Mass Shoot Proliferation of Pistacia khinjuk Stocks Using Temporary Immersion Bioreactor System (TIS)

Abstract

A temporary immersion bioreactor system (TIS), compared to semi-solid systems has many advantages such as (1) the system can be controlled by an automation system; (2) a greater number of explants could be used at the same time and (3) there is no need to use an organic gelling agent. In this study, we aimed to develop a method suitable for mass shoot proliferation of P. khinjuk Stocks. Shoot tips and nodal segments of in vitro grown axenic regenerates derived from seedlings of Pistacia khinjuk Stocks were used as material. In both semi-solid and liquid medium experiments, an MS medium supplemented with 1 mg/L BA and 30 g/L sucrose was used. Various immersion frequencies (2, 4, 8, 16 or 20 h) and immersion times (5, 10 or 15 min) were tested on the TIS system. Repeated trials for immersion frequencies at short time intervals (10 min/2 h, 10 min/4 h etc.) showed hyper hydrated explants up to 100%, while this problem was not observed in immersions with relatively longer time intervals (10 min/20 h). The highest shoot forming capacity index (3.12) and the minimum hyper hydration rate (8%) were obtained in shoot tips with 10 min immersion frequency every 16 h. The protocol presented here, presents the initial stage of mass propagation of P. khinjuk Stocks cultures.