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Abstract：
Dendrobium officinale Kimura et Migo is a tonic plant that has both ornamental and medicinal properties. Terpenoids are significant and diverse secondary metabolites in plants, and are one of the important natural active ingredients in D. officinale. The AP2/ERF gene family plays a major role in primary and secondary metabolism. However, the AP2/ERF transcription factor family has not been identified in D. officinale, and it is unclear if it is involved in the regulation of terpenoid biosynthesis. This study identified a sesquiterpene synthetase-β-patchoulene synthase (DoPAES) using transcriptome and terpenic metabolic profile analyses. A total of 111 members of the AP2/ERF family were identified through the whole genome of D. officinale. The tissue-specific expression and gene co-expression pattern of the DoAP2/ERF family members were analyzed. The results showed that the expression of DoPAES was highly correlated with the expression of DoAP2/ERF89 and DoAP2/ERF47. The yeast one-hybrid (Y1H) assays and dual-luciferase experiments demonstrated that DoAP2/ERF89 and DoAP2/ERF47 could regulate the expression of DoPAES. The transcriptional regulatory effects were examined using homologous transient expression of DoAP2/ERF89 in protocorms of D. officinale. DoAP2/ERF89 positively regulated the biosynthesis of β-patchoulene. This study showed that DoAP2/ERF89 can bind to the promoter region of DoPAES to control its expression and further regulate the biosynthesis of β-patchoulene in D. officinale. These results provide new insights on the regulation of terpenoid biosynthesis.
摘要：
铁皮石斛KimuraetMigo是一种具有观赏和药用特性的补品植物。萜类化合物是植物中重要且多样的次生代谢产物，并且是D.officinale中重要的天然活性成分之一。AP2/ERF基因家族在初级和次级代谢中起主要感化。然而,AP2/ERF转录因子家族尚未在D.officinale中鉴定，目前尚不清楚它是否参与萜类生物合成的调节。这项研究使用转录组和萜类代谢谱分析鉴定了倍半萜合成酶-β-广程序烯合酶（DoPAES）。通过D.officinale的全基因组鉴定了AP2/ERF家族的总共111个成员。分析了DoAP2/ERF家族成员的组织特异性表达和基因共表达模式。结果表明，DoPAES的表达与DoAP2/ERF89和DoAP2/ERF47的表达高度相关。酵母单杂交（Y1H）分析和双荧光素酶实验表明，DoAP2/ERF89和DoAP2/ERF47可以调节DoPAES的表达。使用DoAP2/ERF89在D.officinale原球茎中的同源瞬时表达来检查转录调节作用。DoAP2/ERF89正调控β-广范围香烯的生物合成。这项研究表明，DoAP2/ERF89可以与DoPAES的启动子区结合以控制其表达，并进一步调节β-广贴烯的生物合成。这些结果为萜类化合物生物合成的调控提供了新的见解。
