Callus Maintenance and Cell Line Selection of Grewia Tenax

Abstract

ABSTRACTCallus induction was established on internodal and leaf explants of Grewia tenax cultured on Murashige and Skoog (MS) solid medium supplied with indole-3-butyric acid (IBA), 2,4-dichlorophenoxyacetic acid (2, 4-D) or naphthalene acetic acid (NAA). Yellow and friable callus was obtained from internodal explant on 2.0 mg L−1 NAA supplemented medium with fresh weight (FW) 0.428 g/explant after 28 d of culture. The fresh biomass further improved up to 13 g when callus was sub-cultured on a combination medium of 2.0 mg L−1 NAA + 1.5 mg L−1 benzyladenine (BA). Growth curve was established based on total fresh weight. By the end of stationary phase, the total FW accumulated was 12.09 g in 10 weeks on NAA + BA, and 10.79 g in 14 weeks on NAA cultures. The results from the callus growth kinetics indicated that subculture must be performed by the end of 11th and 9th week for NAA and NAA+BA cultures, respectively. Three cell lines were selected depending on callus color. Yellow-brown line (YB) showed faster growth with a total of 14.06 g FW after 160 d of culture. This indicated that YB line may be able to maintain in culture with high cell viability.