Micropropagation of virus-free plants of Saudi fig (Ficus carica L.) and their identification through enzyme-linked immunosorbent assay methods

In Vitro Cellular & Developmental Biology - Plant - Tập 54 - Trang 626-636 - 2018
Hassan S. M. Al-Zahrani1, Omar A. Almaghrabi2, Michael P. Fuller3, Hemaid I. A. Soliman4,5, Muhammad Farooq6,7,8, Ehab M. R. Metwali2,9
1Biological Science Department, Faculty of Science, King Abdulaziz University, Jeddah, Saudi Arabia
2Biological Science Department, Faculty of Science, University of Jeddah, Jeddah, Saudi Arabia
3School of Biological and Marine Sciences, Faculty of Science and Engineering, University of Plymouth, Plymouth, UK
4Plant Genetic Resources Department, Desert Research Center, Cairo, Egypt
5Tissue Culture and Biotechnology Labs, Maryout Research Station, Desert Research Center, Alexandria, Egypt
6Department of Crop Sciences, College of Agricultural and Marine Sciences, Sultan Qaboos University, Al-Khoud, Oman
7Department of Agronomy, University of Agriculture, Faisalabad, Pakistan
8The UWA Institute of Agriculture, The University of Western Australia, Perth, Australia
9Botany Department, Faculty of Agriculture, Suez Canal University, Ismailia, Egypt

Tóm tắt

Viral infection is one of the most serious biotic stresses, which disturbs the growth and productivity of many horticultural crops, including that of fig (Ficus carica L.). The production of plants free of viruses, such as fig mosaic virus (FMV), has become a priority in many plant breeding programs. In this study, leaves from plants of two fig cultivars, Kodato and Dattora, infected with FMV were collected from both Mecca and Al-Taif, Saudi Arabia. Transmission electron microscopy of ultrathin leaf sections showed double membrane bodies, characteristic of FMV particles, only in the mesophyll cells of infected samples. Protein analysis using sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed the presence of a protein band with a molecular weight of 35 kDa, which corresponded to the viral coat protein; and FMV was confirmed by Western blot and enzyme-linked immunosorbent assay (ELISA) tests. To obtain virus-free plants, apical shoot culture was applied. A comparison of various artificial media with different concentrations of growth regulators was evaluated to optimize shoot formation, shoot multiplication, and root formation, and was followed by plant acclimation ex vitro. Direct ELISA analysis of shoots micropropagated from meristem tip explants indicated that there were virus-free shoots, when compared to infected plants (positive control), while there were no significant differences between these explants and healthy samples (negative control). This study demonstrated that in vitro micropropagation of Saudi F. carica infected with FMV virus led to the successful elimination of the virus.

Tài liệu tham khảo

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