豌豆修尾蚜报警信息素合成与释放相关基因的鉴定与分析

doi:10.6048/j.issn.1001-4330.2025.06.022

新疆农业科学 ›› 2025, Vol. 62 ›› Issue (6): 1496-1506.DOI: 10.6048/j.issn.1001-4330.2025.06.022

豌豆修尾蚜报警信息素合成与释放相关基因的鉴定与分析

李馨妍¹(), 黄天宇²^,³, 王志强², 周洪旭¹, 王冰²(), 王桂荣²^,⁴()

1.青岛农业大学植物医学学院,山东青岛 266109
2.中国农业科学院植物保护研究所/植物病虫害综合治理全国重点实验室,北京 100193
3.中国农业科学院佛山昆鹏现代农业研究所,广东佛山 528200
4.中国农业科学院深圳农业基因组研究所/农业部基因组分析实验室,广东深圳 518120

收稿日期:2024-10-30 出版日期:2025-06-20 发布日期:2025-07-29
通信作者: 王桂荣(1972-),男,安徽宿松人,研究员,研究方向为化学生态学,(E-mail)wangguirong@caas.cn;
王冰(1983-),女,河北宣化人,研究员,研究方向为化学生态学,(E-mail)wangbing02@caas.cn
作者简介:李馨妍(1999-),女,河北唐山人,硕士研究生,研究方向为化学生态学,(E-mail)lixinyan@stu.qau.edu.cn
基金资助:
国家自然科学基金面上项目(32272621);中国烟草总公司重大科技项目(110202201017(LS-01));深圳市科技计划资助(KQTD20180411143628272);中国博士后科学基金(2023M743837);中国农业科学院科技创新工程

Identification and analysis of genes related to the synthesis and release of alarm pheromone of Megoura crassicauda

LI Xinyan¹(), HUANG Tianyu²^,³, WANG Zhiqiang², ZHOU Hongxu¹, WANG Bing²(), WANG Guirong²^,⁴()

1. College of Plant Health and Medicine, Qingdao Agricultural University, Qingdao Shandong 266109, China
2. State Key Laboratory for Biology of Plant Diseases and Insect Pests/Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, China
3. Kunpeng Institute of Modern Agriculture at Foshan, Agricultural Genomics Institute at Shenzhen, Chinese Academy of Agricultural Sciences, Foshan Guangdong 528200, China
4. Shenzhen Institute of Agricultural Genomics, Chinese Academy of Agricultural Sciences/Genome Analysis Laboratory, Ministry of Agriculture, Shenzhen Guangdong 518120, China

Received:2024-10-30 Published:2025-06-20 Online:2025-07-29
Supported by:
National Natural Science Foundation of China(32272621);Major Special Project for Green Pest Control(110202201017(LS-01));Shenzhen S & T Project(KQTD20180411143628272);China Postdoctoral Science Foundation(2023M743837);Agricultural S & T Innovation Program Project

摘要/Abstract

摘要：

【目的】蚜虫腹管是分泌防御性化学物质的重要组织器官,测序与分析腹管转录组,为蚜虫报警信息素合成与释放相关基因的鉴定提供分子基础。【方法】利用Illumina NoveSeq 6000测序平台对豌豆修尾蚜Megoura crassicauda腹管和残体进行转录组测序与比较分析,利用差异基因分析(Differential Expression Analysis,DEG)鉴定参与报警信息素合成与释放相关的候选基因,通过实时荧光定量PCR(Real-time Quantitative PCR,RT-qPCR)技术验证这些基因的表达模式。【结果】共鉴定出2 156个在腹管和残体之间存在显著差异表达的基因(表达差异≥2倍)。分析报警信息素合成相关的细胞色素P450(cytochrome P450 monooxygenase,CYP450)和萜烯合成酶(terpene synthase,TPS)基因家族。其中,3个CYP450s(McraCYP380C、McraCYP4CK1和McraCYP315A1)在腹管中的表达水平显著高于残体。而鉴定到的2个异戊烯基二磷酸合成酶(isoprenyl diphosphate synthase,IDS)基因,即法尼基焦磷酸合酶(farnesyl pyrophosphate synthase,FPPS)和香叶基焦磷酸合酶(geranyl pyrophosphate synthase,GPPS),在组织间的表达水平无显著差异。此外,分析报警信息素释放运输相关的潜在基因,鉴定到2个化学感觉蛋白(chemosensory proteins,CSP)基因在腹管高表达。McraCYP380C、McraCYP4CK1、McraCYP315A1以及McraCSP7基因在腹管中的表达水平显著上调。【结论】鉴定并分析了豌豆修尾蚜M. crassicauda腹管和残体转录组中的差异表达基因。

关键词: 豌豆修尾蚜; 转录组; 腹管; 异戊烯基二磷酸合成酶基因; 化学感受蛋白; 细胞色素P450

Abstract:

【Objective】 The cornicle of aphids is an important organ for secreting defensive chemicals. Cornicle transcriptome sequencing and analysis can provide molecular basis for identification of genes related to aphid alarm pheromone synthesis and release. 【Methods】 Transcriptome sequencing and comparative analysis were performed on the cornicles and residues of Megoura crassicauda using Illumina NoveSeq 6000 sequencing platform. The candidate genes were identified in the synthesis and release of alarm pheromones using analysis of the differentially expressed genes (DEGs), and the expression patterns of these genes were further verified by real-time Quantitative PCR (RT-qPCR). 【Results】 A total of 2156 genes were identified as DEGs between the cornicles and the residues (expression difference ≥2 times). Cytochrome P450 monooxygenase (CYP450) and terpene synthase (TPS) gene families, which might be involved in terpene synthesis alarm pheromone synthesis, were analyzed. Among them, the expression levels of three CYP450s (McraCYP380C, McraCYP4CK1 and McraCYP315A1) in the cornicles were significantly higher than those in the residues. Two isoprenyl diphosphate synthase (IDS) genes, namely farnesyl pyrophosphate synthase (FPPS) and geranyl pyrophosphate synthase (GPPS), were identified. There were no significant differences in the expression levels of FPPS and GPPS between tissues. In addition, potential genes related to alarm pheromone release and transport were analyzed, and two chemosensory protein (CSP) genes were identified as being highly expressed in the cornicle. The expression levels of McraCYP380C, McraCYP4CK1, McraCYP315A1 and McraCSP7 genes in the cornicle were significantly up-regulated. 【Conclusion】 The DEGs in the cornicles and the residue transcriptome of M. crassicauda are successfully identified and analyzed.

Key words: Megoura crassicauda; transcriptome; cornicles; isoprenyl diphosphate synthase genes; chemosensory proteins; cytochrome P450 monooxygenas

中图分类号:

S188

李馨妍, 黄天宇, 王志强, 周洪旭, 王冰, 王桂荣. 豌豆修尾蚜报警信息素合成与释放相关基因的鉴定与分析[J]. 新疆农业科学, 2025, 62(6): 1496-1506.

LI Xinyan, HUANG Tianyu, WANG Zhiqiang, ZHOU Hongxu, WANG Bing, WANG Guirong. Identification and analysis of genes related to the synthesis and release of alarm pheromone of Megoura crassicauda[J]. Xinjiang Agricultural Sciences, 2025, 62(6): 1496-1506.

图/表 10

图1 腹管和残体中差异表达基因的图谱

Fig.1 Expression profiles of DEGs in the cornicles and the residues

图2 豌豆修尾蚜腹管高表达基因的GO功能注释

Fig.2 GO functional annotation of highly expressed genes in the cornicles of M. crassicauda

表1 候选基因在豌豆修尾蚜M. crassicauda腹管和残体中的表达水平

Tab.1 Expression levels of candidate genes in the cornicles and residues of M. crassicauda

基因ID Gene ID	基因名称 Gene Name	FPKM-平均值 FPKM-Mean		对数2倍数变化 log²FoldChange	P-值 P-value	调整后P-值 P-adj	上/下调 Up/Down- Regulation
基因ID Gene ID	基因名称 Gene Name	残体 Residues	腹管 Cornicles	对数2倍数变化 log²FoldChange	P-值 P-value	调整后P-值 P-adj	上/下调 Up/Down- Regulation
MSTRG.2087.1	McraCYP380C	147.453 254 9	2902.503 026	4.299 216 63	3.22E-05	0.000 605 208	Up
MSTRG.6860.1	McraCYP4CK1	5 487.062 689	11 026.344 26	1.007 029 351	1.04E-17	2.15E-15	Up
MSTRG.8070.6	McraCYP315A1	5.033 902 502	518.314 676 4	6.685 247 751	2.80E-06	7.50E-05	Up
MSTRG.11084.1	McraCSP7	2 779.474 791	9 640.821 777	1.794 432 213	2.84E-156	6.04E-152	Up
MSTRG.465.1	McraCSP8	4 680.169 046	18 727.825 01	2.000 517 384	1.69E-34	1.57E-31	Up
MSTRG.11160.2	McraGPPS	5 219	2 429	-0.449 135 111	0.066 290 039	0.066 290 039	-
MSTRG.7985.1	McraFPPS	8 660.666 667	3 161.666 667	-0.597964673	1.48E-07	5.42E-06	-

图3 豌豆修尾蚜M. crassicauda候选基因的表达谱注: Mcra:豌豆修尾蚜;Agos:棉蚜;Apis:豌豆蚜;Pbam:竹茎扁蚜;CYP4CK1分支用红色背景突出显示,CYP315A1分支用橙色背景突出显示,CYP380C分支用蓝紫色背景突出显示

Fig.3 Expression profiles of candidate genes from M. crassicauda Notes: Mcra: M. crassicauda; Agos: A. gossypii; Apis: A. pisum; Pbam: P. bambucicola; The CYP4CK1 clade is highlighted with a red background, the CYP315A1 clade is marked with an orange background, and the CYP380C clade is indicated by a bluish purple background

图4 4种蚜虫的CYP450s基因家族系统发育树注:A. McraCYP380C与ApisCYP380C的氨基酸序列比对;B. McraCYP4CK1与PbamCYP4CK1的氨基酸序列比对;C. McraCYP315A与ApisCYP315A1的氨基酸序列比对;Mcra:豌豆修尾蚜;Apis:豌豆蚜;Pbam:竹茎扁蚜

Fig.4 Phylogenetic tree of the CYP450s gene family of the four aphid species Notes: A. Amino acid sequence alignment between McraCYP380C and ApisCYP380C; B. Amino acid sequence alignment between McraCYP4CK1 and PbamCYP4CK1; C. Amino acid sequence alignment between McraCYP315A and ApisCYP315A1; Mcra: M. crassicauda; Apis: A. pisum; Pbam: P. bambucicola

表2 候选基因的注释信息

Tab.2 Annotated information of candidate genes

参考基因 Unigene reference	基因名 Gene name	长度 Length (nt)	开放阅读框 ORF (aa)	E值 E- value	相似度 Identity (%)	最佳BLASTx匹配 Blastx best hit (Reference/Name/Species)	信号肽 SP (No.)	跨膜结构域 TMD (No.)	全长 Full length	组织 Tissue
MSTRG.2087.1	McraCYP380C	1548	515	0.0	74.37	>XP_060860450.1 PREDICTED: cytochrome P450 4C1-like isoform X1 [Metopolophium dirhodum]	0	1	YES	C&R
MSTRG.6860.1	McraCYP4CK1	1547	515	0.0	89.57	>XP_060860450.1 PREDICTED: cytochrome P450 4C1-like [Metopolophium dirhodum]	0	1	YES	C&R
MSTRG.8070.6	McraCYP315A1	1434	477	0.0	90.78	>XP_001944183.2 PREDICTED: cytochrome P450 315a1, mitochondrial [Acyrthosiphon pisum]	0	0	YES	C
MSTRG.11084.1	McraCSP7	468	155	1e-109	100.00	>ULF48248.1 PREDICTED: chemosensory protein 7 (CSP7) mRNA [Acyrthosiphon pisum]	0	0	YES	C&R
MSTRG.465.1	McraCSP8	219	72	4e-27	100.00	>ULF48249.1 PREDICTED: chemosensory protein 8 (CSP8) mRNA [Acyrthosiphon pisum]	-	-	3’	C&R
MSTRG.11160.2	McraGPPS	930	309	0.0	100.00	>QUH22249.1 PREDICTED: geranyl pyrophosphate synthase mRNA [Megoura viciae]	0	0	YES	C&R
MSTRG.7985.1	McraFPPS	1185	394	0.0	98.98	>AAY33489.2 PREDICTED: putative mitochondrial isoprenyl diphosphate synthase precursor, mRNA [Megoura viciae]	0	0	YES	C&R

图5 豌豆修尾蚜McraCYP450s基因与其他蚜虫同源基因的氨基酸序列比对注: Mcra:豌豆修尾蚜;Agos:棉蚜;Apis:豌豆蚜;Agly:大豆蚜;Mper:桃蚜;Save:麦长管蚜;黑框显示了从豌豆修尾蚜M. crassicauda的腹管中鉴定出的候选CSPs基因。CSP7分支以浅红色背景突出显示,而CSP8分支以紫色背景突出显示

Fig.5 Amino acid sequence alignment of McraCYP450s genes from M. crassicauda with homologous genes of other aphid species Notes: Mcra: M. crassicauda; Agos: A. gossypii; Apis: A. pisum; Agly: A. glycines; Mper: M. persicae; Save: S. avenae; The candidate CSP genes identified from the cornicles of M. crassicauda are indicated by black boxes. The CSP7 clade is highlighted with a light red background, while the CSP8 clade is high-lighted with a purple background

图6 6种蚜虫的CSPs基因家族系统发育树

Fig.6 Phylogenetic tree of the CSPs gene family of the six aphid species

图7 豌豆修尾蚜McraIDSs基因与豌豆蚜ApisIDSs同源基因的氨基酸序列比对注:A. McraGPPS与ApisGPPS的序列比对;B. McraFPPS与ApisFPPS的序列比对;Mcra:豌豆修尾蚜;Apis:豌豆蚜

Fig.7 Amino acid sequence alignment of McraIDS genes from M. crassicauda with homologous genes of A. pisum Notes: A. Sequence alignment between McraGPPS and ApisGPPS; B. Sequence alignment between McraFPPS and ApisFPPS; Mcra: M. crassicauda; Apis: A. pisum

图8 豌豆修尾蚜M. crassicauda 7个候选基因的组织特异性表达水平注:数据以Mean±SE表示(n=3)。通过独立样本T检验分析,星号表示在α=0.05水平存在显著性差异,n.s.表示在α=0.05水平无显著性差异

Fig.8 T Tissue-specific expression levels of seven candidate genes from M. crassicauda Notes: Data are presented as Mean±SE (n=3). The relative expression between cornicles and residues was analyzed by t-test, asterisks indicate a significant difference at the α=0.05 level, while n.s. indicates no significant difference at the α=0.05 level

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豌豆修尾蚜报警信息素合成与释放相关基因的鉴定与分析

Identification and analysis of genes related to the synthesis and release of alarm pheromone of Megoura crassicauda

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