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目的:应用实时定量PCR技术对刺五加鲨烯环氧酶(squalene epoxidase,SE)基因进行表达分析,探讨其与刺五加皂苷含量的关系。方法:以3-磷酸甘油醛脱氢酶(glyceraldehyde-3-phosphate dehydrogenase,GAPDH)基因为内参照基因,通过实时定量PCR检测SE基因的相对表达量,分光光度法测定刺五加皂苷含量。结果:不同产地、器官中的SE表达量存在显著差异(P<0.05),其中,雾灵山产地的表达量最大,达最小值(伊通产地)的4.81倍,各器官中,叶的表达量最大,为根的1.92倍。从叶片萌芽期到衰老期,SE的表达呈先升后降趋势,各发育期之间差异显著(P<0.05),其中盛花期的表达量最高,达萌芽期的3.71倍;茉莉酸甲酯(methyl jasmonate,MeJA)可提高SE的表达量,但未达显著水平。刺五加叶的皂苷含量与SE的表达量之间存在显著的正相关关系(P<0.05)。结论:不同产地、器官及生长发育时期刺五加SE的表达量间差异显著,且SE的表达与刺五加的皂苷含量显著相关。
Objective: To investigate the expression of squalene epoxidase (SE) gene in Acanthopanax senticosus by real-time quantitative PCR. Methods: GAPDH gene was used as an internal reference gene. The relative expression of SE gene was detected by real-time quantitative PCR. The content of Acanthopanax senticosus was determined by spectrophotometry. Results: There were significant differences in the expression of SE in different habitats and organs (P <0.05). The highest expression level of Wuling Mountain was reached 4.81 times of the lowest (Yitong) The largest, 1.92 times the root. From leaf sprouting to senescence, the expression of SE increased first and then decreased, with significant difference between different developmental stages (P <0.05). The expression of SE was the highest at full flowering stage, reaching 3.71 folds of the budding stage. Methyl jasmonate (methylJasmonate, MeJA) can improve the expression of SE, but did not reach the significant level. There was a significant positive correlation between the content of saponin and the expression of SE in Acanthopanax senticosus (P <0.05). Conclusion: The expression of Acanthopanax senticosus SE in different producing areas, organs and growth and development stages is significantly different, and the expression of SE is significantly correlated with the saponin content of Acanthopanax senticosus.