外源诱导子和温度对黑果枸杞悬浮细胞花青素积累的影响

Effects of exogenous elicitors and temperature on anthocyanin content in suspension cells of Lycium ruthenicum

黑果枸杞富含花青素,是一种具有很高开发利用价值的药食同源植物。以黑果枸杞无菌苗的叶片为外植体诱导愈伤组织建立细胞悬浮体系,研究不同外源激素和温度对黑果枸杞悬浮细胞中花青素含量的影响。黑果枸杞愈伤组织诱导的最佳培养体系为WPM+0.5 mg/L激动素+2.0 mg/L 2,4-二氯苯氧乙酸+20.0 g/L蔗糖+6.0 g/L琼脂,悬浮细胞培养的最优体系为WPM+0.5 mg/L激动素+2.0 mg/L 2,4-二氯苯氧乙酸+0.01 mg/L萘乙酸+20.0 g/L蔗糖,其生长曲线呈“S”形,活细胞数量在悬浮培养第18天时达到峰值,为7.93×10^(5)个/mL。30 g/L蔗糖、200 mg/L茉莉酸甲酯、2 mg/L脱落酸及16℃单一处理均能显著促进悬浮细胞花青素产量的提升(P<0.05),2 mg/L脱落酸诱导效果最佳。在16℃和3种外源激素的复合处理下,200 mg/L茉莉酸甲酯+16℃的复合诱导处理能显著促进细胞花青素积累(P<0.05),含量是16℃单因素处理的1.13倍。研究结果为进一步优化黑果枸杞悬浮细胞培养体系、促进花青素积累奠定了基础。

Lycium ruthenicum,rich in anthocyanins,is a medicinal and edible plant with high development and utilization value.In this study,aseptic seedling leaves of Lycium ruthenicum were used as explants to induce callus formation and establish a cell suspension system.The impacts of different exogenous hormones and temperatures on the anthocyanin content in suspension cells were investigated.The optimal culture system for callus induction was WPM+0.5 mg/L KT+2.0 mg/L 2,4-D+20.0 g/L sucrose+6.0 g/L agar,and the optimal system for suspension cell culture was WPM+0.5 mg/L KT+2.0 mg/L 2,4-D+0.01 mg/L NAA+20.0 g/L sucrose.The growth curve exhibited an‘S’-shaped pattern,with the number of viable cells peaking at 7.93×10^(5)cells/mL after 18 days of suspension culture.The treatments with 30 g/L sucrose,200 mg/L methyl jasmonate,2 mg/L abscisic acid and 16℃all can significantly enhance the production of anthocyanin in suspension cells(P<0.05).Among these treatments,the induction effect of 2 mg/L abscisic acid was the best.For the combined treatments of 16℃and 3 exogenous hormones,the combined induction treatment of 200 mg/L methyl jasmonate+16℃could significantly enhance the anthocyanin accumulation in suspension cells(P<0.05),resulting in 1.13 times higher anthocyanin content than that of the 16℃treatment alone.The results laid a foundation for further optimizing the suspension cell culture system and promoting anthocyanin accumulation in Lycium ruthenicum.