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Microbial contamination and browning were the main reason causing the low survival rate of longan tissue culture in vitro.Using the internal tissue of middle stem sections and alternately immersed with multiple disinfectants as pretreatment can decrease contamination and browning.The stem germination numbers varied with seasons, and the stems in summer or autumn was suitable as explants.The rate of responsive explants could reach 73% when they were culture on MS media, and the adventitious stem grew very well.