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    Tissue-specific and stress-inducible promoters establish their suitability for containment of foreign gene(s) expression in transgenic potatoes
    (Springer Heidelberg, 2020) Rahamkulov, Ilhom; Bakhsh, Allah
    The present study was conducted to determine efficiency of green tissue-specific (pRCA) and stress-inducible promoters (pRD29A) to expressE. colibeta-glucuronidase (gusA) gene in transgenic potatoes compared with constitutive promoter (35S CaMV). The promoter fragments were isolated from their original source and cloned upstream togusAin pCAMBIA-1301 binary vector to develop plant expression constructs, i.e., pRCA-pCAMBIA and pRD29A-pCAMBIA.Agrobacteriumstrain GV2260 harboring recombinant plasmids were used to infect leaf discs and internodal explant of Lady Olympia cultivar. GUS histochemical analysis was performed at different stages to determine GUS activity in transgenic plants. To determine activity of stress-inducible promoter (pRD29A), transgenic plants were exposed to heat, drought and combination of both heat and drought stress. The real time (RT-qPCR) and GUS florimetric assays revealed that pRD29A promoter gets more activated under drought, heat and combination of both stresses. GUS expression levels were more than 10 folds high with pRD29A promoter compared to control. Likewise, the reduced transcripts levels ofgusAgene under control of pRCA promoter were found in tuber/roots of transgenic plants compared to 35S promoter. GUS florimetric assays also showed decreased or no GUS expression in tubers. In conclusion, the results encourage the appropriate use of promoters to drive the expression of foreign gene(s) for the development of potato lines tolerant to biotic and abiotic stress while minimizing the risks of transgenic technology in potatoes.
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    Transgenic potato lines expressing CP4-EPSP synthase exhibit resistance against glyphosate
    (Springer, 2020) Bakhsh, Allah; Hussain, Tahira; Rahamkulov, Ilhom; Demirel, Ufuk; Caliskan, Mehmet Emin
    Potato crops are particularly vulnerable to weed competition from emergence to canopy closure and are subject to significant yield loss. Glyphosate is broad spectrum herbicide used to control weeds worldwide. In order to incorporate glyphosate resistant trait in four potato cultivars (Lady Olympia, Desiree, Agria and Granola), an efficient, cost effective, reproducible and stable Agrobacterium-mediated genetic transformation protocol was performed using leaf and internodal explants. Agrobacterium strain LBA4404 harboring newly modified recombinant binary vector pCAMHE-EPSPS containing EPSP synthase gene under the control of Cauliflower mosaic virus 35S promoter was used to infect explants. The overall transformation efficiency was 26.4%. Of the 280 plants transferred to greenhouse, 74 plants were found to be PCR positive with gene of interest. GUS histochemical, Southern blot, RT-qPCR, lateral flow dipstick assays confirmed integration and expression of EPSPS in primary transformants. The putative transgenic plants developed from these cultivars possessed enhanced resistance to glyphosate applications in T0 and first tuber generation. These transgenic potato lines could be used as source of germplasm for an efficient potato breeding program. Key message The article reports the development of potato transgenic lines with enhanced tolerance against glyphosate.