植物生长调节剂对南方红豆杉愈伤组织培养和紫杉醇合成的影响

通过不同种类和水平植物生长调节剂对南方红豆杉(Taxus chinensisvar.mairei)愈伤组织诱导、生长和紫杉醇合成能力影响的研究发现:诱导培养初期,以无植物生长调节剂的MS为基本培养基,在附加不同植物生长调节剂组合作用下愈伤组织产生的时间和生长、在相同植物生长调节剂组合作用下不同外植体愈伤组织的产生时间和生长均表现出较显著差异,2,4-D/NAA高于0.4时,不利于南方红豆杉愈伤组织的诱导。转换到附加不同植物生长调节剂组合的B5培养基上后,随培养继代次数的增加,生长差异逐渐缩小,直至不显著,表明参考不同文献报道最优配方所设计的各植物生长调节剂组合对南方红豆杉愈伤组织的生长均较适宜,有利南方红豆杉愈伤组织生长的植物生长调节剂优化组合没有唯一性。但不同调节剂组合作用下的同源愈伤组织中、相同调节剂作用下不同源愈伤组织中紫杉醇含量均存在着极显著差异,适当水平(2 mg/L)的2,4-D单用,或与适当水平的KT、6-BA、KT GA配合使用,对南方红豆杉愈伤组织紫杉醇的合成较有利,NAA则不太有利,幼茎和叶愈伤组织产紫杉醇的水平较其它愈伤组织为高。

Influence of Plant Growth Regulator on Taxus chinensis var. Mairei Callus Culture and Biosynthesis of Taxol

To explore the influence of different kinds and concentrations of plant growth regulators on induction,growth and biosynthesis ability of taxol of Taxus chinensis var.mairei callus,various explants of T.mairei,including young stem,young stems bark,cambium,immature needle,apical meristem were respectively inoculated on 8 kinds of callus-induction medium containing MS basal salts and vitamins,2% sucrose and different kinds of plant growth regulators with different concentrations,and were induced to produce callus and cultured for 3 ages(about 29 day.age~(-1)),then the calluses were respectively transfered on 8 kinds of callusmultiplication medium containing Gamborg B5 basal salts and vitamins,3% sucrose and different kinds of plant growth regulators with different concentrations.Taxol content of various callus cultures were analysed by HPLC when subcultured for two ages(60 day).The result shows:for initial stages of inducing culture,the differences between the time of producting callus and the growth value of callus were significant when the same callus was cultured in different medium containing various regulator combination,or when different callus were cultured on the same medium containing certain regulator combination.The induction rate of T.mairei callus would be reduced when the concentration of 2,4-D to the concentration of NAA is higher than 0.4.However,for following frequent subculturing,when these calluses were transferred on 8 kinds B5(callusmuliti-)plication medium,the above differences would be gradually deminished till insignificant.These results suggested that all the regulator combinations in callus-multiplication medium determined based on the optimization directions in reference,were beneficial to the growth of T.mairei various callus,i.e..There are many optimized combinations of plant growth regulator suitable for the growth of T.mairei callus.Nevertheless,there were significant difference between taxol content in the calluses,when the same callus was cultured on different medium containing various plant growth regulator combination,or when different callus were cultured on the same medium containing the certain regulator combination.Proper concentration of(2 mg/L)2,4-D used independently,or properly supplemented with KT,6-BA or KT GA,benefit the biosynthesis of taxol in T.mairei callus,but NAA is not.The biosynthesis of taxol in young stem and leaf callus are higher than in other explant calluses.