飞蝗核受体基因LmE75的分子特性和功能分析

doi:10.3864/j.issn.0578-1752.2020.11.008

Abstract

Abstract:

【Objective】Nuclear receptors (NRs) act as ligand-inducible transcription factors, which play an important role during growth and development in insects. Locusta migratoria is an important agricultural pest. The objective of this study is to analyze the molecular characteristics and biological function of the nuclear receptor gene LmE75 in L. migratoria, and to provide a new molecular target for pest control. 【Method】The cDNA sequences of three LmE75 genes were obtained based on the transcriptome database of L. migratoria. The open reading frames, predicted amino acid sequences, physicochemical properties and conserved domains of each gene were analyzed using Blast and ExPASy websites. Using MEGA 6.0 software, the neighbor-joining (NJ) method was used to construct a phylogenetic tree with the homologous sequences of E75 from other insects. Specific primers of three LmE75 isoforms were designed using Primer 3.0 software. Reverse transcription-quantitative PCR (RT-qPCR) was used to analyze the expression characteristics of three LmE75 isoforms in different tissues and developmental days from 4th to 5th instar nymphs. To further test whether LmE75 was regulated by 20-hydroxyecdysone (20E), the expression of LmE75 was detected by RT-qPCR after injected with 20E in vivo and interfered with the 20E receptor gene LmEcR by RNAi. The double-stranded RNA (dsRNA) of the common region of three LmE75 isoforms was synthesized in vitro, and then was injected into the 5th instar nymphs. The total RNA of integument was extracted after 48 h, and the first strand cDNA was synthesized as the template of RT-qPCR. The silencing efficiency of LmE75 was detected by RT-qPCR technology and the phenotype was observed. The effect on the structure of cuticle after injected with dsLmE75 was analyzed by H&E staining method. 【Result】Three cDNA sequences of LmE75 were obtained based on the transcriptome database of L. migratoria. According to structural characteristics, they were named LmE75A, LmE75B and LmE75C, respectively. The GenBank accession numbers were MN584732, MN584733 and MN584734. Each of the three LmE75 isoforms had typical nuclear receptor conserved domains, and DBD (DNA binding domain) and LBD (ligand binding domain) were highly conserved among insects. Phylogenetic tree analysis showed that E75 isoforms of different orders of insects were preferentially clustered into one group. RT-qPCR results showed that three LmE75 isoforms had different tissue and developmental expression characteristics. LmE75A was highly expressed in the integument and muscle. LmE75B was highly expressed in the integument, followed by the foregut. LmE75C was specifically expressed in the integument. The expression level of LmE75A increased first and then decreased at the 4th and 5th instar nymphs. The expression levels of LmE75B were the highest in N4D3 and N5D5, and lower before and after molting. The expression level of LmE75C increased gradually at both the 4th and 5th instar nymphs, with the highest expression before molting. Compared with the control group, the expression level of LmE75 significantly increased by 4.2 and 4.5 times after 20E induction for 3 h and 6 h, respectively. The expression level of LmE75 was significantly down-regulated by 59% after interference with LmEcR for 48 h by RNAi. After injection of dsLmE75 on the 1st day of 5th instar nymphs, L. migratoria exhibited obvious developmental delay phenotype and all L. migratoria could not molt to adults. The H&E staining of the integument found that the cuticle of dsLmE75-injected L. migratoria showed no apolysis. 【Conclusion】There are three LmE75 isoforms in L. migratoria, and they have different tissue and developmental expression characteristics. RNAi results show that LmE75 play an important role in the molting of L. migratoria.

Key words: Locusta migratoria, nuclear receptor (NR), LmE75, 20-hydroxyecdysone (20E), RNA interference (RNAi)

LIU XiaoJian,GUO Jun,ZHANG XueYao,MA EnBo,ZHANG JianZhen. Molecular Characteristics and Function Analysis of Nuclear Receptor Gene LmE75 in Locusta migratoria[J].Scientia Agricultura Sinica, 2020, 53(11): 2219-2231.

Figures/Tables 7

Table 1

Species and GenBank accession number for phylogenetic tree used in this study"

目 Order	物种 Species	基因 Gene	GenBank 登录号 GenBank accession number
半翅目 Hemiptera	绿盲蝽Apolygus lucorum	AlE75A、AlE75B、AlE75C	ATN39778.1、ATN39779.1、ATN39780.1
半翅目 Hemiptera	温带臭虫Cimex lectularius	ClE75X1、ClE75X2、ClE75X3	XP_014240379.1、XP_014240380.1、XP_014240381.1
	褐飞虱Nilaparvata lugens	NlE75X1、NlE75X2	XP_022197917.1、XP_022197918.1
膜翅目 Hymenoptera	哥伦比亚芭切叶蚁Atta colombica	AcE75	KYM84307.1
	黑毛蚁Lasius niger	LnE75	KMQ93219.1
	红胡须蚁Pogonomyrmex barbatus	PbE75X1	XP_011640767.1
	小黄家蚁Monomorium pharaonis	MpE75X1、MpE75X2、MpE75X4	XP_012534660.1、XP_028049966.1、XP_028049973.1
	西方蜜蜂Apis mellifera	AmE75X1	XP_006564380.1
	丽蝇蛹集金小蜂Nasonia vitripennis	NvE75	XP_008204671.1
双翅目 Diptera	黑腹果蝇Drosophila melanogaster	DmE75A、DmE75B、DmE75C、DmE75D	AAN11688.1、AAF49282.3、AAN11687.1、AAN11689.1
	埃及伊蚊Aedes aegypti	AaE75X1、AaE75X2、AaE75X3	XP_021704146.1、XP_011493380.2、XP_001652743.3
	橘小实蝇Bactrocera dorsalis	BdE75	AGF44569.1
鞘翅目 Coleoptera	马铃薯甲虫Leptinotarsa decemlineata	LdE75X1、LdE75X2、LdE75X3	AKN56577.1、AKN56578.1、ALU57795.1
鞘翅目 Coleoptera	赤拟谷盗Tribolium castaneum	TcE75X1、TcE75X2、TcE75X3、TcE75X4、TcE75X5	XP_008197843.1、XP_008197844.1、XP_971362.2、XP_008197845、XP_015838722.1
蜚蠊目 Blattodea	德国小蠊Blattella germanica	BgE75A、BgE75B、BgE75C、BgE75D、BgE75E	CAJ87513.1、CAJ87514.1、CAM97373.1、CAM97374.1、CAM97375.1
蜚蠊目 Blattodea	湿木白蚁Zootermopsis nevadensis	ZnE75X1、ZnE75X2、ZnE75X3	XP_021938067.1、XP_021938068.1、XP_021938069.1
鳞翅目 Lepidoptera	家蚕Bombyx mori	BmE75A、BmE75B、BmE75C、BmE75D	NP_001106079.1、NP_001106080.1、NP_001037042.1、BAR88292.1
直翅目Orthoptera	飞蝗Locusta migratoria	LmE75A、LmE75B、LmE75C	MN584732、MN584733、MN584734

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References 36

[1]	ARANDA A, PASCUAL A . Nuclear hormone receptors and gene expression. Physiological Reviews, 2001,81(3):1269-1304. doi: 10.1152/physrev.2001.81.3.1269
[2]	HELSEN C, CLAESSENS F . Looking at nuclear receptors from a new angle. Molecular and Cellular Endocrinology, 2014,382(1):97-106. doi: 10.1016/j.mce.2013.09.009
[3]	周树堂, 郭伟, 宋佳晟 . 昆虫变态的激素与基因调控. 生物学通报, 2012,47(9):1-6.
	ZHOU S T, GUO W, SONG J S . Insect metamorphosis hormones and gene regulation. Bulletin of Biology, 2012,47(9):1-6. (in Chinese)
[4]	何倩毓, 张原熙, 裴泽华, 李美鑫, 张旭 . 保幼激素对昆虫变态发育调控的分子机制. 昆虫学报, 2017,60(5):594-603.
	HE Q Y, ZHANG Y X, PEI Z H, LI M X, ZHANG X . Molecular mechanisms of juvenile hormone regulation on insect metamorphosis. Acta Entomologica Sinica, 2017,60(5):594-603. (in Chinese)
[5]	GUO X P, HARMON M A, LAUDET V, MANGELSDORF D J, PALMER M J . Isolation of a functional ecdysteroid receptor homologue from the ixodid tick Amblyomma americanum (L.). Insect Biochemistry and Molecular Biology, 1997,27(11):945-962. doi: 10.1016/S0965-1748(97)00075-1
[6]	FAHRBACH S E, SMAGGHE G, VELARDE R A . Insect nuclear receptors. Annual Review of Entomology, 2012,57:83-106. doi: 10.1146/annurev-ento-120710-100607
[7]	BIALECKI M, SHILTON A, FICHTENBERG C, SEGRAVES W A, THUMMEL C S . Loss of the ecdysteroid-inducible E75A orphan nuclear receptor uncouples molting from metamorphosis in Drosophila. Developmental Cell, 2002,3(2):209-220. doi: 10.1016/S1534-5807(02)00204-6
[8]	PIERCEALL W E, LI C, BIRAN A, MIURA K, RAIKHEL A S, SEGRAVES W A . E75 expression in mosquito ovary and fat body suggests reiterative use of ecdysone-regulated hierarchies in development and reproduction. Molecular and Cellular Endocrinology, 1999,150(1/2):73-89. doi: 10.1016/S0303-7207(99)00022-2
[9]	HIRUMA K, RIDDIFORD L M . Differential control of MHR3 promoter activity by isoforms of the ecdysone receptor and inhibitory effects of E75A and MHR3. Developmental Biology, 2004,272(2):510-521. doi: 10.1016/j.ydbio.2004.04.028
[10]	JINDRA M, SEHNAL F, RIDDIFORD L M . Isolation, characterization and developmental expression of the ecdysteroid-induced E75 gene of the wax moth Galleria mellonella. European Journal of Biochemistry, 1994,221(2):665-675. doi: 10.1111/ejb.1994.221.issue-2
[11]	RETNAKARAN A, MACDONALD A, TOMKINS W L, DAVIS C N, BROWNWRIGHT A J, PALLI S R . Ultrastructural effects of a non-steroidal ecdysone agonist, RH-5992, on the sixth instar larva of the spruce budworm, Choristoneura fumiferana. Journal of Insect Physiology, 1997,43(1):55-68.
[12]	SWEVERS L, ITO K, IATROU K . The BmE75 nuclear receptors function as dominant repressors of the nuclear receptor BmHR3A. The Journal of Biological Chemistry, 2002,277(44):41637-41644. doi: 10.1074/jbc.M203581200
[13]	SIAUSSAT D, BOZZOLAN F, QUEGUINER I, PORCHERON P, DEBERNARD S . Effects of juvenile hormone on 20-hydroxyecdysone- inducible EcR, HR3, E75 gene expression in imaginal wing cells of Plodia interpunctella lepidoptera. European Journal of Biochemistry, 2004,271(14):3017-3027. doi: 10.1111/(ISSN)1432-1033
[14]	PAUL R K, TAKEUCHI H, KUBO T . Expression of two ecdysteroid-regulated genes, Broad-Complex and E75, in the brain and ovary of the honeybee (Apis mellifera L.). Zoological Science, 2006,23(12):1085-1092. doi: 10.2108/zsj.23.1085
[15]	PARTHASARATHY R, TAN A, BAI H, PALLI S R . Transcription factor broad suppresses precocious development of adult structures during larval-pupal metamorphosis in the red flour beetle, Tribolium castaneum. Mechanisms of Development, 2008,125(3/4):299-313.
[16]	GUO W C, LIU X P, FU K Y, SHI J F, LÜ F G, LI G Q . Nuclear receptor ecdysone-induced protein 75 is required for larval-pupal metamorphosis in the Colorado potato beetle Leptinotarsa decemlineata (Say). Insect Molecular Biology, 2016,25(1):44-57. doi: 10.1111/imb.12197
[17]	MANE-PADROS D, CRUZ J, VILAPLANA L, PASCUAL N, BELLES X, MARTIN D . The nuclear hormone receptor BgE75 links molting and developmental progression in the direct-developing insect Blattella germanica. Developmental Biology, 2008,315(1):147-160. doi: 10.1016/j.ydbio.2007.12.015
[18]	YANG, Q P, LI Z, CAO J J, ZHANG S D, ZHANG H J, WU X Y, ZHANG Q W, LIU X X . Selection and assessment of reference genes for quantitative PCR normalization in migratory locust Locusta migratoria (Orthoptera: Acrididae). PLoS ONE, 2014,9(6):e98164. doi: 10.1371/journal.pone.0098164
[19]	LIU X J, LI F, LI D Q, MA E B, ZHANG W Q, ZHU K Y, ZHANG J Z . Molecular and functional analysis of UDP-N-acetylglucosamine pyrophosphorylases from the migratory locust, Locusta migratoria. PLoS ONE, 2013,8(8):e71970. doi: 10.1371/journal.pone.0071970
[20]	LIVAK K J, SCHMITTGEN T D . Analysis of relative gene expression data using real-time quantitative PCR and the 2 ^-ΔΔCt method . Methods, 2001,25(4):402-408. doi: 10.1006/meth.2001.1262
[21]	LI D Q, ZHAGN J Q, WANG Y, LIU X J, MA E B, SUN Y, LI S, ZHU K Y, ZHANG J Z . Two chitinase 5 genes from Locusta migratoria: Molecular characteristics and functional differentiation. Insect Biochemistry and Molecular Biology, 2015,58:46-54. doi: 10.1016/j.ibmb.2015.01.004
[22]	LIU X J, SUN Y W, LI D Q, LI S, MA E B, ZHANG J Z . Identification of LmUAP1 as a 20-hydroxyecdysone response gene in the chitin biosynthesis pathway from the migratory locust, Locusta migratoria. Insect Science, 2018,25(2):211-221. doi: 10.1111/1744-7917.12406
[23]	SONG T Q, YANG M L, WANG Y L, LIU Q, WANG H M, ZHANG J, LI T . Cuticular protein LmTwdl1 is involved in molt development of the migratory locust. Insect Science, 2016,23(4):520-530. doi: 10.1111/1744-7917.12342
[24]	LI K, GUO E E, HOSSAIN M S, LI Q R, CAO Y, TIAN L, DENG X J, LI S . Bombyx E75 isoforms display stage- and tissue-specific responses to 20-hydroxyecdysone. Scientific Reports, 2015,5:12114. doi: 10.1038/srep12114
[25]	ESCRIVA H, BERYRAND S, LAUDET V . The evolution of the nuclear receptor superfamily. Essays in Biochemistry, 2004,40(2):11-26. doi: 10.1042/bse0400011
[26]	CACERES L, NECAKOV A S, SCHWARTZ C, KIMBER S, ROBERTS I J, KRAUSE H M . Nitric oxide coordinates metabolism, growth, and development via the nuclear receptor E75. Genes and Development, 2011,25(14):1476-1485. doi: 10.1101/gad.2064111
[27]	JOHNSTON D M, SEDKOV Y, PETRUK S, RILEY K M, FUJILKA M, JAYNES J B, MAZO A . Ecdysone- and no-mediated gene regulation by competing EcR/Usp and E75A nuclear receptors during Drosophila development. Molecular Cell, 2011,44(1):51-61. doi: 10.1016/j.molcel.2011.07.033
[28]	KESHAN B, HIRUMA K, RIDDIFORD L M . Developmental expression and hormonal regulation of different isoforms of the transcription factor E75 in the tobacco hornworm Manduca sexta. Developmental Biology, 2006,295(2):623-632. doi: 10.1016/j.ydbio.2006.03.049
[29]	李凯龙 . 褐飞虱蜕皮及变态信号途径相关基因的功能分析[D]. 北京: 中国农业科学院, 2016.
	LI K L . Functional analysis of the genes involved in molting and metamorphosis signal pathways in Nilaparvata lugens[D]. Beijing: Chinese Academy of Agricultural Sciences, 2016. ( in Chinese)
[30]	ZHAO X M, QIN Z Y, LIU W M, LIU X J, MOUSSIAN B, MA E B, LI S, ZHANG J Z . Nuclear receptor HR3 controls locust molt by regulating chitin synthesis and degradation genes of Locusta migratoria. Insect Biochemistry and Molecular Biology, 2018,92:1-11. doi: 10.1016/j.ibmb.2017.11.001
[31]	ZHAO X M, QIN Z Y, ZHANG J, YANG Y, JIA P, YANG Q, MA E B, ZHANG J Z . Nuclear receptor HR39 is required for locust molting by regulating the chitinase and carboxypeptidase genes. Insect Molecular Biology, 2019,28(4):537-549.
[32]	YU Z T, ZHANG X Y, WANG Y W, MOUSSIAN B, ZHU K Y, LI S, MA E B, ZHANG J Z . LmCYP4G102: An oenocyte-specific cytochrome P450 gene required for cuticular waterproofing in the migratory locust, Locusta migratoria. Scientific Reports, 2016,6:29980. doi: 10.1038/srep29980
[33]	YU Z T, WANG Y W, ZHAO X M, LIU X J, MA E B, MOUSSIAN B, ZHANG J Z . The ABC transporter ABCH-9C is needed for cuticle barrier construction in Locusta migratoria.Insect Biochemistry and Molecular Biology, 2017, 87:90-99. doi: 10.1016/j.ibmb.2017.06.005
[34]	ZHENG J Z, LIU X J, ZHANG J Q, LI D Q, SUN Y, GUO Y P, MA E B, ZHU K Y . Silencing of two alternative splicing-derived mRNA variants of chitin synthase 1 gene by RNAi is lethal to the oriental migratory locust, Locusta migratoria manilensis (Meyen). Insect Biochemistry and Molecular Biology, 2010,40(11):824-833. doi: 10.1016/j.ibmb.2010.08.001
[35]	YU R R, LIU W M, LI D Q, ZHAO X M, DING G W, ZHANG M, MA E B, ZHU K Y, LI S, MOUSSIAN B, ZHANG J Z . Helicoidal organization of chitin in the cuticle of the migratory locust requires the function of the chitin deacetylase 2 enzyme (LmCDA2). The Journal of Biological Chemistry, 2016,291(47):24352-24363.
[36]	ZHAO X M, GOU X, LIU W M, MA E B, MOUSSIAN B, LI S, ZHU K Y, ZHANG J Z . The wing-specific cuticular protein LmACP7 is essential for normal wing morphogenesis in the migratory locust. Insect Biochemistry and Molecular Biology, 2019,112:103206.

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基因Gene	引物序列 Primer sequence (5′-3′)	用途 Application	产物长度 Length of product (bp)
β-actin-F/R	CGAAGCACAGTCAAAGAGAGGTA/GCTTCAGTCAAGAGAACAGGATG	RT-qPCR	156
LmE75A-F/R	TTCCGTGTCCTCCAAGCTAG/ACCTTGCACGGGATGATCTC		111
LmE75B-F/R	ATGATCCTGTCCGACGGC/CCCGTCGAACTCTATGTGGA		150
LmE75C-F/R	ATGACTGTGAAGGACCCGG/ACTCTTCTAGAACCCTGCCT		133
LmE75-F/R	CTGCTCGGTGGTGGTGAT/GTCATCTTGAAGGCGAGCAG		85
LmEcR-F/R	GACAAACTGCTACGGGAAGA/CTATCGTTTCTCCCATACCAG		172
dsLmE75-F/R	taatacgactcactatagggAGCTGGAGGACCAGTCTGC/taatacgactcactatagggGCGAAGTCGAACATGGAGTC	RNAi	460
dsLmEcR-F/R	taatacgactcactatagggCAGCAACGCCGCACCCT/taatacgactcactatagggCACTGGTACACGGCATTT		408
dsGFP-F/R	taatacgactcactatagggTGGAGAGGGTGAAGG/taatacgactcactatagggGGCAGATTGTGTGGAC		712

Molecular Characteristics and Function Analysis of Nuclear Receptor Gene LmE75 in Locusta migratoria

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