Scientia Agricultura Sinica ›› 2014, Vol. 47 ›› Issue (22): 4488-4494.doi: 10.3864/j.issn.0578-1752.2014.22.014

• HORTICULTURE • Previous Articles     Next Articles

Population Structure of ‘Candidatus Liberibacter asiaticus’ in China Revealed by SSR Markers

HUANG Ai-jun1,2, SU Hua-nan3, WANG Xue-feng2, TANG Ke-zhi2, LI Zhong-an2, ZHOU Chang-yong2   

  1. 1College of Plant Protection, Southwest University, Chongqing 400716
    2Citrus Research Institute, Chinese Academy of Agricultural Sciences/Southwest University, Chongqing 400712
    3College of Chemistry & Life Science, Gannan Normal University, Ganzhou 341000, Jiangxi
  • Received:2014-04-08 Revised:2014-06-23 Online:2014-11-16 Published:2014-11-16

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Abstract: 【Objective】 ‘Candidatus Liberibacter asiaticus’ is the pathogeny of Citrus Huanglongbing. The purpose of this research was to investigate the population structure of ‘Candidatus Liberibacter asiaticus’ in China by multi SSR (simple sequence repeat) loci. 【Method】 To identify the polymorphism of SSR loci in the genome of Ca. L. asiaticus, 25 reported SSR primer sets were tested using DNA samples obtained from HLB-affected plants from 8 provinces of China. The selected SSR loci were used to analyze 285 samples by PCR and PAGE electrophoresis. The software Quantity One 4.5.0 was applied to estimate the base pairs of the amplicons. The polymorphism of 285 samples in the selected loci were evaluated via the software PopGen version 1.31, and the clustering analysis was performed by software Powermarker 3.25 and structure 2.3.4. 【Result】 A total of 5 SSR loci were identified, with Nei’s gene diversity index ranging from 0.1542 to 0.9556. The locus LasA showed high diversity. Its effective number of allele (NE) and Nei’s gene diversity (H) were 22.5 and 0.9556, respectively. Analysis on population structure of Ca. L. asiaticus of China from different geographical samples revealed that the isolates from Yunnan province had high diversity, (NE=5.7, H=0.6580). The genetic distance of different geographical populations ranged from 0.0236 to 0.5786 and genetic identity ranged from 0.5607 to 0.9767. The longest genetic distance and lowest genetic identity were identified from the geographical population between Guangxi and Sichuan. The clustering analysis indicated all isolates from China were clustered into two groups. One is from Sichuan and Yunnan, the other is from Fujian, Zhejiang, Guizhou, Guangxi, Guangdong and Jiangxi. Structure analysis also showed the populations of Yunnan and Sichuan were constituted nearly by a single group and the rest of them were made up by mixed groups. 【Conclusion】There are possible two kinds of different population structures of Ca. L. asiaticus in China by using the selected SSR markers and the results of this study will provide better insight on the origin and epidemiology of HLB in China.

Key words: Citrus Huanglongbing, SSR, population structure

[1]    Bové J M. Huanglongbing: a destructive, newly-emerging, century- old disease of citrus. Journal of Plant Pathology, 2006, 88(1): 7-37.
[2]    Sutton B, Duan Y P, Halbert S, Sun X A, Schubert T, Dixon W. Detection and identification of Citrus Huanglongbing (greening) in Florida, USA. Proceedings of the International Citrus Canker and Huanglongbing Research Workshop, Orlando, Florida, 2005: 59.
[3]    Coletta-Filho H, Targon M, Takita M, De Negri J, Pompeu Jr J, Machado M, Do Amaral A, Muller G. First report of the causal agent of Huanglongbing (‘Candidatus Liberibacter asiaticus’) in Brazil. Plant Disease, 2004, 88(12): 1382-1382.
[4]    林孔湘. 柑桔黄梢(黄龙)病研究—病情调查. 植物病理学报, 1956, 2(1): 1-11.
Lin K X. Observation yellow shoot of citrus. Acta Phytopathologica Sinica, 1956, 2(1): 1-11. (in Chinese)
[5]   Sechler A, Schuenzel E L, Cooke P, Donnua S, Thaveechai N, Postnikova E, Stone A L, Schneider W L, Damsteegt V D, Schaad N W. Cultivation of ‘Candidatus Liberibacter asiaticus’, ‘Ca. L. africanus’, and ‘Ca. L. americanus’ associated with Huanglongbing. Phytopathology, 2009, 99(5): 480-486.
[6]    Deng X, Chen J, Feng Z, Shan Z, Guo H, Zhu J, Li H, Civerolo E L. Identification and characterization of the huanglongbing bacterium in pummelo from multiple locations in Guangdong, P. R. China. Plant Disease, 2008, 92(4): 513-518.
[7]    Hu W Z, Wang X F, Zhou Y, Li Z A, Tang K Z, Zhou C Y. Diversity of the omp gene in ‘Candidatus Liberibacter asiaticus’ in China. Journal of Plant Pathology, 2011, 93(1): 211-214.
[8]    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.
[9]    Chen J, Deng X, Sun X, Jones D, Irey M, Civerolo E. Guangdong and Florida populations of ‘Candidatus Liberibacter asiaticus’ distinguished by a genomic locus with short tandem repeats. Phytopathology, 2010, 100(6): 567-572.
[10]   Liu R, Zhang P, Pu X, Xing X, Chen J. 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.
[11]   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.
[12]   Ma W, Liang M, Guan L, Xu M, Wen X, Deng X, Chen J. Population Structures of ‘Candidatus Liberibacter asiaticus’ in Southern China. Phytopathology, 2014, 104(2): 158-162.
[13]   谭锦, 王雪峰, 苏华楠, 李中安, 周常勇. 中国柑橘黄龙病病原菌两个原噬菌体超变异基因遗传多样性. 中国农业科学, 2013, 46(18): 3784-3792.
Tan J, Wang X F, Su H N, Li Z A, Zhou C Y. Genetic Diversity of Two Hypervariable Genes from Prophage Regions of ‘Candidatus Liberibacter asiaticus’ in China. Scientia Agricultura Sinica. (in Chinese), 2013, 46(18): 3784-3792
[14]   Islam M S, Glynn J M, Bai Y, Duan Y P, Coletta-Filho H D, Kuruba G, Civerolo E L, Lin H. Multilocus microsatellite analysis of ‘Candidatus Liberibacter asiaticus’ associated with Citrus Huanglongbing worldwide. BMC Microbiology, 2012, 12:39.
[15]   Katoh H, Subandiyah S, Tomimura K, Okuda M, Su H J, Iwanami T. Differentiation of "Candidatus Liberibacter asiaticus" isolates by variable number tandem repeat analysis. Applied and Environmental Microbiology, 2011, 77(5): 1910-1917.
[16]   Katoh H, Davis R, Smith M W, Weinert M, Iwanami T. Differentiation of Indian, East Timorese, Papuan and Floridian ‘Candidatus Liberibacter asiaticus’ isolates on the basis of simple sequence repeat and single nucleotide polymorphism profiles at 25 loci. Annals of Applied Biology, 2012, 160(3): 291-297.
[17]   苏华楠. 中国柑橘黄龙病病原调查、种群遗传分化和Lysozyme蛋白原核表达[D]. 重庆: 西南大学, 2013.
Su H N. HLB etiology survey, population differentiation of ‘Candidatus Liberibacter asiaticus’ in China and Lysozyme protein prokaryotic expression of its prophage[D]. Chongqing: Southwest University, 2013. (in Chinese)
[18]   Reinking O A. Diseases of economic plants in south China. Philippine Agricalturist, 1919: 109-135.
[19]   Duan Y, Zhou L, Hall D G, Li W, Doddapaneni H, Lin H, Liu L, Vahling C M, Gabriel D W, Williams K P, Dickerman A, Sun Y, Gottwald T. Complete genome sequence of Citrus Huanglongbing bacterium, ‘Candidatus Liberibacter asiaticus’ obtained through metagenomics. Molecular Plant-Microbe Interactions, 2009, 22(8): 1011-1020.
[20]   Glader B E, Backer K, Diamond L K. Elevated erythrocyte adenosine deaminase activity in congenital hypoplastic anemia. New England Journal of Medicine, 1983, 309(24): 1486-1490.
[21]   Cristalli G, Costanzi S, Lambertucci C, Lupidi G, Vittori S, Volpini R, Camaioni E. Adenosine deaminase: functional implications and different classes of inhibitors. Medicinal Research Reviews, 2001, 21(2): 105-128.
[22]   Zhang S, Flores-Cruz Z, Zhou L, Kang B H, Fleites L A, Gooch M D, Wulff N, Davis M J, Duan Y P, Gabriel D W. ‘Ca. 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.
[23]   柏自琴. 中国柑橘黄龙病发生动态及其病原菌亚洲种群分化研究[D]. 重庆: 西南大学, 2012.
Bai Z Q. Occurrence dynamics of Citrus Huanglongbing in P.R. China and population differentiation of ‘Candidatus Liberibacter asiaticus’ [D]. Chongqing: Southwest University, 2012. (in Chinese)
[24]   Gmitter F G, Hu X. The possible role of Yunnan, China, in the origin of contemporary citrus species (Rutaceae). Economic Botany, 1990, 44(2): 267-277.
[25]   Lin H, Islam M S, Bai Y, Wen A, Lan S, Gudmestad N C, Civerolo E L. Genetic diversity of ‘Cadidatus Liberibacter solanacearum’ strains in the United States and Mexico revealed by simple sequence repeat markers. European Journal of Plant Pathology, 2011, 132(2): 297-308.
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