Antibiotic treatment of citrus budwood for the management of huanglongbing disease

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Research Paper 01/07/2019
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Antibiotic treatment of citrus budwood for the management of huanglongbing disease

Muhammad Sarwar Yaqub, Iqrar Ahmad Khan, Rozina Aslam
Int. J. Biosci.15( 1), 99-109, July 2019.
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Control of huanglongbing (HLB) disease is necessary around the world to avoid complete destruction of citrus. Antibiotics were used by many scientists in the form of sprays and trunk injections to control HLB, but the control was not effective for longer time and disease symptoms appeared again after some time because of uneven distribution of pathogen in the host plant. Tetracycline and penicillin antibiotics were used in this study to eliminate Candidatus Liberibacter bacterium from sweet orangebudwood with the objective that the pathogen elimination from the budwood could be comparatively more effective. Tetracycline doses used were 250ppm, 500ppm, 750ppm, 1000ppm and 1250ppm. Penicillin doses were 25ppm, 50ppm, 75ppm, 100ppm and 125ppm. Kinnow mandarin (Citrus reticulata Blanco) nurtured in screen house were used as indicator plant. These plants were confirmedHLB negative before grafting. Budwood was obtained from HLB positive plants confirmed by conventional PCR using 16S rDNA primers OI1/OI2C and rplKAJL-rpoBC operon primers A2/J5. Budsticks treated with the above mentioned doses of antibiotics were grafted on the indicator plants. Tetracycline at 1000 ppm with mean Ct value of 38.40 and penicillin antibiotic at 75ppm dose with mean Ct value of 38.61 were found effective to eliminate Candidatus Liberibacter asiaticus from budwood to stop further spread of the disease through budding/grafting.


Aslam R, Khan IA, Rahman KU, Asghar M, Yaqub MS. 2017a. Expression profiling of huanglongbing disease in kinnow (Citrus reticulata) and succari (Citrus sinensis) leaves. International Journal of Agriculture and Biology 19, 1187‒1192.

Aslam R, Khan IA, Rahman KU, Asghar M. 2017b. Detection of ‘Candidatus Liberibacter asiaticus’ the causal organism of huanglongbing, in mandarin group of citrus. International Journal of Agriculture and Biology 19, 255‒258. 17957/IJAB/15.0271.

Aubert B. 1987. Trioza erytreae Del Guercio and Diaphorina citri Kuwayama (Homoptera: Psylloidea), the two vectors of citrus greening disease: biological aspects and possible control strategies. Fruits 42, 149-162.

Bove JM. 2006. Huanglongbing: a destructive, newly emerging, century old disease of citrus. Journal of Plant Pathology 88, 7-37.

Chohan SN, Qamar R, Sadiq I, Azam M, Holford P, Beattie A. 2007. Molecular evidence for the presence of huanglongbing in Pakistan. Australian Plant Disease Note 2, 37-38.

Cochran LC. 1976. Theoccurrence of greening disease in Pakistan. p. 21. In: E.C.Calavan (ed.) Proc. 7th Conf. IOVC, Riverside, University of California.

Da Graca JV. 1991. Citrus greening disease. Annual Review of Phytopathology 29, 109–36.

Hall DG, Richardson ML, Ammar E, Halbert SE. 2012. Asian citrus psyllid, Diaphorina citri, vector of citrus huanglongbing disease. Entomologia Experimentalis et Applicata.146, 207-223.

Hocquellet A, Toorawa P, Bove JM, Garnier M. 1999. Detection and identification of two Candidatus liberobacter species associated citrus huanglongbing by PCR amplification of ribosomal protein genes of the beta operon. Molecular and Cellular Probes 13, 373-379.

Hoffman MT, Doud MS, Williams L, Zhang MQ, Ding F, Stover E, Hall D, Zhang S,  Jones L, Gooch M, Fleites L, Dixon W, Gabriel D, Duan YP. 2013. Heat treatment eliminates ‘Candidatus Liberibacter asiaticus’ from infected citrus trees under controlled conditions. Phytopathology 103, 15-22.

Hussain MA, Nath D. 1927. The citrus psylla (Diaphorina citri Kuwayama) [Psyllidae:Homoptera]. Memories of the Department of Agriculture in India, Entomological Series 10, 1-27.

Jagoueix S, Bove JM, Garnier M. 1994. The phloem-limited bacterium of greening is a member of the alpha subdivision of the proteobacteria. International Journal of Systematic Bacteriology 44, 379-386.

Jagoueix S, Bove JM, Garnier M. 1996. PCR detection of two candidatus Liberibacter species associated with greening disease of citrus. Molecular and Cellular Probes 10, 43-50.

Kapur S, Kapur SP, Kang SS. 1996. Chemo-Trunk injection for the control of citrus greening. Indian Journal of Virology 12, 55-57.

Li W, Hartung JH, Levy L. 2006. Quantitative real time PCR for detection and identification of Candidatus Liberibacter species associated with citrus huanglongbing. Journal of Microbiological Methods 66, 104-115.

Li W, Cong Q, Pei J, Kinch LN, Grishin NV. 2012. The ABC transporters in Candidatus Liberibacter asiaticus. Proteins 80, 2614-2628.

Lin KH. 1956. Observation on yellow shoot on citrus. Etiological studies of yellow shoot on citrus. Acta Phytopathologica Sinica 2, 1-42.

Nariani TK, Ghosh SK, Kumar D, Raychaudhuri SP, Wishwanath SM. 1975. Detection and possibilities of therapeutic control of the greening disease of citrus caused by mycoplasma. Proceeding of Indian Natural Science Academic Series B 41, 334-339.

Nariani TK, Raychaudhuri SP, Wishwanath SM. 1971. Response of greening pathogen of citrus to certain tetracycline antibiotics. Current Science 20, 552.

Teixeira DC, Danet JL, Eveillard S, Martins EC, Junior WCJ, Yamamoto PT,   Lopes SA, Bassanezi RB, Ayers AJ, Sailard C, Bove JM. 2005. 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.

Yaqub MS, Khan IA, Usman M, Rana IA. 2017. Molecular detection of Candidatus Liberibacter asiaticus, the causal organism of huanglongbing (citrus greening) in Faisalabad, Pakistan for huanglongbing management. Pakistan Journal of Agricultural Sciences 54, 21-26.

Zhang M, Duan Y, Zhou L, Turechek WW, Stover E, Powell CA. 2010. Screening molecules for control of citrus huanglongbing using an optimized regeneration system for Candidatus Liberibacter asiaticus infected periwinkle (Catharanthus roseus) cuttings. Phytopathology 100, 239-245.

Zhang M, Powell CA, Guo Y, Doud MS, Duan Y. 2012. A graft-based chemotherapy method for screening effective molecules and rescuing huanglongbing-affected citrus plants. Phytopathology 102, 567-574.

Zhang M, Guo Y, Powell CA, Doud MS, Yang C, Duan Y. 2014. Effective antibiotics against Candidatus Liberibacter asiaticus in HLB-affected citrus plants identified via the graft-based evaluation. PLoS One 9(11), e111032.

Zhang M, Yang C, Powell CA. 2015. Application of antibiotics for control of citrus huanglongbing. Advances in Antibiotics and Antibodies 1, 1.