PJB-2019-265
Effective regeneration and transformation of potato variety Desiree with nucleotide diphosphate kinase 2 using IAA and BAP
Samiha Rashid, Muhammad Saad, Sofia Baig, Mohammad Maroof Shah, Ismat Nawaz and Ayesha Baig
Abstract
The main objective of the present study was to investigate the effect of different combinations of two growth regulators IAA and BAP on the In vitro regeneration of potato for successful transformation. The best optimized concentrations for potato regeneration was used for Agrobacterium mediated transformation with Arabidopsis Nucleotide Diphosphate Kinase2 (NDPK2). Potato is vulnerable to a number of biotic and abiotic stresses which limit its production. The effective In vitro regeneration of potato was carried out using Indol acetic acid (IAA) and Benzyl aminopurine (BAP) for transformation with (NDPK2) known to be involved in various environmental stresses. The internode explants of potato Desiree variety were cultured on MS media augmented with different concentrations of IAA i.e. (1.2, 1.5, 1.7, 2.0, 2.5 mg.L-1) in combination with BAP 1 mg.L-1. Data was collected after 15, 30 and 45 days of culturing and analyzed for statistical significance using analysis of variance (ANOVA). Fisher least significance using R- program was used to compare means at p = 0.05 level of significance. Transformation with NDPK2 was carried out using optimized regeneration protocol. The results indicated that high concentration of IAA i.e. 2.0 mg.L-1 with 1 mg.L-1 of BAP proved to be best for root regeneration. In all other parameters like shoot length, number of leaves, stem diameter and leaf area and potato regeneration, 1:1.5 mg.L-1 of BAP: IAA produced best result and thus was transformed with NDPK2 that generated 7 confirmed transgenic potato plants on PCR based sequencing. The findings demonstrate that the combination of BAP:IAA of 1:1.5 mg.L-1 can be regarded as the best optimized concentration for regeneration for stress related NDPK2 Agrobacterium mediated transformation of Desiree variety in potato.
To Cite this article:
Rashid, S., M. Saad, S. Baig, M.M. Shah, I. Nawaz and A. Baig. 2021. Effective regeneration and transformation of potato variety Desiree with nucleotide diphosphate kinase 2 using IAA and BAP. Pak. J. Bot., 53(1): DOI: http://dx.doi.org/10.30848/PJB2021-1(22)
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