PJB-2020-297
Influence of pH of co-cultivation medium, claforan® concentration, co-cultivation period, pre-selection duration and selection regime on tomato transformation
Sabir Hussain Shah, Shaukat Ali, Sher Muhammad, Shehla Shoukat, Zahid Hussain, Muhammad Amir Zia, Arshad Iqbal, Siffat Ullah Khan and Muhammad Latif
Abstract
Tomato fruit is enriched with lycopene and β-carotene which is an initiator of vitamin A, whereas lycopene has a shielding effect against prostate cancer. The tomato crop is very sensitive to cold stress which is a great threat to the yield of cultivated tomato. Genetic transformation offers the basis of foreign genes integration in the plant for the improvement of crop. Therefore, the present study was carried out to optimize different parameters of gene transformation in three tomato genotypes. These three tomato genotypes were transformed for cold tolerance via DREB1A gene with inducible promoter (Lip9). Optimal conditions to achieve 14.16-16.45% transformation efficiency (TE) was found as pH ranged between 5.6-5.7 for co-cultivation media, 500 mg/l claforan®, 6-d pre-selection period, 48-h co-cultivation duration and 35 mg/l hygromycin. Specific gene primers were used for screening and real time semi quantitative PCR assay that proved the insertion of DREB1A gene and its expression in T0 and T1 transformed lines. During the cold stress period, the increase in expression pattern was confirmed by the results of semi-quantitative PCR. Under normal growth settings, there were no considerable differences among transformants and non transformants based on morphological characteristics. Convincingly, DREB1A gene over expression in transgenic tomato plants increased the protection and delivered cold tolerance
To Cite this article:
Shah, S.H., S. Ali, S. Muhammad, S. Shoukat, Z. Hussain, M.A. Zia, A. Iqbal, S.U. Khan and M. Latif. 2022. Influence of pH of co-cultivation medium, claforan® concentration, co-cultivation period, pre-selection duration and selection regime on tomato transformation. Pak. J. Bot., 54(3): DOI: http://dx.doi.org/10.30848/PJB2022-3(11)
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