Serological and Molecular Characterisation of Virus Infecting Watermelon (Citrullus lanatus) in Adim-Biase, Cross River State, Nigeria

Main Article Content

O. I. Eyong
E. E. Ekpiken
G. M. Ubi
A. O. Alobi

Abstract

Watermelon, Citrullus lanatus, (Thunb.) is an annual crop in the Cucurbitaceae family grown for it fruits. It nutritional and medical uses helps in combating some illnesses and maintaining good health. The production of this crop in Nigeria is seriously under threat by virus disease. A survey of some farms in Adim, Biase Local Government Area of Cross River State, Nigeria during the 2019 planting season revealed mosaic and chlorotic symptoms. Mechanical inoculation was performed on young seedlings of Cucumeropsis mannii from watermelon samples showing viral symptoms and latter tested against the universal potyvirus antiserum and further detected by RT-PCR assay using cylindrical inclusion (CI) primer. The amplified cDNA was cloned and the nucleotide sequence determined. The serology result showed that there was positive reaction against the potyvirus antiserum confirming it to be a potyvirus. The gene sequence analysis revealed 86% sequence identity with Algerian watermelon mosaic virus. This is the first report of Algerian watermelon mosaic virus strain infecting watermelon in Nigeria.

Keywords:
ACP-ELISA, RT-PCR, watermelon, AWMV, Nigeria.

Article Details

How to Cite
Eyong, O. I., Ekpiken, E. E., Ubi, G. M., & Alobi, A. O. (2020). Serological and Molecular Characterisation of Virus Infecting Watermelon (Citrullus lanatus) in Adim-Biase, Cross River State, Nigeria. Annual Research & Review in Biology, 35(11), 66-72. https://doi.org/10.9734/arrb/2020/v35i1130300
Section
Original Research Article

References

Bailey LB. Three discussions in Cucurbitaceae. Gentes Herbarum. 2016;2: 175–186.

Dane F, Liu J. Diversity and origin of cultivated and citron type watermelon (Citrullus lanatus). Genetic Resources and Crop Evolution. 2007;54:1255-1265.

Rimando AM, Perkins-Veazie PM. Determination of citrulline in watermelon rind. Journal of Chromatography. 2005;1078:196–200.

Zhe C, Zhang M, Zhou X, Wu J. Development and detection application of monoclonal antibodies against Zucchini yellow mosaic virus. Journal of Intergrative Agriculture. 2017;16(1):115–124.

Chomicki G, Renner SS. Watermelon origin solved with molecular phylogenetics including Linnaean material: Another example of museomics. New Phytologist. 2014;205(2):526–32.

Renner SS, Sousa A, Chomicki G. Chromosome numbers, Sudanese wild forms and classification of the watermelon genus Citrullus, with 50 names allocated to seven biological species. Taxon. 2017;66(6):1393-1405.

Anonymous. TurkStat, Agricultural Structure, Production, Price, Value; 2009. (Accessed: 15 December 2013) Available:http://tuikapp.tuik.gov.tr/bitkiselapp/bitkisel.zul

Lecoq H, Desbiez C. Cucurbit viruses in the Mediterranean Region: An ever-changing picture. Advances in Virus Research. 2012;84:67-126.

Romay G, Lecoq H, Geraud-Pouey F, Chirinos DT, Desbiez C. Current status of cucurbit viruses in Venezuela and characterization of Venezuelan isolates of Zucchini yellow mosaic virus. Plant Pathology. 2014;63:78-87.

Eyong OI, Owolabi AT, Mofunanya AAJ, Ekpiken EE. Biological, serological and molecular characterisation of a new virus species infecting Telfairia occidentalis in Calabar, Southern Nigeria. Journal of Experimental Agriculture International. 2020a;42(1):23-33.

Atiri GI. An isolate of Cucumber mosaic virus from fluted pumpkin in Nigeria. Journal of Phytopathology. 1985;3:4-5.

Ayo-John EI, Hughes JD. Identification of Cucumber mosaic virus infecting Musa spp. and vegetable crops in Nigeria. International Journal of Virology. 2014;10: 204-210.

Owolabi AT, Rabenstein F, Ehrig F. A strain of Papaya ringspot virus naturally infecting cucumber (Cucumis sativus L.) in Calabar, South Eastern Nigeria. Nigerian Journal of Botany. 2008;21(1):97-108.

Owolabi AT, Rabenstein F, Taiwo MA, Ehrigh F, Maiss E. Characterization and identification of a potyvirus causing mosaic disease of Cucurbita moschata Duch Ex. Poir in Calabar, South East Nigeria. International Journal of Plant Pathology. 2011;2(4):165-176.

Eyong OI, Owolabi AT, Ekpiken EE, Effa EA. First report of Algerian watermelon mosaic virus infecting Cucumis sativus L in South South Nigeria. Journal of Biotechnology and Biochemistry. 2020b;42(1):23-33.

Kumar. Methods for the diagnosis of plant virus disease. IITA Ibadan, Laboratory Manual; 2009.

Abarshi MM, Mohammed IU, Wasswa P, Hillocks RJ, Holt J, Legg JP, Seal SE, Maruthi MN. Optimization of diagnostic RT-PCR protocols and sampling procedures for the reliable and cost-effective detection of Cassava brown streak virus. Journal of Virology Methods. 2010;163(2):353- 359.

Pappu S, Brand R, Pappu H, Rybicki E, Gough K, Frenkel M, Niblett C. A polymerase chain reaction method adapted for selective amplification and cloning of 3 sequence of potyviral genomes: Application to Dasheen mosaic virus. Journal of Virological Methods. 1993;41:9-20.

Camacho C, Coulouris G, Avagyan V, Ma N, Papadopoulos J, Bealer K, Madden TL. BLAST+: Architecture and applications. BMC Bioinformatics. 2008;10:421.

Shang H, Xie Y, Zhou X, Qian Y, Wu J. Monoclonal antibody-based serological methods for detection of Cucumber green mottle mosaic virus. Virology of Journal. 2011;8:228.

Wulandari DR, Ermayanti TM. Detection of potyvirus using RT-PCR and ACP-ELISA of Dioscorea species and in vitro shoot multiplication of the virus free plants. Annales Bogoriences. 2011;15(2):56- 78.

Wu JX, Ni YQ, Liu H, Ding M, Zhou XP. Monoclonal antibody-based serological assays and immunocapture RT-PCR for detecting Rice dwarf virus in field rice plants and leafhopper vectors. Journal of Virological Methods. 2014;195:134–140.

Shukla DD, Ward CW. Amino acid sequence homology of coat proteins as a for identification and classification of the potyvirus group. Journal of Genetic Virology. 1988;69:2703-2710.

King AM, Lefkowitz E, Adams MJ, Carstens EB. Virus taxonomy: 9th report of the international committee on taxonomy of viruses. Elsevier Academic Press, San Diego; 2011.

Frenkle MJ, Ward CW, Shukla DD. The use of 3’ non-coding nucleotide sequences in the taxonomy of potyviruses: Application to watermelon mosaic virus 2 and soybean mosaic virus-N. Journal of General Virology. 1989;70:2775-2783.

Yakoubi S, Desbiez C, Fakhfakh H, Wipf-Scheibel C, Marrakchi M, Lecoq H. Biological characterization and complete nucleotide sequence of a Tunisian isolate of Moroccan watermelon mosaic virus. Archives of Virology. 2008;153:117–125.

Lecoq H, Jane R. Zucchini yellow mosaic virus. Plant Pathology Journal. 2001;46: 809-829.