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Tissue-specific and stress-inducible promoters establish their suitability for containment of foreign gene(s) expression in transgenic potatoes

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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.

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Acknowledgements

The present study is a part of research project supported by TÜBİTAK (Grant No. 215O520), where we used pRCA promoter to drive the expression of EcR dsRNAs in green part of potato plants. The authors are highly thankful to Tübitak for providing fellowship to Mr. Ilhom Rahamkulov from the project budget. Authors are grateful to Prof. Dr. Mehmet Emin Çalışkan for providing seed tuber of Lady Olympia used in the study. Special thanks to Dr. Emre Aksoy for his technical help during experiments and interpretation of results. We are also thankful to Dr. Halil Toktay for providing access to microscope facility.

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  1. Department of Agricultural Genetic Engineering, Faculty of Agricultural Sciences and Technologies, Nigde Omer Halisdemir University, 51240, Nigde, Turkey

    Ilhom Rahamkulov & Allah Bakhsh

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  1. Ilhom Rahamkulov
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  2. Allah Bakhsh
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The data presented in the manuscript is MS thesis work of IR who completed his studies under the supervision of AB.

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Correspondence to Allah Bakhsh.

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Rahamkulov, I., Bakhsh, A. Tissue-specific and stress-inducible promoters establish their suitability for containment of foreign gene(s) expression in transgenic potatoes. 3 Biotech 10, 426 (2020). https://doi.org/10.1007/s13205-020-02350-x

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