目的 探讨不同肥料配施对北苍术Atractylodes chinensis(DC.) Koidz.3种倍半萜成分苍术酮、白术内酯II和β-桉叶醇含量及2种生物合成关键酶3-羟基-3-甲基戊二酸单酰辅酶A还原酶（3-hydroxy-3-methylglutarate monoacylase A reductase，HMGR）和法呢基焦磷酸合酶基因（farnesyl pyrophosphate synthase，FPPS）活性及基因表达的影响。方法 在不同的肥料配施下，采用高效液相色谱法测定北苍术3种倍半萜类成分的含量，采用试剂盒法（双抗体夹心法）测定北苍术根茎中HMGR和FPPS的活性；采用实时荧光定量PCR（qRT-PCR）法测定北苍术HMGR和FPPS的表达量；采用SPSS软件对生物合成关键酶活性和基因表达量进行相关性分析。结果 适宜的氮磷钾配施，可促进北苍术倍半萜类成分的积累，并显著提高其生物合成关键酶活性及基因表达。该试验区所代表的土壤肥力状况下有利于倍半萜类成分积累的最优施肥方案为氮肥（N）180 kg/hm²，磷肥（P₂O₅）225 kg/hm²，钾肥（K₂O）105 kg/hm²。不同肥料配施下，关键酶基因HMGR与苍术酮、β-桉叶醇呈显著正相关（P＜0.05）；关键酶基因FPPS与白术内酯II、β-桉叶醇呈极显著正相关（P＜0.01），与苍术酮显著相关（P＜0.05）。结论 优化氮磷钾配施对北苍术倍半萜类成分苍术酮、白术内酯II和β-桉叶醇积累及生物合成关键酶HMGR和FPPS活性及基因表达作用显著，北苍术根茎中倍半萜类成分苍术酮、白术内酯II和β-桉叶醇的生物合成受到多种基因及酶的调控，其中关键酶FPPS发挥了较大的调控作用。

Objective To investigate the effects of different fertilizer combinations on the contents of sesquiterpene components in Atractylodes chinensis, including atractylone, atractylenolide-II and β-eudesmol, and the activities and gene expression of the key enzymes for biosynthesis of 3-hydroxy-3-methylglutarate monoacylase A reductase (HMGR) and farnesyl pyrophosphate synthase (FPPS). Methods Under different fertilizer combinations, the contents of three sesquiterpenes in A. chinensis were determined by HPLC, and the activities of HMGR and FPPS in rhizomes of A. chinensis were determined by kit method (double antibody sandwich method). qRT-PCR was used to determine the expression levels of HMGR and FPPS in A. chinensis. SPSS software was used to analyze the correlation between the activities of key enzymes in biosynthesis and gene expression quantity. Results The suitable combination of nitrogen, phosphorus and potassium could promote the accumulation of sesquiterpenes in A. chinensis, and significantly improved the activity and gene expression of key enzymes in biosynthesis. The optimal fertilization scheme for sesquiterpene accumulation under the soil fertility condition represented by the test area was N 180 kg/hm², P₂O₅ 225 kg/hm² and K₂O 105 kg/hm². Under different fertilizer combinations, the key enzyme gene HMGR was significantly positively correlated with atractylone and β-eudesmol (P < 0.05). The key enzyme gene FPPS was significantly positively correlated with atractylenolide II and β-eudesmol (P < 0.01), and significantly correlated with atractylone (P< 0.05). Conclusion The optimization of nitrogen, phosphorus and potassium fertilizer application has an effect on the accumulation of atractylone, atractylenolide II and β-eudesmol in A. chinensis, and also has significant effects on the activities and gene expression of key biosynthesis enzymes of HMGR and FPPS. The biosynthesis of sesquiterpenes atractylone, atractylenolide II and β-eudesmol in rhizomes of Atractylodes. chinensis is regulated by a variety of genes and enzymes, and the key enzyme FPPS plays a greater regulatory role.

R286.2

国家自然科学基金资助项目(81803649);吉林省科技厅科技项目(20191102035YY);吉林省中医药管理局科技项目(2020047)