Plant Tissue Cult. & Biotech. 33(1): 37-46, 2023 (June)

General

Agrobacterium-mediated Genetic Transformation of two Peanut (Arachis hypogaea L.) Varieties using Salinity Tolerant PDH45 Gene

Ishita Haider^1*, R. H. Sarker and M. I. Hoque

Plant Breeding and Biotechnology Laboratory, Department of Botany, University of Dhaka, Dhaka‐1000, Bangladesh

Key words: Peanut, genetic transformation, pea DNA helicase (PDH45), salinity tolerance

Abastract

An efficient Agrobacterium-mediated genetic transformation protocol has been developed to integrate salinity tolerant pea DNA helicase (PDH45) gene in two peanut varieties, Dhaka-1 and BARI Chinabadam-8. Decapitated half-embryo explants from these two varieties of peanut were also used to develop a transformation compatible in vitro regeneration system. MS medium supplemented with 5.0 mg/l BAP and 0.5 mg/l Kn showed maximum responses towards shoot induction in both the varieties. Half-strength of MS medium supplemented with 0.2 mg/l IBA showed better responses for root induction in case of these varieties. Genetic transformation experiments were performed using Agrobacterium strain LBA4404 containing pCAMBIA1301-PDH45 conferring gus reporter gene, hygromycin resistance hpt gene, and salinity tolerant gene pdh45. The transformation variables were optimized, OD₆₀₀ 1.4 of the bacterial suspension with 45 minutes of incubation period, and three days of co-culture showed the best performance towards transformation. The transformed shoots were selected using 30.0 mg/l hygromycin. The overall transformation frequency was found to be very low, where 0.10% shoots of Dhaka-1 and 0.07% shoots of BARI Chinabadam-8 survived through required selection pressure. The stable integration of the transgene in the transformed shoots was confirmed through PCR analysis.

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