H2O2对远志愈伤组织中总酚总黄酮含量及相关酶活性的影响

Effect of Hydrogen Peroxide（ H2O2） on Content of Total Phenol Flavonoids and Activity of Related Enzymes in Polygala tenuifolia Callus

目的:研究远志愈伤组织在加有不同浓度H2O2的MS培养基上总酚总黄酮含量变化及抗氧化酶活性变化,从细胞水平探讨其适应H2O2环境胁迫的生理机制。方法:以远志愈伤组织为实验材料,H2O2浓度设置为0,5,10,15,20 mmol·L-15个梯度加至MS培养基中,分别在培养5,10,15,20,25 d后取样测定愈伤组织总酚、总黄酮含量及抗氧化酶活性。结果:H2O2浓度在5 mmol·L-1下远志愈伤组织15 d时总酚、总黄酮含量最高。当远志愈伤组织培养5 d,外源H2O2浓度为5 mmol·L-1时的超氧化物歧化酶(SOD)活性最高。培养25 d,外源H2O2浓度为5 mmol·L-1时的过氧化物酶(POD)活性最高。培养25 d,外源H2O2浓度为15 mmol·L-1时的过氧化氢酶(CAT)活性最高。结论:远志愈伤组织具有适应一定浓度H2O2环境胁迫的能力,在其受到H2O2环境胁迫时,远志愈伤组织能通过调节自身的次生代谢物、保护酶系统来缓解带来的伤害。对次生代谢物总酚、总黄酮的含量在5~10 mmol·L-1时有显著促进作用;在5 d,外源H2O2浓度为5 mmol·L-1时可显著提高SOD活性;在25 d,外源H2O2浓度为5 mmol·L-1时可显著提高POD活性;在25 d,外源H2O2浓度为15 mmol·L-1时可显著提高CAT活性。

Objective:To study the changes in total phenolic and flavonoid content and antioxidant enzyme activity of Polygala tenuifolia callus in MS medium with different concentrations of H2O2,in order to explore the physiological mechanism of Polygala tenuifolia callus in adapting to H2O2 environmental stress at the cellular level.Method:Five gradients of 0,5,10,15,20 mmol·L-1 were set for H2O2 concentration and added to MS medium,with P.tenuifolia callus as the experimental material.Total phenols,total flavonoids and antioxidant enzyme activities of callus were determined after 5,10,15,20,25 d of culture,respectively.Result:The contents of total phenols and flavonoids were the highest when the concentration of H2O2 was5 mmol·L-1 for 15 d.The SOD activity was the highest when the callus was cultured for 5 d and the exogenous H2O2 concentration was 5 mmol·L-1.POD activity was the highest at 25 d and 5 mmol·L-1 concentration of exogenous H2O2.CAT activity was the highest at 25 d and 15 mmol·L-1 concentration of exogenous H2O2.Conclusion:P.tenuifolia callus has the ability to adapt to the environmental stress of H2O2 at a certain concentration.When it is subjected to the environmental stress of H2O2,P.tenuifolia callus can alleviate the damage by regulating its secondary metabolites and protecting enzyme system.It can significantly promote the content of total phenols and flavonoids in secondary metabolites at 5-10 mmol·L-1.SOD activity was significantly increased at 5 d and the concentration of exogenous H2O2 of 5 mmol·L-1.POD activity was significantly increased at 25 d and the concentration of exogenous H2O2 of 5 mmol·L-1.CAT activity was significantly increased at 25 d and concentration of exogenous H2O2 of 15 mmol·L-1.