Plant Tissue Cult. & Biotech. 25(2): 155-164, 2015 (December)

Optimization of Factors Influencing Agrobacterium mediated Genetic Transformation in Eclipta alba (L.) Hassk.

Shruti Bardar, S.L. Kothari¹and Sumita Kachhwaha*

Department of Botany, University of Rajasthan, Jaipur, India-302 004, India

Key words: Eclipta alba, Nodal segments, Genetic transformation

Abastract

An efficient and reproducible protocol for in vitro plant regeneration and Agrobacterium-mediated genetic transformation was developed for Eclipta alba (L.) Hassk. using nodal explants. Several parameters affecting Agrobacterium-mediated gene delivery were investigated and optimized. These include antibiotic concentration, preculture period, infection time, use of acetosyringone, co-cultivation period and temperature and increased level of copper in induction, co-cultivation and regeneration medium. Incorporation of elevated level of CuSO₄ led to significant improvements in plant regeneration and recovery of transformed plants. The highest frequency of transformation was observed when explants were infected and co-cultivated at 23^oC. Higher transformation frequency was attained by augmenting the medium with 1.0

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