诱导子对雷公藤悬浮细胞中雷公藤甲素积累的影响  

Effects of Elicitors on the Accumulation of Triptolide in Cell Suspension Culture of Tripterygium wilfordii

作　　者：张强 宋萍[1] 朱留刚[1] 封磊[2] 洪伟[1,3] 吴承祯 ZHANG Qiang;SONG Ping;ZHU Liugang;FENG Lei;HONG Wei;WU Chengzhen(College of Forestry,Fujian Agriculture and Forestry University,Fuzhou 350002 China;College of Resources and Environment,Fujian Agriculture and Forestry University,Fuzhou 350002,China;Fujian Provincial Key Laboratory of Forest Ecosystem Processing and Management,Fuzhou 350002,China;Wuyi University,Wuyishan,Fujian 354300,China)

机构地区：[1]福建农林大学林学院,福建福州350002 [2]福建农林大学资源与环境学院,福建福州350002 [3]福建省高校森林生态系统过程与经营重点实验室,福建福州350002 [4]武夷学院,福建武夷山354300

出　　处：《江西农业大学学报》2021年第5期1149-1158,共10页Acta Agriculturae Universitatis Jiangxiensis

基　　金：国家自然科学基金项目(31470577);福建省高校产学合作科技重大项目(2019N5006);福建农林大学科技创新专项基金项目(CXZX2016061,CXZX2018105,CXZX2019078G)。

摘　　要：【目的】雷公藤甲素是雷公藤主要药用活性成分之一,具有抗肿瘤、免疫抑制和抗炎等多种作用。应用植物细胞培养技术生产雷公藤甲素具有重要意义,而诱导子是提高植物细胞培养中次生代谢产物积累的有效途径之一。研究通过探究非生物和生物诱导子对雷公藤悬浮细胞中雷公藤甲素积累的影响,以期提高雷公藤甲素产量,为雷公藤悬浮细胞规模化生产雷公藤甲素等药用活性物质提供基础。【方法】以水杨酸(SA)、LaCl_(3)以及雷公藤内生真菌NS-5、NS-17作为诱导子,研究不同类型及不同浓度诱导子添加对雷公藤悬浮细胞中雷公藤甲素积累及细胞活力、细胞生长、总糖消耗、PAL活性的影响。【结果】SA、LaCl_(3)和内生真菌NS-5、NS-17诱导子均可促进雷公藤悬浮细胞培养物中雷公藤甲素的积累。非生物诱导子的效应大小受其浓度的影响,0.1 mg/L的SA诱导作用最好,可提高细胞活力,促进细胞生长,总糖消耗增加,升高PAL活性,雷公藤甲素产量最高是对照的3.5倍。随着浓度的降低,LaCl_(3)对雷公藤甲素的诱导作用增强,20 mg/L LaCl_(3)诱导下的最高含量是对照的2.0倍,同时20 mg/L LaCl_(3)也提高了PAL活性和总糖消耗,但对细胞活力和细胞生长影响不大。内生真菌诱导子的效应大小即与诱导子种类和添加时间有关。细胞培养第4天添加NS-5、NS-17菌丝体提取物,第8天添加任一内生真菌诱导子均提高了雷公藤甲素的积累,其中NS-5菌丝体提取物的诱导效果相对最强,细胞培养第8天添加时雷公藤甲素产量最高,是对照的2.4倍,但雷公藤甲素的积累与细胞生长及PAL活性无明显的关联。【结论】诱导子的添加能够提高雷公藤悬浮细胞中雷公藤甲素的积累,诱导效应的大小与诱导子种类、添加浓度及添加时间有关。本研究中的0.1 mg/L SA、20 mg/L LaCl_(3)及内生真菌NS-5菌丝体提取物显示了良好的诱导[Objective]Triptolide is one of the main medicinal active ingredients of Tripterygium wilfordii,which has many effects such as resistance against tumor,immunosuppression and resistance against inflammation.It is of great significance to produce triptolide by plant cell culture technology,and the use of elicitor is one of the effective ways to improve the accumulation of secondary metabolites in plant cell culture.This study was to explore the effects of abiotic and biotic elicitors on the accumulation of triptolide in T.wilfordii suspension cells,so as to increase the production of triptolide and provide a basis for the large-scale production of triptolide and other medicinal active substances in T.wilfordii cell suspension culture.[Method]Salicylic acid(SA),LaCl_(3),and endophytic fungi NS-5 and NS-17 from T.wilfordii were used as elicitors to study the effects of different types and different concentrations of elicitors on the accumulation of triptolide,cell viability,cell growth,total sugar consumption and PAL activity in cell suspension culture of T.wilfordii.[Result]SA,LaCl_(3) and the elicitors of endophytic fungi NS-5 and NS-17 could promote the accumulation of triptolide in suspension cells of T.wilfordii.The effect of abiotic elicitor was affected by its concentration.SA of 0.1 mg/L had the greatest induction effect,which improved cell viability,promoted cell growth,increased total sugar consumption and PAL activity.The highest triptolide yield inducted by 0.1 mg/L SA was 3.5 times that of the control.With the decrease of the concentration,the inducing effect of LaCl_(3) on the triptolide increased.The highest content of triptolide induced by 20 mg/L LaCl_(3) was 2.0 times that of the control.At the same time,20 mg/L LaCl_(3) also increased PAL activity and total sugar consumption,but had little effect on cell viability and cell growth.The effect of endophytic fungal elicitor was related to the type of elicitor and aslo affected by the time of addition.The addition of NS-5 and NS-17 mycelium extracts on

关 键 词：雷公藤甲素 非生物诱导子 内生真菌诱导子 悬浮细胞 雷公藤 

分 类 号：Q813.1[生物学—生物工程]