摘要
目的研究生态因子和人参皂苷合成关键酶表达对人参皂苷合成、积累的影响。方法以人工栽培的4年生人参为研究对象,用实时荧光定量PCR法对在不同生长时期根组织中8个关键酶基因(HMGR、FPS、SS、SE、DS、β-AS、CYP82D47、CYP716A47)表达量进行测定;采用HPLC法测定根中8种单体皂苷(人参皂苷Rg_1、Re、Rf、Rb_1、Rb_2、Rb_3、Rc、Rd)的量;以小型气象站对人参样地气象数据进行采集;采用SPSS软件进行相关性分析。结果人参皂苷合成关键酶基因在人参皂苷积累的重要时期(开花期至果熟期)的表达高于果后参根生长期和枯萎期,人参皂苷合成关键酶基因的表达相互影响,协同增减,人参根组织中关键酶基因的表达对人参皂苷积累多表现出了正相关关系;人参根中人参皂苷Rg_1、Rb_1、Re、Rc量较高,8种单体皂苷量动态变化趋势并不相同;温度、光合有效辐射、土壤水势是影响根部人参皂苷合成的重要生态因子,温度与人参皂苷Rb_1、Rd显著负相关(P<0.05),光合有效辐射对人参皂苷Rg_1的生成有显著的促进作用(P<0.05),土壤水势与人参皂苷Rb_1显著负相关(P<0.05);灰色关联度分析结果表明温度、光合有效辐射、相对湿度是影响人参皂苷量的主导生态因子,人参皂苷合成关键酶基因的表达是次要因子,在生态因子主导下人参皂苷合成关键酶基因调控人参皂苷的合成与积累。生态因子与关键酶基因的表达共同调控了人参皂苷的合成,对人参皂苷的积累有重要影响。结论明确了人参皂苷合成关键酶基因表达和人参皂苷量在人参中的动态变化,为人参皂苷合成生理生态机制的明晰及对人参药材质量的调控提供了理论依据。
Objective To explore the effect of ecological factors and the expression of key enzyme genes on the synthesis and accumulation of ginsenosides. Methods Cultivated four-year-old ginseng were used as test materials, the expression of key enzyme genes（HMGR, FPS, SS, SE, DS, β-AS, CYP82 D47, CYP716 A47） in the biosynthesis of ginsenosides of roots in different growth periods was determined by real-time quantitative PCR, determination of the content of eight ginsenosides（ginsenoside Rg1, Re, Rf, Rb1, Rb2, Rb3, Rc, Rd） in roots by HPLC, the meteorological data were collected by a small weather station, correlation analysis was performed with SPSS. Results The expression of key enzyme genes in the period of flowering to fruit ripening was higher than the root growing after fruit period and the withering period, the expression of key enzyme genes involved in the synthesis of ginsenosides was influenced by each other, and the expression of key enzyme genes in ginseng roots showed a positive correlation with the accumulation of ginsenosides; The contents of ginsenoside Rg1, Rb1, Re, and Rc were higher in the roots of ginseng, eight kinds of monomer ginsenosides content dynamic changes trend is different; Temperature, photosynthetically active radiation, soil water potential are important ecological factors for ginsenosides synthesis in roots, temperature was significantly negatively correlated with ginsenoside Rb1 and Rd（P〈0.05）, PAR can significantly promote the formation of ginsenoside Rg1（P〈0.05）, soil water potential was significantly negatively correlated with ginsenoside Rb1（P〈0.05）; Grey correlation analysis results showed that the major ecological factors that influenced ginsenosides content in ginseng roots were temperature, PAR and relative humidity, the grey correlation between the expression of the key enzyme genes with the content of ginsenosides is less than ecological factors with the content of ginsenosides, under the guidance of ecological factors, the expression of the key enzyme genes regulate the synthesis and accumulation of ginsenosides. Conclusion The dynamic changes of the expression of key enzyme genes and the content of ginsenosides in ginseng were determined, it provides a theoretical basis for elucidating the physiological and ecological mechanism of ginsenoside synthesis and the quality control of Radix Ginseng.
作者
杨林林
张涛
杨利民
韩梅
YANG Lin-lin ZHANG Tao YANG Li-min HAN Mei(College of Chinese Materials, Jilin Agricultural University, Changchun 130118, China)
出处
《中草药》
CAS
CSCD
北大核心
2017年第20期4296-4305,共10页
Chinese Traditional and Herbal Drugs
基金
国家自然科学基金资助项目(31270371)
国家中药材产业技术体系(CARS-21)
吉林省重大科技成果转化项目(20170307009YY)
