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为了探明刺五加糖基转移酶基因(glycosyltransferase,GT)的表达特点及其对刺五加总皂苷含量的影响。以GAPDH基因为内参照基因,采用实时定量PCR技术分析了不同产地、不同生长发育时期、不同器官刺五加中GT基因的表达规律及茉莉酸甲酯(MeJA)对其表达的影响。并采用浓硫酸-香草醛比色法测定刺五加总皂苷的含量,利用SPPS17.0软件进行相关性分析。结果表明,GT基因在根、幼茎、叶片和叶柄中均有表达,但表达量差异不显著。刺五加的GT基因在叶片完全展开期的表达量最高,盛花期和果实基本成熟期的表达量最低,仅为叶片完全展开期表达量的66.9%。MeJA处理可显著提高GT基因的表达量(p<0.05)。不同产地刺五加的GT基因表达量和总皂苷含量差异显著,但GT基因的表达量与总皂苷含量同升同降,存在极显著的正相关关系(p<0.01)。初步判定,GT基因的表达量决定了刺五加中总皂苷的含量,GT是决定刺五加中总皂苷含量的关键酶。
In order to investigate the expression of glycosyltransferase (GT) and its effect on the content of total saponins in Acanthopanax senticosus. Using GAPDH gene as an internal reference gene, real-time quantitative PCR was used to analyze the expression of GT gene and the effect of methyl jasmonate (MeJA) on the expression of GT gene in different organs, growth and development stages. The content of total saponins in Acanthopanax senticosus was determined by concentrated sulfuric acid-vanillin colorimetric method. The correlation analysis was carried out by SPPS17.0 software. The results showed that the GT gene was expressed in roots, young stems, leaves and petioles, but the expression level was not significantly different. The expression of GT gene of Acanthopanax senticosus was the highest during the full developmental period of leaf and lowest at full flowering and fruit ripening stage, which was only 66.9% of that of full development of leaf. MeJA treatment significantly increased GT gene expression (p <0.05). GT gene expression and total saponin content of Acanthopanax senticosus from different areas were significantly different, but there was a significant positive correlation between GT gene expression and total saponin content (p <0.01). Preliminary determination, GT gene expression determines the acanthopanax total saponin content, GT is to determine the acanthopanax total saponin content of key enzymes.