Establishment of High-Efficiency Agro bacterium-Mediated Transformation of Callus Derived From Sehima Nervosum, An Important Range Grass Species
Journal Title: International Journal of Agricultural Science and Research (IJASR) - Year 2017, Vol 7, Issue 6
Abstract
Sehima Nervosum is a natural range grass found inherently rich in precursors, for several industrially important biomolecules. Production of nutraceuticals (prebiotics xylo-oligosaccharides) from this grass is promising. It reproduces predominantly through apomixes, which provides a means of clonal propagation through seeds. The lack of sexual reproduction in S. Nervosum limits the possibilities of genetic improvement by hybridization. Development of an efficient gene delivery system would be very useful for maintenance, multiplication and genetic improvement of this grass and to enhance nutraceuticals production. A callus inducing and transformation system was developed utilizing embryogenic callus derived from seed explants of this grass on Murashige and Skoog medium with various concentrations of 2,4-D followed by infection of embryogenic calli with A. tumefaciens strain EHA105 harbouring binary vector p CAMBIA1305.1 with GUS gene. The result showed that, the number of calli expressing GUS gene first increased with inoculation time and bacterial density (OD600) and then, dramatically decreased with the increment of both factors. The highest percentage (90%) of transformation efficiency was obtained in the calli, inoculated with Agrobacterium for up to 30 minutes at OD600 of 1.0, followed by 83% and 81% at OD600 of 0.6 and 1.0, respectively. Our result suggests that infection time and Agrobacterium density affects transformation efficiency. To the best of our knowledge efficient in-vitro callus induction from mature seed explants, callus transformation and transformation efficiency in Sehima is reported here for the first time.
Authors and Affiliations
Krishna Kumar Dwivedi, R. Katiyar, A Radhakrishna, M. K. Srivastava, P. Kaushal
Keywords
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