雷帕霉素靶蛋白基因调控逆境胁迫下华山松大小蠹能量代谢的功能分析

doi:10.11707/j.1001-7488.LYKX20260030

摘要/Abstract

摘要：

目的: 分析华山松大小蠹雷帕霉素靶蛋白基因（DaTOR）的功能，揭示DaTOR在饥饿、高温和低温胁迫下调控能量代谢的作用，为华山松大小蠹的代谢靶向防控提供新思路。方法: 通过基因克隆技术获得DaTOR基因全长序列，利用实时荧光定量PCR技术检测DaTOR时空表达模式及其在饥饿和温度胁迫下的表达变化，应用RNA干扰技术探究其在逆境胁迫下调控脂和糖代谢的功能以及在抵御饥饿、高温和低温中的作用。结果: 1）华山松大小蠹DaTOR氨基酸序列符合TOR特征，与山松大小蠹DpTOR基因相似度达96.57%。2）华山松大小蠹DaTOR在蛹和羽化后迁飞雌虫中表达量最高，在幼虫中表达量最低，不同组织中脂肪体表达量最高。3）饥饿和低温胁迫下随着胁迫时间延长DaTOR表达量下调；高温胁迫下DaTOR表达量先短暂上升，在12 h表达量达到最高，随后下降。4）对华山松大小蠹幼虫和雌雄成虫进行RNA干扰后，DaTOR表达量显著降低；饥饿和高温胁迫下幼虫死亡率均高于雌雄成虫，分别达73.33%和71.67%；低温胁迫下雄性成虫死亡率最高，为76.67%。5）华山松大小蠹雌性成虫、雄性成虫和幼虫注射dsTOR后，饥饿、高温和低温胁迫下甘油三酯、糖原和海藻糖含量均呈下降趋势，游离脂肪酸含量均呈上升趋势。结论: DaTOR在华山松大小蠹不同发育阶段和不同组织中存在差异表达，饥饿、高温和低温胁迫下呈现不同的响应模式，RNA干扰后华山松大小蠹能量代谢发生变化。

关键词: 华山松大小蠹, 雷帕霉素靶蛋白, 饥饿与温度胁迫, RNA干扰, 能量代谢

Abstract:

Objective: This study aims to analyze the function of the rapamycin target protein gene (DaTOR) in Dendroctonus armandi, and elucidate its regulatory role in energy metabolism under starvation, heat stress, and cold stress, so as to provide new insights into metabolism-targeted control strategies against D. armandi. Method: The full-length sequence of DaTOR gene was obtained through gene cloning. Quantitative real-time PCR was used to determine its spatiotemporal expression patterns and assess expression changes under starvation and temperature stresses. RNA interference was applied to investigate the role of DaTOR in regulating lipid and carbohydrate metabolism under different stresses, as well as its contribution to survival under starvation, heat stress, and cold stress. Result: 1) The amino acid sequence encoded by DaTOR contained conserved TOR domains and shared 96.57% similarity with that encoded by DpTOR from Dendroctonus ponderosae. 2) The expression level of DaTOR in D. armandi was the highest in pupae and emerged female adults, the lowest in larvae, and highest in fat bodies among different tissues. 3) Under starvation and cold stress, DaTOR expression showed a progressive down-regulation with increasing stress duration. Under heat stress, DaTOR expression increased transiently, reaching its peak at 12 h, and then declined. 4) RNA interference of DaTOR in larvae and adult males and females of D. armandi resulted in a significant reduction in DaTOR transcript levels. Under starvation and heat stress, post-interference mortality in larvae exceeded that in adult males and females, reaching 73.33% and 71.67%, respectively. Under cold stress, adult males exhibited the highest mortality rate, reaching 76.67%. 5) After injection of dsTOR, the contents of triacylglycerol, glycogen and trehalose in female adults, male adults and larvae of D. armandi showed a decreasing trend, while the content of free fatty acids showed an increasing trend under starvation, heat and cold stresses. Conclusion: DaTOR exhibits differential expression across developmental stages and tissues of D. armandi, and displays distinct response patterns under starvation, heat stress, and cold stress. RNAi-mediated knockdown alters energy metabolism, confirming the involvement of DaTOR in regulating energy homeostasis under these stresses.

Key words: Dendroctonus armandi, target protein of rapamycin, starvation and temperature stresses, RNA interference, energy metabolism

中图分类号:

S763.3

王临君,毛新杰,陈辉. 雷帕霉素靶蛋白基因调控逆境胁迫下华山松大小蠹能量代谢的功能分析[J]. 林业科学, 2026, 62(8): 1-10.

Linjun Wang,Xinjie Mao,Hui Chen. Functional Analysis of the Rapamycin Target Protein Gene in Regulating Energy Metabolism of Dendroctonus armandi under Stress Conditions[J]. Scientia Silvae Sinicae, 2026, 62(8): 1-10.

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基因Gene	物种名称 Species	名称 Name	登录号 Accession No.	相似性 Identity (%)
DaTOR	华山松大小蠹 Dendroctonus ponderosae	TOR	XP_048517977.1	96.57
	墨西哥棉铃象甲 Anthonomus grandis grandis	TOR	XP_050295995.1	80.15
	四粒方胸小蠹 Euwallacea similis	TOR	XP_066249755.1	74.39

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