Protocorm wounding enhanced Agrobacteriummediated transformation of Hygrochilus parishii (Veitch & Rchb.f.) Pfit
Journal Title: Advances in Agriculture & Botanics - International Journal of the Bioflux Society - Year 2010, Vol 2, Issue 3
Abstract
This study was aimed to encourage the use of protocorm wounding as a substitute of acetosyringone in an attempt to reduce cocultivation time and to increase possibility of successful genetic transformation of Hygrochilus parishii (Veitch & Rchb.f.) Pfitz. Agrobacterium tumefaciens strain LBA4404 harboring the plasmid pBI121 was employed as a transgene carrier. The plasmid pBI121 contained β-glucuronidase (gus) and kanamycin resistance (nptII) genes, both of which were driven by the CaMV35Spromoter. It was found that wounded protocorms cocultivated with A. tumefaciens for 30 min and acetosyringone-treated protocorms cocultivated for 30 min exhibited high gene expressions of 75 and 80%, respectively, while normal protocorms (control) showed gene expression of 60% when cocultivated with A. tumefaciens for 45 min. The presence of CaMV35S promoter and NOS terminator detected by PCR technique confirmed a successful T-DNA integration. This suggested that protocorm wounding could be used as a substitute of acetosyringone in enhancement of genetic modification.
Authors and Affiliations
Witaya Pimda, Sumontip Bunnag
Keywords
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