Journal of Bioscience and Agriculture Research |
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RESEARCH ARTICLE:
Diversity analysis of begomovirus in Golden dewdrop (Duranta erecta) through PCR-RFLP
Maaz Ahmad (1), Luqman Amrao (1), Shahab Habib (1), Muhammad Zeshan Ahmed (1), Adnan Ahmed (2) and Salman Ghuffar (1)
1Dept. of Plant Pathology, University of Agriculture Faisalabad, Pakistan
2Dept. of Plant Pathology, PMAS Arid Agriculture University, Rawalpindi, Pakistan
Article info.
Received: 16.07.17, Revised: 24.09.17, Available online: 14 October 2017.
J. Bios. Agric. Res. | Volume 15, Issue 01, pp. 1260-1265
Crossref: https://doi.org/10.18801/jbar.150117.155
Diversity analysis of begomovirus in Golden dewdrop (Duranta erecta) through PCR-RFLP
Maaz Ahmad (1), Luqman Amrao (1), Shahab Habib (1), Muhammad Zeshan Ahmed (1), Adnan Ahmed (2) and Salman Ghuffar (1)
1Dept. of Plant Pathology, University of Agriculture Faisalabad, Pakistan
2Dept. of Plant Pathology, PMAS Arid Agriculture University, Rawalpindi, Pakistan
Article info.
Received: 16.07.17, Revised: 24.09.17, Available online: 14 October 2017.
J. Bios. Agric. Res. | Volume 15, Issue 01, pp. 1260-1265
Crossref: https://doi.org/10.18801/jbar.150117.155
FULL TEXT PDF:
| 155.17.15.1_diversity_analysis_of_begomovirus_in_golden_dewdrop__duranta_erecta__through_pcr-rflp.pdf |
Title: Diversity analysis of begomovirus in Golden dewdrop (Duranta erecta) through PCR-RFLP
Abstract: Golden dewdrop (Duranta erecta) is an ornamental plant which is commonly grown in Pakistan due to its natural beauty and medicinal properties. It is mainly grown in tropical and sub-tropical region. Golden dewdrop is known to suffer many diseases. Among all golden dewdrop leaf curl disease is caused by the begomovirusis very common. This study analyzed the DNA- A component of begomovirusin by using PCR-RFLP. For this purpose survey was conducted from ten different locations in UAF (University of Agriculture Faisalabad) from golden dewdrop plant according to the typical symptoms such as upward curling of leaves, darkening and thickening of veins, stunting of plant size. Infected samples were brought into Virology lab in CABB (Centre of Agricultural Biochemistry and Biotechnology). For DNA extraction CTAB method was used but DNA was not showed due to low concentration after quantification through spectrophotometer which was less than 100ng/µl. Therefore Rolling Circle Amplification method (RCA) was used for amplification of DNA-A component which only amplified two samples (D1 and D2) successfully. For polymerase chain reaction universal primers β01 and β02 was performed on RCA products for amplification of DNA-A component of begomoviruses which amplified the DNA at 2.8 kbp. For diversity analysis three commonly restriction enzyme Pst1, Sac1 and BamH1was used which restrict the DNA at different bp. Final conclusion of the analysis describe about the diversity of begomoviruses in golden dewdrop which emerging as an alternate host for begomoviruses.
Key Words: Begomovirus, Golden dewdrop, PCR, RCA and Restriction enzyme
Abstract: Golden dewdrop (Duranta erecta) is an ornamental plant which is commonly grown in Pakistan due to its natural beauty and medicinal properties. It is mainly grown in tropical and sub-tropical region. Golden dewdrop is known to suffer many diseases. Among all golden dewdrop leaf curl disease is caused by the begomovirusis very common. This study analyzed the DNA- A component of begomovirusin by using PCR-RFLP. For this purpose survey was conducted from ten different locations in UAF (University of Agriculture Faisalabad) from golden dewdrop plant according to the typical symptoms such as upward curling of leaves, darkening and thickening of veins, stunting of plant size. Infected samples were brought into Virology lab in CABB (Centre of Agricultural Biochemistry and Biotechnology). For DNA extraction CTAB method was used but DNA was not showed due to low concentration after quantification through spectrophotometer which was less than 100ng/µl. Therefore Rolling Circle Amplification method (RCA) was used for amplification of DNA-A component which only amplified two samples (D1 and D2) successfully. For polymerase chain reaction universal primers β01 and β02 was performed on RCA products for amplification of DNA-A component of begomoviruses which amplified the DNA at 2.8 kbp. For diversity analysis three commonly restriction enzyme Pst1, Sac1 and BamH1was used which restrict the DNA at different bp. Final conclusion of the analysis describe about the diversity of begomoviruses in golden dewdrop which emerging as an alternate host for begomoviruses.
Key Words: Begomovirus, Golden dewdrop, PCR, RCA and Restriction enzyme
HOW TO CITE THIS ARTICLE?
APA (American Psychological Association)
Ahmad, M., Amrao, L., Habib, S., Ahmed, M. Z., Ahmed, A. and Ghuffar, S. (2017). Diversity analysis of begomovirus in golden dewdrop (Duranta erecta) through PCR-RFLP. Journal of Bioscience and Agriculture Research,15(01), 1260-1265.
MLA (Modern Language Association)
Ahmad, M., Amrao, L., Habib, S., Ahmed, M. Z., Ahmed, A. and Ghuffar, S. “Effect of seed priming on growth and physiological traits of five Jordanian wheat (Triticum aestivum L.) landraces under salt stress”. Journal of Bioscience and Agriculture Research, 15.01(2017), 1260-1265.
Chicago and orTurabian
Ahmad, M., Amrao, L., Habib, S., Ahmed, M. Z., Ahmed, A. and Ghuffar, S. “Effect of seed priming on growth and physiological traits of five Jordanian wheat (Triticum aestivum L.) landraces under salt stress”. Journal of Bioscience and Agriculture Research, 15. no. 01(2017), 1260-1265.
APA (American Psychological Association)
Ahmad, M., Amrao, L., Habib, S., Ahmed, M. Z., Ahmed, A. and Ghuffar, S. (2017). Diversity analysis of begomovirus in golden dewdrop (Duranta erecta) through PCR-RFLP. Journal of Bioscience and Agriculture Research,15(01), 1260-1265.
MLA (Modern Language Association)
Ahmad, M., Amrao, L., Habib, S., Ahmed, M. Z., Ahmed, A. and Ghuffar, S. “Effect of seed priming on growth and physiological traits of five Jordanian wheat (Triticum aestivum L.) landraces under salt stress”. Journal of Bioscience and Agriculture Research, 15.01(2017), 1260-1265.
Chicago and orTurabian
Ahmad, M., Amrao, L., Habib, S., Ahmed, M. Z., Ahmed, A. and Ghuffar, S. “Effect of seed priming on growth and physiological traits of five Jordanian wheat (Triticum aestivum L.) landraces under salt stress”. Journal of Bioscience and Agriculture Research, 15. no. 01(2017), 1260-1265.
- Briddon, R. W. and Stanley, J. (2006). Sub-viral agents associated with plantinfecting single-stranded DNA viruses. Virology, 344, 198-210. https://doi.org/10.1016/j.virol.2005.09.042
- Dean, F. B., Nelson, J. R., Giesler, T. L. and Lasken, R. S. (2001). Rapid Amplification of Plasmid and Phage DNA Using Phi29 DNA Polymerase and Multiply-Primed Rolling Circle Amplification. Genome Research, 11(6), 1095–1099. https://doi.org/10.1101/gr.180501
- Doyle, J. J. and Doyle, J. L. (1990). Isolation of plant DNA from fresh tissue. Focus. 12, 13-15.
- Gusev, Y., Sparkowski, J., Raghunathan, A., Ferguson, H., Montano, J., Bogdan, N. and Wheeler, V. (2001). Rolling Circle Amplification: A New Approach to Increase Sensitivity for Immunohistochemistry and Flow Cytometry. The American Journal of Pathol, 159(1), 63–69.
- Lee, I. M. D., Gundersen-Rindal, E., Davis, R. E. and Bartoszyk, I. M. (2012). Revised classification scheme of phytoplasmas based onRFLP analyses of 16S rRNA and ribosomal protein genesequences. Int J Syst Bacteriol, 48, 1153-1169. https://doi.org/10.1099/00207713-48-4-1153
- Mansoor, S., Briddon, R. W., Bull, S. E., Bedford, I. D., Bashir, A., Hussain, M., Saeed, M., Zafar, M. Y., Malik, K. A., Fauquet, C. and Markham, P. G. (2003). Cotton leaf curl disease is associated with multiple monopartite begomoviruses supported by single DNA. Archives of Virol, 148, 1969-1986.
- Patil, B. L. and Dasgupta, I. (2006). Defective Interfering DNAs of Plant Viruses. Critical Reviews in Plant Sci, 25, 47–64. https://doi.org/10.1080/07352680500391295
- Relman, D. A., Schmidt, T. M., Gajadhar, A., Sogin, M., Cross, J., Yoder, K., Sethabutr, O. and echeverria, P. (1996). Molecular phylogenetic analysis of Cyclospora, the human intestinal pathogen, suggests that it is closely related to Eimeria species. J. Infect. Dis, 173, 440-445.
- 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. J. of General Virol, 81, 1839-1849.
- Saunders, K., Norman, A., Gucciardo, S. and Stanley, J. (2004). The DNA β satellite component associated with ageratum yellow vein disease encodes an essential pathogenicity protein (βC1). Virol, 324, 37-47. https://doi.org/10.1016/j.virol.2004.03.018
- Sunter, G., Hartitz, M. D., Hormuzdi, S. G., Brough, C. L. and Bisaro, D. M. (1990). Genetic analysis of tomato golden mosaic virus: ORF AL2 is required for coat protein accumulation while ORF AL3 is necessary for efficient DNA replication. Virol, 179, 69–77.
- Seal, S. E., van den Bosch, F. and Jeger, M. J. (2006). Factors influencing begomovirus evolution and their increasing global significance: implications for sustainable control. Critical Reviews in Plant Sciences, 25, 23-46. https://doi.org/10.1080/07352680500365257
- Whistler, W. A. (2000). Tropical ornamentals, a guide. Timber Press, Inc., Portland. p. 542.
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