Authors: Valentine Otang Ntui Raham Sher khan Dong Poh Chin Ikuo Nakamura Masahiro Mii
Publish Date: 2010/05/09
Volume: 103, Issue: 1, Pages: 15-22
Abstract
Cotyledonary explants of two “Egusi” genotypes ‘Ejagham’ and NHC1130 were cocultivated with Agrobacterium tumefaciens strain EHA101 carrying either plasmid pIG121Hm harbouring genes coding for betaglucuronidase gus hygromycin phosphotransferase hpt and neomycin phosphotransferase II nptII or plasmid pBBRacdS harbouring these same genes along with a gene coding for 1aminocyclopropane1carboxylate ACC deaminase Six weeks after cocultivation more than 35 of explants produced shoots in both cultivars A DNA fragment corresponding to the gus gene or the selection marker nptII was amplified from genomic DNA extracted from leaves of regenerated plant clones rooted on hormonefree MS medium containing 100 mg/l kanamycin suggesting their transgenic nature Southern blot analysis confirmed successful integration of one to three copies of the gus gene Transformation efficiencies of cultivar NHC1130 with EHA101pIG121Hm and EHA101pIG121Hm pBBRacdS were 38 and 10 respectively which were higher than those obtained for cultivar ‘Ejagham’ of 24 and 57 respectively Cocultivation medium containing 5 mg/l BA was effective for obtaining high transformation efficiency for both cultivars as compared with that without it
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