目的 分析不同石杉碱甲（Hup A）含量蛇足石杉叶状体（离体培养物，分别为株系A、B、C）的蛋白差异，探究可能与Hup A积累合成的相关酶。方法 利用定量蛋白质组学串联质谱标签（tandem mass tag，TMT）技术对不同株系蛇足石杉叶状体进行定量蛋白质组学检测和鉴定，再对差异蛋白进行GO和KEGG等生物信息学分析。结果 株系B的Hup A含量最低，株系C的Hup A含量最高，是B的2倍，株系B和C两者有78个差异蛋白，且差异蛋白GO富集分析，分子功能（MF）所占比例为28.75%；通过不同株系间蛋白质差异表达的比对，分析得到这3个株系在以氨基酸为起始的生物碱代谢通路中共有2种差异蛋白（P93541、Q8RXU4）。P93541蛋白在Hup A低含量的株系B中表达下调，在相对高含量的株系A和C中均上调；而Q8RXU4蛋白在低含量的株系B中表达上调，在相对高含量的株系A和C中均表达下调。结论 Hup A含量差异与蛋白表达差异呈正相关，分析得到2种可能与Hup A积累相关的酶P93541、Q8RXU4，为Hup A生物合成生物信息学分析提供了依据。

Objective To analyze the differences in protein of thallus of Huperzia serrata (in vitro cultures, strains A, B, and C, respectively) with different Hup A content, and to explore related enzymes that may be synthesized with Hup A accumulation. Methods Quantitative proteomics was performed in vitro cultures of Huperzia serrata with different Hup A content using quantitative proteomics tandem mass tag (TMT) techniques, followed by differential protein analysis:GO, KEGG and other biological information analysis. Results Strain B hadthe lowest Hup A content and strain C had the highest Hup A content, which was twice than that of B. There were 78 differential proteins between the strain B and C. Analysis of differential protein GO enrichment showed that MF accounted for 28.75%; Analysis of differential protein expression showed that three strains shared two differential proteins (P93541, Q8RXU4) in the alkaloid metabolic pathway starting from amino acids. P93541 protein was down-regulated in the low-yield strain B and up-regulated in the relatively high-yield strains A and C. The Q8RXU4 protein was up-regulated in the low-yield strain B and down-regulated in the relatively high-yield strains A and C. Conclusion This study found that the difference in Hup A content was positively correlated with the protein expression. Two enzymes P93541 and Q8RXU4 that may be related to Hup A accumulation were analyzed, providing a basis for bioinformatics analysis of Hup A biosynthesis.

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国家自然科学基金资助项目（81660597）；国家自然科学基金资助项目（81360614）；江西省自然科学基金项目（20132BAB204023）