Detection of Cotton Leaf Curl Disease using Betasatellite-based Molecular Marker: Detection of Cotton Leaf Curl Disease

Umer Sohail Meer

doi:10.54393/fbt.v3i02.45

Authors

Umer Sohail Meer International School Lahore, Pakistan

DOI:

https://doi.org/10.54393/fbt.v3i02.45

Keywords:

Betasatellites, Cotton Leaf Curl Disease, Begomoviruses, Geminiviruses, PCR

Abstract

Cotton leaf curl disease (CLCuD) caused by whitefly-transmitted begomoviruses has hampered cotton production across the Punjab and Sindh provinces of Pakistan and northeastern India. Eight species of begomoviruses in association with a single betasatellite “Cotton leaf curl Multan betasatellite (CLCuMB)” have been reported to cause CLCuD. Objective: To detect early and efficiently Cotton leaf curl disease (CLCuD) using betasatellite-based molecular marker. Methods: 3-7 samples leaves were collected from symptomatic cotton fields in selected five areas of cotton production in Pakistan. Total DNA was extracted from collected leaves using the Cetyl trimethylammonium bromide (CTAB) method. Primers were designed by MUSCLE alignment tool and target region was amplified by PCR and amplification confirmed by performing gel electrophoresis. After DNA sequencing Phylogenetic analysis of the was carried out using software MEGA-X. Results: Amplified target region of483bp was observed by running 1% agarose gel. Comparison of DNA sequences revealed two nucleotide substitutions in DNA sequence from samples collected from Multan, Sakrand, Rahim Yar Khan and, while four nucleotide substations in sample collected from Vehari. High nucleotide substation in DNA sequence from Vehari as compared to other regions. Conclusions: In conclusion all of eight distinct begomoviruses causing CLCuD with CLCuMB is indicative of the fact that CLCuMB based molecular marker can be developed for detection of the disease. Early detection of disease will help the breeders and farmers to manage the disease.

References

Gill RJ and Brown JK. Systematics of Bemisia and Bemisia relatives: can molecular techniques solve the Bemisia tabaci complex conundrum–a taxonomist’s viewpoint. Bemisia: Bionomics and Management of a Global Pest. 2010: 5-29. doi: 10.1007/978-90-481-2460-2_1.

Briddon RW, Bull SE, Amin I, Mansoor S, Bedford ID, Rishi N et al. Diversity of DNA 1: a satellite-like molecule associated with monopartite begomovirus–DNA β complexes. Virology. 2004 Jul; 324(2): 462-74. doi: 10.1016/j.virol.2004.03.041.

Idris AM, Shahid MS, Briddon RW, Khan AJ, Zhu JK, Brown JK. An unusual alphasatellite associated with monopartite begomoviruses attenuates symptoms and reduces betasatellite accumulation. Journal of General Virology. 2011 Mar; 92(3): 706-17. doi: 10.1099/vir.0.025288-0.

Mansoor S, Briddon RW, Bull SE, Bedford ID, Bashir A, Hussain M et al. Cotton leaf curl disease is associated with multiple monopartite begomoviruses supported by single DNA β. Archives of Virology. 2003 Oct; 148: 1969-86. doi: 10.1007/s00705-003-0149-y.

Saunders K, Briddon RW, Stanley J. Replication promiscuity of DNA-β satellites associated with monopartite begomoviruses; deletion mutagenesis of the Ageratum yellow vein virus DNA-β satellite localizes sequences involved in replication. Journal of General Virology. 2008 Dec; 89(12): 3165-72. doi: 10.1099/vir.0.2008/003848-0.

Briddon RW and Markham PG. Complementation of bipartite begomovirus movement functions by topocuviruses and curtoviruses. Archives of Virology. 2001 Sep; 146: 1811-9. doi: 10.1007/s007050170067

Brown JK, Frohlich DE, Rosell RC. The sweetpotato or silverleaf whiteflies: biotypes of Bemisia tabaci or a species complex? Annual Review of Entomology. 1995 Jan; 40(1): 511-34. doi: 10.1146/annurev.en.40.010195.002455.

Mansoor S, Zafar Y, Briddon RW. Geminivirus disease complexes: the threat is spreading. Trends in Plant Science. 2006 May; 11(5): 209-12. doi: 10.1016/j.tplants.2006.03.003.

Karthikeyan A, Shobhana VG, Sudha M, Raveendran M, Senthil N, Pandiyan M et al. Mungbean yellow mosaic virus (MYMV): a threat to green gram (Vigna radiata) production in Asia. International Journal of Pest Management. 2014 Oct; 60(4): 314-24. doi: 10.1080/09670874.2014.982230.

Idris AM and Brown JK. Molecular analysis of Cotton leaf curl virus-Sudan reveals an evolutionary history of recombination. Virus Genes. 2002 Jun; 24: 249-56. doi: 10.1023/A:1015380600089.

Tahir MN, Amin I, Briddon RW, Mansoor S. The merging of two dynasties-identification of an African cotton leaf curl disease-associated begomovirus with cotton in Pakistan. PLoS One. 2011 May; 6(5): doi: 10.1371/journal.pone.0020366.

Sattar MN, Kvarnheden A, Saeed M, Briddon RW. Cotton leaf curl disease–an emerging threat to cotton production worldwide. Journal of General Virology. 2013 Apr; 94(4): 695-710. doi: 10.1099/vir.0.049627-0.

Rajagopalan PA, Naik A, Katturi P, Kurulekar M, Kankanallu RS, Anandalakshmi R. Dominance of resistance-breaking cotton leaf curl Burewala virus (CLCuBuV) in northwestern India. Archives of Virology. 2012 May; 157: 855-68. doi: 10.1007/s00705-012-1225-y.

Zaffalon V, Mukherjee SK, Reddy VS, Thompson JR, Tepfer M. A survey of geminiviruses and associated satellite DNAs in the cotton-growing areas of northwestern India. Archives of Virology. 2012 Mar; 157(3): 483-95. doi: 10.1007/s00705-011-1201-y.

Doyle JJ and Doyle JL. Isolation of plant DNA from fresh tissue. Focus.1990; (12)13-15. doi: 10.2307/2419362.

Tamura K and Nei M. Estimation of the number of nucleotide substitutions in the control region of mitochondrial DNA in humans and chimpanzees. Molecular Biology and Evolution. 1993 May 1;10(3):512-26.

Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: molecular evolutionary genetics analysis version 6.0. Molecular Biology and Evolution. 2013 Dec; 30(12): 2725-9. doi: 10.1093/molbev/mst197.

Haider MS, Tahir M, Saeed A, Ahmed S, Parveen R, Rashid N. First report of a begomovirus infecting the ornamental plant Vinca minor L. Australasian Plant Disease Notes. 2008 Nov; 3(1): 150-1. doi: 10.1007/BF03211274.

Hussain K, Hussain M, Mansoor S, Briddon RW. Complete nucleotide sequence of a begomovirus and associated betasatellite infecting croton (Croton bonplandianus) in Pakistan. Archives of Virology. 2011 Jun; 156: 1101-5. doi: 10.1007/s00705-011-0993-0.

Shahid MS, Mansoor S, Briddon RW. Complete nucleotide sequences of cotton leaf curl Rajasthan virus and its associated DNA [beta] molecule infecting tomato. Archives of Virology. 2007 Oct; 152(11): 2131. doi: 10.1007/s00705-007-1043-9.

Ashraf KH and Hanif M. Screening of cotton germplasm against cotton leaf curl virus. Pakistan Journal of Botany. 2010; 42(5): 3327-42.

Cui X, Tao X, Xie Y, Fauquet CM, Zhou X. A DNAβ associated with Tomato yellow leaf curl China virus is required for symptom induction. Journal of Virology. 2004 Dec; 78(24). doi: 10.1128/JVI.78.24.13966-13974.2004.

Sharma D, Kulshreshtha A, Roshan P, Hallan V. Molecular characterization and infectivity analysis of a bipartite begomovirus associated with cotton leaf curl Multan betasatellite naturally infecting Rumex nepalensis in northern India. Journal of Plant Pathology. 2019 Nov; 101: 935-41. doi: 10.1007/s42161-019-00295-6.

Cai JH, Xie K, Lin L, Qin BX, Chen BS, Meng JR, Liu YL. Cotton leaf curl Multan virus newly reported to be associated with cotton leaf curl disease in China. Plant Pathology. 2010; 59(4): 794-5. doi: 10.1111/j.1365-3059.2010.02266.x.