双斑长跗萤叶甲雌虫、雄虫触角转录组及差异表达基因分析

doi:10.6048/j.issn.1001-4330.2024.04.023

新疆农业科学 ›› 2024, Vol. 61 ›› Issue (4): 984-995.DOI: 10.6048/j.issn.1001-4330.2024.04.023

• 植物保护·微生物·农业装备工程与机械化 • 上一篇下一篇

双斑长跗萤叶甲雌虫、雄虫触角转录组及差异表达基因分析

何婉洁(), 孟涵颖, 支梦婷, 陈静()

石河子大学农学院/新疆绿洲农业病虫害治理与植保资源利用重点实验室,新疆石河子 832003

收稿日期:2023-08-19 出版日期:2024-04-20 发布日期:2024-05-31
通信作者: 陈静(1979-),女,陕西人,教授,研究方向为农业昆虫与害虫防治,(E-mail)Chj_agr@shzu.edu.cn
作者简介:何婉洁(1997-),女,新疆人,硕士研究生,研究方向为农业昆虫与害虫防治,(E-mail)1287336486@qq.com
基金资助:
新疆维吾尔自治区重点研发任务专项(2022B02043-2);国家自然科学基金项目(31960541);国家自然科学基金项目(31460473);新疆生产建设兵团指导性科技计划项目(2022ZD010)

Analysis of the antennal transcriptome and differentially expressed genes of female and male Monolepta signata

HE Wanjie(), MENG Hanying, ZHI Mengting, CHEN Jing()

Key Laboratory for Oasis Agricultural Pest Management and Plant Protection Resource Utilization / Agricultural College of Shihezi University, Shihezi Xinjiang 832003, China

Received:2023-08-19 Published:2024-04-20 Online:2024-05-31
Correspondence author: CHEN Jing (1979-), female, from Shaanxi,professor, research direction: agricultural entomology and pest control,(E-mail)Chj_agr@shzu.edu.cn.
Supported by:
the Key R& D Task Special Project of Xinjiang Uygur Autonomous Region(2022B02043-2);National Natural Science Foundation of China(31960541);National Natural Science Foundation of China(31460473);The Guiding Science and Technology Program Project of Xinjiang Production an Construction Corps(2022ZD010)

摘要/Abstract

摘要：

【目的】建立双斑长跗萤叶甲成虫触角转录组数据库,筛选雌虫、雄虫触角中的差异表达基因并解析其涉及的生理功能,为进一步研究双斑长跗萤叶甲的基因功能分析及嗅觉感受机制奠定分子基础。【方法】采用Illumina高通量测序技术对双斑长跗萤叶甲雌虫、雄虫触角进行转录组测序,对获得的unigenes进行功能注释、通路富集等生物信息学分析,利用RT-qPCR技术检测气味结合蛋白基因在触角中的表达水平。【结果】双斑长跗萤叶甲雌、雄成虫触角转录组测序共获得73 050条unigenes,Q30碱基百分比均在93.23%以上,34 233条unigenes至少在1个数据库中获得注释。与双斑长跗萤叶甲雌虫触角相比,雄虫触角中检测到395个差异表达基因,其中288个基因上调表达,107个基因下调表达。【结论】在GO注释库中,差异表达基因富集最显著的功能为气味结合、嗅觉受体活性和嗅觉感知。气味结合蛋白基因在雌虫、雄虫触角中的相对表达水平趋势与转录组测序结果相符合,获得了双斑长跗萤叶甲成虫触角转录组数据库。

关键词: 双斑长跗萤叶甲; 触角转录组; 功能注释; 差异表达基因

Abstract:

【Objective】 To establish the transcriptome database of adult Monolepta signata antennae, screen the differentially expressed genes in the antennae of female and male M. signata and analyze the physiological functions of these genes. 【Methods】 In this study, Illumina high-throughput sequencing technology was used to conduct transcriptome sequencing on the antennae of male and female M. signata and analyze the functional annotation and pathway enrichment of unigenes, and after that the expression level of odorant binding protein genes in antennae was detected by RT-qPCR. 【Results】 A total of 73,050 unigenes were obtained from the antennae transcriptome sequencing of female and male of adult M. signata, with the percentage of Q30 bases above 93.23, and 34,233 unigenes were annotated in at least one database (46.86%). A total of 395 differentially expressed genes were detected in the antennae of male M. signata compared with that of female M. signata. Among them, 288 genes were up-regulated and 107 genes were down-regulated.【Conclusion】 In the GO database, the most significant function of enrichment of these differentially expressed genes were odorant binding, olfactory receptor activity, and sensory perception of smell. RT-qPCR results showed that the relative expression levels of odorant binding protein genes in antennae of female and male M. signata were consistent with the transcriptome sequencing results. The transcriptome database of the antenna of adult M. signata is successfully obtained.

Key words: Monolepta signata; antennae transcriptome; functional annotation; differentially expressed genes

中图分类号:

S433

何婉洁, 孟涵颖, 支梦婷, 陈静. 双斑长跗萤叶甲雌虫、雄虫触角转录组及差异表达基因分析[J]. 新疆农业科学, 2024, 61(4): 984-995.

HE Wanjie, MENG Hanying, ZHI Mengting, CHEN Jing. Analysis of the antennal transcriptome and differentially expressed genes of female and male Monolepta signata[J]. Xinjiang Agricultural Sciences, 2024, 61(4): 984-995.

图/表 13

参考文献 45

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基因ID Gene ID	基因名称 Gene name	上游引物(5’-3’) Forward	下游引物(3’-5’) Reverse	扩增片段长度 Product length
Cluster-12689.30761	MsigOBP2	TGGTTTCAGCGAACGCAATC	TCCTTCATGACTGTCTGGTTCC	152
Cluster-12689.31568	MsigOBP3	TAGAGGCAAATGCACCACCAT	TAGAGGCAAATGCACCACCAT	194
Cluster-12689.26420	MsigOBP5	AAATGCGGATGGCTCCATGA	TCCCAGTGCTCGTCCTTCTA	199
Cluster-12689.31318	MsigOBP6	GCAACCCATTTGTTGCCTCA	TGTACGTGTGTCGGACAGTG	165
Cluster-12689.8427	MsigOBP8	CATCCATGCGCCTTTCAAGA	GCCAAAGAAGAGTGTCAAGCTG	199
Cluster-12689.26435	MsigOBP15	TGGGAATCTTAGCTGGTGGC	CACACCAATTTCATCGAAAACGC	191
Cluster-12689.34706	MsigOBP18	AGGACTCCAAAATGAAGACGGT	ACAAGGACACGTTTGGGGTT	189
Cluster-12689.31354	MsigOBP20	ACGGTTGCGTCGTCTATTCC	AAGATCCAGAAACACGAGCGG	181
Cluster-12689.30900	MsigOBP21	TGGACACCAAGGCATCTCAC	TCTGAGGCGCCTGAATAACA	196

基因ID Gene ID	基因名称 Gene name	上游引物(5’-3’) Forward	下游引物(3’-5’) Reverse	扩增片段长度 Product length
Cluster-12689.30761	MsigOBP2	TGGTTTCAGCGAACGCAATC	TCCTTCATGACTGTCTGGTTCC	152
Cluster-12689.31568	MsigOBP3	TAGAGGCAAATGCACCACCAT	TAGAGGCAAATGCACCACCAT	194
Cluster-12689.26420	MsigOBP5	AAATGCGGATGGCTCCATGA	TCCCAGTGCTCGTCCTTCTA	199
Cluster-12689.31318	MsigOBP6	GCAACCCATTTGTTGCCTCA	TGTACGTGTGTCGGACAGTG	165
Cluster-12689.8427	MsigOBP8	CATCCATGCGCCTTTCAAGA	GCCAAAGAAGAGTGTCAAGCTG	199
Cluster-12689.26435	MsigOBP15	TGGGAATCTTAGCTGGTGGC	CACACCAATTTCATCGAAAACGC	191
Cluster-12689.34706	MsigOBP18	AGGACTCCAAAATGAAGACGGT	ACAAGGACACGTTTGGGGTT	189
Cluster-12689.31354	MsigOBP20	ACGGTTGCGTCGTCTATTCC	AAGATCCAGAAACACGAGCGG	181
Cluster-12689.30900	MsigOBP21	TGGACACCAAGGCATCTCAC	TCTGAGGCGCCTGAATAACA	196

样品 Sample	原始读数 Raw reads	筛选后读数 Clean reads	总核酸数(G) Clean bases	错误率 Error rate (%)	Q₂₀ (%)	Q₃₀ (%)	GC比值 GC content (%)
FA₁	21 774 061	20 648 522	6.2	0.03	97.79	93.3	34.16
FA₂	20 757 300	20 132 668	6.0	0.03	97.88	93.58	36.30
FA₃	22 134 770	21 179 365	6.4	0.03	98.07	93.99	35.45
MA₁	21 323 904	20 130 023	6.0	0.03	97.82	93.43	35.49
MA₂	20 885 385	20 454 004	6.1	0.03	97.75	93.23	33.98
MA₃	21 288 624	20 829 381	6.2	0.03	97.83	93.42	34.95

样品 Sample	原始读数 Raw reads	筛选后读数 Clean reads	总核酸数(G) Clean bases	错误率 Error rate (%)	Q₂₀ (%)	Q₃₀ (%)	GC比值 GC content (%)
FA₁	21 774 061	20 648 522	6.2	0.03	97.79	93.3	34.16
FA₂	20 757 300	20 132 668	6.0	0.03	97.88	93.58	36.30
FA₃	22 134 770	21 179 365	6.4	0.03	98.07	93.99	35.45
MA₁	21 323 904	20 130 023	6.0	0.03	97.82	93.43	35.49
MA₂	20 885 385	20 454 004	6.1	0.03	97.75	93.23	33.98
MA₃	21 288 624	20 829 381	6.2	0.03	97.83	93.42	34.95

长度范围 Length range (bp)	Transcripts		Unigenes
长度范围 Length range (bp)	数量 Number (个)	百分比 Percentage (%)	数量 Number (个)	百分比 Percentage (%)
300~500	53 300	33.39	29 009	39.71
500~1k	42 661	26.72	21 009	28.76
1k~2k	31 535	19.75	12 325	16.87
>2k	32 162	20.14	10 707	14.66
总数Total number	159 658		73 050
总长度Total length	216 704 872		83 267 839
平均长度Mean length	1 357		1 140
N₅₀长度N₅₀ length	2 339		1 896
N₉₀长度N₉₀ length	516		440

双斑长跗萤叶甲雌虫、雄虫触角转录组及差异表达基因分析

Analysis of the antennal transcriptome and differentially expressed genes of female and male Monolepta signata

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Metrics

数据库 Database	基因数量 Number of unigenes (个)	百分比 Percentage (%)
NR	29 403	40.25
NT	10 676	14.61
KEGG	10 014	13.70
Swiss-Prot	15 487	21.20
PFAM	19 137	26.19
GO	19 135	26.19
KOG	7 554	10.34
所有数据库均有注释 Annotated in all databases	3 475	4.75
至少有一个数据库注释 Annotated in at least one database	34 233	46.86
总基因数 Total unigenes	73 050

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