芳樟醇、桉叶油素取食胁迫下香樟齿喙象（鞘翅目：象甲科）幼虫的生长发育及抗性基因的转录表达

doi:10.11707/j.1001-7488.LYKX20220093

摘要/Abstract

摘要：

目的: 探究香樟齿喙象与香樟互作中的寄主选择和适应机制，明确香樟齿喙象幼虫对芳樟醇、桉叶油素胁迫生境下的生长发育适应性及其解毒代谢方式，以阐明香樟齿喙象专性为害香樟的潜在机制。方法: 采用人工饲料混药法，测定香樟齿喙象初孵幼虫取食不同质量分数芳樟醇和桉叶油素3天后的死亡率，明确香樟齿喙象幼虫对2种萜类物质的敏感性。分别设定10.0 、19.7 mg·g^?1的芳樟醇及5.0、15.2 mg·g^?1的桉叶油素进行长期取食处理，测定香樟齿喙象幼虫阶段的存活曲线、发育历期、体重等生长发育指标，明确香樟齿喙象幼虫对2种萜类物质的生物学响应。利用qRT-PCR技术，测定香樟齿喙象4龄幼虫取食芳樟醇和桉叶油素后的解毒酶基因（细胞色素P450酶、羧酸酯酶、UDP-葡萄糖醛酸转移酶、谷胱甘肽-S-转移酶和ABC转运蛋白）和表皮蛋白基因的相对表达量变化。结果: 芳樟醇和桉叶油素在2种处理质量分数条件长期胁迫下，对香樟齿喙象幼虫的生长发育产生显著的抑制作用，并且抑制作用随质量分数增加而增强，即发育历期延长、体重降低以及死亡率提高。此外，还表现出幼虫在蜕皮期前后出现高频死亡的特征。相较于桉叶油素，香樟齿喙象幼虫对芳樟醇表现出更强的耐受性。qRT-PCR结果显示，芳樟醇胁迫能够显著地诱导香樟齿喙象幼虫体内细胞色素P450酶和表皮蛋白的基因表达，而桉叶油素取食胁迫下仅有1条UDP-葡萄糖醛酸转移酶的基因表达量显著上调。结论: 香樟齿喙象幼虫对2种寄主植物次生物质的取食胁迫产生不同生理适应性。同时，其幼虫的解毒酶相关基因和表皮蛋白基因对2种萜类物质胁迫的表达响应模式存在差异性。香樟齿喙象幼虫能够有目的地调控不同生理适应机制，以克服不同的寄主次生物质防御，以实现寄主定殖和种群暴发。

关键词: 香樟齿喙象, 寄主次生物质, 解毒酶, 表皮蛋白, 生理适应性

Abstract:

Objective: Pagiophloeus tsushimanus (Coleoptera: Curculionidae) is a serious wood borer on Cinnamomum camphora, which has rapidly outbroken in Shanghai . Linalool and eucalyptol are important terpene metabolites in C. camphora, and also constitute the chemical defense system in C. camphora. This study aims to explore the host selection and adaptation mechanism in the interaction between P. tsushimanus and C. camphora and clarify the development adaptability and detoxification mechanism that P. tsushimanus copes with the linalool and eucalyptol, so as to analyze the potential mechanism that P. tsushimanus obligates to C. camphora. Method: The newly hatched P. tsushimanus larvae were fed with linalool and eucalyptol for 3 days using residual film method and the sensitivity of P. tsushimanus larvae to the two terpenoids was determined. Linalool (10.0 mg·g^–1 and 19.7 mg·g^–1) and eucalyptol (5.0 mg·g^–1 and 15.2 mg·g^–1) were set for the long term feeding treatment, to measure the survival curve, developmental duration, body weight and other growth and development indexes of P. tsushimanus larvae in order to evaluate the phenotypic response of P. tsushimanus to the two terpenoids. The qRT PCR technique was used to determine the relative expressions of detoxification enzyme genes (CYP450, carboxylesterase, UDP-glucuronosyltransferase, Glutathione S-transferase and ATP binding cassette transporter) and insect cuticle protein genes in the 4^th instar larvae of P. tsushimanus. Result: Linalool and eucalyptol significantly inhibited growth and development of P. tsushimanus larvae under the long-term feeding of two mass fraction of the terpenoids and this inhibitory effect was correlated with the dose, that is, prolonged the developmental duration, reduced the weight and showed high mortality. Moreover, the larvae also exhibited high-frequency mortality before and after the molting period. Compared with eucalyptol, P. tsushimanus larvae showed stronger tolerance to linalool. The qRT-PCR results revealed that the expression of four cytochrome P450 genes were significantly up-regulated, four cuticle protein genes were up-regulated in P. tsushimanus treated with linalool and one UGT gene was significantly up-regulated in P. tsushimanus larvae treated with eucalyptol. Conclusion: The larvae of P. tsushimanus show different physiological adaptability to the two secondary metabolites. Meanwhile, the expression patterns of detoxification enzyme genes and cuticle protein genes in P. tsushimanus larvae are different under two secondary metabolites. The result also suggests that P. tsushimanus larvae could detoxify different plant secondary metabolites through different mechanism so as to achieve host colonization and population outbreak.

Key words: Pagiophloeus tsushimanus, host plant secondary metabolite, detoxification enzyme, cuticle protein, physiological adaptability.

中图分类号:

S718.7

王璟廷,李寿银,左壮,徐文轩,郝德君. 芳樟醇、桉叶油素取食胁迫下香樟齿喙象（鞘翅目：象甲科）幼虫的生长发育及抗性基因的转录表达[J]. 林业科学, 2023, 59(5): 109-120.

Jingting Wang,Shouyin Li,Zhuang Zuo,Wenxuan Xu,Dejun Hao. Growth, Development and the Resistance Gene Transcriptional Expression of Pagiophloeus tsushimanus（Coleoptera: Curculionidae） Larvae after Feeding on Linalool and Eucalyptol[J]. Scientia Silvae Sinicae, 2023, 59(5): 109-120.

图/表 8

表1

所选香樟齿喙象有关解毒酶基因和表皮蛋白基因转录组基因序列引物"

基因 Gene	引物序列（5'?3'） Primer sequence（5'?3'）	Tm/℃	产物大小 Product size/bp	扩增效率 Amplification efficiency（%）
RPS3	AACGATTGGGCATCTTCAT	54.0	139	113.7
RPS3	CACAGGATCATTTGCTGGA	56.0	139	113.7
18S RNA	GGCATCGTCCTAACACCCA	60.0	124	91.6
18S RNA	CCGCTAGAGGCGTTTCATC	60.0	124	91.6
CYP-1	CATCTGCTTCTTCGGACTG	58.0	95	107.9
CYP-1	TTATGGGTCTCATTTGGGT	54.0	95	107.9
CYP-2	AATCAGGAGGGCTCTACAT	56.0	116	101.9
CYP-2	GAAATACTCATAATCCGTTG	49.3	116	101.9
CYP-3	GTCCTTGGTCCCGATGCTC	62.0	89	111.7
CYP-3	ACGCCGCCGAATCTTACTC	60.0	89	111.7
CYP-4	TCCTTGGTCCCGATGCTC	58.0	86	106.4
CYP-4	CGCCGCCGAATCTTACTC	58.0	86	106.4
CYP-5	TTCTCCAGGGAACAAGCAC	58.0	115	98.1
CYP-5	TGGCGTCACCACAGATGTA	58.0	115	98.1
GST-1	GCTGGTGGGTCAACAGATCA	57.7	82	92.5
GST-1	AGCGTCTGGCTTCCTTTCAA	56.9	82	92.5
GST-2	CTTCTATGAGCCCCTACTGCC	57.6	137	109.2
GST-2	ATTCTCCCCGTTCAGCTCG	57.6	137	109.2
GST-3	ACGAAGCGTGTGGAATGAGAT	56.5	144	95.6
GST-3	TGATCTGTTGACCCACCAGC	57.7	144	95.6
GST-4	AACTTGGCACATGCCACAAC	56.8	129	99.3
GST-4	CGTACCGAGAGCATGTTGGA	57.3	129	99.3
GST-5	GAGTTTCTCAAAGCGTCGCC	57.0	118	93.5
GST-5	TCGTAATCGCCCAGAGCATC	57.5	118	93.5
COE-1	AGGAGCAAGCAGATTCACGA	56.4	105	94.0
COE-1	AGCTTCCAAATCCTCATCCCT	55.8	105	94.0
COE-2	CCCCGAACAATCAACGCTTC	57.2	136	97.6
COE-2	TGCCTCCTTCCATAACGGAT	55.8	136	97.6
COE-3	GGGATCGGGTTCGGAACAAA	58.0	139	94.0
COE-3	CGCATTTCCGGGAGTTTCCA	58.5	139	94.0
COE-4	GATCTTTGGGCAAAGTGCCG	57.5	140	91.3
COE-4	CTTGACGTGCCCTTGTTGTC	57.1	140	91.3
COE-5	GACCAGTTTCCTGCTAGCGAT	57.3	93.0	97.3
COE-5	TGGTTCGGGAGTTGGGTTTC	57.9	93.0	97.3
UGT-1	GAATGCGGCCAAAGCTGAAA	56.8	123.0	99.8
UGT-1	ACCTGTGCTTTGTATCGGGG	57.8	123.0	99.8
UGT-2	TGATCAGCCCATGAGTTCCT	55.8	97.0	105.6
UGT-2	ATCAACGGCAGCATAGCGTA	56.8	97.0	105.6
UGT-3	CCCCGATACAAAGCACAGGT	57.8	135.0	97.1
UGT-3	ACTGGGAGATTGCAGATGCC	57.9	135.0	97.1
UGT-4	CCTCCGAAGAAACTTCCGCA	57.8	88.0	104.4
UGT-4	TCAAGTTCGACCCCATGCTG	57.9	88.0	104.4
UGT-5	GCCGGCATTAGATTGCCTTG	57.4	133.0	93.7
UGT-5	TAGTCCTCGTGCTTCTGCAA	56.1	133.0	93.7
ABC-1	GCCAACACTCCGCAAACATT	56.7	147.0	98.0
ABC-1	TTGTCCAGCTCGTTTCTGCT	56.9	147.0	98.0
ABC-2	TCTCGTGCTGTTAGGGGAGA	57.8	102.0	94.6
ABC-2	AGTTGAGGTAGGGTGCCTGA	58.2	102.0	94.6
ABC-3	ATTCCTCCTCTTCTCCGGCT	58.1	132.0	100.9
ABC-3	GCCCTGTTGAAGCCATAGGT	57.9	132.0	100.9
ABC-4	GAGGATGCGCATAGCGTTAG	56.2	95.0	98.0
ABC-4	CGCAAGCGTCCAAATCCAAA	56.7	95.0	98.0
ABC-5	TGTGGGACCTTCTGTTGAGC	57.6	133.0	100.4
ABC-5	GGCGTTCCGTGGGAGTTTAT	58.0	133.0	100.4
CP-1	AGCGGTGGGTTCTTTGTGTA	56.0	95.0	100.9
CP-1	CCCTGGTTGATTCTGACGGA	56.0	95.0	100.9
CP-2	ACTCCAGTCGTCCTCCAAG	60.0	88.0	102.9
CP-2	AACCGTTCTCGTCTGCTAC	58.0	88.0	102.9
CP-3	CTACCCACAACAAGGCGGAT	60.0	83.0	114.9
CP-3	ATCCTGCACTTGGGGGAATG	56.0	83.0	114.9
CP-4	CAAGAATCTCGTCGTGGTG	58.0	93.0	115.6
CP-4	TGGGTCAGCGGTGTAATC	56.0	93.0	115.6
CP-5	TGGGTCAGCGGTGTAATC	56.3	198.0	90.6
CP-5	CTACGACAGCGTTGAAGC	56.0	198.0	90.6

表1

表2

香樟齿喙象有关解毒酶基因和表皮蛋白基因转录组基因序列同源性分析"

昆虫种类 Species	GenBank 登录号 GenBank accession No.	基因 Gene	匹配度 Identity(%)	长度 Length/bp
中欧山松大小蠹 Dendroctonus ponderosae	XP_019761906.1	RPS3	48.78	370
红棕象甲 Rhynchophorus ferrugineus	KAF7284967.1	18S RNA	88.2	230
中欧山松大小蠹 Dendroctonus ponderosae	AFI45038.1	CYP-1	45.62	488
华山松大小蠹 Dendroctonus armandi	ALD15919.1	CYP-2	54.83	507
米象 Sitophilus oryzae	XP_030748038.1	CYP-3	47.64	509
米象 Sitophilus oryzae	XP_030758350.1	CYP-4	42.92	506
中欧山松大小蠹 Dendroctonus ponderosae	AFI45002.1	CYP-5	85.29	697
中欧山松大小蠹 Dendroctonus ponderosae	AFE62899.1	GST-1	68.23	207
中欧山松大小蠹 Dendroctonus ponderosae	XP_019759096.1	GST-2	95	377
中欧山松大小蠹 Dendroctonus ponderosae	AEE62899.1	GST-3	68.09	207
米象 Sitophilus oryzae	XP_030752825.1	GST-4	95.64	378
香樟齿喙象 Pagiophloeus tsushimanus	QCX41803.1	GST-5	94.44	126
红棕象甲 Rhynchophorus ferrugineus	KAF7281696.1	COE-1	60.65	553
中欧山松大小蠹 Dendroctonus ponderosae	ERL87113.1	COE-2	81.75	481
中欧山松大小蠹 Dendroctonus ponderosae	ENN80857.1	COE-3	61.11	188
中欧山松大小蠹 Dendroctonus ponderosae	XP_019756056.1	COE-4	66.04	559
华山松大小蠹 Dendroctonus armandi00	AYN64429.1	COE-5	78.19	552
中欧山松大小蠹 Dendroctonus ponderosae	XP_019756751.1	UGT-1	84.35	525
稻水象甲 Lissorhoptrus oryzophilus	AVT42216.1	UGT-2	65.24	524
中欧山松大小蠹 Dendroctonus ponderosae	XP_019756751.1	UGT-3	84	525
中欧山松大小蠹 Dendroctonus ponderosae	XP_019771423.1	UGT-4	68.38	519
中欧山松大小蠹 Dendroctonus ponderosae	AEE61792.1	UGT-5	73.91	523
米象 Sitophilus oryzae	XP_030757858.1	ABC-1	96.19	609
米象 Sitophilus oryzae	XP_030757858.2	ABC-2	94.01	609
中欧山松大小蠹 Dendroctonus ponderosae	XP_019764834.1	ABC-3	87.78	687
米象 Sitophilus oryzae	XP_030758286.1	ABC-4	93.75	621
玉米根萤叶甲 Diabrotica virgifera virgifera	XP_028153387.1	ABC-5	73.58	400
中欧山松大小蠹 Dendroctonus ponderosae	XP_019767588.1	CP-1	93.55	229
中欧山松大小蠹 Dendroctonus ponderosae	KAH1004877.1	CP-2	82.81	132
中欧山松大小蠹 Dendroctonus ponderosae	XP_019761931.1	CP-3	74.43	211
中欧山松大小蠹 Dendroctonus ponderosae	XP_019767588.1	CP-4	93.44	229
中欧山松大小蠹 Dendroctonus ponderosae	KAH1013867.1	CP-5	66.9	178

表2

图1

图2

表1

图3

表2

图4

参考文献 0

	陈澄宇, 康志娇, 史雪岩, 等. 昆虫对植物次生物质的代谢适应机制及其对昆虫抗药性的意义. 昆虫学报, 2015, 58 (10): 1126- 1139.
	Chen C Y, Kang Z J, Shi X Y, et al. Metabolic adaptation mechanisms of insects to plant secondary metabolites and their implications for insecticide resistance of insects. Acta Entomologica Sinica, 2015, 58 (10): 1126- 1139.
	陈高满, 陈展博, 葛　辉, 等. 苹果蠹蛾细胞色素P450基因CYP332A19和CYP337B19的克隆及表达分析. 昆虫学报, 2020, 63 (8): 941- 951.
	Chen G M, Chen Z B, Ge H, et al. Cloning and expression analysis of cytochrome P450 Genes CYP332A19 and CYP337B19 in the codling moth, Cydia pomonella (Lepedotera: Tortricidae) . Acta Entomologica Sinica, 2020, 63 (8): 941- 951.
	黄俊浩, 吴时英, 高　磊, 等. 中国新记录种——香樟齿喙象的鉴别与为害. 浙江农林大学学报, 2014, 31 (5): 764- 767.
	Huang J H, Wu S Y, Gao L, et al. Diagnosis and damage of weevil pest Pagiophloeus tsushimanus on camphor tree . Journal of Zhejiang AF University, 2014, 31 (5): 764- 767.
	李寿银, 陈　聪, 李　慧, 等. 取食含不同植物源成分的饲料对香樟齿喙象幼虫生长发育及体内解毒酶活性的影响. 昆虫学报, 2019, 62 (11): 1286- 1296.
	Li S Y, Chen C, Li H, et al. Effects of feeding on diets containing components of different plants on the development and detoxifying enzyme activities in Pagiophloeus tsushimanus (Coleptera: Curculionidae) larvae . Acta Entomologica Sinica, 2019, 62 (11): 1286- 1296.
	梁　欣, 陈　斌, 乔　梁. 昆虫表皮蛋白基因研究进展. 昆虫学报, 2014, 57 (09): 1084- 1093.
	Liang X, Chen B, Qiao L. Research progress in insect cuticle protein genes. Acta Entomologica Sinica, 2014, 57 (09): 1084- 1093.
	刘兴平, 陈春平, 王国红, 等. 2003. 我国松树诱导抗虫性研究进展. 林业科学, 39(5): 119−128.
	Liu X P, Chen C P, Wang G H, et al. 2003. Progress in induced resistance of pines. Scientia Silvae Sinicae, 39(5) : 119−128.［in Chinese］
	孙雅雯, 郑　彬. 昆虫表皮与化学杀虫剂抗性机制关系的研究进展. 中国病原生物学杂志, 2015, 10 (11): 1055- 1059.
	Sun Y W, Zheng B. Advances in the study of the relationship between insect cuticle proteins and insecticide resistance. Journal of Pathogen Biology, 2015, 10 (11): 1055- 1059.
	谢　辉, 王　燕, 刘银泉, 等. 2012. 植物组成型防御对植食性昆虫的影响, 植物保护, 38(1): 1−5.
	Xie H, Wang Y, Liu Y Q, et al. 2012. The influence of plant constitutive defense system on phytophagous insects. Plant Protection, 38(1): 1−5.［in Chinese］
	张　峰, 毕良武, 赵振东. 樟树植物资源分布及化学成分研究进展. 天然产物研究与开发, 2017, 29 (3): 517- 531.
	Zhang F, Bi L W, Zhao Z D. Review on plant resources and chemical composition of camphor tree. Natural Product Research and Development, 2017, 29 (3): 517- 531.
	Adamski Z, Korolczuk D, Purgiel M, et al. Effect of fenitrothion on Spodoptera exigua larval development and ultrastructure of follicle cells . Biologia, 2009, 64 (1): 197- 202. doi: 10.2478/s11756-009-0022-x
	Awolola T S, Oduola O A, Strode C, et al. Evidence of multiple pyrethroid resistance mechanisms in the malaria vector Anopheles gambiae sensu stricto from Nigeria . Transactions of the Royal Society of Tropical Medicine & Hygiene, 2009, 103 (11): 1139- 1145. doi: 10.1016/j.trstmh.2008.08.021
	Balabanidou V, Grigoraki L, John V. Insect cuticle: a critical determinant of insecticide resistance. Current Opinion in Insect Science, 2018, 27, 68- 74. doi: 10.1016/j.cois.2018.03.001
	Birnbaum S S, Abbot P. Gene expression and diet breadth in plant-feeding insects: summarizing trends. Trends in Ecology & Evolution, 2020, 35, 259- 277. doi: 10.1016/j.tree.2019.10.014
	Blomquist G J, Tittiger C, Maclean M, et al. Cytochromes P450: terpene detoxification and pheromone production in bark beetles. Current Opinion in Insect Science, 2021, 43, 97- 102. doi: 10.1016/j.cois.2020.11.010
	Charles J P. The regulation of expression of insect cuticle protein genes. Insect Biochemistry Molecular Biology, 2010, 40, 205- 213. doi: 10.1016/j.ibmb.2009.12.005
	Chen C, Li S, Zhu H, et al. Identification and evaluation of reference genes for gene expression analysis in the weevil pest Pagiophloeus tsushimanus using RT-qPCR . Journal of Asia-Pacific Entomology, 2020, 23 (2): 336- 344. doi: 10.1016/j.aspen.2020.01.010
	Chen C, Zhang C, Li S, et al. Biological traits and life history of Pagiophloeus Tsushimanus (Coleoptera: Curculionidae), a weevil pest on camphor trees in China . Journal of Forestry Research, 2021a, 32 (5): 1979- 1988. doi: 10.1007/s11676-020-01227-2
	Chen C, Zhu H, Li S, et al. Insights into chemosensory genes of Pagiophloeus tsushimanus adults using transcriptome and QRT-PCR analysis . Comparative Biochemistry and Physiology Part D:Genomics and Proteomics, 2021b, 37, 100785. doi: 10.1016/j.cbd.2020.100785
	Chiu C C, Keeling C I, Bohlmann J. The cytochrome P450 CYP6DE1 catalyzes the conversion of alpha-pinene into the mountain pine beetle aggregation pheromone trans-verbenol. Science Report, 2019a, 9, 1477. doi: 10.1038/s41598-018-38047-8
	Chiu C C, Keeling C I, Bohlmann J. Functions of mountain pine beetle cytochromes P450 CYP6DJ1, CYP6BW1 and CYP6BW3 in the oxidation of pine monoterpenes and diterpene resin acids. Plos On, 2019b, 14 (5): e0216753. doi: 10.1371/journal.pone.0216753
	Dai L, Ma M, Wang C, et al. Cytochrome P450s from the Chinese white pine beetle, Dendroctonus armandi (Curculionidae: Scolytinae): expression profiles of different stages and responses to host allelochemicals . Insect Biochemistry and Molecular Biology, 2015, 65, 35- 46. doi: 10.1016/j.ibmb.2015.08.004
	Després L, David J P, Gallet C. The evolutionary ecology of insect resistance to plant chemicals. Trends in Ecology and Evolution, 2007, 22 (6): 298- 307. doi: 10.1016/j.tree.2007.02.010
	Fang F J, Wang W J, Zhang D H, et al. The cuticle proteins: a putative role for deltamethrin resistance in Culex pipiens pallens . Parasitology Research, 2015, 114, 4421- 4429. doi: 10.1007/s00436-015-4683-9
	Halon E, Eakteiman G, Moshitzky P, et al. Only a minority of broad-range detoxification genes respond to a variety of phytotoxins in generalist Bemisia tabaci species. Scientific Reports, 2015, 5, 17975. doi: 10.1038/srep17975
	Heidel-Fischer H, Vogel H. Molecular mechanisms of insect adaptation to plant secondary compounds. Current Opinion in Insect Science, 2015, 8, 8- 14. doi: 10.1016/j.cois.2015.02.004
	Hu B, Zhang S H, Ren M M, et al. The expression of Spodoptera exigua P450 and UGT genes: tissue specificity and response to insecticides . Insect Science, 2019, 26 (2): 199- 216. doi: 10.1111/1744-7917.12538
	Huang X Z, Xiao Y T, Köllner T G, et al. The terpene synthase gene family in Gossypium hirsutum harbors a linalool synthase GhTPS12 implicated in direct defence responses against herbivores . Plant, Cell & Environment, 2017, 41 (1): 261- 274. doi: 10.1111/pce.13088
	Li S Y, Chen C, Jia Z Y, et al. Offspring performance and female preference of Pagiophloeus tsushimanus (Coleoptera: Curculionidae) on three Lauraceae tree species: a potential risk of host shift caused by larval experience . Journal of Applied Entomology, 2021, 145 (6): 530- 542. doi: 10.1111/jen.12865
	Li S Y, Wang J T, Chen C, et al. Tolerance, biochemistry and related gene expression in Pagiophloeus tsushimanus (Coleoptera: Curculionidae) exposed to chemical stress from headspace host-plant volatiles . Agricultural and Forest Entomology, 2022, 24 (2): 189- 203. doi: 10.1111/afe.12482
	Lilly D G, Latham S L, Webb C E, et al. Cuticle thickening in a pyrethroid-resistant strain of the common bed bug, Cimex lectularius L . (Hemiptera:Cimicidae). Plos One, 2016, 11 (4): e0153302. doi: 10.1371/journal.pone.0153302
	Loivamaki M, Mumm R, Dicke M, et al. Isoprene interferes with the attraction of body-guards by herbaceous plants. Proceedings of the National Academy of Sciences of the United States of America, 2008, 105, 17430- 17435. doi: 10.1073/pnas.0804488105
	Matthews B B, dos Santos G, Crosby M A, et al. Gene Model Annotations for Drosophila melanogaster: Impact of High-Throughput Data . G3-Genes Genomes Genetics, 2015, 5 (8): 1721- 1736. doi: 10.1534/g3.115.018937
	Maurya A K, Patel R C, Frost C J. Acute toxicity of the plant volatile indole depends on herbivore specialization. Journal of Pest Science, 2020, 93, 1107- 1117. doi: 10.1007/s10340-020-01218-6
	Mccallum E J, Cunningham J P, Lucker J, et al. Increased plant volatile production affects oviposition, but not larval development, in the moth Helicoverpa armigera . Journal of Experimental Biology, 2011, 214 (21): 3672- 3677. doi: 10.1242/jeb.059923
	Mithofer A, Bloand W. Plant defense against herbivores: chemical aspects. Annual Review Plant Biology, 2012, 63, 431- 450. doi: 10.1146/annurev-arplant-042110-103854
	Moussian B, 2010. Recent advances in understanding mechanisms of insect cuticle differentiation. Insect Biochemistry and Molecular Biology, 40(5): 363-375. DOI: 10.1016/j.cois.2017.11.009
	Nishida R. Chemical ecology of insect-plant interactions: ecological significance of plant secondary metabolites. Bioscience Biotechnology and Biochemistry, 2014, 78 (1): 1- 13. doi: 10.1080/09168451.2014.877836
	Pan C, Yun Z, Mo J. The clone of laccase gene and its potential function in cuticular penetration resistance of Culex pipiens pallens to fenvalerate . Pesticide Biochemistry and Physiology, 2009, 93 (3): 105- 111. doi: 10.1016/j.pestbp.2008.12.003
	Pinto L, Fiuza L M. PCR and bioassays screening of bacillus thuringiensis isolates from rice-fields of Rio Grande Do Sul, specific to Lepidopterans and Coleopterans . Brazilian Journal of Microbiology, 2003, 34 (4): 305- 310. DOI: 10.1590/S1517-83822003000400003.
	Richards S, Gibbs R A, Weinstock G M, et al. The genome of the model beetle and pest Tribolium castaneum . Nature, 2008, 452 (7190): 949- 955. doi: 10.1038/nature06784
	Sabina B, Wannes D, Robert G, et al. Transcriptome profiling of a Spirodiclofen susceptible and resistant strain of the european red mite Panonychus ulmi using strand-specific RNA-seq . BMC Genomics, 2015, 16, 974. doi: 10.1186/s12864-015-2157-1
	Seybold S J, Huber D, Lee J. Pine monoterpenes and pine bark beetles: a marriage of convenience for defense and chemical communication. Phytochemical Review, 2006, 5, 143- 178. doi: 10.1007/s11101-006-9002-8
	Sfara V, Zerba E N, Alzogaray R A. Fumigant insecticidal activity and repellent effect of five essential oils and seven monoterpenes on first-instar nymphs of Rhodnius prolixus . Journal of Medical Entomology, 2009, 46 (3): 511- 515. doi: 10.1603/033.046.0315
	Song G P, Hu D K, Tian H, et al. 2016. Synthesis and larvicidal activity of novel thenoylhydrazide derivatives. Scientific Reports, 6: 22977. DOI: 10.1038/srep22977
	Soto I M, Carreira V P, Corio C, et al. Differences in tolerance to host cactus alkaloids in Drosophila koepferae and Dbuzzatii . PLoS One, 2014, 9 (2): e88370. doi: 10.1371/journal.pone.0088370
	Wei C, Zhou S, Li W, et al. Chemical composition and allelopathic, phytotoxic and pesticidal activities of Atriplex cana ledeb . (Amaranthaceae) essential oil. Chemistry & Biodiversity, 2019, 16 (4): e1800595.
	Willoughby L, Chung H, Lumb C, et al. A comparison of Drosophila melanogaster detoxification gene induction responses for six insecticides, caffeine and phenobarbital . Insect Biochemistry & Molecular Biology, 2006, 36 (12): 934- 942. doi: 10.1016/j.ibmb.2006.09.004
	Yactayo-Chang J P, Tang H V, Mendoza J, et al. 2020. Plant defense chemicals against insect pests. Agronomy. 10(8): 1156. DOI: 10.3390/agronomy10081156
	Yahouedo G A, Chandre F, Rossignol M, et al. 2017. Contributions of cuticle permeability and enzyme detoxification to pyrethroid resistance in the major malaria vector Anopheles gambiae. Scientific Reports. 7: 11091. DOI: 10.1038/s41598-017-11357-z

药剂 Pesticides	质量分数 Mass fraction/(mg·g^?1)	体重 Weight/mg
		虫龄 Instar				蛹
		1st-2nd	2nd-3rd	3rd-4th	4th-5th	Pupation
对照组 Control	—	4.84±0.15a	11.18±0.60a	22.53±1.26a	38.42±2.55a	126.04±1.85a
芳樟醇 Linalool	10.0	4.15±0.12b	7.97±0.33b	17.04±1.17b	24.01±1.89b	103.01±4.08bc
芳樟醇 Linalool	19.7	3.31±0.06c	6.26±0.32c	10.09±0.61c	16.41±1.17c	87.16±2.72c
桉叶油素 Eucalyptol	5.0	4.32±0.20b	8.36±0.36b	17.66±1.08b	26.65±2.14b	118.64±3.03ab
桉叶油素 Eucalyptol	15.2	4.02±0.10b	8.17±0.30b	14.86±0.82c	21.61±0.98b	96.05±4.49c

药剂 Pesticides	质量分数 Mass fraction/(mg·g^?1)	虫龄 Instar
药剂 Pesticides	质量分数 Mass fraction/(mg·g^?1)	1st	2nd	3rd	4th	5th
对照组Control	—	0.476	0.708	0.984	1.185	1.187
芳樟醇Linalool	10.0	0.330	0.404	0.837	0.617	0.865
芳樟醇Linalool	19.7	0.086	0.326	0.355	0.467	0.800
桉叶油素Eucalyptol	5.0	0.297	0.423	0.657	0.728	1.098
桉叶油素Eucalyptol	15.2	0.158	0.356	0.581	0.613	0.673

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