Study of different factors affecting Agrobacterium- mediated genetic transformation in lentil (Lens culinaris Medik.)

S. K. Das; M. I. Hoque; R. H. Sarker

doi:10.30574/gscbps.2018.5.1.0087

Authors

S. K. Das Department of Botany, University of Barisal, Barisal-8200, Bangladesh.
M. I. Hoque Department of Botany, University of Dhaka, Dhaka-1000, Bangladesh.
R. H. Sarker Department of Botany, University of Dhaka, Dhaka-1000, Bangladesh.

DOI:

https://doi.org/10.30574/gscbps.2018.5.1.0087

Keywords:

Lentil, Bacterial density, Incubation, Co-cultivation, Agrobacterium, Transformation

Abstract

The present work was aimed to study the various factors affecting Agrobacterium tumefaciens mediated genetic transformation of three BARI masur variety of lentil (Lens culinaris Medik.), namely, BM-4, BM-5 and BM-6. Agrobacterium strain LBA4404 containing plasmid pBI121 which carrying gus and nptII as screenable and selectable marker genes, respectively, was used for transformation. Factors affecting transformation efficiency, such as, type of explants, bacterial density, incubation time and co-cultivation period were studied. The result showed that decapitated embryo (DE) explants of BM-6 variety of lentil inoculated in Agrobacterium suspensions at density of 1.0 (OD₆₀₀) for 45 minutes and co-cultivated on MS medium with hormones in the dark for 3 days gave the highest transient expression of gus gene (86.90%). The lowest percentage (15.00%) of GUS +ve explants observed in cotyledonary node (CN) of BM-5 variety of lentil when they were inoculated in Agrobacterium suspensions at density of 0.5 (OD₆₀₀) for 15 minutes. Using this protocol agronomical and economically important trait could be transfer to the locally grown lentil varieties.

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