J. Biosci. Agric. Res. | Volume 20, Issue 01, 1664-1670| https://doi.org/10.18801/jbar.200119.202
Article type: Research article|Received: 14.01.2019, Revised: 04.02.2019, Date of Publication: 14 April 2019.
Article type: Research article|Received: 14.01.2019, Revised: 04.02.2019, Date of Publication: 14 April 2019.
Identification of beta satellite component in (Hibiscus rosa-sinensis L.) through PCR-RFLP from Faisalabad, Pakistan
Shahab Habib 1, Luqman Amrao 1, Muhammad Zeshan Ahmed 1, Maaz Ahmad 1, Shanam Naz 1, Muhammad Ahmad Zeshan 4, Hafiz Muhammad Asadullah 2, Abdul Manan Hamza 3 and Salman Ghuffar 1
1 Dept. of Plant Pathology, University of Agriculture Faisalabad, Pakistan
2 Pulses Research Program, NARC Islamabad, Pakistan
3 Dept. of Entomology, PMAS-Arid Agriculture University, Rawalpinidi, Pakistan
4 Dept. of Plant Pathology, University of Sargodha, Sargodha, Pakistan.
1 Dept. of Plant Pathology, University of Agriculture Faisalabad, Pakistan
2 Pulses Research Program, NARC Islamabad, Pakistan
3 Dept. of Entomology, PMAS-Arid Agriculture University, Rawalpinidi, Pakistan
4 Dept. of Plant Pathology, University of Sargodha, Sargodha, Pakistan.
Abstract
Shoe flower (Hibiscus rosa-sinensis L.) is an ornamental plant belongs to the family Malvaceae. Due to its natural beauty and medicinal properties it is mainly grown in regions with tropical and subtropical environment. Shoe flower is known to infect by many diseases. Among all diseases Hibiscus leaf curl disease is caused by the begomovirus transmitted through Bemisia tabaci is very common and also an alternative host of cotton leaf curl disease due to the similar symptomology. In present study the identification of begomovirus in shoe flower was analyzed by using PCR-RFLP (Polymerase chain reaction- Restriction fragment length polymorphism). For this purpose, survey was conducted from seven different locations in university of agriculture Faisalabad. Plants were observed with virus symptoms of Hibiscus leaf curl disease such as upward curling of leaves, darkening and thickening of veins, stunting of plant size as well as leaf enation. For further molecular studies, DNA was isolated from all infected samples by using CTAB method and confirmed by Gel electrophoresis. PCR reaction amplified three DNA samples having beta satellite component, labeled (S3, S4 and S6) at 1.4 kbp. According to the (PCR-RFLP) analysis of three amplified samples, In BamH1 restriction enzyme, all the samples were restricted into 625 bp. In second restriction enzyme Pst1, DNA samples were restricted into 1.4 kbp. While in the case of ECOR1, fragment was restricted into 1.3 kbp.
Key Words: Shoe flower, DNA extraction, PCR, PFLP, Beta satellite and Begomovirus
Shoe flower (Hibiscus rosa-sinensis L.) is an ornamental plant belongs to the family Malvaceae. Due to its natural beauty and medicinal properties it is mainly grown in regions with tropical and subtropical environment. Shoe flower is known to infect by many diseases. Among all diseases Hibiscus leaf curl disease is caused by the begomovirus transmitted through Bemisia tabaci is very common and also an alternative host of cotton leaf curl disease due to the similar symptomology. In present study the identification of begomovirus in shoe flower was analyzed by using PCR-RFLP (Polymerase chain reaction- Restriction fragment length polymorphism). For this purpose, survey was conducted from seven different locations in university of agriculture Faisalabad. Plants were observed with virus symptoms of Hibiscus leaf curl disease such as upward curling of leaves, darkening and thickening of veins, stunting of plant size as well as leaf enation. For further molecular studies, DNA was isolated from all infected samples by using CTAB method and confirmed by Gel electrophoresis. PCR reaction amplified three DNA samples having beta satellite component, labeled (S3, S4 and S6) at 1.4 kbp. According to the (PCR-RFLP) analysis of three amplified samples, In BamH1 restriction enzyme, all the samples were restricted into 625 bp. In second restriction enzyme Pst1, DNA samples were restricted into 1.4 kbp. While in the case of ECOR1, fragment was restricted into 1.3 kbp.
Key Words: Shoe flower, DNA extraction, PCR, PFLP, Beta satellite and Begomovirus
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MLA
Habib, et al. “Identification of beta satellite component in (Hibiscus rosa-sinensis L.) through PCR-RFLP from Faisalabad, Pakistan.” Journal of Bioscience and Agriculture Research 20(01) (2019): 1664-1670.
APA
Habib, S., Amrao, L., Ahmed, M. Z., Ahmad, M., Naz, S., Zeshan, M. A., Asadullah, H. M., Hamza, A. M. and Ghuffar, S. (2019). Identification of beta satellite component in (Hibiscus rosa-sinensis L.) through PCR-RFLP from Faisalabad, Pakistan. Journal of Bioscience and Agriculture Research, 20(01), 1664-1670.
Chicago
Habib, S., Amrao, L., Ahmed, M. Z., Ahmad, M., Naz, S., Zeshan, M. A., Asadullah, H. M., Hamza, A. M. and Ghuffar, S. “Identification of beta satellite component in (Hibiscus rosa-sinensis L.) through PCR-RFLP from Faisalabad, Pakistan.” Journal of Bioscience and Agriculture Research 20(01) (2019): 1664-1670.
Harvard
Habib, S., Amrao, L., Ahmed, M. Z., Ahmad, M., Naz, S., Zeshan, M. A., Asadullah, H. M., Hamza, A. M. and Ghuffar, S. 2019. Identification of beta satellite component in (Hibiscus rosa-sinensis L.) through PCR-RFLP from Faisalabad, Pakistan. Journal of Bioscience and Agriculture Research, 20(01), pp. 1664-1670.
Vancouver
Habib S, Amrao L, Ahmed MZ, Ahmad M, Naz S, Zeshan MA, Asadullah HM, Hamza AM and Ghuffar S. Identification of beta satellite component in (Hibiscus rosa-sinensis L.) through PCR-RFLP from Faisalabad, Pakistan. Journal of Bioscience and Agriculture Research. 2019 April 20(01): 1664-1670.
Habib, et al. “Identification of beta satellite component in (Hibiscus rosa-sinensis L.) through PCR-RFLP from Faisalabad, Pakistan.” Journal of Bioscience and Agriculture Research 20(01) (2019): 1664-1670.
APA
Habib, S., Amrao, L., Ahmed, M. Z., Ahmad, M., Naz, S., Zeshan, M. A., Asadullah, H. M., Hamza, A. M. and Ghuffar, S. (2019). Identification of beta satellite component in (Hibiscus rosa-sinensis L.) through PCR-RFLP from Faisalabad, Pakistan. Journal of Bioscience and Agriculture Research, 20(01), 1664-1670.
Chicago
Habib, S., Amrao, L., Ahmed, M. Z., Ahmad, M., Naz, S., Zeshan, M. A., Asadullah, H. M., Hamza, A. M. and Ghuffar, S. “Identification of beta satellite component in (Hibiscus rosa-sinensis L.) through PCR-RFLP from Faisalabad, Pakistan.” Journal of Bioscience and Agriculture Research 20(01) (2019): 1664-1670.
Harvard
Habib, S., Amrao, L., Ahmed, M. Z., Ahmad, M., Naz, S., Zeshan, M. A., Asadullah, H. M., Hamza, A. M. and Ghuffar, S. 2019. Identification of beta satellite component in (Hibiscus rosa-sinensis L.) through PCR-RFLP from Faisalabad, Pakistan. Journal of Bioscience and Agriculture Research, 20(01), pp. 1664-1670.
Vancouver
Habib S, Amrao L, Ahmed MZ, Ahmad M, Naz S, Zeshan MA, Asadullah HM, Hamza AM and Ghuffar S. Identification of beta satellite component in (Hibiscus rosa-sinensis L.) through PCR-RFLP from Faisalabad, Pakistan. Journal of Bioscience and Agriculture Research. 2019 April 20(01): 1664-1670.
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