Evaluation of shake flask and bioreactor systems on cell growth and regeneration of Musa Acuminata Cv. berangan cell suspension culture / Wendy Chin Yi Wen

Wendy, Chin Yi Wen (2014) Evaluation of shake flask and bioreactor systems on cell growth and regeneration of Musa Acuminata Cv. berangan cell suspension culture / Wendy Chin Yi Wen. Masters thesis, University of Malaya.

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Abstract

In this study, a semi automated cell suspension culture protocol for Musa acuminata cv. ‘Berangan’ using bioreactor has been established. Flower clusters at the positions of 8 to 10 were the most responsive (70 %) towards embryogenesis. After eight months of culture, embryogenic calli generated from male inflorescences were transferred to liquid suspension medium. Embryogenic cell suspension culture was successfully established after two months of culture in liquid M2 medium. Cell suspension cultures showed optimum cell growth when cultured in the M2 modified liquid medium containing 2 % sucrose with an initial inoculum density of 2 % settled cell volume (SCV). The yield of cell suspension cultures was increased to 165 % and 210 %, respectively, when inoculated in 5 l balloon type bubble column bioreactors (BTBCBs) without pH control and with pH maintained at 5.7, over 14 days of culture. The results showed that the growth yield was 3-, 2.5- and 2.2- fold higher than initial culture (day 1) when cultured in pH-controlled BTBCB, non-pH-controlled BTBCB and shake flask respectively.In all growth vessels tested, catalase (CAT) activity was found to be correlated with hydrogen peroxide (H2O2) concentration, suggesting its effective scavenging activity, whereas no significant difference was observed for superoxide dismutase (SOD). More than 60 % of the embryos started to form shoots after 2 weeks of culture on M4 medium for all growth vessels tested. Abnormalities in plantlets were less than 20 % in all three types of culture vessels. Potentially, this protocol could be used to mass produce disease-free and high yielding banana as it provides a comparable cell growth rate and number of regenerants which phenotypically were identical to the donor plants compared to conventional shake flask cultures.

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