Open Access Open Access  Restricted Access Subscription Access
Cover Image

Detection of candidatus liberibacter asiaticus associated with citrus greening (huanglongbing) of mandarin by template preparation

G.P. Jagtap, M.V. Mahajan, D. Utpal


The polymerase ChainReaction (PCR) diagnosis is more reliable and sensitive diagnostic tool forgreening bacterium than other conventional approaches like Electron microscopy,DNA-DNA hybridization and immunofluorescence (IF) for detection of citrusgreening. During experiment, it was observed that sodium sulphite method of DNAisolation provided higher yield and better quality DNA than other methods.Primer C (450 bp) was more efficient in amplifying the DNA of greeningbacterium even at a very low concentration of 0.1 pg. To confirm thereliability of PCR, the greening bacterium was also detected ingraft-inoculated plants, which showed typical greening bacterium was alsodetected in graft-inoculated plants, which showed typical greening symptoms.Results showed amplification of 450 bp in PCR suggesting sampling in March ismore suitable for PCR detection of greening bacterium.


Ahlawat, Y.S., 1997. Viruses, greening, bacterium and viroids associated with citrus (Citrus sp.) decline in India. Indian J. Agril. Sci. 67, 51-57.

Ahlawat, Y.S., Pant, R.P., 2003. Major virus and virus-like diseases of citrus in India, their diagnosis and management. Annual Rev. Plant Pathol. 2, 447-474.

Ahlawat, Y.S., Baranwal, V.K., Mazumdar, S., 2003. First report of citrus greening disease and associated bacterium. Candidatus Liberibacter from Bhutan. Plant Dis. 87, 448.

Baranwal, V.K., Majumdar, S., Ahlawat, Y.S., Singh, R.P., 2003. Sodium sulphite yields improved DNA of higher stability for PCR detection of citrus yellow mosaic virus from citrus leaves. J. Virol. Meth. 112, 153-156.

Bove, J.M., Garnier, M., Ahlawat, Y.S., Chakraborty, N.K., Verma, A., 1993. Detection of Asian strains of greening BLO by DNA-DNA hybridization in Indian orchards trees and Malysian Diaphorina citri psyllids pp, 258-263 In Porc. 12th Conf. IOCV, IOCV Riverside, California.

Fraser, L.R., Singh, D., Capoor, S.P., Nariani, T.K., 1966. Greening virus, the likely cause of citrus dieback in India. FAO Plant Prot. Bull. 14, 127-130.

Harakava, R., Morais, L.J., Ochasou, J., Manjunath, K.L., Febres, V.J., Lee, R.F., Niblett, G.L., 2000. Improved sensitivity in the detection and differentiation of citrus Huanglongbing bacteria from South Africa and the Phillippines. In. Proc., 14th Conf., IOCV. IOCV. Riverside, California. Pp, 195-199

Hocquellet, A., Bove, J.M., Garnier, M., 2000. Isolation of Candidatus liberibacter genes by RAPD and new PCR detection technique. In : Proc. 14th Conf. IOCV, IOCV Riverside, California. PP, 363-368

Jagouiex, S. Bove J.M., Grnier, M., 1996. PCR detection of the two candidates liberobacter species associated with greening diseases of citrus. Molecul. Cellul. Prob. 10, 43-50.

Korsten, L.G., Sanders, M., Su, H.J., Garnier, M. Bove, J.M. and Gotze, J.M. (1993). Detection of citrus greening infected citrus in South Africa using DNA probes and monoclonal antibodies. In Proc. 12th Conf. IOCV, IOCV, Riverside. pp : 224-232

Planet, P.S., Jagoueix, Bove, J.M., Garnier, M., 1995. Detection and characterization of the African citrus greening Liberobacter by amplification, cloning and sequencing of rplKJL-ropBC operon. Curr. Microbiol. 30 : 137-141.

Singh, R.P. and Nie, X. (2003). Multiple virus and viriod detection and strain seperation via multiplex riverse transcription-polymerase chain reaction. Can J. Plant. Path. 25, 127-134.

Tian, Y., Ke, S., Ke, C., 1996. Polymerase chain reaction for detection and quantification of liberiobacter asiaticus, the bacterium associated with Huangolongbing (Greening) of citrus in China. In Proc. 13th Conf., IOCV. IOCV Riverside, California. Pp, 252-257

Varma, A., Ahlawat, Y.S., Chakraborty, N.K., Garnier, M., Bove, J.M., 1993. Detection of greening BLO by electron Microscopy, DNA hybridization in citrus leaves with and without mottle from various regions in India. In Proc. 12th Conf., IOCV Riverside, California. Pp, 280-285.

Full Text: PDF


  • There are currently no refbacks.