香梨优斑螟脂肪酸Δ9去饱和酶基因序列及不同虫态的表达分析

doi:10.6048/j.issn.1001-4330.2020.05.011

新疆农业科学 ›› 2020, Vol. 57 ›› Issue (5): 877-887.DOI: 10.6048/j.issn.1001-4330.2020.05.011

香梨优斑螟脂肪酸Δ9去饱和酶基因序列及不同虫态的表达分析

王德钢^1,2, 马光皇^1,3, 贺鹏鹏^1,3, 熊仁次^2,3, 曹玉^1,3, 张萍^1,3, 韩旭^1,3, 杨明禄^1,2,3

1.塔里木大学植物科学学院,新疆阿拉尔 843300;
2.塔里木大学南疆特色果树高效优质栽培与深加工技术国家地方联合工程实验室,新疆阿拉尔,843300;
3.农业部阿拉尔作物有害生物科学观测站,新疆阿拉尔 843300

收稿日期:2019-09-24 发布日期:2020-04-23
通信作者: 杨明禄(1976-),男,副教授,硕士生导师,研究方向为农业昆虫与害虫防治,(E-mail)ymlzkytd@163.com
熊仁次(1970-),男,教授,硕士,研究方向为果树生理与高效栽培技术,(E-mail)xrcqwb@qq.com
作者简介:王德钢(1994-),男,硕士研究生,研究方向为果树病虫害与防治,(E-mail)18509998929@163.com
基金资助:
国家自然科学基金“香梨优斑螟滞育特征及FADs基因表达规律研究(31560512)”;兵团国际科技合作计划“果树食心虫扩散与防治(2017BC004)”

Gene Sequences and Expression Analysis of Fatty Acid Δ9 Desaturase of Euzophera pyriella during Different Stages

WANG Degang^1,2, MA Guanghuang^1,3, HE Pengpeng^1,3, XIONG Renci^2,3, CAO Yu^1,3, ZHANG Ping^1,3, HAN Xu^1,3, YANG Minglu^1,2,3

1.College of Plant Science,Tarim University, Alar Xinjiang 843300, China;
2.The National and Local Joint Engineering Laboratory of High Efficiency and Superior-Quality Cultivation and Fruit Deep Processing Technology of Characteristic Fruit Trees in Southern Xinjiang, Alar Xinjiang 843300, China;
3.Scientific Observing and Experiment Station of Crop Pests in Alar, Ministry of Agriculture, Alar Xinjiang 843300, China

Received:2019-09-24 Published:2020-04-23
Correspondence author: YANG Minglu (1976-), male, Master, associate professor, research field: Agricultural Entomology and Pest Control, (E-mail) ymlzkytd@163.com
XIONG Renci (1970-), male, Master, professor, research field: Physiology and efficient cultivation techniques of fruit trees, (E-mail) xrcqwb@qq.com
Supported by:
Supported by the National Natural Science Foundation of China ( 31560512) and the International Scientific and Technological Cooperation Program of XPCC ( 2017BC004)

摘要/Abstract

摘要： 【目的】研究Δ9去饱和酶基因特性及其在不同虫态下的表达,研究EpFAD9基因功能及潜在应用提供数据,为该香梨优玫玉暝、饱和脂肪酸积累中的作用提供参考。【方法】采用生物信息学软件鉴定、分析和预测Δ9去饱和酶(EpFAD9)的核苷酸和氨基酸序列,研究蛋白质结构和功能,并利用qRT-PCR方法测定△9去饱和酶基因在不同虫态下的表达水平。【结果】EpFAD9基因开放阅读框由1 056 bp组成,含352个氨基酸,起始密码子ATG位于获得序列的第1 280碱基上,终止密码子TAA位于该序列第2 338碱基。推测该蛋白无信号肽,共4个跨膜结构域,整体表现为亲水性蛋白,主要在内质网中发挥生物学作用。EpFAD9基因在幼虫期表达量较高,分别是成虫期和蛹期的17.75和29.58倍。【结论】EpFAD9基因基因在香梨优斑螟不同虫态下存在表达差异,在幼虫体内的表达量最高,成虫和蛹期的表达很低,尤其是虫蛹,其表达量更低于成虫,虫蛹和成虫状态下的香梨优斑螟对低温抵抗能力都低于幼虫。

关键词: 香梨优斑螟; 脂肪酸去饱和酶; 生物信息学; 基因表达

Abstract: 【Objective】 The Δ9 desaturase gene in Euzophera pyriella plays an important role in the accumulation of unsaturated fatty acids, so this project aims to clarify the characteristics of the gene and its expression in different insect states. 【Method】This study aimed to identify, analyze, and predict the nucleotide and amino acid sequences of Δ9 desaturase in Euzophera pyriella(EpFAD9 ) using bioinformatics software, and to deduce the structure and function of the protein. The mRNA expression of the EpFAD9 gene in different stages of E. pyriella was also determined by RT-PCR.【Result】The open reading frame of EpFAD9 gene consisted of 1,056 bp and 352 amino acids. The start codon ATG was located at the 1,280 base of the obtained sequence, and the stop codon TAA was located at the 2,338 base of the sequence. It was speculated that the protein had no signal peptide but had four transmembrane domains, which behaved as a hydrophilic protein and played a biological role in the endoplasmic reticulum. The relative expression level of EpFAD9 gene was rich in larval stage, which was 17.75 times and 29.58 times higher than those in adult and sputum stages, respectively.【Conclusion】 The expression of EpFAD9 gene is different in different Euzophera pyriella states, and the expression level in larva was the highest, while that in adult and pupa was low, the pupa stage is the lowest, this shows that the resistance of pupa and adults to low temperature is lower than that of larvae.

Key words: Euzophera pyriella; fatty acid desaturase; bioinformatics; gene expression

中图分类号:

S188

王德钢, 马光皇, 贺鹏鹏, 熊仁次, 曹玉, 张萍, 韩旭, 杨明禄. 香梨优斑螟脂肪酸Δ9去饱和酶基因序列及不同虫态的表达分析[J]. 新疆农业科学, 2020, 57(5): 877-887.

WANG Degang, MA Guanghuang, HE Pengpeng, XIONG Renci, CAO Yu, ZHANG Ping, HAN Xu, YANG Minglu. Gene Sequences and Expression Analysis of Fatty Acid Δ9 Desaturase of Euzophera pyriella during Different Stages[J]. Xinjiang Agricultural Sciences, 2020, 57(5): 877-887.

参考文献

[1] 杨集昆.库尔勒香梨优斑螟新种记述[J].华中农业大学学报,1994,13(6):560-562.
YANG Jikun. Description of a new pear pest from kurla (lepidoptera: pyralidae) [J]. Journal of Huazhong Agricultural University, 1994, 13(6): 560-562.
[2] 庞华,杨梅.香梨优斑螟的发生规律及综合防控措施[J].植物医生,2015,28(3):11-12.
PANG Hua, YANG Mei. Occurrence Regularity and Integrated Control of Euzophera pyriella[J]. Plant Doctor, 2015,28(3):11-12.
[3] 陈丽君, 杜孟芳, 安世恒, 等. 棉铃虫酰基辅酶 AΔ9 去饱和酶基因的分子鉴定[J].河南农业大学学报, 2011, 45(5): 552-557.
CHEN Lijuan, DU Mengfang, AN Shiheng, et al. Molecular identification of Helicoverpa armigera acyl-CoA delta-9 desaturase gene [J]. Journal of Henan Agricultural University, 2011, 45(5): 552-557.
[4] Pereira S L, Leonard A E, Mukerji P. Recent advances in the study of fatty acid desaturases from animals and lower eukaryotes [J]. Prostaglandins Leukotrienes & Essential Fatty Acids, 2003, 68(2):97-106.
[5] 张燕. 橘林油脂酵母△9-和△15-脂肪酸去饱和酶基因的克隆及功能验证[D]. 武汉:华中农业大学硕士论文,2011.
ZHANG Yan. Clone and Functional Verification of △9-and △15-Fatty Acid Desaturase Gene from Lipomyces kononenkoae [D]. Wuhan: Huazhong Agricultural University, 2011.
[6] Los, D. A. ＆ Murata, N. Structure and expression of fatty acid desaturases[J]. Biochimica et Biophysica Acta (BBA)-Lipids and Lipid Metabolism, 1998, 1394(1): 3-15.
[7] 郭兵. 两株海洋微藻中Δ4-脂肪酸去饱和酶基因和Δ5-脂肪酸延长酶基因的克隆以及功能验证[D]. 北京:中国农业科学院硕士论文, 2011.
GUO Bing. Cloning and Functional Charaeterication of Δ4-Fatty Acid Desaturase Gene and Δ5-Fatty Acid Elongase Gene from Two Marine Microalgae [D]. Beijing: Chinese Academy of Agricultural Sciences, 2011.
[8] Aitzetmüller, K. ＆Tsevegsüren, N. Seed fatty acids, 《front-end》-desaturases and chemotaxonomy-a case study in the Ranunculaceae [J]. Journal of Plant Physiology, 1994, 143(4-5): 538-543.
[9] Eigenheer, A. L, Young, S, Blomquist, G. J, Borgeson, C. E. Tillman, J. A. Tittiger, C. et al. Isolation and molecular characterization of Musca domestica delta‐9 desaturase sequences[J]. Insect Molecular Biology, 2002, 11(6): 533-542.
[10] Rodríguez, S, Hao, G. Liu, W. Pia, B, Rooney, A. P, Camps, F, Roelofs W. L, Fabriàs, G. et al. Expression and evolution of Δ9 and Δ11 desaturase genes in the moth Spodoptera littoralis[J]. Insect Biochemistry and Molecular Biology, 2004, 34(12): 1315-1328.
[11] Badouin, H, Belkhir, K, Gregson, E, Galindo, J. Sundström, L, Martin, S. J, Butlin, R. K, Smadja, C. M. et al. Transcriptome characterisation of the ant Formica exsecta with new insights into the evolution of desaturase genes in social hymenoptera [J]. PloS One, 2013, 8(7): e68200.
[12] Chen, Q. M, Cheng, D. J, Liu, S. P, Ma, Z. G. Tan, X. Zhao, P. et al. Genome-wide identification and expression profiling of the fatty acid desaturase gene family in the silkworm, Bombyx mori, Genet [J]. Mol. Res, 2014, 13: 3747-3760.
[13] Kayukawa, T. Chen, B. Hoshizaki, S. Ishikawa, Y. et al. Upregulation of a desaturase is associated with the enhancement of cold hardiness in the onion maggot, Delia antiqua[J]. Insect Biochemistry and Molecular Biology, 2007, 37(11): 1160-1167.
[14] Rinehart, J. P. Robich, R. M. Denlinger, D. L. et al. Isolation of diapause-regulated genes from the flesh fly, Sarcophaga crassipalpis by suppressive subtractive hybridization [J]. Journal of Insect Physiology, 2010, 56(6): 603-609.
[15] Reynolds, J. A. Poelchau, M. F. Rahman, Z. Armbruster, P. A. Denlinger, D. L. et al.Transcript profiling reveals mechanisms for lipid conservation during diapause in themosquito, Aedes albopictus. Journal of Insect Physiology, 2012, 58(7): 966-973.
[16] Waagner, D. Holmstrup, M. Bayley, M. Sørensen, J. G. et al. Induced cold tolerance mechanisms depend on duration of acclimation in the chill sensitive Folsomia candida (Collembola) [J]. Journal of Experimental Biology, jeb-079814.
[17]马光皇,符平,孙妍,等.香梨优斑螟不同虫态内参基因稳定性分析[J].农业科学, 2019, 9(6): 427-431.
Ma Guang-huang, Fu Ping, Sun Yan. et al. Expression Stability Analysis of Internal Reference Genes in Insect States of Euzophera pyriella[J]. Hans Journal of Agricultural Sciences, 2019, 9(6): 427-431.
[18]王海鸿,雷仲仁. 用荧光定量RT-PCR方法分析B型和ZHJ-1型烟粉虱热休克基因hsp70和hsp90的表达[J]. 中国农业科学,2008, (11): 3636-3644.
WANG Haihong, Lei Zhongren. Detection of hsp 70 and hsp90 transcription in bemisia tabaci by fluorescence quantitative RT-PCR [J]. Scientia Agricultura Sinica, 2008, (11): 3636-3644.
[19] 庞竟公. 香梨优斑螟越冬幼虫的生化指标动态变化研究[D]. 阿拉尔:塔里木大学硕士论文, 2018.
PANG Jinggong. Study on the dynamic change of biochemical indicators of Euzophera pyriella Yang over wintering larvae [D]. Alar: Tarim University, 2018.
[20] 易杰群, 邹志文, 张古忍, 等.昆虫脂肪酸及其脱饱和酶与耐寒性的关系概述[J]. 环境昆虫学报,2015, 37(1): 155-162.
YI Jiequn, ZOU Zhiwen, ZHANG Guren, et al. The relationship between the fatty acids and those desaturases and the cold tolerance in the insects [J]. Journal of Environmental Entomology, 2015, 37(1): 155-162.
[21] 宋淑香. 凤丹脂肪酸去饱和酶基因PoFAD2的克隆及其表达分析[D]. 济南:山东农业大学硕士论文, 2016.
SONG Shuxiang. Isolation and Expression Analysis of PoFAD2 Gene in Paeonia ostia [D]. Tai’an: Shandong Agricultural University, 2016.
[22] Albre, J. Liénard, M. A. Sirey, T. M. Schmidt, S. Tooman, L. K. Carraher, C. Greenwood, D. R. L fstedt, C. Newcomb, R. D. et al. Sex pheromone evolution is associated with differential regulation of the same desaturase gene in two genera of leafroller moths [J]. PLoS genetics, 2012, 8(1): e1002489
[23] Gu, S. H. Wu, K. M. Guo, Y. Y. Pickett, J. A. Field, L. M. Zhou, J. J. Zhang, Y. J. et al. Identification of genes expressed in the sex pheromone gland of the black cutworm Agrotis ipsilon with putative roles in sex pheromone biosynthesis and transport [J]. BMC Genomics, 2013, 14(1): 636.
[24] 郝攀. 弓形虫硬脂酰辅酶A去饱和酶的鉴定及功能研究[D]. 北京:中国农业大学博士论文, 2016.
HAO Pan. Identification and functionality of Toxoplasma gondii stearoyl-coenzyme A desaturase [D]. Beijing: China Agricultural University, 2016.
[25] 曹威荣. 猪硬脂酰辅酶A去饱和酶蛋白生物信息学分析[J]. 畜牧与饲料科学, 2014,35(11):5-7.
CAO Weirong. Bioinformatics Analysis of the SCD Protein in Pig[J]. Animal Husbandry and Feed Science,2014, 35(11): 5-7.
[26] Man W.C. Miyazaki M. Chu K. Ntambi J.M. et al. Membrane topology of mouse stearoyl-CoA desaturase 1[J]. Journal of Biological Chemistry, 2006, 281(2): 1251-1260.
[27] 张敏, 张古忍. 大头金蝇酰基辅酶AΔ9去饱和酶cDNA克隆与原核表达[J]. 环境昆虫学报,2018, 40(6): 1306-1315.
ZHANG Min, ZHANG Guren. Cdna cloning and prokaryotic expression of acylcoa delta 9 desaturase from chrysomya megacephala(fabricius)[J]. Journal of Environmental Entomology, 2018, 40(6): 1306-1315.
[28] Gupta R, Brunak S. Prediction of glycosylation across the human proteome and the correlation to protein function.[J]. Pac Symp Biocomput, 2002, 7(3):310-322.
[29] 刘贵芬, 房刚奇, 詹帅, 等.亮斑扁角水虻脂肪酸去饱和酶基因的鉴定及其在双翅目中的进化格局[J]. 昆虫学报, 2018, 61(8): 905-913.
LIU Guifang, FANG Gangqi, ZHAN Shuai, et al. Identification of fatty acid desaturase genes in hemetia illucens (diptera: stratiomyidae) and their evolutionary pattern in diptera [J]. Acta Entomologica Sinica, 2018, 61(8): 905-913.
[30] 陆承志.香梨优斑螟发生规律与综合防治[J].中国果树,2004, (3):41-42.
LU Chengzhi. Control of the occurrence of Euzophera pyriella and integrated control [J]. China Fruits,2004,(3):41-42.

香梨优斑螟脂肪酸Δ9去饱和酶基因序列及不同虫态的表达分析

Gene Sequences and Expression Analysis of Fatty Acid Δ9 Desaturase of Euzophera pyriella during Different Stages

PDF下载

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

[1]	邵盘霞, 赵准, 邵武奎, 郝晓燕, 高升旗, 李建平, 胡文冉, 黄全生. 玉米ZmCDPK22基因在干旱胁迫下的表达分析[J]. 新疆农业科学, 2023, 60(6): 1372-1378.
[2]	唐碧徽, 张俐华, 李海英, 张冲, 蒋廷浩, 赵晓钰, 蒋腾, 丁雅文, 吴盈萍, 赵全庄. 伊犁鹅和霍尔多巴吉鹅在繁殖性能、血清激素水平和基因表达量的比较[J]. 新疆农业科学, 2023, 60(5): 1271-1280.
[3]	商靖, 逄洪波, 王兰兰, 李雪梅, 王艳秋, 李玥莹. 生长素与高粱杂种优势关系分析[J]. 新疆农业科学, 2023, 60(4): 841-846.
[4]	刘晨曦, 朱雨婷, 周强, 陈瑾, 赵文杰, 郑凯. 海岛棉β-胡萝卜素异构酶GbD27-6基因克隆及表达分析[J]. 新疆农业科学, 2023, 60(12): 2869-2877.
[5]	胡文冉, 赵准, 邵武奎, 黄全生. 陆地棉TCP家族基因鉴定及组织表达分析[J]. 新疆农业科学, 2023, 60(11): 2627-2637.
[6]	孙丰磊, 任姣姣, 雷斌, 高文伟, 曲延英. 基于BSA分析技术对陆地棉抗旱基因的鉴定[J]. 新疆农业科学, 2023, 60(10): 2341-2351.
[7]	玛依拉·玉素音, 阳妮, 徐海江, 杨延龙, 张大伟, 李春平, 石必显, 赖成霞. 受棉铃虫诱导的棉花糖基转移酶UGT基因家族的生物信息学及表达分析[J]. 新疆农业科学, 2023, 60(10): 2396-2403.
[8]	才羿, 戴冬洋, 谭海, 王迪, 杨芬, 王岭, 盛云燕. 甜瓜AMS基因结构分析及基因编辑载体构建[J]. 新疆农业科学, 2023, 60(1): 61-68.
[9]	李敏, 马英, 呼尔查, 何文文, 史倩云, 阿力木江·加帕尔, 蒋倩, 巴音查汗. 边缘革蜱Dm 86基因表达及免疫原性分析[J]. 新疆农业科学, 2022, 59(9): 2324-2332.
[10]	戴麒, 徐瑞强, 李宁, 王柏柯, 王娟, 黄少勇, 帕提古丽·艾斯木托拉, 余庆辉. 潘那利番茄GRF基因家族全基因组鉴定分析[J]. 新疆农业科学, 2022, 59(10): 2456-2465.
[11]	支昊宇, 王智慧, 王兴仔, 丁新华, 贾尊尊, 郭文超, 姜卫华. 亚洲玉米螟的抗药性监测及双酰胺类药剂抗性靶标分子检测[J]. 新疆农业科学, 2021, 58(4): 690-699.
[12]	李建平, 足木热木·吐尔逊, 常晓春, 郝晓燕, 陈果, 高升旗, 孙良斌, 黄全生. 玉米钙依赖蛋白激酶38(ZmCDPK38)的生物信息学及表达特性分析[J]. 新疆农业科学, 2021, 58(1): 49-55.
[13]	杨帅, 任明见, 李振华, 李鲁华, 徐如宏. 小麦ZY96-3籽粒灌浆过程中籽粒发育相关基因的表达分析[J]. 新疆农业科学, 2020, 57(7): 1187-1194.
[14]	龚林涛, 苏秀娟, 尹松松, 孙明辉, 闫博文, 阿迪莱·阿布都热依木, 陈全家. 薰衣草DXS基因的克隆、表达分析及原核表达[J]. 新疆农业科学, 2020, 57(7): 1233-1242.
[15]	徐航, 全绍文, 马丽, 周丽, 牛建新. 库尔勒香梨PsiERF基因的克隆和表达分析[J]. 新疆农业科学, 2018, 55(9): 1616-1625.