中华蜜蜂细胞色素P450基因及其全长转录本的鉴定及分析

doi:10.16380/j.kcxb.2023.01.002

昆虫学报 ›› 2023, Vol. 66 ›› Issue (1): 11-18.doi: 10.16380/j.kcxb.2023.01.002

中华蜜蜂细胞色素P450基因及其全长转录本的鉴定及分析

蔡宗兵^1,#, 王紫馨^1,#, 吴鹰¹, 王思懿¹, 钱加珺¹, 胡颖¹, 张凯遥¹, 顾小雨¹, 徐细建³，骆群³，陈大福^1,2,*, 郭睿^1,2,*

(1. 福建农林大学动物科学学院(蜂学学院), 福州 350002; 2. 福建省蜂疗研究所, 福州 350002; 3. 江西省养蜂研究所，南昌 330000)

出版日期:2023-01-20 发布日期:2023-03-04

Identification and analysis of cytochrome P450 genes and their full-length transcripts in Apis cerana cerana (Hymenoptera: Apidae)#br#

CAI Zong-Bing^1,#, WANG Zi-Xin^1,#, WU Ying¹, WANG Si-Yi¹, QIAN Jia-Jun¹, HU Ying¹, ZHANG Kai-Yao¹, GU Xiao-Yu¹, XU Xi-Jian³, LUO Qun³, CHEN Da-Fu^1,2,*, GUO Rui^1,2,*

(1. College of Animal Sciences (College of Bee Science), Fujian Agriculture and Forestry University, Fuzhou 350002, China; 2. Apitherapy Research Institute of Fujian Province, Fuzhou 350002, China; 3. Apicultural Research Institute of Jiangxi Province, Nanchang 330000, China)

Online:2023-01-20 Published:2023-03-04

摘要/Abstract

摘要： 【目的】利用纳米孔(nanopore)测序技术鉴定中华蜜蜂Apis cerana cerana工蜂幼虫肠道中细胞色素P450(cytochrome P450, CYP450)基因及其全长转录本，为后续功能研究提供参考信息和基础。【方法】通过Nanopore PromethION平台对中华蜜蜂工蜂4-6日龄幼虫肠道进行转录组测序。利用Guppy软件对原始读段(raw reads)进行质控以得到有效读段(clean reads)。通过识别两端引物鉴定全长转录本序列。使用BLAST工具将上述全长转录本的序列比对到Nr和GO数据库以鉴定CYP450基因及其全长转录本。采用Astalavista软件鉴定基因的可变剪接(alternative splicing, AS)事件。通过RTPCR验证不同类型AS事件的可靠性。【结果】在中华蜜蜂工蜂4-6日龄幼虫肠道中分别测得7 338 627, 7 003 419和7 434 233条原始读段，经质控得到的有效读段数分别为7 289 494, 6 959 880和7 387 756条。鉴定到的非冗余全长转录本总数为48 200条。共鉴定到47个CYP450基因和265条CYP450基因全长转录本。共鉴定到CYP450基因的90次AS事件，包括36次外显子跳跃事件、20次可变5′端剪接位点事件、17次内含子保留事件、9次可变3′端剪接位点事件及8次外显子互斥事件。RTPCR结果证实随机选取的3种AS事件类型真实可靠。【结论】鉴定了中华蜜蜂的CYP450基因及其全长转录本，补充了东方蜜蜂参考基因组的相关注释，并揭示中华蜜蜂CYP450基因可通过多种AS类型产生丰富的剪接体。

关键词: 中华蜜蜂, 幼虫, 第三代测序技术, 细胞色素P450, 全长转录本, 可变剪接

Abstract: 【Aim】To identify cytochrome P450 (CYP450) genes and their full-length transcripts in the larval guts of Apsi cerana cerana workers using nanopore sequencing technology and offer reference information and basis for further functional study.【Methods】The 4-6-day-old larval gut transcriptomes of A. cerana cerana workers were sequenced by Nanopore PromethION platform. Quality control of raw reads was performed using Guppy software to gain clean reads. Full-length transcript sequences were detected through recognizing primers at both ends. The sequences of the above-mentioned full-length transcripts were aligned to the Nr and GO databases with BLAST tool to identify CYP450 genes and their full-length transcripts. Astalavista software was used to identify alternative splicing (AS) events of genes. RT-PCR was used to validate the reliability of AS events of various types. 【Results】In the 4-6-day-old larval guts of A. cerana cerana workers, 7 338 627, 7 003 419 and 7 434 233 raw reads were obtained, respectively, and after quality control, 7 289 494, 6 959 880 and 7 387 756 clean reads were gained. The total number of identified non-redundant fulllength transcripts was 48 200. Forty-seven CYP450 genes and their 265 full-length transcripts were identified. A total of 90 AS events of CYP450 genes were identified, including 36 exon skipping events, 20 alternative 5′ splicing site events, 17 intron retention events, nine alternative 3′ splicing site events and eight mutually exclusive exon events. The RT-PCR result confirmed the authenticity of randomly selected three types of AS events. 【Conclusion】 The findings provide CYP450 genes and their full-length transcripts in A. cerana cerana, supplement the annotation of A. cerana reference genome, and reveal that CYP450 genes of A. cerana cerana can generate abundant isoforms via multiple AS types.

Key words: Apis cerana cerana, larvae, third-generation sequencing technique, cytochrome P450, full-length transcripts, alternative splicing

蔡宗兵, 王紫馨, 吴鹰, 王思懿, 钱加珺, 胡颖, 张凯遥, 顾小雨, 徐细建, 骆群, 陈大福, 郭睿. 中华蜜蜂细胞色素P450基因及其全长转录本的鉴定及分析[J]. 昆虫学报, 2023, 66(1): 11-18.

CAI Zong-Bing, WANG Zi-Xin, WU Ying, WANG Si-Yi, QIAN Jia-Jun, HU Ying, ZHANG Kai-Yao, GU Xiao-Yu, XU Xi-Jian, LUO Qun, CHEN Da-Fu, GUO Rui. Identification and analysis of cytochrome P450 genes and their full-length transcripts in Apis cerana cerana (Hymenoptera: Apidae)#br#[J]. Acta Entomologica Sinica, 2023, 66(1): 11-18.

[1]	王紫馨, 许雅静, 张文德, 张凯遥, 吴鹰, 刘佳美, 朱乐冉, 牛庆生, 赵红霞, 陈大福, 郭睿. ame-miR-14在意大利蜜蜂工蜂幼虫肠道发育过程的调控作用[J]. 昆虫学报, 2023, 66(1): 1-10.
[2]	曹红妹, 胡桂萍, 石旭平, 王军文, 蔡翔, 胡丽春, 王丰. 幼虫密度对桑螟生长发育和繁殖的影响[J]. 昆虫学报, 2023, 66(1): 77-84.
[3]	静大鹏, 黄晓丹, 张天涛, 王振营. 桃蛀螟CYP6AE76基因参与对Cry1Ab蛋白的解毒作用[J]. 昆虫学报, 2022, 65(9): 1136-1143.
[4]	李秋方, 高艳, 梁立强, 李正汉卿, 朱雅楠, 张娟, 杨尚宁, 傅云熙, 苏松坤, 聂红毅. 西方蜜蜂采集蜂上颚腺中高表达基因的筛选与表达分析[J]. 昆虫学报, 2022, 65(8): 937-948.
[5]	祝智威, 王杰, 隆琦, 许雅静, 冯睿蓉, 刘佳美, 赵浩东, 朱乐冉, 侯海青, 陈大福, 郭睿. 过表达和敲减ame-miR-13b对意大利蜜蜂幼虫肠道内基因表达的影响[J]. 昆虫学报, 2022, 65(4): 460-468.
[6]	成付平, 袁芳, 席芳贵, 秦凯鑫, 胡小芬, 王子龙. 中华蜜蜂精巢和卵巢差异表达基因分析[J]. 昆虫学报, 2022, 65(2): 167-175.
[7]	蔡宗兵, 张文德, 隆琦, 余岢骏, 孙明会, 吴鹰, 许雅静, 刘佳美, 郭意龙, 徐细建, 陈大福, 郭睿. 基于PacBio测序数据的蜜蜂球囊菌SNP与InDel位点发掘及分析[J]. 昆虫学报, 2022, 65(2): 187-196.
[8]	蔡宗兵, 王思懿, 王紫馨, 赵萧, 张凯遥, 郭意龙, 姚雨彤, 钱加珺, 胡颖, 付中民, 陈大福, 郭睿. 过表达和敲减ace-miR-3720对中华蜜蜂工蜂幼虫肠道的靶基因表达和重量的影响[J]. 昆虫学报, 2022, 65(10): 1247-1255.
[9]	张凯遥, 刘佳美, 张文德, 胡颖, 康育欣, 王紫馨, 王思懿, 钱加珺, 赵萧, 张佳欣, 陈大福, 郭睿, 付中民. ame-miR-79负调控意大利蜜蜂工蜂幼虫肠道中靶基因CYP450和FG的表达[J]. 昆虫学报, 2022, 65(10): 1256-1265.
[10]	寇若玫, 李月, 窦飞越, 周泽扬, 黄敦元. 大分舌蜂不同龄期幼虫及滞育预蛹肠道细菌群落多样性及其差异[J]. 昆虫学报, 2021, 64(6): 682-693.
[11]	杜宇, 蒋海宾, 王杰, 范小雪, 王秀娜, 冯睿蓉, 张文德, 隆琦, 熊翠玲, 郑燕珍, 陈大福, 郭睿. 蜜蜂球囊菌菌丝和孢子中全长转录本的差异表达分析 [J]. 昆虫学报, 2021, 64(3): 363-373.
[12]	朱江, 邱星辉. 昆虫抗药性相关细胞色素P450基因的表达调控机制[J]. 昆虫学报, 2021, 64(1): 109-120.
[13]	彭竹, 黄丽, 赵淑果, 吕建华, 赵慧婷. 中华蜜蜂气味结合蛋白AcerOBP14的表达及配体结合特性分析[J]. 昆虫学报, 2021, 64( 2): 178-186.
[14]	陈高满, 陈展博, 葛辉, 杨雪清, 王小奇. 苹果蠹蛾细胞色素P450基因CYP332A19和CYP337B19的克隆及表达分析[J]. 昆虫学报, 2020, 63(8): 941-951.
[15]	韩宝珠, 车林茸, 陈晓洁, 闫振天, 乔梁, 陈斌. 细胞色素P450基因AsCYP6Z2在中华按蚊溴氰菊酯抗性维持中的作用[J]. 昆虫学报, 2020, 63(8): 961-972.

中华蜜蜂细胞色素P450基因及其全长转录本的鉴定及分析

Identification and analysis of cytochrome P450 genes and their full-length transcripts in Apis cerana cerana (Hymenoptera: Apidae)#br#

PDF (PC)

赞

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

Metrics

本文评价

推荐阅读 0