Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (18): 3784-3792.doi: 10.3864/j.issn.0578-1752.2013.18.006

• PLANT PROTECTION • Previous Articles     Next Articles

Genetic Diversity of Two Hypervariable Genes from Prophage Regions of ‘Candidatus Liberibacter asiaticus’ in China

 TAN  Jin-12, WANG  Xue-Feng-2, SU  Hua-Nan-12, LI  Zhong-An-2, ZHOU  Chang-Yong-2   

  1. 1.College of Plant Protection, Southwest University, Chongqing 400716
    2.Citrus Research Institute, Chinese Academy of Agricultural Sciences, Chongqing 400712
  • Received:2013-02-26 Online:2013-09-15 Published:2013-06-04

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Abstract: 【Objective】 Hypervariable genomic loci of prophage region were used to study population differentiation and genetic diversity of ‘Candidatus Liberibacter asiaticus’, the putative casual agent of citrus Huanglongbing (HLB). 【Method】 Two primer sets (Lap-TJ-F/Lap-TJ-R1 and Lap-TJ-F/Lap-TJ-R2), targeted hypervariable genomic regions of two prophage types (SC1 and SC2), respectively, were used for PCR detection and sequence analysis of 224 ‘Ca. L. asiaticus’ strains from different citrus producing areas in China. 【Result】 PCR amplicon polymorphisms and four amplion types (SC1-1, SC1-2, SC2-1 and SC2-2) were observed, SC1-1 and SC2-1 were the predominant types in Southwestern region and Guangdong-Guangxi region, respectively, and no dominant types were found in Fujian-Jiangxi-Zhejiang region. The sequence differences in SC1-1/SC1-2 were mainly caused by different combinations of two kinds of tandem repeats with 132 bp satellite sequence and 32 bp mini-satellite sequence, respectively. The diversity in SC2-1/SC2-2 likely derived from prophage gene rearrangement. 【Conclusion】Significant diversity in the prophage regions was investigated from ‘Ca. L. asiaticus’ strains collected from different geographical origins. Further studies on these genomic regions would be helpful for deciphering the genetic diversity of ‘Ca. L. asiaticus’ populations in China.

Key words: citrus Huanglongbing (HLB) , prophage , tandem repeat , genetic diversity

[1]范国成, 刘波, 吴如健, 李韬, 蔡子坚, 柯冲. 中国柑橘黄龙病研究30年. 福建农业学报, 2009, 24(2): 183-190.

Fan G C, Liu B, Wu R J, Li T, Cai Z J, Ke C. Thirty years of research on citrus Huanglongbing in China. Fujian Journal of Agricultural Sciences, 2009, 24(2):183-190. (in Chinese)

[2]Jagoueix S, Bové J M, Garnier M. The phloem-limited bacterium of greening disease of citrus is a member of the alpha subdivision of the proteobacteria. International Journal of Systematic Bacteriology, 1994, 44(3): 379-386.

[3]Teixeira do D C, Saillard C, Eveillard S, Danet J L, da Costa P I, Ayres A J, Bové J. ‘Candidatus Liberibacter americanus’, associated with citrus Huanglongbing (greening disease) in São Paulo State, Brazil. International Journal of Systematic and Evolutionary Microbiology, 2005, 55(5): 1857-1862.

[4]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橘黄龙病病原. 植物病理学报, 1996, 26(3): 243-250.

Tian Y N, Ke S, Ke C. Detection and quantitation of citrus Huanglongbing pathogen by polymerase chain reaction. Acta Phopathologica Sinica, 1996, 26(3): 243-250. (in Chinese)

[5]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. 植物病理学报, 2000, 30(1): 71-75.

Kong W W, Deng X L, Liang Z H, Tang W W. Cloning and sequencing of the citrus Huanglongbing pathogen DNA. Acta Phopathologica Sinica, 2000, 30(1): 71-75. (in Chinese)

[6]刘利华, 姚锦爱, 种藏文, 谢荔岩. 柑桔黄龙病亚洲种病原的PCR-SSCP分析. 福建农业学报, 2005, 20(2): 77-79.

Liu L H, Yao J A, Chong C W, Xie L Y. Analysis of citrus Huanglongbing pathogen from different places by PCR-SSCP assay. Fujian Journal of Agricultural Sciences, 2005, 20(2): 77-79. (in Chinese)

[7]姚锦爱, 谢荔岩, 郑璐平, 胡奇勇. 柑桔黄龙病亚洲种病原 (Candidatus Liberobacter asiaticus) 16S rDNA的PCR-SSCP分析. 福建农业学报, 2008, 23(3): 331-333.

Yao J A, Xie L Y, Zheng L P, Hu Q Y. PCR-SSCP analysis on 16S rDNA of citrus Huanglongbing pathogen (Candidatus Liberobacter asiaticus). Fujian Journal of Agricultural Sciences, 2008, 23(3): 331-333. (in Chinese)

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

Ding F, Hong N, Zhong Y, Yi G J, Wang G P. Studies on 16S rDNA sequence of citrus Huanglongbing bacteria in China. Acta Horticulturae Sinica, 2008, 35(5): 649-654. (in Chinese)

[9]Subandiyah S, Iwanami T, Tsuyumu S, Ieki H. Comparison of 16S rDNA and 16S/23S intergenic region sequences among citrus greening organisms in Asia. Plant Disease, 2000, 84(1): 15-18.

[10]Okuda M, Matsumoto M, Tanaka Y, Subandiyah S, Iwanami T. Characterization of the tuf B-sec E-nus G-rpl KAJL-rpo B gene cluster of the citrus greening organism and detection by loop-mediated isothermal amplification. Plant Disease, 2005, 89(7): 705-711.

[11]娄兵海. 中国八省柑桔黄龙病病原菌种类和种内分化的初步研究[D]. 重庆: 西南大学, 2008.

Lou B H. Primary Study on species and intraspecific differentiations of HLB pathogens in eight provinces of China[D]. Chongqing: Southwest University, 2008. (in Chinese)

[12]Bastianel C, Garnier-Semancik M, Renaudin J, Bové J M, Eveillard S. Diversity of Candidatus Liberibacter asiaticus, based on the omp gene sequence. Applied and Environmental Microbiology, 2005, 71(11): 6473-6478.

[13]王中康, 周彬彬, 田圣超, 石小刚, 贤家旭, 陈世伟, 殷幼平. 中国不同地区亚洲韧皮杆菌遗传多样性分析. 植物病理学报, 2009, 39(6): 593-599.

Wang Z K, Zhou B B, Tian S C, Shi X G, Xian J X, Chen S W, Yin Y P. Genetic diversity of Candidatus Liberibacter asiaticus isolates from different geographical regions in China. Acta Phytopathologica Sinica, 2009, 39(6): 593-599. (in Chinese)

[14]Hu W Z, Wang X F, Zhou Y. Diversity of the omp gene in Candidatus Liberibacter asiaticus in China. Journal of Plant Pathology, 2011, 93(1): 211-214.

[15]Duan Y P, Zhou L J, Hall D G, Li W B, Doddapaneni H, Lin H, Liu L, Vahling C M, Gabriel D W, Williams K P, Dickerman A, Sun Y J, Gottwald T. Complete genome sequence of citrus Huanglongbing bacterium, ‘Candidatus Liberibacter asiaticus’ obtained through metagenomics. Molecular Plant-Microbe Interactions, 2009, 22(8): 1011-1020.

[16]Zhang S J, Flores-Cruz Z, Zhou L J, Kang B H, Fleites L A, Gooch M D, Wulff N A, Davis M J, Duan Y P, Gabriel D W. ‘Candidatus Liberibacter asiaticus’ carries an excision plasmid prophage and a chromosomally integrated prophage that becomes lytic in plant infections. Molecular Plant-Microbe Interactions, 2011, 24(4): 458-468.

[17]Tomimura K, Miyata S, Furuya N, Kubota K, Okuda M, Subandiyah S, Hung T H, Su H J, Iwanami T. Evaluation of genetic diversity among 'Candidatus liberibacter asiaticus' isolates collected in southeast Asia. Phytopathology, 2009, 99(9): 1062-1069.

[18]Liu R, Zhang P, Pu X L, Chen J , Deng X L. Analysis of a prophage gene frequency revealed population variation of ‘Candidatus Liberibacter asiaticus’ from two citrus-growing provinces in China. Plant Disease, 2011, 95(4): 431-435.

[19]Wang X F, Zhou C Y, Deng X L, Su H N, Chen J C. Molecular characterization of a mosaic locus in the genome of ‘Candidatus Liberibacter asiaticus’. BMC Microbiology, 2012, 12(1): 12-18.

[20]Zhou L J, Powell C A, Hoffman M T, Li W B, Fan G C, Liu B, Lin H, Duan Y P. Diversity and plasticity of the intracellular plant pathogen and insect symbiont, ‘Candidatus Liberibacter asiaticus’, revealed by hyper variable prophage genes with intragenic tandem repeats. Applied and Environmental Microbiology, 2011, 77(18): 6663-6673.

[21]Murray M G, Thompson W F. Rapid isolation of high molecular weight plant DNA. Nucleic Acids Research, 1980, 8(19): 4321-4326.

[22]Jagoueix S, Bové J M, Garnier M. PCR detection of the two ‘Candidatus Liberobacter’ species associated with greening disease of citrus. Molecular and Cellular Probes, 1996, 10(1): 43-50.

[23]Tamura K, Dudley J, Nei M, Kumar S. MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Molecular Biology and Evolution, 2007, 24(8): 1596-1599.

[24]Benson G. Tandem repeats finder: a program to analyze DNA sequences. Nucleic Acids Research, 1999, 27(2): 573-580.
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