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    Development of Insect Resistant Potato Transplastomic Lines.
    (Springer, 2019) Bakhsh, Allah; Joyia, Faiz Ahmad; Hossain, Md. Jakir; Aksoy, Emre; Gokce, Neslihan Zahide Ozturk; Mustafa, Ghulam; Khan, Muhammad Sarwar
    [Abstract Not Available]
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    Engineering of insecticidal hybrid gene into potato chloroplast genome exhibits promising control of Colorado potato beetle, Leptinotarsa decemlineata (Coleoptera: Chrysomelidae)
    (Springer, 2023) Hossain, Md. Jakir; Bakhsh, Allah; Joyia, Faiz Ahmad; Aksoy, Emre; Gokce, Neslihan Zahide Ozturk; Khan, Muhammad Sarwar
    The potato chloroplast was transformed with codon optimized synthetic hybrid cry gene (SN19) to mitigate crop losses by Colorado potato beetle (CPB). The bombarded explants (leaves and internode) were cultured on MS medium supplemented with BAP (2.0 mg/l), NAA (0.2 mg/l), TDZ (2.0 mg/l) and GA3 (0.1 mg/l); spectinomycin 50 mg/l was used as a selection agent in the medium. Leaf explants of cultivar Kuroda induced highest percentage (92%) of callus where cultivar Santae produced the highest percentage (85.7%) of transplastomic shoots. Sante and Challenger showed 9.6% shoot regeneration efficiency followed by cultivar Simply Red (8.8%). PCR amplification yielded 16 postive transplastomic plantlets out of 21 spectinomycin resistant ones. Target gene integration was confirmed by PCR and Southern blot, whereas RT-qPCR was used to assess the expression level of transgene. The localization of visual marker gene gfp was tracked by laser scanning confocal microscopy which confirmed its expression in chloroplasts of leaf cells. The transplastomic plants ensured high mortality to both larvae and adult CPB. Foliage consumption and weight gain of CPB fed on transplastomic leaves were lower compared to the control plants. Sucessful implementation of current research findings can lead to a viable solution to CPB mediated potato losses globally.
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    Rapid and efficient in vitro regeneration of transplastomic potato (Solanum tuberosum L.) plants after particle bombardment
    (Tubitak Scientific & Technological Research Council Turkey, 2021) Hossain, Md Jakir; Aksoy, Emre; Gokce, Neslihan Zahide Ozturk; Joyia, Faiz Ahmad; Khan, Muhammad Sarwar; Bakhsh, Allah
    In contrast with traditional nuclear gene transformation, transplastomic technology has opened a new horizon for the transgenic plant research that offers several beneficial aspects including the convenient use of transgene stacking and the generation of high expression levels of foreign proteins. However, this technology has been well adopted and established in tobacco, the introduction and adoption of cost-effective, swift, and reproducible protocol for in vitro regeneration of transplastomic potato is challenging and laborious. The present research aimed to develop such prompt and efficient protocol to instigate and revive the regeneration potential with the combinations of different plant growth regulators (PGRs). Leaves and internodal explants from four potato cultivars were transformed with chloroplast transformation vector via particle bombardment and cultured on MS media supplemented with suitable PGRs and selection agents. Leaf explants of cultivar Kuroda induced highest (92%) number of calli where cultivar Sante produced the highest (85.7%) transplastomic shoots. Thidiazuron was found more proficient (41%) for shoot regeneration. Finally, within only seven weeks, we got 21 spectinomycin resistant shoot, and 16 of those showed integration of target genes into the plastome in PCR screening.