扁穗牛鞭草组织培养中褐化控制技术初探

Evaluation of three antioxidants to reduce browning in Hemarthria compressa callus

以雅安’扁穗牛鞭草(Hemarthria compressa cv.Ya’an)幼茎诱导的愈伤组织为试验材料,探讨了3种抗褐化剂[活性炭(AC)、聚乙烯吡咯烷酮(PVP)、抗坏血酸(Vc)],以及不同继代培养周期和培养温度对扁穗牛鞭草愈伤组织褐化控制及愈伤组织生长状况的影响。结果表明,向培养基中添加AC和低浓度的PVP能有效降低褐化率并促进愈伤组织的生长,其中以2.0g.L-1 AC的效果最好,1.0g.L-1 AC和1.0g.L-1 PVP的效果次之,Vc抗褐化性能较差;以10d为一个继代培养周期,愈伤组织的褐化率最低;培养室温度控制在(23±2)℃左右,既可有效降低愈伤组织的褐化率,又不影响愈伤组织的生长。

Browning and subsequent cell death in Hemarthria compressa callus is a serious problem reducing the effectiveness of this culturing system.This study evaluated three antioxidants（activated carbon,polyvinyl pyrrolidone,and vitamin C） for their potential to reduce browning.These agents were added into subculture media and incubated under different subculture cycles（10,15,20,30 d） and temperatures（21,23,25,27 ℃）.Results showed that adding activated carbon（2.0 g·L-1）,and incubating at（23±2）℃ for 10 d reduced the browning rate significantly and did not decrease callus growth.This culturing technique will allow more routine use of H.compressa callus in tissue culture and gene transformation experiments.