管纹艳虎天牛UDP-葡萄糖基转移酶基因的鉴定及表达特征分析 *

应用昆虫学报, 2020, 57(4): 898-910 doi: 10.7679/j.issn.2095-1353.2020.092

研究论文

管纹艳虎天牛UDP-葡萄糖基转移酶基因的鉴定及表达特征分析 *

王政全,**, 尹宁娜, 赵宁, 刘乃勇,***

西南林业大学,云南省森林灾害预警与控制重点实验室,昆明 650224

Identification and expression characterization of UDP-glucosyltransferase genes in Rhaphuma horsfieldi

WANG Zheng-Quan,**, YIN Ning-Na, ZHAO Ning, LIU Nai-Yong,***

Key Laboratory of Forest Disaster Warning and Control of Yunnan Province, Southwest Forestry University, Kunming 650224, China

通讯作者: *** E-mail:Naiyong_2013@163.com

**第一作者First author,E-mail: 1835949501@qq.com

收稿日期: 2019-11-11   接受日期: 2020-02-12   网络出版日期: 2020-07-27

基金资助: *云南省科技厅青年项目.  2017FD101
云南省“高层次人才支持培养计划”青年拔尖人才项目.  YNWR-QNBJ-2019-057
云南省教育厅科学研究基金项目.  2019Y0141

Corresponding authors: *** E-mail:Naiyong_2013@163.com

Received: 2019-11-11   Accepted: 2020-02-12   Online: 2020-07-27

摘要

【目的】 昆虫UDP-葡萄糖基转移酶(UGT)在其内源性和外源性有毒化合物的解毒代谢过程中起着重要作用,但在管纹艳虎天牛Rhaphuma horsfieldi中UGT基因的研究尚属空白。【方法】 采用转录组学、生物信息学、进化和表达谱分析、蛋白同源建模等技术研究了管纹艳虎天牛的UGT基因家族。【结果】 从管纹艳虎天牛转录组中一共鉴定到36个RhorUGTs基因,其中17个具有全长序列。鞘翅目不同种间UGT基因数量的比较表明,管纹艳虎天牛具有中等数量的UGT基因。进化分析结果表明,RhorUGTs分布在10个亚家族中,其中UGT352亚家族为天牛科昆虫所特有,且该亚家族中RhorUGT2/7/10/16/18/27可能通过基因的复制产生。表达谱结果表明,大部分RhorUGTs基因在检测的所有组织中均有表达,部分基因呈现触角或跗节特异或高表达的特点,暗示它们具有嗅觉或触觉等功能。三级结构分析发现,RhorUGT17的α3、α4、β4和β5主要参与UDP-葡萄糖的结合。【结论】 本研究明确了管纹艳虎天牛UGT基因的数量、序列特征、进化关系及组织表达特征,研究结果为该种天牛解毒代谢机制的阐明奠定基础。

关键词: 管纹艳虎天牛 ; UDP-葡萄糖基转移酶 ; 进化分析 ; 表达谱 ; 同源建模

Abstract

[Objectives] UDP-glucosyltransferase (UGT) in insects is of particular importance for the detoxification of endogenous and exogenous toxic compounds. However, no information about RhorUGT genes of Rhaphuma horsfieldi is available. [Methods] In this study, we characterized the UGT gene family from R. horsfieldi using transcriptomics, bioinformatics, phylogenic and expression profiling analyses, and homology modeling approaches. [Results] Based on the transcriptomic data, totally 36 RhorUGT genes were identified from this beetle, 17 of which were full-length sequences. A comparative study on UGT gene numbers among different coleopteran species revealed that R. horsfieldi harbored a medium UGT number. Phylogenetic analysis indicated that RhorUGTs were clustered into ten sub-families, with a cerambycid-specific one of UGT352. Notably, these members formed by RhorUGT2/7/10/16/18/27 from UGT352 were possibly derived from gene duplications. Expression profiles showed that most of RhorUGTs were detected in tested tissues and some were specifically or highly expressed in antennae or tarsi, suggesting their diverse functions including olfaction and touch. Results by three-dimensional structures showed that α3, α4, β4 and β5 of RhorUGT17 were mainly involved in the binding of UDP-glucose. [Conclusion] This study has identified the numbers, sequence characteristics, phylogenic relationships and tissue expression characteristics of R. horsfieldi UGTs, and thus provides reference data for addressing the detoxification mechanisms of this beetle.

Keywords: Rhaphuma horsfieldi ; UDP-glycosyltransferase ; phylogenetic analysis ; expression profile ; homology modeling

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王政全, 尹宁娜, 赵宁, 刘乃勇. 管纹艳虎天牛UDP-葡萄糖基转移酶基因的鉴定及表达特征分析 *. 应用昆虫学报[J], 2020, 57(4): 898-910 doi:10.7679/j.issn.2095-1353.2020.092

WANG Zheng-Quan, YIN Ning-Na, ZHAO Ning, LIU Nai-Yong. Identification and expression characterization of UDP-glucosyltransferase genes in Rhaphuma horsfieldi. Chinese Journal of Applied Entomology[J], 2020, 57(4): 898-910 doi:10.7679/j.issn.2095-1353.2020.092

管纹艳虎天牛Rhaphuma horsfieldi是隶属于鞘翅目Coleoptera天牛科Cerambycidae艳虎天牛属Rhaphuma的一种重要蛀干害虫,主要为害香须树Albizia odoratissima、木姜子属Litsea、合欢属Albizia、胡桃属Juglans等树木。该种害虫在国内主要分布于四川和云南,国外主要分布在印度、缅甸、越南和老挝等地区(云南省林业厅和中国科学院动物研究所,1987)。管纹艳虎天牛主要以幼虫钻蛀到树干木质部进行为害,直至化蛹;每年5-7月份,蛹羽化为成虫,随即进行交配和产卵(尹宁娜等,2019)。目前,针对管纹艳虎天牛尚未有可行且有效的防治措施。

UDP-葡萄糖基转移酶(UDP- glycosyltransferase,UGT)是一类广泛存在于脊椎动物、昆虫、植物、真菌以及细菌中的多功能超家族酶(Mackenzie et al.,1997;Meech and Mackenzie,1997;Bock,2016)。昆虫的UGT主要由一个可变的N端和一个高度保守的C端组成,在N端通常具有20个氨基酸左右的信号肽序列,其主要负责结合糖苷配基;C端具有参与UDP-葡萄糖结合的两个供体结合域、多个关键氨基酸位点、一个跨膜区域和一个胞内尾区(Meech and Mackenzie,1997;黄飞飞等,2009;Ahn et al.,2012;Wang et al.,2018)。与植物UGT作为胞质蛋白不同(Osmani et al.,2009),昆虫UGT是一种特异性的膜结合蛋白,位于内质网膜上,其蛋白质的主体部分镶嵌在内质网腔内,使得UGT的纯化较为困难(娄琳琳,2014;张书恒,2016)。

昆虫UGT基因具有多样性的组织和发育表达特征,暗示其功能的多样性。家蚕Bombyx moriHuang et al.,2008;Ahn et al.,2012)和黑腹果蝇Drosophila melanogasterAhn et al.,2012;Younus et al.,2014)的UGT基因在成虫和幼虫的不同组织中均有表达。此外,研究表明昆虫的部分UGT基因具有触角特异或高表达的特点,说明其在昆虫的嗅觉感受中具有重要作用。在黑腹果蝇中,4个UGT基因(DmelUGT35aUGT35bUGT36BcUGT86Da)在成虫触角高表达(Wang et al.,1999;Younus et al.,2014);类似的结果在家蚕(Huang et al.,2008)、二点委夜蛾Athesis lepigoneZhang et al.,2017)和小菜蛾Plutella xylostellaHe et al.,2017)等鳞翅目昆虫中也有发现。值得注意的是,海灰翅夜蛾Spodoptera littoralis在分别经性信息素、植物气味和杀虫剂处理后,其触角高表达的SlitUGT40R3SlitUGT46A6基因的表达水平显著上调,暗示它们在其嗅觉感受和杀虫剂降解方面具有重要作用(Bozzolan et al.,2014)。在同属的甜菜夜蛾S. exigua中,脂肪体中多个SexiUGTs基因的表达呈现杀虫剂依赖性(Hu et al.,2019)。在鞘翅目昆虫中,暗黑鳃金龟Holotrichia parallelaHparUGTs基因在包括触角在内的多个组织中有表达(Wang et al.,2018);采用RNAi技术证实马铃薯甲虫Leptinotarsa decemlineata对吡虫啉的抗性与LdecUGT2基因有关(Kaplanoglu et al.,2017)。

管纹艳虎天牛是一种重要的蛀干害虫,Wickham等(2014)采用10种已知的天牛性信息素作为诱芯在云南林间进行诱捕试验,发现2,3-Hexanediol和(2R*,3R*)-2,3-Octanediol能够诱捕到大量管纹艳虎天牛。截至目前,针对该种害虫的研究主要集中在外部形态特征及感器类型的描述(蒋书南等,1985;尹宁娜等,2019),其与寄主互作或自身生理特性的研究仍属空白。本研究基于测序的转录组数据,采用生物信息学、转录组学、进化分析和蛋白同源建模等方法研究了管纹艳虎天牛中参与有毒化合物降解的重要解毒酶:UGT。研究结果不仅可以明确管纹艳虎天牛中参与化合物解毒代谢的UGT基因,还可为该种天牛的防治提供理论基础。

1 材料与方法

1.1 供试昆虫

管纹艳虎天牛采自云南省楚雄州大姚县三台乡(N26°00′01.6″,E101°04′04.7″),海拔1 999 m。在野外采集带有天牛产卵刻痕的受害核桃木段至实验室内,木段两端用海绵包裹,并使海绵始终处于湿润状态,以保持受害核桃木段水分。待翌年成虫羽化后,分开收集雌雄成虫触角、跗节和不含触角和跗节的身体,液氮速冻后存于﹣80 ℃用于转录组测序。

1.2 实验方法

1.2.1 总RNA的提取 采用TRIzol试剂盒(Ambion, Life Technologies, Carlsbad, CA)提取管纹艳虎天牛各组织总RNA,具体操作步骤见TRIzol试剂盒说明书。提取的总RNA用焦炭酸二乙酯(Diethyl pyrocarbonate,DEPC)水溶解后,取2 μL用NanoDrop 1000分光光度计(Thermo Fisher Scientific, San Jose, CA)测定RNA浓度和质量。每个组织样品取至少2 μg送至北京诺禾致源科技股份有限公司进行转录组测序,每个样品测序2次重复。

1.2.2 UGT基因的鉴定和比较 基于测序的管纹艳虎天牛转录组数据,采用BLAST同源搜索的方法鉴定其UGT基因。首先,从NCBI(National Center for Biotechnology Information)数据库及已发表文献中收集赤拟谷盗Tribolium castaneum、光肩星天牛Anoplophora glabripennis、马铃薯甲虫等鞘翅目昆虫的UGT蛋白序列。其次,将已测序的转录组导入BioEdit软件中,采用BLAST同源搜索的方法鉴定候选的RhorUGTs基因。然后,将已鉴定的所有UGT基因在NCBI NR蛋白序列数据库(Non- redundant protein sequence database)中进行BLAST比对验证,进一步确定候选的RhorUGTs基因是否属于UGT基因家族。

基于已发表的鞘翅目昆虫的UGT基因,收集并比较鞘翅目昆虫UGT不同亚家族间基因数量及其差异,其中鞘翅目昆虫UGT不同亚家族根据赤拟谷盗的UGT进行划分(Ahn et al.,2012)。

1.2.3 UGT基因的表达水平分析 以拼接得到的转录组作为参考序列,采用Bowtie软件将测序获得的Clean reads比对到Unigenes数据库,使用RSEM软件得到每个样品比对到每个基因上的Read count数目,并对其进行FPKM(Fragments Per Kilobase of transcript sequence per Millions base pairs sequenced)值转换,据此分析RhorUGTs基因的表达水平(Trapnell et al.,2010)。根据转录组中RhorUGTs基因的编号,在FPKM值数据表格中找到对应的基因和该基因在同一组织中2次重复的FPKM值,然后用2次重复的FPKM平均值绘制不同RhorUGTs基因的HeatMap表达谱。

1.2.4 UGT基因的序列及进化分析 利用NCBI上的Open reading frame finder(ORF finder)预测RhorUGTs的开放阅读框(https://www.ncbi.nlm.nih.gov/orffinder/)。理论等电点(Isoelectric point,pI)和分子量(Molecular weight,Mw)由在线工具Compute pI/Mw(https://web.expasy.org/compute_pi/)计算。采用SignalP 4.1 Server在线软件预测UGT的信号肽(Petersen et al.,2011)。采用NetNGlyc 1.0 Server在线软件(http://www.cbs.dtu.dk/services/netnglyc/)预测UGT的N-糖基化位点(N-glycosylation predicted sites,NPS)。UGT序列间的氨基酸一致性用GeneDoc进行计算。用MAFFT v7.388进行UGT的氨基酸多序列比对(Katoh and Standley,2013)。在进化树分析中,选取光肩星天牛、中欧山松大小蠹Dendroctonus ponderosae、赤拟谷盗和管纹艳虎天牛的UGT序列,利用PhyML 3.0软件最大似然法(Maximum-likelihood method)构建进化树(Guindon et al.,2010)。

1.2.5 RhorUGT17蛋白的同源建模 首先,选取具有完整ORF的17个RhorUGTs(RhorUGT1-17),采用ClustalW2进行序列比对后截取RhorUGTs的C端保守区域(Larkin et al.,2007),采用GeneDoc分别计算它们与人类Homo sapiens HsapUGT2B7序列的氨基酸一致性;然后,以HsapUGT2B7蛋白的晶体结构(PDB:2O6L)(Miley et al.,2007)为模板,选取一致性最高的RhorUGT17构建其三级结构。蛋白三级结构同源构建采用MODELLER 9v7软件进行(Sali and Blundell,1993),三级结构编辑和可视化采用PyMOL软件(https://pymol.org/)进行。

2 结果与分析

2.1 管纹艳虎天牛UGT基因的鉴定

基于管纹艳虎天牛的转录组,通过BLAST同源搜索的方法共鉴定到36个UGT基因。其中,17个UGT基因(RhorUGT1-17)具有完整的ORF,编码514-534个氨基酸,其余基因为片段,编码184-505个氨基酸。信号肽预测结果表明,17个具有全长ORF的UGT蛋白在N端均具有信号肽序列,长度为15-24个氨基酸。等电点和分子量预测结果表明,除RhorUGT15为中性外,其余16个RhorUGTs均偏碱性;RhorUGT1-17分子量大小在58.12-60.82 ku之间。NCBI BLAST比对结果显示,所有RhorUGTs均比对到鞘翅目其他昆虫的UGT,且具有较高的氨基酸一致性(52%- 82%),其中34个RhorUGTs比对到光肩星天牛的AglaUGTs;RhorUGT17与光肩星天牛AglaUGT2C1- like的氨基酸一致性最高,为82%,其次是RhorUGT35与AglaUGT2B31,为77%(表1)。

表1   管纹艳虎天牛UGT基因信息

Table 1  Information for UGT genes in Rhaphuma horsfieldi

新窗口打开| 下载CSV


基于已发表的鞘翅目昆虫的UGT基因,比较了不同物种间UGT亚家族基因的数量差异。结果表明,不同物种间UGT基因数量差异较大,其中光肩星天牛的UGT基因数量最多,为58个;黄曲条跳甲Phyllotreta striolata最少,仅有8个;管纹艳虎天牛具有中等数量的UGT基因,为36个。在不同亚家族基因数量的比较中,所有物种均只有一个UGT50基因;管纹艳虎天牛的UGT基因数量在不同亚家族中的比例与光肩星天牛相似,两个物种在UGT352亚家族中均具有最多的UGT基因,而在UGT331亚家族中则丢失了该类基因(表2)。

表2   鞘翅目昆虫UGT不同亚家族基因的数量比较

Table 2  Comparison of UGT gene numbers of different sub-families in Coleoptera

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2.2 管纹艳虎天牛UGT基因的序列及进化分析

与其他昆虫的UGT相似,管纹艳虎天牛的UGT也包括两个主要的功能区域:一个可变的N端和一个高度保守的C端。N端区域主要包括信号肽序列以及起催化作用的组氨酸(H)和天冬氨酸(D);C端区域主要由两个供体结合区域(Donor binding regions,DBRs)和一个高度保守的特征性基序组成。此外,在C端末区还具有一个由15个氨基酸组成的跨膜结构域和一段细胞质尾区,在跨膜域前具有一个带有负电荷的天冬氨酸;在DBRs中具有8个与糖源相互作用的关键氨基酸残基,包括与核苷酸互作的丝氨酸(S)、色氨酸(W)、谷氨酰胺(Q)和谷氨酸(E),与含磷化合物互作的苏氨酸(T)/S和Q/H以及与糖苷互作的D和Q/H(图1)。

图1

图1   管纹艳虎天牛UGT的氨基酸序列比对

三角形表示起催化作用的组氨酸(H)和天冬氨酸(D);1、2和3分别表示与核苷酸、含磷化合物和糖苷作用的氨基酸。

Fig. 1   Multiple alignment of UGT amino acid sequences in Rhaphuma horsfieldi

Triangle represents key catalytic residues (H, histidine and D, aspartic acid). 1, 2 and 3 represent nucleotide, phosphate and glucoside interacting residues, respectively.


选取管纹艳虎天牛、光肩星天牛、中欧山松大小蠹和赤拟谷盗共4种鞘翅目昆虫的UGT构建进化树。结果表明,鞘翅目昆虫的UGT可划分为10个亚家族:UGT50、UGT311/312、UGT319/320/321、UGT323、UGT324、UGT325、UGT326/327/347、UGT328、UGT331和UGT352,其中UGT311/312、UGT319/320/321和UGT326/327/347三个亚家族由混合UGT组成;UGT352仅包括天牛科昆虫的UGT;4个物种在UGT50中各自有1个UGT。除UGT331外,RhorUGTs聚类到其余9个亚家族中,其中UGT352、UGT319/320/321和UGT324数量最多,分别为有10个、9个和8个;其余亚家族UGT数量较少,仅为1-2个。此外,进化分析发现UGT成员主要以种特异性的形式聚类,包括UGT311/312、UGT323、UGT324、UGT325、UGT326/327/347、UGT328和UGT352(图2)。

图2

图2   鞘翅目昆虫UGT的进化树

Fig. 2   Phylogenetic tree of UGTs in Coleoptera


2.3 管纹艳虎天牛UGT基因的组织表达谱

根据转录组测序的FPKM值,研究了36个RhorUGTs基因在雌雄虫触角、跗节以及不含触角和跗节的身体等组织中的表达情况。结果表明,所有RhorUGTs基因在检测的一至多个组织中有表达;大部分基因(21个)在检测的所有组织中均有不同程度的表达;部分基因呈现组织特异表达的特点,如RhorUGT1RhorUGT2RhorUGT4RhorUGT6基因在雌虫跗节特异表达,RhorUGT8RhorUGT9RhorUGT15RhorUGT18RhorUGT30基因仅在雌雄虫跗节有表达;部分基因呈现组织高表达的特点,如RhorUGT23基因在雌虫跗节高表达,RhorUGT33基因在雌虫不含触角和跗节的身体高表达。另外,部分基因在雌雄虫触角中具有较高表达,包括RhorUGT11RhorUGT12RhorUGT14RhorUGT28RhorUGT32RhorUGT36基因(图3)。

图3

图3   管纹艳虎天牛RhorUGTs基因在成虫不同组织的表达谱

FA:成虫触角;FTa:雌虫跗节;FB:不含触角和跗节的身体;MA:雄虫触角;MTa:雄虫跗节;MB:不含触角和跗节的身体。

Fig. 3   Expression pattern of RhorUGT genes in different tissues of Rhaphuma horsfieldi adults

FA: Female antennae; FTa: Female tarsi; FB: Female bodies without antennae and tarsi; MA: Male antennae; MTa: Male tarsi; MB: Male bodies without antennae and tarsi.


2.4 RhorUGT17三级结构建模及分析

以HsapUGT2B7的晶体结构为模板,采用同源建模的方法构建了RhorUGT17的三级结构。首先,根据HsapUGT2B7晶体结构的氨基酸序列和其与RhorUGT17的序列比对结果,截取RhorUGT17相应C端区域的170个氨基酸,氨基酸序列比对结果表明两者的一致性为42%。两个供体结合域(DBR1和DBR2)具有较高的保守性。除与糖苷互作的Q和H不同外,其余7个关键氨基酸位点完全相同(图4:A)。

图4

图4   管纹艳虎天牛RhorUGT17的三级结构

A. 管纹艳虎天牛RhorUGT17与人HsapUGT2B7的氨基酸序列比对;B. 管纹艳虎天牛RhorUGT17的三级结构(左边)及其与HsapUGT2B7的三级结构叠加(右边)。两个蛋白的关键氨基酸(S、W、Q、T、H、E、D和H/Q)和UDP-葡萄糖结合腔在结构中被标示;供体结合区域(DBR1和DBR2)用黄色标示;Nt、Ct、α1-7和β1-6:分别表示氨基酸端、羧基端、α螺旋1至7和β折叠1至6。

Fig. 4   Three-dimensional (3D) structure of Rhaphuma horsfieldi UGT17

A. Alignment of RhorUGT17 and HsapUGT2B7 amino acid sequences; B. 3D model of RhorUGT17 (left) and superimposition of RhorUGT17 and HsapUGT2B7 structures (right). Key residues (S, W, Q, T, H, E, D and H/Q) and UDP-glucose binding cavities of RhorUGT17 and HsapUGT2B7 are labeled on the structures, respectively. Donor binding domains (DBR1 and DBR2) are highlighted in yellow. Nt, Ct, α1-7 and β1-6 represent N-terminus, C-terminus, seven α-helixes and six β-sheets, respectively.


同源建模结果表明,RhorUGT17由N端、C 端、loop环、7个α螺旋(α1-7)和6个β折叠(β1-6)组成。此外,为了比较HsapUGT2B7和RhorUGT17的三级结构差异,进一步将它们的结构进行叠加。结果发现,两个蛋白的α2、α4、α5、β3、β6、β1-α2间loop、β2-β3间loop和β6-α6间loop差异较大;RhorUGT17的UDP-葡萄糖结合腔主要由α3、α4、β4和β5构成,与核酸、含磷化合物和糖苷作用的关键氨基酸均位于结合腔附近(图4:B)。

3 结论与讨论

管纹艳虎天牛是三台核桃上新发现的一种重要蛀干害虫,该种天牛的卵、幼虫、蛹和成虫部分时期都生活在树皮下或是树干内,因此其长期遭受各种有毒化合物(包括内源性化合物)的侵袭;此外,化学农药的使用进一步加剧了该种天牛的生存环境,增加了其与各种有毒化合物接触的几率。在长期的自然选择压力和人类活动的影响下,管纹艳虎天牛已进化出一套特有的解毒代谢酶系统,UGT基因家族即为该系统中一类重要的解毒代谢酶家族,主要参与外源性和内源性有毒化合物的生物转化过程,在昆虫生存和生境适应性中具有重要作用。基于测序的转录组数据,从管纹艳虎天牛中一共鉴定到36个UGT基因,其数量与粪金龟UGT基因数量(38个)相近,明显多于中欧山松大小蠹(21个)(Keeling et al.,2013)、暗黑鳃金龟(20个)(Wang et al.,2018)、圣甲虫(16个)(Meyer et al.,2016)、黄曲条跳甲(8个)(Wu et al.,2016)等鞘翅目昆虫和二点委夜蛾(20个)(Zhang et al.,2017)、海灰翅夜蛾(11个)(Bozzolan et al.,2014)等非鞘翅目昆虫,但是少于光肩星天牛(58个)(McKenna et al.,2016)、蜂房小甲虫(52个)(Evans et al.,2018)、玉米根萤叶甲(50个)、赤拟谷盗(47个)(Ahn et al.,2012)等鞘翅目昆虫以及家蚕(45个)(Huang et al.,2008;Ahn et al.,2012)、棉铃虫(46个)(Ahn et al.,2012;Pearce et al.,2017)和豌豆蚜Acyrthosiphon pisum(72个)(Ahn et al.,2012;McKenna et al.,2016)等非鞘翅目昆虫,这可能与不同昆虫转录组的测序组织数量、类型、测序技术和深度不同有关;而数量较多的物种的UGT则从基因组中鉴定。

在序列比对和分析中,超过90%的RhorUGTs比对到光肩星天牛的AglaUGTs(McKenna et al.,2016),且均具有较高的氨基酸一致性(>50%),说明UGT在同科不同种间具有较高的保守性,其中以UGT50亚家族成员间一致性最高(平均值为72%),暗示其功能最为保守(Ahn et al.,2012)。与其他昆虫的UGT类似(Ahn et al.,2012;Zhang et al.,2017;Wang et al.,2018),管纹艳虎天牛的UGT在C端也呈现出较高的氨基酸一致性,尤其是在供体结合区域内的关键氨基酸高度保守,进一步支持UGT基因功能的保守性。在进化分析中,UGT352亚家族仅含有天牛科昆虫的UGT,说明该分支为天牛科昆虫特有;进一步分析发现,聚类到该亚家族的光肩星天牛的AglaUGTs主要分布在2个Scaffold上(Scaffold 72和Scaffold 772),且呈邻近排列和物种特异聚类的特点,暗示这些基因很可能起源于基因的复制(McKenna et al.,2016)。在管纹艳虎天牛中,6个RhorUGTs(RhorUGT2/7/10/16/18/27)单独聚类到UGT352亚家族的一个小分支,说明它们可能在同一条染色体上邻近排列,且通过基因的复制产生。

基因的表达谱在一定程度上可以反映其特定的功能,是筛选组织特异或决定功能研究候选基因的关键步骤。与其他大部分昆虫UGT基因的表达特征类似(Huang et al.,2008;Ahn et al.,2012;Younus et al.,2014;Zhang et al.,2017;Wang et al.,2018),管纹艳虎天牛的大部分RhorUGTs基因也呈现多样性的组织表达谱,说明其具有多种功能。在表达谱的研究中,虽然并未发现触角特异的RhorUGTs基因,但是部分RhorUGTs基因在触角中具有较高的表达水平,暗示它们主要具有嗅觉方面的功能。类似的UGT基因触角高表达的特征在黑腹果蝇(Wang et al.,1999;Younus et al.,2014)、海灰翅夜蛾(Bozzolan et al.,2014)、暗黑鳃金龟(Wang et al.,2018)和小菜蛾(He et al.,2017)中也有发现。值得注意的是,在管纹艳虎天牛的跗节上发现多个RhorUGTs基因呈现特异或高表达的特点,考虑到鞘翅目昆虫的跗节上也具有大量的感器和经常接触植物表面(Betz,2003;吉帅帅等,2017;尹宁娜等,2019),说明这些UGT基因可能与触角表达的UGT基因功能类似,即具有嗅觉或触觉方面的功能。

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DOI:10.1097/00008571-199708000-00001      URL     PMID:9295054      [本文引用: 1]

This review represents an update of the nomenclature system for the UDP glucuronosyltransferase gene superfamily, which is based on divergent evolution. Since the previous review in 1991, sequences of many related UDP glycosyltransferases from lower organisms have appeared in the database, which expand our database considerably. At latest count, in animals, yeast, plants and bacteria there are 110 distinct cDNAs/genes whose protein products all contain a characteristic 'signature sequence' and, thus, are regarded as members of the same superfamily. Comparison of a relatedness tree of proteins leads to the definition of 33 families. It should be emphasized that at least six cloned UDP-GlcNAc N-acetylglucosaminyltransferases are not sufficiently homologous to be included as members of this superfamily and may represent an example of convergent evolution. For naming each gene, it is recommended that the root symbol UGT for human (Ugt for mouse and Drosophila), denoting 'UDP glycosyltransferase,' be followed by an Arabic number representing the family, a letter designating the subfamily, and an Arabic numeral denoting the individual gene within the family or subfamily, e.g. 'human UGT2B4' and 'mouse Ugt2b5'. We recommend the name 'UDP glycosyltransferase' because many of the proteins do not preferentially use UDP glucuronic acid, or their nucleotide sugar preference is unknown. Whereas the gene is italicized, the corresponding cDNA, transcript, protein and enzyme activity should be written with upper-case letters and without italics, e.g. 'human or mouse UGT1A1.' The UGT1 gene (spanning > 500 kb) contains at least 12 promoters/first exons, which can be spliced and joined with common exons 2 through 5, leading to different N-terminal halves but identical C-terminal halves of the gene products; in this scheme each first exon is regarded as a distinct gene (e.g. UGT1A1, UGT1A2, ... UGT1A12). When an orthologous gene between species cannot be identified with certainty, as occurs in the UGT2B subfamily, sequential naming of the genes is being carried out chronologically as they become characterized. We suggest that the Human Gene Nomenclature Guidelines (http://www.gene.acl.ac.uk/nomenclature/guidelines.html++ +) be used for all species other than the mouse and Drosophila. Thirty published human UGT1A1 mutant alleles responsible for clinical hyperbilirubinemias are listed herein, and given numbers following an asterisk (e.g. UGT1A1*30) consistent with the Human Gene Nomenclature Guidelines. It is anticipated that this UGT gene nomenclature system will require updating on a regular basis.

McKenna DD, Scully ED, Pauchet Y, Hoover K, Kirsch R, Geib SM, Mitchell RF, Waterhouse RM, Ahn SJ, Arsala D, Benoit JB, Blackmon H, Bledsoe T, Bowsher JH, Busch A, Calla B, Chao H, Childers AK, Childers C, Clarke DJ, Cohen L, Demuth JP, Dinh H, Doddapaneni H, Dolan A, Duan JJ, Dugan S, Friedrich M, Glastad KM, Goodisman MA, Haddad S, Han Y, Hughes DS, Ioannidis P, Johnston JS, Jones JW, Kuhn LA, Lance DR, Lee CY, Lee SL, Lin H, Lynch JA, Moczek AP, Murali SC, Muzny DM, Nelson DR, Palli SR, Panfilio KA, Pers D, Poelchau MF, Quan H, Qu J, Ray AM, Rinehart JP, Robertson HM, Roehrdanz R, Rosendale AJ, Shin S, Silva C, Torson AS, Jentzsch IM, Werren JH, Worley KC, Yocum G, Zdobnov EM, Gibbs RA, Richards S , 2016.

Genome of the Asian longhorned beetle ( Anoplophora glabripennis), a globally significant invasive species, reveals key functional and evolutionary innovations at the beetle-plant interface

Genome Biol., 17(1):227.

DOI:10.1186/s13059-016-1088-8      URL     PMID:27832824      [本文引用: 4]

BACKGROUND: Relatively little is known about the genomic basis and evolution of wood-feeding in beetles. We undertook genome sequencing and annotation, gene expression assays, studies of plant cell wall degrading enzymes, and other functional and comparative studies of the Asian longhorned beetle, Anoplophora glabripennis, a globally significant invasive species capable of inflicting severe feeding damage on many important tree species. Complementary studies of genes encoding enzymes involved in digestion of woody plant tissues or detoxification of plant allelochemicals were undertaken with the genomes of 14 additional insects, including the newly sequenced emerald ash borer and bull-headed dung beetle. RESULTS: The Asian longhorned beetle genome encodes a uniquely diverse arsenal of enzymes that can degrade the main polysaccharide networks in plant cell walls, detoxify plant allelochemicals, and otherwise facilitate feeding on woody plants. It has the metabolic plasticity needed to feed on diverse plant species, contributing to its highly invasive nature. Large expansions of chemosensory genes involved in the reception of pheromones and plant kairomones are consistent with the complexity of chemical cues it uses to find host plants and mates. CONCLUSIONS: Amplification and functional divergence of genes associated with specialized feeding on plants, including genes originally obtained via horizontal gene transfer from fungi and bacteria, contributed to the addition, expansion, and enhancement of the metabolic repertoire of the Asian longhorned beetle, certain other phytophagous beetles, and to a lesser degree, other phytophagous insects. Our results thus begin to establish a genomic basis for the evolutionary success of beetles on plants.

Meech R, Mackenzie PI , 1997.

Structure and function of uridine diphosphate glucuronosyltransferases

Clin. Exp. Pharanacol. Physiol., 24(12):907-915.

[本文引用: 2]

Meyer JM, Markov GV, Baskaran P, Herrmann M, Sommer RJ, Rödelsperger C , 2016.

Draft genome of the scarab beetle Oryctes borbonicus on La Réunion Island

Genome Biol. Evol., 8(7):2093-2105.

DOI:10.1093/gbe/evw133      URL     PMID:27289092      [本文引用: 1]

Beetles represent the largest insect order and they display extreme morphological, ecological and behavioral diversity, which makes them ideal models for evolutionary studies. Here, we present the draft genome of the scarab beetle Oryctes borbonicus, which has a more basal phylogenetic position than the two previously sequenced pest species Tribolium castaneum and Dendroctonus ponderosae providing the potential for sequence polarization. Oryctes borbonicus is endemic to La Reunion, an island located in the Indian Ocean, and is the host of the nematode Pristionchus pacificus, a well-established model organism for integrative evolutionary biology. At 518 Mb, the O. borbonicus genome is substantially larger and encodes more genes than T. castaneum and D. ponderosae We found that only 25% of the predicted genes of O. borbonicus are conserved as single copy genes across the nine investigated insect genomes, suggesting substantial gene turnover within insects. Even within beetles, up to 21% of genes are restricted to only one species, whereas most other genes have undergone lineage-specific duplications and losses. We illustrate lineage-specific duplications using detailed phylogenetic analysis of two gene families. This study serves as a reference point for insect/coleopteran genomics, although its original motivation was to find evidence for potential horizontal gene transfer (HGT) between O. borbonicus and P. pacificus The latter was previously shown to be the recipient of multiple horizontally transferred genes including some genes from insect donors. However, our study failed to provide any clear evidence for additional HGTs between the two species.

Miley MJ, Zielinska AK, Keenan JE, Bratton SM, Radominska- Pandya A, Redinbo MR , 2007.

Crystal structure of the cofactor-binding domain of the human phase II drug-metabolism enzyme UDP-glucuronosyltransferase 2B7

J. Mol. Biol., 369(2):498-511.

DOI:10.1016/j.jmb.2007.03.066      URL     PMID:17442341      [本文引用: 1]

Human UDP-glucuronosyltransferases (UGT) are the dominant phase II conjugative drug metabolism enzymes that also play a central role in processing a range of endobiotic compounds. UGTs catalyze the covalent addition of glucuronic acid sugar moieties to a host of therapeutics and environmental toxins, as well as to a variety of endogenous steroids and other signaling molecules. We report the 1.8-A resolution apo crystal structure of the UDP-glucuronic acid binding domain of human UGT isoform 2B7 (UGT2B7), which catalyzes the conjugative elimination of opioid, antiviral, and anticancer drugs. This is the first crystal structure of any region of a mammalian UGT drug metabolism enzyme. Designated UGT2B7 mutants at residues predicted to interact with the UDP-glucuronic acid cofactor exhibited significantly impaired catalytic activity, with maximum effects observed for amino acids closest to the glucuronic acid sugar transferred to the acceptor molecule. Homology modeling of UGT2B7 with related plant flavonoid glucosyltransferases indicates human UGTs share a common catalytic mechanism. Point mutations at predicted catalytic residues in UGT2B7 abrogated activity, strongly suggesting human UGTs also utilize a serine hydrolase-like catalytic mechanism to facilitate glucuronic acid transfer.

Osmani SA, Bak S, Moller BL , 2009.

Substrate specificity of plant UDP-dependent glycosyltransferases predicted from crystal structures and homology modeling

Phytochemistry, 70(3):325-347.

DOI:10.1016/j.phytochem.2008.12.009      URL     [本文引用: 1]

AbstractPlant family 1 UDP-dependent glycosyltransferases (UGTs) catalyze the glycosylation of a plethora of bioactive natural products. In Arabidopsis thaliana, 120 UGT encoding genes have been identified. The crystal-based 3D structures of four plant UGTs have recently been published. Despite low sequence conservation, the UGTs show a highly conserved secondary and tertiary structure. The sugar acceptor and sugar donor substrates of UGTs are accommodated in the cleft formed between the N- and C-terminal domains. Several regions of the primary sequence contribute to the formation of the substrate binding pocket including structurally conserved domains as well as loop regions differing both with respect to their amino acid sequence and sequence length. In this review we provide a detailed analysis of the available plant UGT crystal structures to reveal structural features determining substrate specificity. The high 3D structural conservation of the plant UGTs render homology modeling an attractive tool for structure elucidation. The accuracy and utility of UGT structures obtained by homology modeling are discussed and quantitative assessments of model quality are performed by modeling of a plant UGT for which the 3D crystal structure is known. We conclude that homology modeling offers a high degree of accuracy. Shortcomings in homology modeling are also apparent with modeling of loop regions remaining as a particularly difficult task.Graphical abstractPlant UGTs glycosylate a vide range of acceptor molecules using several different sugars. This review summarizes current knowledge of the relationship between 3D structure and substrate specificity and examines the accuracy of plant UGT structures derived from homology modeling by comparison to the crystal structures currently available.]]>

Pearce SL, Clarke DF, East PD, Elfekih S, Gordon KHJ, Jermiin LS, McGaughran A, Oakeshott JG, Papanikolaou A, Perera OP, Rane RV, Richards S, Tay WT, Walsh TK, Anderson A, Anderson CJ, Asgari S, Board PG, Bretschneider A, Campbell PM, Chertemps T, Christeller JT, Coppin CW, Downes SJ, Duan G, Farnsworth CA, Good RT, Han LB, Han YC, Hatje K, Horne I, Huang YP, Hughes DST, Jacquin-Joly E, James W, Jhangiani S, Kollmar M, Kuwar SS, Li S, Liu NY, Maibeche MT, Miller JR, Montagne N, Perry T, Qu J, Song SV, Sutton GG, Vogel H, Walenz BP, Xu W, Zhang HJ, Zou Z, Batterham P, Edwards OR, Feyereisen R, Gibbs RA, Heckel DG, McGrath A, Robin C, Scherer SE, Worley KC, Wu YD , 2017.

Genomic innovations, transcriptional plasticity and gene loss underlying the evolution and divergence of two highly polyphagous and invasive Helicoverpa pest species

BMC Biol., 15:63.

DOI:10.1186/s12915-017-0402-6      URL     PMID:28756777      [本文引用: 1]

BACKGROUND: Helicoverpa armigera and Helicoverpa zea are major caterpillar pests of Old and New World agriculture, respectively. Both, particularly H. armigera, are extremely polyphagous, and H. armigera has developed resistance to many insecticides. Here we use comparative genomics, transcriptomics and resequencing to elucidate the genetic basis for their properties as pests. RESULTS: We find that, prior to their divergence about 1.5 Mya, the H. armigera/H. zea lineage had accumulated up to more than 100 more members of specific detoxification and digestion gene families and more than 100 extra gustatory receptor genes, compared to other lepidopterans with narrower host ranges. The two genomes remain very similar in gene content and order, but H. armigera is more polymorphic overall, and H. zea has lost several detoxification genes, as well as about 50 gustatory receptor genes. It also lacks certain genes and alleles conferring insecticide resistance found in H. armigera. Non-synonymous sites in the expanded gene families above are rapidly diverging, both between paralogues and between orthologues in the two species. Whole genome transcriptomic analyses of H. armigera larvae show widely divergent responses to different host plants, including responses among many of the duplicated detoxification and digestion genes. CONCLUSIONS: The extreme polyphagy of the two heliothines is associated with extensive amplification and neofunctionalisation of genes involved in host finding and use, coupled with versatile transcriptional responses on different hosts. H. armigera's invasion of the Americas in recent years means that hybridisation could generate populations that are both locally adapted and insecticide resistant.

Petersen TN, Brunak S, von Heijne G, Nielsen H , 2011.

SignalP 4. 0: Discriminating signal peptides from transmembrane regions

Nat. Methods, 8(10):785-786.

URL     PMID:21959131      [本文引用: 1]

Sali A, Blundell TL , 1993.

Comparative protein modelling by satisfaction of spatial restraints

J. Mol. Biol., 234(3):779-815.

DOI:10.1006/jmbi.1993.1626      URL     [本文引用: 1]

Trapnell C, Williams B, Pertea G, Mortazavi A, Kwan G, van BMJ, Salzberg SL, Wold BJ, Pachter L , 2010.

Transcript assembly and quantification by RNA-Seq reveals unannotated transcripts and isoform switching during cell differentiation

Nat. Biotechnol., 28(5):511-515.

DOI:10.1038/nbt.1621      URL     PMID:20436464      [本文引用: 1]

High-throughput mRNA sequencing (RNA-Seq) promises simultaneous transcript discovery and abundance estimation. However, this would require algorithms that are not restricted by prior gene annotations and that account for alternative transcription and splicing. Here we introduce such algorithms in an open-source software program called Cufflinks. To test Cufflinks, we sequenced and analyzed >430 million paired 75-bp RNA-Seq reads from a mouse myoblast cell line over a differentiation time series. We detected 13,692 known transcripts and 3,724 previously unannotated ones, 62% of which are supported by independent expression data or by homologous genes in other species. Over the time series, 330 genes showed complete switches in the dominant transcription start site (TSS) or splice isoform, and we observed more subtle shifts in 1,304 other genes. These results suggest that Cufflinks can illuminate the substantial regulatory flexibility and complexity in even this well-studied model of muscle development and that it can improve transcriptome-based genome annotation.

Wang Q, Hasan G, Pikielny CW , 1999.

Preferential expression of biotransformation enzymes in the olfactory organs of Drosophila melanogaster, the antennae

J. Biol. Chem., 274(15):10309-10315.

DOI:10.1074/jbc.274.15.10309      URL     PMID:10187818      [本文引用: 2]

Biotransformation enzymes have been found in the olfactory epithelium of vertebrates. We now show that in Drosophila melanogaster, a UDP-glycosyltransferase (UGT), as well as a short chain dehydrogenase/reductase and a cytochrome P450 are expressed specifically or preferentially in the olfactory organs, the antennae. The evolutionarily conserved expression of biotransformation enzymes in olfactory organs suggests that they play an important role in olfaction. In addition, we describe five Drosophila UGTs belonging to two families. All five UGTs contain a putative transmembrane domain at their C terminus as is the case for vertebrate UGTs where it is required for enzymatic activity. The primary sequence of the C terminus, including part of the transmembrane domain, differs between the two families but is highly conserved not only within each Drosophila family, but also between the members of one of the Drosophila families and vertebrate UGTs. The partial overlap of the conserved primary sequence with the transmembrane domain suggests that this part of the protein is involved in specific interactions occurring at the membrane surface. The presence of different C termini in the two Drosophila families suggests that they interact with different targets, one of which is conserved between Drosophila and vertebrates.

Wang S, Liu Y, Zhou JJ, Yi JK, Pan Y, Wang J, Zhang XX, Wang JX, Yang S, Xi JH , 2018.

Identification and tissue expression profiling of candidate UDP-glycosyltransferase genes expressed in Holotrichia parallela motschulsky antennae

Bull. Entomol. Res., 108(6):807-816.

DOI:10.1017/S0007485318000068      URL     PMID:29397056      [本文引用: 5]

It is difficult to control Holotrichia parallela Motschulsky with chemical insecticides due to the larvae's soil-living habit, thus the pest has caused great economic losses in agriculture. In addition, uridine diphosphate-glycosyltransferases (UGTs) catalyze the glycosylation process of a variety of small lipophilic molecules with sugars to produce water-soluble glycosides, and play multiple roles in detoxification, endobiotic modulation, and sequestration in an insect. Some UGTs were found specifically expressed in antennae of Drosophila melanogaster and Spodoptera littoralis, and glucurono-conjugated odorants could not elicit any olfactory signals, suggesting that the UGTs may play roles in odorant inactivation by biotransformation. In the current study, we performed a genome-wide analysis of the candidate UGT family in the dark black chafer, H. parallela. Based on a UGT gene signature and the similarity of these genes to UGT homologs from other organisms, 20 putative H. parallela UGT genes were identified. Bioinformatics analysis was used to predict sequence and structural features of H. parallela UGT proteins, and revealed important domains and residues involved in sugar donor binding and catalysis by comparison with human UGT2B7. Phylogenetic analysis of these 20 UGT protein sequences revealed eight major groups, including both order-specific and conserved groups, which are common to more than one order. Of these 20 UGT genes, HparUGT1265-1, HparUGT3119, and HparUGT8312 were highly (>100-fold change) expressed in antennae, suggesting a possible role in olfactory tissue, and most likely in odorant inactivation and olfactory processing. The remaining UGT genes were expressed in all tissues (head, thorax, abdomen, leg, and wing), indicating that these UGTs likely have different biological functions. This study provides the fundamental basis for determining the function of UGTs in a highly specialized olfactory organ, the H. parallela antenna.

Wickhamm JD, Harrison RD, Lu W, Guo Z, Millar JG, Hanks LM, Chen Y , 2014.

Generic lures attract cerambycid beetles in a tropical montane rain forest in southern China

J. Econ. Entomol., 107(1):259-267.

DOI:10.1603/EC13333      URL     [本文引用: 1]

The attraction of cerambycid beetles to 10 known cerambycid pheromones was tested in a tropical montane rain forest in southern China. From 28 May to 25 June 2010, 1,526 cerambycids representing 71 species were captured in pheromone-baited traps, with 14 species accounting for 92% of the specimens. Test compounds with a 3-hydroxyalkan-2-one or 2,3-alkanediol motif attracted significant numbers of both sexes for eight species in the subfamily Cerambycinae, including species of Demonax, Rhaphuma, and Xylotrechus. Rhaphuma horsfieldi (White) was the only species that was strongly attracted to more than one test compound, with significant attraction to both (2R*, 3R*)-2,3-hexanediol and (2R*, 3R*)-2,3-octanediol. Within the Lamiinae, males and females of five species were significantly attracted to 2-(undecyloxy) ethanol, including Acalolepta formosana (Breuning), Monochamus bimaculatus Gahan, Pharsalia subgemmata (Thomson), Pseudomacrochenus antennatus (Gahan), and Xenohammus bimaculatus Schwarzer. Only male Megopis costipennis White (Prioninae) were significantly attracted to (2R*,3S*)-2,3-octanediol, suggesting that this compound may be a sex pheromone component for this species. To date, 2,3-octanediols have only been reported as aggregation pheromone components for cerambycids in the subfamily Cerambycinae. Our results support the hypothesis that both closely related (congeners) and more distantly related cerambycid species (different tribes and even subfamilies) may share pheromone components. Our results also demonstrate that traps baited with even a limited number of different classes of pheromones may be useful tools for surveying cerambycid diversity, as well as for detecting and monitoring particular species, especially those that have the potential to be invasive pests in other parts of the world.

Wu Z, Bin S, He H, Wang Z, Li M, Lin J , 2016.

Differential expression analysis of chemoreception genes in the striped flea beetle Phyllotreta striolata using a transcriptomic approach

PLoS ONE, 11(4):e0153067.

DOI:10.1371/journal.pone.0153067      URL     PMID:27064483      [本文引用: 1]

Olfactory transduction is a process by which olfactory sensory neurons (OSNs) transform odor information into neuronal electrical signals. This process begins with the binding of odor molecules to receptor proteins on olfactory receptor neuron (ORN) dendrites. The major molecular components involved in olfaction include odorant-binding proteins (OBPs), chemosensory proteins (CSPs), odorant receptors (ORs), gustatory receptors (GRs), ionotropic receptors (IRs), sensory neuron membrane proteins (SNMPs) and odorant-degrading enzymes (ODEs). More importantly, as potential molecular targets, chemosensory proteins are used to identify novel attractants or repellants for environmental-friendly pest management. In this study we analyzed the transcriptome of the flea beetle, Phyllotreta striolata (Coleoptera, Chrysomelidae), a serious pest of Brassicaceae crops, to better understand the molecular mechanisms of olfactory recognition in this pest. The analysis of transcriptomes from the antennae and terminal abdomens of specimens of both sexes identified transcripts from several key molecular components of chemoreception including 73 ORs, 36 GRs, 49 IRs, 2 SNMPs, 32 OBPs, 8 CSPs, and four candidate odorant degrading enzymes (ODEs): 143 cytochrome P450s (CYPs), 68 esterases (ESTs), 27 glutathione S-transferases (GSTs) and 8 UDP-glycosyltransferases (UGTs). Bioinformatic analyses indicated that a large number of chemosensory genes were up-regulated in the antennae. This was consistent with a potential role in olfaction. To validate the differential abundance analyses, the expression of 19 genes encoding various ORs, CSPs, and OBPs was assessed via qRT-PCR between non-chemosensory tissue and antennae. Consistent with the bioinformatic analyses, transcripts for all of the genes in the qRT-PCR subset were elevated in antennae. These findings provide the first insights into the molecular basis of chemoreception in the striped flea beetle.

Yin NN, Zhao N, Liu NY , 2019.

Morphological characteristics of Rhaphuma horsfieldi and ultrastructure of its antennae and tarsi

Journal of Southwest Forestry University (Natural Sciences), 39(3):132-140.

[本文引用: 3]

[ 尹宁娜, 赵宁, 刘乃勇 , 2019.

管纹艳虎天牛形态特征与触角及跗节的超微结构

西南林业大学学报(自然科学), 39(3):132-140.]

[本文引用: 3]

Younus F, Chertemps T, Pearce SL, Pandey G, Bozzolan F, Coppin CW, Russell RJ, MaibecheCoisne M, Oakeshott JG , 2014.

Identification of candidate odorant degrading gene/enzyme systems in the antennal transcriptome of Drosophila melanogaster

Insect Biochem. Mol. Biol., 53:30-43.

DOI:10.1016/j.ibmb.2014.07.003      URL     PMID:25038463      [本文引用: 4]

The metabolism of volatile signal molecules by odorant degrading enzymes (ODEs) is crucial to the ongoing sensitivity and specificity of chemoreception in various insects, and a few specific esterases, cytochrome P450s, glutathione S-transferases (GSTs) and UDP-glycosyltransferases (UGTs) have previously been implicated in this process. Significant progress has been made in characterizing ODEs in Lepidoptera but very little is known about them in Diptera, including in Drosophila melanogaster, a major insect model. We have therefore carried out a transcriptomic analysis of the antennae of D. melanogaster in order to identify candidate ODEs. Virgin male and female and mated female antennal transcriptomes were determined by RNAseq. As with the Lepidoptera, we found that many esterases, cytochrome P450 enzymes, GSTs and UGTs are expressed in D. melanogaster antennae. As olfactory genes generally show selective expression in the antennae, a comparison to previously published transcriptomes for other tissues has been performed, showing preferential expression in the antennae for one esterase, JHEdup, one cytochrome P450, CYP308a1, and one GST, GSTE4. These largely uncharacterized enzymes are now prime candidates for ODE functions. JHEdup was expressed heterologously and found to have high catalytic activity against a chemically diverse group of known ester odorants for this species. This is a finding consistent with an ODE although it might suggest a general role in clearing several odorants rather than a specific role in clearing a particular odorant. Our findings do not preclude the possibility of odorant degrading functions for other antennally expressed esterases, P450s, GSTs and UGTs but, if so, they suggest that these enzymes also have additional functions in other tissues.

Zhang SH , 2016.

The effect of insecticide on expression levels of UDP-glycosyltransferase in Spodoptera exigua.

Master dissertation. Nanjing: Nanjing Agricultural University.

[本文引用: 1]

[ 张书恒 , 2016.

杀虫剂处理对甜菜夜蛾尿苷二磷酸葡萄糖基转移酶基因表达的影响

硕士学位论文. 南京: 南京农业大学.]

[本文引用: 1]

Zhang YN, Ma JF, Xu L, Dong ZP, Xu JW, Li MY, Zhu XY , 2017.

Identification and expression patterns of UDP- glycosyltransferase (UGT) genes from insect pest Athetis lepigone(Lepidoptera: Noctuidae)

J. Asia-Pac. Entomol., 20(1):253-259.

DOI:10.1016/j.aspen.2017.01.008      URL     [本文引用: 4]

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