Tissue-specific and stress-inducible promoters establish their suitability for containment of foreign gene(s) expression
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ORIGINAL ARTICLE
Tissue‑specific and stress‑inducible promoters establish their suitability for containment of foreign gene(s) expression in transgenic potatoes Ilhom Rahamkulov1 · Allah Bakhsh1 Received: 26 May 2020 / Accepted: 20 July 2020 © King Abdulaziz City for Science and Technology 2020
Abstract The present study was conducted to determine efficiency of green tissue-specific (pRCA) and stress-inducible promoters (pRD29A) to express E. coli beta-glucuronidase (gusA) gene in transgenic potatoes compared with constitutive promoter (35S CaMV). The promoter fragments were isolated from their original source and cloned upstream to gusA in pCAMBIA-1301 binary vector to develop plant expression constructs, i.e., pRCA-pCAMBIA and pRD29A-pCAMBIA. Agrobacterium strain 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 of gusA gene 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. Keywords Promoter efficiency · Potato · Transgene development · Targeted expression
Introduction Potato (Solanum tuberosum L.) is one of the most important cultivated plants in the world in terms of its uses as both industrial raw material and direct food. Potato has the potential to solve the insufficient problems of food resources for the increasing population because of its richness in nutrient composition (Alisdair et al. 2001). Besides that, eight species of Solanum genus are suitable for consumption as Electronic supplementary material The online version of this article (https://doi.org/10.1007/s13205-020-02350-x) contains supplementary material, which is available to authorized users. * Allah Bakhsh [email protected]; [email protected] 1
Department of Agricultural Genetic Engineering, Faculty of Agricultural Sciences and Technologies, Nigde Omer Halisdemir University, 51240 Nigde, Turkey
human food; however, the most widely known and produced species is Solanum tuberosum (Rowe 1993). The commercialized transgenic crops express insect or herbicide resistant gene(s) under the control of 35S cauliflower mosaic virus that is cons
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