Establishment of in vitro culture to produce friable callus from leaf of Camellia sinensis (L.)

Abstract

Attempts were made to produce in vitro callus from tea (Camellia sinensis L.) "leaves. Unfolded leaves were collected and surface sterilized in various concentrations of Clorox™ (15% -75%) in combination with various exposure times (15-60 min) to obtain optimal concentration and exposure time for sterilization of field grown leaves. Results indicated that 50% and 58% aseptic cultures-were achieved in 60% and 75% solutions of Cloroxm in a soaking period of 30 min respectively. Futher, sterilized mature zygotic embryos were cultured on MS media containing 1 to 10 mg/l BAP in combination with 0.1 mg/l NAA. to obtain the suitable concentration ofBAPfor the establishment ofin vitro micro shoots. The result showed that 5 mg/l concentration of BAP would be suitable for the initiation of in-vitro micro shoot cultures. At 12'h week, plantlets regenerated in BAP at 5 mg/l were subcultured in the presence of 3 mg/l BAP and 0.1 mg/l NAA. Multiplication rate of first two subcultures was 3.6 ±0.2. Further leafsegments at 2nd, 3rd and 4'!\ subculture periods were cultured on callus medium to determine the competence of friable callus initiation on leaves of newly establishing in vitro micro shoots. Results revealed that'initiation of friable callus was fairly better on leaves obtained at 4th subculture among the tested, treatments. Moreover, in vitro and field grown leaves were compared on the efficiency of callus initiation. A significant high frequency of callus induction (79.2%) was achieved from in vitro leaf explants, which were collected at 5lh subculture.