白背飞虱海藻糖合成酶基因调控几丁质合成的功能

doi:10.3864/j.issn.0578-1752.2019.19.007

中国农业科学 ›› 2019, Vol. 52 ›› Issue (19): 3357-3366.doi: 10.3864/j.issn.0578-1752.2019.19.007

白背飞虱海藻糖合成酶基因调控几丁质合成的功能

张道伟¹,余亚娅²,潘碧莹³,康奎¹,曾伯平¹,陈静²,唐斌^1,³()

¹ 遵义师范学院生物与农业科技学院/赤水河流域动物资源保护与应用研究重点实验室,贵州遵义 563006
² 遵义医科大学基础医学院,贵州遵义 563006
³ 杭州师范大学生命与环境科学学院,杭州 310036

收稿日期:2019-05-05 接受日期:2019-06-10 出版日期:2019-10-01 发布日期:2019-10-11
通讯作者: 唐斌
作者简介:张道伟,E-mail：zhangdw1000@163.com。
基金资助:
国家自然科学基金(31560511);国家自然科学基金(31672081);贵州省科学技术基金黔科合(JZ字[2014]2014号)

Regulation Function of Trehalose-6-phosphate Synthase Genes on Chitin Synthesis in Sogatella furcifera

ZHANG DaoWei¹,YU YaYa²,PAN BiYing³,KANG Kui¹,ZENG BoPing¹,CHEN Jing²,TANG Bin^1,³()

¹ College of Biology and Agriculture, Zunyi Normal University/Key Laboratory of Protection and Utilization of Animal Resource in Chishui River Basin, Zunyi 563006, Guizhou
² College of Basic Medical Science, Zunyi Medical University, Zunyi 563006, Guizhou
³ College of Life and Environmental Science, Hangzhou Normal University, Hangzhou 310036

Received:2019-05-05 Accepted:2019-06-10 Online:2019-10-01 Published:2019-10-11
Contact: Bin TANG

摘要/Abstract

摘要：

【背景】 海藻糖合成酶（trehalose-6-phosphate synthase,TPS）在海藻糖合成中起着重要作用,其能够介导海藻糖代谢调控几丁质合成及昆虫发育。【目的】 本研究通过抑制白背飞虱（Sogatella furcifera）TPS的表达,检测RNAi沉默SfTPS效果,观察白背飞虱蜕皮状况,测定几丁质含量及几丁质合成酶（chitin synthase,CHS）基因的定量表达,探究SfTPS在白背飞虱几丁质合成中的潜在调控作用。【方法】 利用注射法RNAi技术,以实验室饲养多年的白背飞虱种群为试验材料,体外合成两个SfTPS（SfTPS1和SfTPS2）与GFP的双链RNA（dsRNA）后,分别注射到白背飞虱体内抑制TPS。首先,在dsRNA注射后48 h采用Trizol法提取白背飞虱的总RNA,反转录并合成第一链DNA后,采用实时荧光定量PCR（qRT-PCR）技术检测TPS表达沉默情况,以确定RNAi的效果;其次,测定dsRNA注射后48 h和72 h白背飞虱整体几丁质含量并对翅发育畸形虫体进行拍照;最后,采用qRT-PCR技术检测白背飞虱SfCHS在mRNA水平上的相对表达量变化,分析SfTPS1和SfTPS2在几丁质合成调控中的作用。【结果】 与注射dsGFP相比较,dsSfTPS1和dsSfTPS2的RNA注射后,能够促进SfCHS表达量上升,几丁质含量增加,白背飞虱成虫翅出现畸形。qRT-PCR结果显示,单个SfTPS dsRNA注射后本基因的表达能够被极显著抑制,与注射dsGFP相比,不足对照组表达量的30%,且单个SfTPS的dsRNA注射后,另外一个SfTPS表达同样显著下降;dsSfTPS1和dsSfTPS2注射后,白背飞虱成虫翅均为长翅,出现一定比例的翅卷曲等畸形情况,其后48 h和72 h产生一定的死亡率;几丁质含量检测发现,SfTPS1和SfTPS2的dsRNA注射后72 h,几丁质含量显著上升。与注射dsGFP对照组相比较,SfCHS1与SfCHS1a表达量在dsSfTPS1注射后72 h极显著上升,在dsSfTPS2注射后48 h和72 h时极显著上升,且dsSfTPS1和dsSfTPS2注射后SfCHS1b的表达极显著增加。【结论】 SfTPS能够通过调控白背飞虱几丁质合成酶基因的表达来控制几丁质的合成,研究结果有助于评价SfTPS在白背飞虱等昆虫中的调控作用并作为潜在控害靶标,为进一步开展和筛选有效的海藻糖合成酶抑制剂控制白背飞虱等害虫提供理论依据。

关键词: 白背飞虱, 海藻糖合成酶, RNAi, 几丁质合成, 实时荧光定量PCR

Abstract:

【Background】 It is well known that trehalose-6-phosphate synthase (TPS) plays an important role in trehalose synthesis, which can mediate trehalose metabolism to regulate chitin synthesis and insect development. 【Objective】 In this study, the effect of silencing SfTPS was detected, the molting status of Sogatella furcifera was observed and the content of chitin and the quantitative expression of chitin synthase (CHS) gene were determined through inhibiting the expression of TPS by RNAi in S. furcifera. The purpose is to explore the potential regulatory effects of SfTPS on the chitin synthesis in S. furcifera.【Method】 The S. furcifera population, which has been fed in laboratory for many years, was used as experimental material. The double-stranded RNA (dsRNA) of SfTPS and GFP were synthesized in vitro in order to study the potential function of TPSs, the RNA of two dsSfTPSs (dsSfTPS1 and dsSfTPS2) was injected into S. furcifera, respectively. Firstly, after dsRNA injection, the total RNA of S. furcifera was extracted by Trizol method at 48 h, and the first strand DNA was synthesized as the template of quantitative real-time PCR (qRT-PCR). As well as qRT-PCR was used to detect the silencing of SfTPS expression for the effect of RNAi. Secondly, the whole chitin of S. furcifera was detected at 48 and 72 h after dsRNA injection, and photographs were taken of the winged developmental malformations. Finally, the relative expression level of SfCHS in S. furcifera was detected by qRT-PCR, and the roles of SfTPS1 and SfTPS2 in the regulation of chitin synthesis were analyzed.【Result】 Compared with dsGFP injection, the expression of SfCHS increased after dsSfTPS1 and dsSfTPS2 injection, as well as the chitin content increased, which lead to the wing deformity of S. furcifera. The qRT-PCR results showed that the expression of SfTPS was significantly inhibited after dsRNA injection of a single SfTPS, which was less than 30% of that in the control group injected with dsGFP. In addition, the expression of another SfTPS also decreased significantly after a single SfTPS dsRNA injection. The adult wings of S. furcifera were all long wings after dsSfTPS1 and dsSfTPS2 injection. Some deformities such as wing curl in a certain ratio had been found in these long wings, and a certain ratio of mortality was found at 48 and 72 h. The chitin content increased significantly at 72 h after these two dsSfTPSs RNA injection. Compared with the control group, the expression of SfCHS1 and SfCHS1a increased significantly at 72 h after dsSfTPS1 injection, followed it increased significantly at 48 and 72 h after dsSfTPS2 injection. In the same time, the expression of SfCHS1b increased significantly after dsSfTPS1 and dsSfTPS2 injection.【Conclusion】 The SfTPS can control the synthesis of chitin through the regulation of chitin synthase gene in S. furcifera. The results are helpful to evaluate the regulatory role of SfTPS in S. furcifera and other insects and as the potential pest control target. It provides a theoretical basis for further development and screening of effective trehalose-6-phosphate synthase inhibitors to control S. furcifera pests and so on.

Key words: white-backed planthopper (Sogatella furcifera), trehalose-6-phosphate synthase (TPS), RNA interference, chitin synthesis, quantitative real-time PCR (qRT-PCR)

张道伟,余亚娅,潘碧莹,康奎,曾伯平,陈静,唐斌. 白背飞虱海藻糖合成酶基因调控几丁质合成的功能[J]. 中国农业科学, 2019, 52(19): 3357-3366.

ZHANG DaoWei,YU YaYa,PAN BiYing,KANG Kui,ZENG BoPing,CHEN Jing,TANG Bin. Regulation Function of Trehalose-6-phosphate Synthase Genes on Chitin Synthesis in Sogatella furcifera[J]. Scientia Agricultura Sinica, 2019, 52(19): 3357-3366.

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参考文献 39

[1]	BARRION A T, LITSINGER J A . Taxonomy of rice insect pests and their arthropod parasites and predators//HEINRICHS E A. Biology and Management of Rice Insects. Manila, Philippines: Wiley Eastern Ltd., India and IRRI, 1994: 13-362.
[2]	赵梦, 欧阳芳, 张永生, 李魏, 曹婧, 戈峰 . 2000-2010年我国水稻病虫害发生与为害特征分析. 生物灾害科学, 2014,37(4):275-280.
	ZHAO M, OUYANG F, ZHANG Y S, LI W, CAO J, GE F . Characteristics of occurrence and damage from diseases and insect pests in rice production in China during 2000-2010. Biological Disaster Science, 2014,37(4):275-280. (in Chinese)
[3]	赵颖, 黄凤宽, 童晓立, 庞雄飞 . 水稻品种对褐飞虱不同生物型抗性的HPLC分析. 华南农业大学学报, 2005,26(2):52-55.
	ZHAO Y, HUANG F K, TONG X L, PANG X F . HPLC analysis of rice variety resistance to different biotypes of Nilaparvata lugens. Journal of South China Agricultural University, 2005,26(2):52-55. (in Chinese)
[4]	WANG Y C, TANG M, HAO P Y, YANG Z F, ZHU L L, HE G C . Penetration into rice tissues by brown planthopper and fine structure of the salivary sheaths. Entomologia Experimentalis et Applicata, 2008,129(3):295-307.
[5]	XI Y, PAN P L, YE Y X, YU B, XU H J, ZHANG C X . Chitinase-like gene family in the brown planthopper,Nilaparvata lugens. Insect Molecular Biology, 2015,24(1):29-40.
[6]	XI Y, PAN P L, ZHANG C X . The β-N-acetylhexosaminidase gene family in the brown planthopper, Nilaparvata lugens. Insect Molecular Biology, 2015,24(6):601-610.
[7]	GHAFFAR M B, PRITCHARD J, FORD-LLOYD B . Brown planthopper (N. lugens Stål) feeding behavior on rice germplasm as an indicator of resistance. PLoS ONE, 2011,6(7):e22137.
[8]	唐斌, 魏苹, 陈洁, 王世贵, 张文庆 . 昆虫海藻糖酶的基因特性及功能研究进展. 昆虫学报, 2012,55(11):1315-1321.
	TANG B, WEI P, CHEN J, WANG S G, ZHANG W Q . Progress in gene features and functions of insect trehalases. Acta Entomologica Sinica, 2012,55(11):1315-1321. (in Chinese)
[9]	唐斌, 徐青叶, 赵丽娜, 王世贵, 张帆 . 昆虫海藻糖及其合成酶基因的特性与功能研究进展. 应用昆虫学报, 2014,51(6):1397-1405.
	TANG B, XU Q Y, ZHAO L N, WANG S G, ZHANG F . Progress in research on the characteristics and functions of trehalose and the TPS gene in insects. Chinese Journal of Applied Entomology, 2014,51(6):1397-1405. (in Chinese)
[10]	SHUKLA E, THORAT L J, NATH B B, GAIKWAD S M . Insect trehalase: Physiological significance and potential applications. Glycobiology, 2015,25(4):357-367.
[11]	唐斌, 张露, 熊旭萍, 汪慧娟, 王世贵 . 海藻糖代谢及其调控昆虫几丁质合成研究进展. 中国农业科学, 2018,51(4):697-707. doi: 10.3864/j.issn.0578-1752.2018.04.009
	TANG B, ZHANG L, XIONG X P, WANG H J, WANG S J . Advances in trehalose metabolism and its regulation of insect chitin synthesis. Scientia Agricultura Sinica, 2018,51(4):697-707. (in Chinese) doi: 10.3864/j.issn.0578-1752.2018.04.009
[12]	CHEN J, TANG B, CHEN H, YAO Q, HUANG X, CHEN J, ZHANG D, ZHANG W Q . Different functions of the insect soluble and membrane-bound trehalase genes in chitin biosynthesis revealed by RNA interference. PLoS ONE, 2010,5(4):e10133.
[13]	张文庆, 陈晓菲, 唐斌, 田宏刚, 陈洁, 姚琼 . 昆虫几丁质合成及其调控研究前沿. 应用昆虫学报, 2011,48(3):475-479.
	ZHANG W Q, CHEN X F, TANG B, TIAN H G, CHEN J, YAO Q . Insect chitin biosynthesis and its regulation. Chinese Journal of Applied Entomology, 2011,48(3):475-479. (in Chinese)
[14]	TANG B, WANG S, WANG S G, WANG H J, ZHANG J Y, CUI S Y . Invertebrate trehalose-6-phosphate synthase gene: Genetic architecture, biochemistry, physiological function, and potential applications. Frontiers in Physiology, 2018,9:30.
[15]	ZHU K Y, MERZENDORFER H, ZHANG W Q, ZHANG J Z, MUTHUKRISHNAN S . Biosynthesis, turnover, and functions of chitin in insects. Annual Review of Entomology, 2016,61:177-196.
[16]	CHEN X F, TIAN H G, ZOU L Z, TANG B, HU J, ZHANG W Q . Disruption of Spodoptera exigua larval development by silencing chitin synthase gene A with RNA interference. Bulletin of Entomological Research, 2008,98(6):613-619.
[17]	ZHAO L N, YANG M M, SHEN Q D, LIU X J, SHI Z K, WANG S G, TANG B . Functional characterization of three trehalase genes regulating the chitin metabolism pathway in rice brown planthopper using RNA interference. Scientific Reports, 2016,6:27841.
[18]	陈静, 张道伟 . 德国小蠊两个海藻糖合成酶基因的克隆、组织分布及温度诱导表达分析. 昆虫学报, 2015,58(10):1046-1053.
	CHEN J, ZHANG D W . Molecular cloning, tissue distribution and temperature-induced expression of two trehalose-6-phosphate synthase genes in Blattella germanica(Blattodea: Blattellidae). Acta Entomologica Sinica, 2015,58(10):1046-1053. (in Chinese)
[19]	YANG M M, ZHAO L N, SHEN Q D, XIE G Q, WANG S G, TANG B . Knockdown of two trehalose-6-phosphate synthases severely affects chitin metabolism gene expression in the rice brown planthopper Nilaparvata lugens. Pest Management Science, 2017,73(1):206-216.
[20]	陈静 . 褐飞虱遗传多样性及其海藻糖合成酶基因功能的分析[D]. 广州: 中山大学, 2010.
	CHEN J . Genetic diversity and function analysis of a trehalose phosphate synthase gene of Nilaparvata lugens[D]. Guangzhou:Sun Yat-Sen University, 2010. ( in Chinese)
[21]	唐斌, 沈祺达, 曾伯平, 肖仲久, 邱玲玉, 潘碧莹, 李昆, 张道伟 . 褐飞虱一个新的海藻糖合成酶基因的特性、发育表达及RNAi效果分析. 中国农业科学, 2019,52(3):466-477. doi: 10.3864/j.issn.0578-1752.2019.03.007
	TANG B, SHEN Q D, ZENG B P, XIAO Z J, QIU L Y, PAN B Y, LI K, ZHANG D W . Characteristics, developmental expression and RNAi effect analysis of a novel trehalose-6-phosphate synthase gene in Nilaparvata lugens. Scientia Agricultura Sinica, 2019,52(3):466-477. (in Chinese) doi: 10.3864/j.issn.0578-1752.2019.03.007
[22]	TANG B, CHEN J, YAO Q, PAN Z Q, XU W H, WANG S G, ZHANG W Q . Characterization of a trehalose-6-phosphate synthase gene from Spodoptera exigua and its function identification through RNA interference. Journal of Insect Physiology, 2010,56(7):813-821.
[23]	张道伟, 陈静, 郭玉双 . 白背飞虱海藻糖合成酶基因的克隆及序列分析. 黑龙江农业科学, 2012(5):14-19.
	ZHANG D W, CHEN J, GUO Y S . Cloning and sequence analysis oftrehalose phosphate synthase gene from Sogatella furcifera. Heilongjiang Agricultural Sciences, 2012(5):14-19. (in Chinese)
[24]	张露, 朱世城, 郑好, 沈祺达, 王世贵, 唐斌 . 褐飞虱海藻糖酶基因在表皮几丁质代谢中的调控作用. 中国农业科学, 2017,50(6):1047-1056. doi: 10.3864/j.issn.0578-1752.2017.06.006
	ZHANG L, ZHU S C, ZHENG H, SHEN Q D, WANG S G, TANG B . Regulatory function of trehalase genes on chitin metabolism in the cuticle of Nilaparvata lugens. Scientia Agricultura Sinica, 2017,50(6):1047-1056. (in Chinese) doi: 10.3864/j.issn.0578-1752.2017.06.006
[25]	曹传旺, 高彩球 . 昆虫生化与分子生物学实验技术. 哈尔滨: 东北林业大学出版社, 2009: 24-26.
	CAO C W, GAO C Q. Experimental Techniques of Insect Biochemistry and Molecular Biology. Harbin: Northeast Forestry University Press, 2009: 24-26. (in Chinese)
[26]	LIVAK K J, SCHMITTGEN T D . Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods, 2001,25(4):402-408.
[27]	WANG Y, FAN H W, HUANG H J, XUE J, WU W J, BAO Y Y, XU H J, ZHU Z R, CHENG J A, ZHANG C X . Chitin synthase 1 gene and its two alternative splicing variants from two sap-sucking insects,Nilaparvata lugens and Laodelphax striatellus(Hemiptera: Delphacidae). Insect Biochemistry and Molecular Biology, 2012,42(9):637-646.
[28]	THOMPSON S N . Trehalose - The insect ‘blood’ sugar. Advances in Insect Physiology, 2003,31:205-285.
[29]	AVONCE N, MENDOZA-VARGAS A, MORETT E, ITURRIAGA G . Insights on the evolution of trehalose biosynthesis. BMC Evolutionary Biology, 2006,6:109.
[30]	LU X Y, LI J Q, YANG J H, LIU X N, MA J . De novo transcriptome of the desert beetle Microdera punctipennis(Coleoptera: Tenebrionidae) using illumine RNA-seq technology. Molecular Biology Reports, 2014,41(11):7293-7303.
[31]	FIRE A, XU S Q, MONTGOMERY M K, KOSTAS S A, DRIVER S E, MELLO C C . Potent and specific genetic interference by double-stranded RNA in Caenorhabditis elegans. Nature, 1998,391:806-811.
[32]	MA Z, ZHU P, SHI H, GUO L, ZHANG Q, CHEN Y, CHEN S, ZHANG Z, PENG J, CHEN J . PTC-bearing mRNA elicits a genetic compensation response via Upf3a and COMPASS components. Nature, 2019,568(7751):259-263.
[33]	ZHANG L, QIU L Y, YANG H L, WANG H J, ZHOU M, WANG S G, TANG B . Study on the effect of wing bud chitin metabolism and its developmental network genes in the brown planthopper,Nilaparvata lugens, by knockdown of TRE gene. Frontiers in Physiology, 2017,8:750.
[34]	ZHU Q S, ARAKANE Y, BEEMAN R W, KRAMER K J, MUTHUKRISHNAN S . Functional specialization among insect chitinase family genes revealed by RNA interference. Proceedings of the National Academy of Sciences of the United States of America, 2008,105(18):6650-6655.
[35]	BELLES X . Beyond Drosophila: RNAi in vivo and functional genomics in insects. Annual Review of Entomology, 2010,55:111-128.
[36]	SCOTT J G, MICHEL K, BARTHOLOMAY L C, SIEGFRIED B D, HUNTER W B, SMAGGHE G, ZHU K Y, DOUGLAS A E . Towards the elements of successful insect RNAi. Journal of Insect Physiology, 2013,59(12):1212-1221.
[37]	CHEN X F, YANG X, KUMAR N S, TANG B, SUN X J, QIU X M, HU J, ZHANG W Q . The class A chitin synthase gene of Spodoptera exigua: Molecular cloning and expression patterns. Insect Biochemistry and Molecular Biology, 2007,37(5):409-417.
[38]	TANG B, YANG M M, SHEN Q D, XU Y X, WANG H J, WANG S G . Suppressing the activity of trehalase with validamycin disrupts the trehalose and chitin biosynthesis pathways in rice brown planthopper,Nilaparvata lugens. Pesticide Biochemistry and Physiology, 2017,137:81-90.
[39]	CHEN Q W, JIN S, ZHANG L, SHEN Q D, WEI P, WEI Z M, WANG S G, TANG B . Regulatory functions of trehalose-6-phosphate synthase in the chitin biosynthesis pathway in Tribolium castaneum(Coleoptera: Tenebrionidae) revealed by RNA interference. Bulletin of Entomological Research, 2018,108(3):388-399.

引物名称 Primer name	上游引物 Forward primer (5′-3′)	下游引物 Reverse primer (5′-3′)	产物长度 Length (bp)
dsSfTPS1	CCCGTTGTGGTGAGAAATA	CAAGGTGGGAATGGAATG	473
dsSfTPS1-T7	T7-CCCGTTGTGGTGAGAAATA	T7-CAAGGTGGGAATGGAATG	523
dsSfTPS2	CGCATAGACCGCAACAAC	TCGCAACGGAGTAACCAG	459
dsSfTPS2-T7	T7-CGCATAGACCGCAACAAC	T7-TCGCAACGGAGTAACCAG	509
dsGFP	AAGGGCGAGGAGCTGTTCACCG	CAGCAGGACCATGTGATCGCGC	688
dsGFP-T7	T7-AAGGGCGAGGAGCTGTTCACCG	T7-CAGCAGGACCATGTGATCGCGC	738

引物名称 Primer name	上游引物 Forward primer (5′-3′)	下游引物 Reverse primer (5′-3′)	产物长度 Length (bp)
Q-SfTPS1	CCGATTCGCTACATCTACG	GACAAACTCTTTCGCCACTAA	123
Q-SfTPS2	GATGCTGAGGGCAAAGAC	TGTGGAAGCCGACAAAGT	226
Q-SfCHS1	GATTGGTCATTGGCTTCAGA	GTAATGTCTTGCTTCGTCAG	151
Q-SfCHS1a	CTTCGGTGTTTGGTTTCTT	TGGGTAACATCATCATAGGA	136
Q-SfCHS1b	GAGAAGGCGAGAATAGCA	GCAGCAAGAACACGATTA	103
Q-18S rRNA	GCCCCGTAATCGGAATGAGT	GACAAGACGTCCCGCAAAAC	205

白背飞虱海藻糖合成酶基因调控几丁质合成的功能

Regulation Function of Trehalose-6-phosphate Synthase Genes on Chitin Synthesis in Sogatella furcifera

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