Cloning, Expression and Protein Purification of AV1 Gene of Okra Leaf Curl Virus Egyptian Isolate and Genetic Diversity between Whitefly and Different Plant Hosts
Authors: Dalia. G. Aseel
Begomoviruses are economically important plant viruses that infect dicotyledonous plants and exclusively transmitted by the whitefly Bemisia tabaci. Here, replicative form was isolated from Okra, Cotton, Tomato plants and whitefly infected with Begomoviruses. Using coat protein specific primers (AV1), the viral infection was verified with amplicon at 450 bp. The sequence of OLCuV-AV1 gene was recorded and received an accession number (FJ441605) from Genebank. The phylogenetic tree of OLCuV was closely related to Okra leaf curl virus previously isolated from Cameroon and USA with nucleotide sequence identity of 92%. The protein purification was carried out using His-Tag methodology by using Affinity Chromatography. The purified protein was separated on SDS-PAGE analysis and an enriched expected size of band at 30 kDa was observed. Furthermore, RAPD and SDS-PAGE were used to detect genetic variability between different hosts of okra leaf curl virus (OLCuV), cotton leaf curl virus (CLCuV), tomato yellow leaf curl virus (TYLCuV) and the whitefly vector. Finally, the present study would help to understand the relationship between the whitefly and different economical crops in Egypt.
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 Sopid, S. (2009). The complete nucleotide sequence of Squash leaf curl China virus-(Wax gourd) and its phylogenetic relationship to other geminiviruses. Science Asia, 35: 131–136.
 Varsani, A., Navas-Castillo, J. Moriones, E. Hernández-Zepeda, C. Idris, A. Brown, J. K. and Martin, D. P. (2014). Establishment of three new genera in the family Geminiviridae: Becurtovirus, Eragrovirus and Turncurtovirus. Archives of virology, 1-11.
 Fauquet, C. M, Briddon RW, Brown JK, Moriones E, Stanley J, Zerbini M, Zhou X. (2008). Geminivirus strain demarcation and nomenclature. Archives of Virology, 153:783-821
 Brown, J. K. (1994). Current status of Bemisia tabaci as a plant pest and virus vector in agro-ecosystems worldwide. FAO Plant Protection Bulletin.
 Moffat, A. S (1999). Geminiviruses emerge as serious crop threat. Science, 286:1835-1835.
 Nawaz-ul-Rehman, M. S. and Fauquet, C. M. (2009). Evolution of geminiviruses and their satellites. FEBS Letters., 583, 1825–1832.
 Jeske, H. Gotthardt, D. and Kober, S. (2010). In planta cloning of geminiviral DNA: The true Sidamicrantha mosaic virus. Journal of Virology Methods, 163, 301–308.
 Zhou X. (2013). Advances in understanding begomovirus satellites Annual Review of Phytopathology, 51: 357-81.
 Fernandes, F. R. de Albuquerque, L. C. and Inoue-Nagata, A. K. (2010). Development of a species-specific detection method for three Brazilia tomato begomoviruses. Tropical Plant Pathology, vol. 35, 1, 043-047.
 Sharma, P. and Ikegami, M. (2009). Characterization of signals that dictate nuclear/nucleolar and cytoplasmic shuttling of the capsid protein of Tomato leaf curl Java virus associated with DNAβsatellite.Virus Research 144(1-2), 145-153.
 Hanley-Bowdoin, L., Settlage, S. B., Orozco, B. M., Nagar, S. & Robertson, D. (1999). Geminviruses: models for plant DNA replication, transcription, and cell cycle regulation. Critical Reviews in Plant Sciences 18, 71-106.
 Yadava, P., Suyal, G. and Mukherjee, S. K. (2010). Begomovirus DNA replication and pathogenicity. Current Science 98(3), 360-368.
 Rojas, M. R., Hagen, C., Lucas, W. J. and Gilbertson, R. L. (2005). Exploiting chinks in the plant's armor: Evolution and emergence of geminiviruses. Annual Review of Phytopathology 43, 361-394.
 Shih, S. L. Tsai, W. S. Green, S. K. and Singh, D. (2006). First report of Tomato leaf curl Joydebpur virus infecting chilli in India. New Disease Reports, 14 17.
 Simón, B. J. Cenis, L. Beitia, F. Khalid, S. Moreno, I. M. Fraile, A. and García-Arenal, F. (2003). Genetic Structure of Field Populations of Begomoviruses and of Their Vector Bemisia tabaci in Pakistan. Virology 93: 1422-1429.
 Dellaporta, S. L. Wood, J. Hicks, J. B. (1983). A plant DNA minipreparation: version II. Plant Molecular Biology Reports, 1:19-21.
 Nadeem, A. (1995). Molecular characterization of two cotton geminiviruses, PhD Dissertation. University of Arizona, Tucson.
 NCBI-BLAST. National Center for Biotechnology International (http://bast.ncbi.nlm.nih.gov/Blast.cgi).
 Tamura, K., Dudley, J., Nei, M. and Kumar, S. (2007). MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0, Mol. Biol. Evol., 24: 1596-1599.
 Aseel, D. G. Molecular Genetical Studies on Cotton Leaf Curl Virus in Egypt (2018). ISBN: 978-613-8-38657-5, Project ID: # 173299, LAMBERT Academic Publishing.
 Ni, M., Dehesh, K., Tepperman, J. M., and Quail, P. H. (1996). GT-2: In vivo transcriptional activation activity and definition of novel twin DNA binding domains with reciprocal target sequence selectivity. Plant Cell 8:1041-1059.
 Sattar, M. N. Kvarnheden, A. Saeed, M. and Briddon R. W. (2013). Cotton leaf curl disease an emerging threat to cotton production worldwide. Journal of General Virology, 94, 695–710.
 Leke, W. N. Mignouna, D. B. Brown, J. K. and Kvarnheden, A. (2015). Begomovirus disease complex: emerging threat to vegetable production systems of West and Central Africa. Agriculture & Food Security; 4:1.
 Saleem, H. Khan, A. A. Azhar, M. T. and Nawaz-ul-Rehman, M. S. (2017). Molecular Screening of Cotton Germplasm for Cotton Leaf Curl Disease Caused by Viral Strains. International journal of Agriculture and Biology, 19: 125‒130.
 Navot, N. Pichersky, E. Zeidan, M. Zamir, D. and Czosnek, H. (1991). Tomato Yettow Leaf Curl Virus: A Whitefly-Transmitted Geminivirus with a Single Genomic Component. Virology, 185, 151.-161.
 Basak, J. (2016). Tomato Yellow Leaf Curl Virus: A Serious Threat to Tomato Plants World.
 Ghanim, M. Morin, S. Zeidan, M. and Czosnek, H. (1998). Evidence for Transovarial Transmission of Tomato Yellow Leaf Curl Virus by Its Vector, the Whitefly Bemisiatabaci. Virology, 240, 295–303.
 Marwal, A. Kumar Sahu, A. and Kumar Gaur, R. (2013). Molecular Characterization of Begomoviruses and DNA Satellites Associated with a New Host Spanish Flag (Lantana camara) in India. ISRN Virology, 1- 5.
 Alhudiab, K. Alaraby, W. and Rezk, A. (2014). Molecular characterization of tomato yellow leaf curl disease associated viruses in Saudi Arabia. Int. J. Virol.,10:192-203.
 Gul-Seker, M. Ekinci, H. Ozturk, C. and Elibuyuk, I. O. (2015). Current Situation of Tomato Yellow Leaf Curl Disease (TYLCD) in Antalya, Turkey. Plant protection science,51 (4), 208–213.
 Zhou, X. Liu, Y. Robinson, D.J. and Harrison, B. D. (1998). Four DNA-A variants among Pakistani isolates of Cotton leaf curl virus and their affinities to DNA-A of geminivirus isolates from Okra. J. Gen. Virol., 79 (4), pp. 915-923.
 Sharma, P. Rishi, N. and Malathi, V.G. (2005). Molecular cloning of coat protein gene of an Indian cotton leaf curl virus (CLCuV-HS2) isolate and its phylogenetic relationship with other members of Geminiviridae. Virus Genes, 30, pp. 85-91.
 Sohrab, S. S. Azhar, E. I. Kamal, M. A. Bhattacharya, P. S. and Rana, D. (2014). Genetic variability of Cotton leaf curl betasatellite in Northern India. audi J Biol Sci; 21(6): 626–631.
 Farrag, A. A. El-Attar, A. K. El-Banna, O. M. Ibrahim, A. I. and H. M. Mazyad (2014). Molecular and Biological Characterization of Squash Leaf Curl Virus (SLCV) Affecting Common Beans in Egypt. Egyptian J. Virol, Vol. 11 (2):14-27.
 Reynoso, D. L. G. Valverde, R. A. and Murai, N. (2015). Sweet Potato Leaf Curl Virus: Coat Protein Gene Expression in Escherichia coli and Product Identification by Mass Spectrometry. American Journal of Plant Sciences, 6, 3013-3024.
 Hasan, H. S. (2006). Survey of Bemisiatabaci (Gennadius) (Homopetra: Aleyrodidae) Biotypes in Jordan using RAPD marker Journal of Entomology. 4 290-297.
 Ali, M., Ahmad, Z., Tanveer, M., and Mahmood, T. (1995). Cotton leaf curl virus in the Punjab: Current situation and review of work. Central Cotton Research Institute/Ministry of Food, Agriculture and Livestock, Government of Pakistan/Asian Development Bank, Multan, Pakistan.
 Briddon, R. W., and Markham, P. G. (2000). Cotton leaf curl virus disease. Virus Res. 71:151-159.
 Khalid, S., Soomro, M. H., and Ahmad, I. (1997). Occurrence of cotton leaf curl virus (CLCuV) in Sindh. Pakistan journal of Botany, 29:173-174.
 Sanz, A. I., Fraile, A., Gallego, J. M., Malpica, J. M., and García- Arenal, F. (1999). Genetic variability of natural populations of cotton leaf curl geminivirus, a single-stranded DNA virus. Journal of Molecular evolution, 49: 672- 681.
 Sanz, A. I., Fraile, A., García-Arenal, F., Zhou, X., Robinson, D. J., Khalid, S., Butt, T., and Harrison, B. D. (2000). Multiple infection recombination and genome relationships among begomovirus isolates found in cotton and other plants in Pakistan. Journal of General Virology, 8,1839-49.
 Sharma, R. K. Gupta, V. K. Jindal, J. and Dilawari, V. K. (2008). Host associated genetic variation in whitefly, Bemisiatabaci (Genn). Indian Journal of Biotechnology 7: 366:370.
 Bisaro, D. M. (2006). Silencing suppression by geminivirus proteins. Virology 344, 158-168.
 Vanitharani, R., Chellappan, P. and Fouquet, C. M. (2005). Geminiviruses and RNA silencing. Trends in Plant Science 10, 144-151.
 Rojas, M. R. Jiang, H. Salati, R. Xoconostle-Casares, B. Sudarshana, M. R. Lucas, W. J. and Gilbertson, R. L. (2001). Functional analysis of proteins involved in movement of the monopartite Begomovirus, Tomato yellow leaf curl virus. Virology, 291, 110–125.
 Lazarowitz S. G. (1992). Geminiviruses: genome structure and gene function. Critical Reviews in Plant Sciences, 11: 327–349.