Plant Tissue Cult. & Biotech. 28(1): 125-140, 2018 (June)

In vitro Regeneration and Over Expression of Pea DNA Helicase 45 (PDH45) Gene into the Local Cultivars of Chickpea (Cicer arietinum L.) through Agrobacterium-mediated Genetic Transformation

Nuram Mubina¹, M.I. Hoque and R.H. Sarker^*

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

Key words: Chickpea, In vitro regeneration, Genetic transformation, Salinity tolerance, PDH45 gene

Abastract

In vitro regeneration studies compatible to Agrobacterium-mediated genetic transformation were carried out using two different types of zygotic embryo derived explants namely, decapitated embryo (DE) and decapitated embryo with single cotyledon disc (DEC) from three varieties of chickpea (Cicer arietinum L.) such as BARI chhola-4, -5 and -9 cultivated in Bangladesh. The best responses towards in vitro shoot regeneration was obtained from decapitated embryo with DEC on MS containing 0.5 mg/l BAP, 0.5 mg/l Kn and 0.2 mg/l NAA. Healthy and effective roots from the regenerated shoots were developed on MS supplemented with 0.2 mg/l IBA. Genetic transformation was carried out with Agrobacterium strain LBA4404 containing the binary plasmid pCAMBIA1301-PDH45 to integrate salt tolerant PDH45 gene in locally grown varieties of chickpea. The transformed plantlets were successfully established in soil following adequate hardening. Integration of salt tolerant PDH45 gene within the genomic DNA was confirmed through GUS histochemical assay and PCR analysis.

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