雷瑞祥 杨冰月 胡本祥 高静 魏艳妮 曹福麟 罗露 李洁 张岗 刘清 彭亮

摘要 [目的]研究添加不同濃度的水杨酸（SA）对秦艽愈伤组织生长量、总黄酮、总酚含量及抗氧化酶活性的影响。[方法]以秦艽种子为原材料诱导出秦艽愈伤组织，在秦艽愈伤组织的继代培养中加入0、10、50、100、150、200 μmol/L的SA，暗培养25 d后，测定秦艽愈伤组织的生长量、抗氧化酶活性和总黄酮、总酚含量。[结果]与对照组相比，当SA浓度为150 μmol/L时秦艽愈伤组织鲜重达到最大值，SA浓度为100 μmol/L时秦艽愈伤组织干重达到最大值，分别为2.863 5、0.201 7 g，显著高于对照组的鲜重和干重（P<0.05）。秦艽愈伤组织SOD活性在SA浓度为150 μmol/L时达到最大值，为183.95 U/g，是对照组的2.44倍;CAT活性在SA浓度为200 μmol/L时达到最大值，为22.44 U/mg，是对照组的2.56倍;当SA浓度为200 μmol/L时，POD活性虽有所上升，但仍然显著低于对照组（P<0.05），为61.16 U/mg，是对照组的0.40倍。SA浓度为100 μmol/L时，秦艽愈伤组织总黄酮含量达到最大值，为0.77 mg/g，是对照组的1.35倍;SA浓度为50 μmol/L时，总酚含量达到最大值，为0.98 mg/g，是对照组的1.31倍。[结论]一定浓度的SA处理下，有利于秦艽愈伤组织的生长和干物质的积累;适当浓度的SA对秦艽愈伤组织SOD、CAT活性有促进作用，而添加SA会对POD活性有抑制作用;当添加SA浓度为100、50 μmol/L时，分别有利于秦艽愈伤组织中总黄酮、总酚含量的积累。

关键词 秦艽;愈伤组织;水杨酸;总酚;总黄酮;抗氧化酶活性

中图分类号 R284文献标识码 A文章编号 0517-6611（2021）02-0165-05

doi：10.3969/j.issn.0517-6611.2021.02.045

开放科学（资源服务）标识码（OSID）：

Effects of Salicylic Acid on Total Phenol， Total Flavonoids Content and Antioxidant Enzyme Activities in Callus of Gentiana macrophylla

LEI Ruixiang，YANG Bingyue，HU Benxiang et al

（School of Pharmacy， Shaanxi University of Traditional Chinese Medicine，Qinling Chinese Herbal Medicine Application Development Engineering Technology Research Center， Xianyang，Shaanxi 712046）

Abstract [Objective]To study the effects of different concentrations of salicylic acid （SA） on callus growth， total flavonoids， total phenolic content and antioxidant enzyme activities of Gentiana macrophylla.[Method]The callus of Gentiana macrophylla was induced by the seeds of Gentiana macrophylla. The SA of 0， 10， 50， 100， 150， 200 μmol/L was added to the subculture of Gentiana macrophylla callus. Growth， antioxidant enzyme activity and total flavonoids， total phenolic content was determined after 25 days of dark culture. [Result]Compared with the control group， when the concentration of SA was 150 μmol/L， the fresh weight of callus reached the maximum. When the concentration of SA was 100 μmol/L， the dry weight of callus reached the maximum， which was 2.863 5 and 0.201 7 g， respectively. Significantly （P < 0.05） higher than the fresh weight and dry weight of the control group. The SOD activity of Gentiana macrophylla callus reached a maximum of 183.95 U/g at a concentration of 150 μmol/L， which was 2.44 times that of the control group. The CAT activity reached a maximum at a concentration of 200 μmol/L， which was 22.44 U/mg. The group was 2.56 times;when the concentration of SA was 200 μmol/L， the activity of POD increased， but it was still significantly lower than that of the control group （P<0.05）， which was 61.16 U/mg， which was 0.40 times of the control group. When the concentration of SA was 100 μmol/L， the total flavonoid content of callus reached the maximum， which was 0.77 mg/g， which was 1.35 times of the control group. When the concentration of SA was 50 μmol/L， the total phenolic content reached the maximum. 0.98 mg/g was 1.31 times that of the control group.[Conclusion]A certain concentration of SA treatment is beneficial to the growth and dry matter accumulation of callus in Gentiana macrophylla. The appropriate concentration of SA can promote the activity of SOD and CAT in callus of Gentiana macrophylla， and the addition of SA can inhibit the activity of POD. When the concentration of SA was 100 and 50 μmol/L， it was beneficial to the accumulation of total flavonoids and total phenols in callus of Gentiana macrophylla.

Key words Gentiana macrophylla;Callus;Salicylic acid;Total phenol;Total flavonoids;Antioxidant enzyme activity

中药材秦艽（Gentiana macrophylla Pall.）为龙胆科龙胆属植物，既是我国的传统中药材，也是国家重点保护的中药材品种之一，主产于陕西、甘肃、四川、内蒙古等地[1]。秦艽始载于《神农本草经》，列为中品，《神农本草经》曰：“（秦艽）主寒热邪气，寒湿风痹，肢节痛，下水利小便。”[2]性苦平，微寒，归胃、肝、胆经，以根入药，具有祛风除湿、活血舒筋、清热利尿的功效，其药理研究主要集中在治疗风湿痹痛、筋脉拘挛、骨蒸潮热、湿热黄疸等病症[3]。近年来，由于人们对于野生秦艽药材的过度采挖，导致秦艽的野生资源分布大大减少[4]，虽然秦艽的人工种植已有研究报道[5]，但是由于秦艽的种质资源混乱，造成了秦艽的种质退化，药材质量参差不齐，极大地影响了秦艽的产业化发展。植物组织培养技术能以少量的组织材料在短时间内进行大量增殖，且不受季节天气的影响[6]，已经被用于多种药用植物的快速增殖之中，如天山雪莲[7]、南方红豆杉[8]等。……