUEIX S, BOVE J M, et al. Detection and characterization of the Africa citrus greening Liberobacter by amplication,cloning, and sequenceing of the rpIKAJL- rpoBC operon[J]. Current Microbiology. 1995, 26(3) : 243 - 250.
  • 3JAGOUEJX S, BOVE J M, GAMIER M. PCR detection of the two Candidatus Liberobacter species associated with greening disease of citrus[J]. Mol Cell Probes, 1996, 10(1)-43-50.
  • 4HOCQUELLET A, TOORAWA P, BOVE J M, et al. Detection and identificatioinof the two Candidatus Liberobacter spevies associated with citrus huanglongbing by PCR amplification of ribosomal: Protein genes of the β- operon[J]. Molecular and Cellular Probes, 1999,13 : 373 - 379.
  • 5刘新华,管宽生,侯郁辉.沙田柚黄龙病的发生及防治[J].植物检疫,1997,11(5):269-270. 被引量:5
  • 6田亚南,柯穗,李韬,柯冲.琯溪蜜柚黄龙病病原的克隆及序列分析[J].福建农业学报,2002,17(1):11-15. 被引量:4
  • 7孔维文,邓晓玲,梁志慧,唐伟文.柑桔黄龙病病原DNA片段的克隆及序列分析[J].植物病理学报,2000,30(1):71-75. 被引量:28
  • 8MURRAY M G, THOMPSON W F. Rapid isolation of high molecular weight plant DNA[J]. Nucleic Acids Res, 1980, 8(12): 4321- 4324.
  • 9MAMBROOK T, FRITSCH E F, MANIATIS T. Molecular cloning(2nd edition)[M]. New York: Cold Spring Harbor Laboratory Press,1989, 183 - 257.
  • 10SAMBROOK J. FRITSCH E F, MANIATIS T. Molecular cloning: a laboratory manual(2nd ed)[M]. New York: Cold Spring Harbor Laboratory Press, 1989. 16- 34.

二级参考文献11

  • 1田亚南,柯穗,柯冲.应用多聚酶链式反应(PCR)技术检测和定量分析柑桔黄龙病病原[J].植物病理学报,1996,26(3):243-250. 被引量:93
  • 2邓晓玲,唐伟文.应用PCR技术检测柑桔黄龙病病原的研究[J].华南农业大学学报,1996,17(3):119-120. 被引量:21
  • 3卢圣栋,现代分子生物学实验技术,1993年,662页
  • 4柯冲 林先沾 陈辉 等.柑桔黄龙病与类立克次体及线状病毒的初步研究[J].科学通报,1979,10:493-496.
  • 5Jagoueix S, Bove J M, Garnier M. The phloem-limited bacterium of greening disease of citrus is a member of the alpha subdivision of proteobacteria[J]. International Journal of Systematic Bacteriology, 1994, 44:397.
  • 6Murray R G E, Schleifer K H. Taxonomic notes: a proposal for recording the properties of putative taxa of procaryotes[J]. International Journal of Systematic Bacteriology, 1999, 44:174-176.
  • 7Villechanoux S, Garnier M, Laigret F, et al. The genome of the non-cultured, bacterial-like organism associated with citrus greening disease contains the nus G-rplKAJL-rpoBC gene cluster and the gene for a bacteriophage type DNA polymerase[J]. Current Microbiology, 1993, 26:161-166.
  • 8Planet P, Jagoueix S, Bove J M,et al.Detection and characterization of the african citrus greening liberobacter by amplification, cloning and sequancing of the rplKAJL-rpoBC operon[J]. Current Microbiology, 1995, 30 : 137-141.
  • 9万仁秀,蒋元晖,戴胜根,邹俊渝,段景陶,徐忠强.我国柑桔苗木生产现状剖析[J].中国南方果树,1997,26(3):13-14. 被引量:1
  • 10孔维文,邓晓玲,梁志慧,唐伟文.柑桔黄龙病病原DNA片段的克隆及序列分析[J].植物病理学报,2000,30(1):71-75. 被引量:28

共引文献32

同被引文献77

引证文献4

二级引证文献160

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部