Agrobacterium-Mediated Genetic Transformation Involving Hygromycin Phosphotransferase to Impart Herbicide Tolerance in Indica Rice

International Journal of Research and Scientific Innovation (IJRSI) | Volume V, Issue V, May 2018 | ISSN 2321–2705

Agrobacterium-Mediated Genetic Transformation Involving Hygromycin Phosphotransferase to Impart Herbicide Tolerance in Indica Rice

 Aparna Maiti^{1, 3}, Chiranjib Mandal², Sourav Dutta², Pranit Mukherjee² and Asit B. Mandal^{1, 2*}

¹Biotechnology Section, Central Agricultural Research Institute, Port Blair 744101, A&N Islands, India
²Biotechnology Unit, Division of Crop Improvement, ICAR-Central Research Institute for Jute and Allied Fibres, Barrackpore 700120, WB, India
³Department of Orthopedic Surgery, Virginia Commonwealth University, USA

Abstract: – Hygromycin  phosphotransferase (hpt) gene has been  successfully intruded in an indica rice variety-IET 13856 through Agrobacterium-mediated genetic transformation to impart herbicide-tolerance. Transgenic plants were regenerated from in vitro developed embryogenic calli of IET 13856 through co-cultivation with Agrobacterium tumifaciens strain LBA 4404 (pTOK 233). The  hpt gene was housed under the transcriptional control of CaMV 35S promoter and Nos polyadenylation signal in the super binary vector pTOK233. This protocol of Agrobacterium -mediated genetic transformation was found to be very efficient as evident from high (51.4% transient and 13.51% stable) transformation efficiency. Transient expression was confirmed through GUS histochemical assay, while stable transgene integration at the genomic level was confirmed by PCR, slot blot and Southern blot hybridization. Inheritance of the transgene was worked out through seed germination test of the seeds at T₁ generation on hygromycin supplemented MS medium. It showed typical Mendelian ratio for monogenic inheritance (dominant transgene) in the progenies of maximum transgenic lines except a few deviations, which might be due to epistatic interactions or small sample size (less number of seeds used in seed germination test). This study prospects amply, use of transgenic indica rice harbouring hpt to enhance productivity in rice by saving the crops from weed menace in Andaman and Nicobar Islands, India, endowed with prolonged high rain fall with rampant weed infestation and other rice growing areas elsewhere.

Key words: Agrobacterium-mediated genetic transformation, GUS, Hygromycin phosphotransferase, Glyphosate, Indica rice, Andaman and Nicobar Islands.

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