Developing of specific antibody against chickpea chlorotic dwarf virus (CpCDV) through recombinant coat protein

Safarnejad, Mohammad Reza; Bananej, Kaveh; Sokhansanj, Yalda

doi:10.48311/jcp.2019.1426

Developing of specific antibody against chickpea chlorotic dwarf virus (CpCDV) through recombinant coat protein

https://doi.org/10.48311/jcp.2019.1426

Volume 8, Issue 2
June 2019

Pages 179-190

Document Type : Original Research

Authors

M. R. Safarnejad ¹

K. Bananej ¹

Y. Sokhansanj ²

¹ Iranian Research Institute of Plant Protections, Agricultural Research, Education and Extension Organization (AREEO), Tehran, Iran.

² Department of Plant Pathology, Faculty of Agriculture and Natural Resources, Tehran Science and Research Branch, Islamic Azad University, Tehran, Iran.

Abstract

The legume crops such as chickpea and lentils are mainly cultivated in semi-arid tropical lands. Chickpea chlorotic dwarf virus (CpCDV) causes major losses to legumes throughout the world. Producing of specific antibody against this virus is crucial for surveys of disease in the fields and assessment of vial resistance in plant cultivars. Present article describes developing of specific antibody against the CpCDV virus by applying recombinant protein. In this study, coat protein of CpCDV was selected as a target for detection and preparation of polyclonal antibody. To achieve this aim CP gene encoding coat protein of CpCDV was initially PCR-amplified and inserted into bacterial expression vector. Expression of recombinant protein was performed in Bl21 strain of Escherichia coli. Purification was carried out under native conditions and the accuracy of recombinant protein production was confirmed by electrophoresis. The purified recombinant coat protein of CpCDV was used for immunization of rabbit. Purification of immunoglobulin molecules was performed by affinity chromatography using protein A column followed by conjugating of IgG to alkaline phosphatase enzyme. The capability of purified antibodies and conjugates for efficient detection of infected plants was assessed by double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), western blotting and dot immunosorbent assay (DIBA). These results proved that prepared IgG and conjugate are able to distinguish with high efficiency CpCDV infected plants. To the best of our knowledge, this is the first report for production of anti-CpCDV antibodies raised through recombinant protein technology.

Keywords

Antibody

Chickpea

CpCDV

ELISA

Recombinant Protein

20.1001.1.22519041.2019.8.2.8.0

Subjects

Plant Virology

Abou-Jawdah, Y., Sobh, H., Cordahi, N., Kawtharani, H., Nemer, G., Maxwell, D. P. and Nakhla, M. K. 2004. Immunodiagnosis of Prune dwarf virus using antiserum produced to its recombinant coat protein. Journal of Virological Methods, 121(1), 31-38.
Akhtar, S., Khan, A. and Briddon, R. 2014. A distinct strain of Chickpea chlorotic dwarf virus infecting pepper in Oman. Plant Disease, 98(2), 286-286.
Al-Moslih, M. 2012. Serological Diagnosis of Certain Human, Animal and Plant Disease. InTech.
Ali, M., Kumari, S., Makkouk, K. and Hassan, M. 2004. Chickpea chlorotic dwarf virus (CpCDV) naturally infects Phaseolus bean and other wild species in the Gezira region of Sudan. Arab Journal of Plant Protection, 22, 96.
Astier, S., Albouy, J., Maury, Y. and Lecoq, H. 2001. Principles of plant virology: genome, pathogenicity, virus ecology Institut National de la Recherche Agronomique.
Ausubel, F. M., Brent , R. and Kingstone, R. E. 1995. Current Protocols in Molecular Biology. New York, Wiley Interscience.
Bosque-Perez, N. and Buddenhagen, I. 1990. Studies on epidemiology of virus disease of chickpea in California. Plant Disease, 74(5), 372-378.
Cerovska, N., Moravec, T., Plchova, H., Hoffmeisterova, H. and Dedic, P. 2012. Production of polyclonal antibodies to the recombinant Potato virus M (PVM) Non‐structural triple gene block protein 1 and coat protein. Journal of Phytopathology, 160(5), 251-254.
Čeřovská, N., Moravec, T., Rosecka, P., Dědič, P. and Filigarova, M. 2003. Production of Polyclonal Antibodies to a Recombinant Coat Protein of Potato moptop virus. Journal of Phytopathology, 151(4), 195-200.
Clark, M. F. and Adams, A. 1977. Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant viruses. Journal of General Virology, 34(3), 475-483.
Galluzzi, L., Magnani, M., Saunders, N., Harms, C. and Bruce, I. J. 2007. Current molecular techniques for the detection of microbial pathogens. Science Progress, 90(1), 29-50.
Hema, M., Kirthi, N., Sreenivasulu, P. and Savithri, H. 2003. Development of recombinant coat protein antibody based IC-RT-PCR for detection and discrimination of sugarcane streak mosaic virus isolates from Southern India. Archives of Virology, 148(6), 1185-1193.
Horn, N., Reddy, S. and Reddy, D. 1995. Assessment of yield losses caused by chickpea chlorotic dwarf geminivirus in chickpea (Cicer arietinum) in India. European Journal of Plant Pathology, 101(2), 221-224.
Horn, N., Reddy, S., Roberts, I. and Reddy, D. 1993. Chickpea chlorotic dwarf virus, a new leafhopper‐transmitted geminivirus of chickpea in India. Annals of Applied Biology, 122(3), 467-479.
Horn, N., Reddy, S. and Vandenheuvel, J. 1996. Survey of chickpea (Cicer arietinum L.) for chickpea stunt disease and associated viruses in India and Pakistan. Viruses involved in chickpea stunt, 23.
Iracheta‐Cárdenas, M., Sandoval‐Alejos, B., Román‐Calderón, M., Manjunath, K., Lee, R. and Rocha‐Peña, M. 2008. Production of polyclonal antibodies to the recombinant coat protein of Citrus tristeza virus and their effectiveness for virus detection. Journal of Phytopathology, 156(4), 243-250.
Jain, R., Pandey, A. N., Krishnareddy, M. and Mandal, B. 2005. Immunodiagnosis of groundnut and watermelon bud necrosis viruses using polyclonal antiserum to recombinant nucleocapsid protein of Groundnut bud necrosis virus. Journal of Virological Methods, 130(1), 162-164.
Koolivand, D., Bashir, N. S., Behjatnia, S. A. and Joozani, R. J. 2016. Production of Polyclonal Antibody against Grapevine fanleaf virus Movement Protein Expressed in Escherichia coli. The Plant Pathology Journal, 32(5), 452.
Korimbocus, J., Preston, S., Danks, C., Barker, I., Coates, D. and Boonham, N. 2002. Production of monoclonal antibodies to Sugarcane yellow leaf virus using recombinant readthrough protein. Journal of Phytopathology, 150(8‐9), 488-494.
Kraberger, S., Harkins, G. W., Kumari, S. G., Thomas, J. E., Schwinghamer, M. W., Sharman, M., Collings, D. A., Briddon, R. W., Martin, D. P. and Varsani, A. 2013. Evidence that dicot-infecting mastreviruses are particularly prone to inter-species recombination and have likely been circulating in Australia for longer than in Africa and the Middle East. Virology, 444(1), 282-291.
Kraberger, S., Kumari, S. G., Hamed, A. A., Gronenborn, B., Thomas, J. E., Sharman, M., Harkins, G. W., Muhire, B. M., Martin, D. P. and Varsani, A. 2015. Molecular diversity of Chickpea chlorotic dwarf virus in Sudan: High rates of intra-species recombination–a driving force in the emergence of new strains. Infection, Genetics and Evolution, 29, 203-215.
Kumari, S., Makkouk, K. and Attar, N. 2006. An improved antiserum for sensitive serologic detection of Chickpea chlorotic dwarf virus. Journal of Phytopathology, 154(3), 129-133.
Kumari, S., Makkouk, K., Attar, N., Ghulam, W. and Lesemann, D.-E. 2004. First report of Chickpea chlorotic dwarf virus infecting spring chickpea in Syria. Plant Disease, 88(4), 424-424.
Kumari, S., Makkouk, K., Katul, L. and Vetten, H. 2001. Polyclonal antibodies to the bacterially expressed coat protein of Faba bean necrotic yellows virus. Journal of Phytopathology, 149(9), 543-550.
Lee, S.-C. and Chang, Y.-C. 2008. Performances and application of antisera produced by recombinant capsid proteins of Cymbidium mosaic virus and Odontoglossum ringspot virus. European Journal of Plant Pathology, 122(2), 297-306.
Makkouk, K.M., Hsu, H.T. and Kumari, S.G., 1993. Detection of Three Plant Viruses by Dot‐Blot and Tissue‐Blot Immunoassays Using Chemiluminescent and Chromogenic Substrates 1. Journal of Phytopathology, 139(2), pp.97-102.
Makkouk, K., Bahamish, H., Kumari, S. and Lotf, A. 1998. Major viruses affecting faba bean (Vicia faba L.) in Yemen. Arab Journal of Plant Protection, 16(2), 98-101.
Makkouk, K., Bashir, M., Jones, R. and Kumari, S. 2001. Survey for viruses in lentil and chickpea crops in Pakistan. Zeitschrift für Pflanzenkrankheiten und Pflanzenschutz, 108(3), 258-268.
Makkouk, K., Kumari, S., Shahraeen, N., Fazlali, Y., Farzadfar, S., Ghotbi, T. and Mansouri, A. R. 2003. Identification and seasonal variation of viral diseases of chickpea and lentil in Iran. Journal of Plant Diseases and Protection, 157-169.
Manzoor, M. T., Ilyas, M., Shafiq, M., Haider, M. S., Shahid, A. A. and Briddon, R. W. 2014. A distinct strain of chickpea chlorotic dwarf virus (genus Mastrevirus, family Geminiviridae) identified in cotton plants affected by leaf curl disease. Archives of Virology, 159(5), 1217-1221.
Muhire, B., Martin, D. P., Brown, J. K., Navas-Castillo, J., Moriones, E., Zerbini, F. M., Rivera-Bustamante, R., Malathi, V., Briddon, R. W. and Varsani, A. 2013. A genome-wide pairwise-identity-based proposal for the classification of viruses in the genus Mastrevirus (family Geminiviridae). Archives of Virology, 158(6), 1411-1424.
Ouattara, A., Tiendrébéogo, F., Lefeuvre, P., Hoareau, M., Claverie, S., Traoré, E. V., Barro, N., Traoré, O., Varsani, A. and Lett, J.-M. 2017. New strains of chickpea chlorotic dwarf virus discovered on diseased papaya and tomato plants in Burkina Faso. Archives of Virology, 162(6), 1791-1794.
Raikhy, G., Hallan, V., Kulshrestha, S. and Zaidi, A. 2007. Polyclonal antibodies to the coat protein of Carnation etched ring virus expressed in bacterial system: production and use in immunodiagnosis. Journal of Phytopathology, 155(10), 616-622.
Reddy, M., Nene, Y. and Verma, J. 1979. Pea leaf roll virus causes chickpea stunt. International Chickpea Newsletter, 1, 8-8.
Safarnejad, M. R., Fischer, R. and Commandeur, U. 2008. Generation and characterization of functional recombinant antibody fragments against Tomato yellow leaf curl virus replication-associated protein. Comm Agri Appl Biol Sci, 73, 311-323.
Sambrook, J., F., F. E. and Maniatis, T. 1996. Molecular Cloning - A Laboratory Manual. New York, Cold Spring Harbor Laboratory.
Shahmirzaie, M., Safarnejad, M. R., Rakhshandehroo, F., Safarpour, H., Rabbani, H., Zamanizadeh, H. R. and Elbeaino, T. 2019. Production of a polyclonal antiserum against recombinant nucleocapsid protein and its application for the detection of fig mosaic virus. Journal of Virological Methods 265:22-25.
Wisdom, G. B. 2005. Conjugation of antibodies to alkaline phosphatase. Immunochemical Protocols, 123-126.

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