Detection of Candidatus Liberibacter asiaticus causing Citrus Huanglongbing disease
Article Sidebar
Main Article Content
Abstract
Detection of Candidatus Liberibacter spp. causing citrus Huanglongbing (HLB) disease was facilitated by taking samples from three different citrus orchards located in Chiang Mai, Chiang Rai, and Phrae provinces. A polymerase chain reaction (PCR) with specific primers OI1/OI2c and A2/J5 were used for detection and they produced specific band of 1160 bp and 703 bp, respectively. These were amplified from diseased leaves whereas no product from healthy citrus plants. To improve the method of detection for HLB disease, this study were a utilized five methods for pathogen DNA extraction from disease leaves and applied to a PCR; protocol three and four were most effective. When primers A2/J5 were used to identify the Liberibacter species, a 703 bp band was observed from all infected plants. The sequence of the bacterial ribosomal protein genes of samples from Chiang Mai (HLB-CM), Chiang Rai (HLBCR), and Phrae (HLB-P) were analysed. From these results, it is apparent that the causal agent of citrus HLB disease, in these provinces, is Candidatus Liberibacter asiaticus.
Article Details
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
References
Dellaporta, S.L., Wood, J. and Hicks, J.B. (1983). A Plant DNA minipreparation : version II. Plant Molecular Reporter 4: 19-20.
Hocquellet, A., Toorawa, P., Bové, J.M. and Garniner, M. (1999). Detection and identification of the two Candidatus Liberobacter species associated citrus huanglongbing by PCR amplification of ribosomal protein genes of the operon. Molecular and Cellular Probes 13: 373-379.
Huang, CH. (1979). Distribution of likubin pathogen in likubin affected citrus plant. Journal of Agricultural Research of China 28: 29-33.
Hung, T.H., Wu, M.L. and Hong-J, S. (1999). Development of a rapid method for the diagnosis of Citrus Greening Disease using the Polymerase Chain Reaction. Journal Phytopathology 147: 599-604.
Jagoueix, S., Bové, J.M. and Garnier, M. (1994). The phloem-limited bacterium of Greening is a member of the alpha subdivision of the proteobacteria. International Journal of Systemic Bacteriology 44: 379-386.
Jagoueix, S., Bové, J.M. and Garnier, M. (1996). PCR detection of the two ‘Candidatus’ liberobacter species associated with greening disease of citrus. Molecular and Cellular Probes 10: 43-50.
McClean, A.P.D. (1970). Greening disease of sweet orange: its transmission in propagative parts and distribution in partially disease trees. Phytophylactica 2: 263-268.
Murray, R.G.E. and Schleifer, K.H. (1994). Taxonomic notes: a proposal for recording the properties of putative taxa of procaryotes. International Journal of Systematic Bacteriology 44: 174-176.
Nakashima, K., Promintara, M., Ohtsu, Y. and Kano, T. (1995). Improve detection of 16S rDNA of the Thai Isolate of GO. In: Development of Methods for the Diagnosis and Control of Citrus Greening Disease in Thailand. Under the cooperation research program between Thailand and Japan: 33-46.
Teixeira, D.D.C., Danetb, J.L., Eveillardb, S., Martinsa, E.C., Juniora, W.C.J., Yamamotoa, P.T., Lopesa, S.A., Bassanezia, R.B., Ayresa, A.J., Saillardb, C. and Bove, J.M. (2004). Citrus huanglongbing in Sao Paulo State, Brazil: PCR detection of the “Candidatus” Liberibacter species associated with the disease. Molecular and Cellular Probes 19:173–179.
Timmer, L.W., Garnsey, S.M. and Broadbent, P. (2003). Diseases of Citrus. In: Disease of Tropical Fruit Crops (ed. R.C. Ploetz). APS Press, USA: 163-195.
