Q型烟粉虱芳香基硫酸酯酶B基因的克隆及表达分析
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摘要
为明确烟粉虱Bemisia tabaci取食感染番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)的番茄植株后,其体内的芳香基硫酸酯酶B基因(arylsulfatase B,ARSB)是否能够做出应答反应,基于Q型烟粉虱基因组数据克隆得到ARSB基因cDNA全长,采用生物信息学方法分析其序列特征,并通过实时荧光定量PCR技术测定ARSB基因在Q型烟粉虱不同发育阶段、不同组织及携带TYLCV前后的表达量变化情况。结果显示:Q型烟粉虱ARSB基因的cDNA全长为1 731 bp,编码576个氨基酸,分子量为64.89 kD,具有ARSB的保守结构域。ARSB基因在Q型烟粉虱不同发育阶段均有表达,在卵期表达量最高,成虫期表达量最低;该基因在Q型烟粉虱头胸部的表达量显著高于腹部;Q型烟粉虱获取TYLCV 72 h后其体内ARSB基因表达量显著提高。表明ARSB基因在Q型烟粉虱不同龄期、不同组织内存在差异表达,并可能参与Q型烟粉虱对TYLCV的响应和传毒过程。
In order to investigate whether the arylsulfatase B(ARSB) gene of Bemisia tabaci Q biotype can respond to Tomato yellow leaf curl virus(TYLCV) on tomatoes, the full-length cDNA of ARSB gene was cloned from B. tabaci Q biotype based on the genomic data; the sequence characteristics of the gene were analyzed with bioinformatics, and the expression levels of ARSB gene in different developmental stages, different tissues and under TYLCV treatment were detected in B. tabaci Q biotype by using quantitative real-time PCR. The results showed that the cDNA length of ARSB gene was 1 731 bp,encoding 576 amino acids, with a molecular weight of 64.89 kD. It had the conserved domain of ARSB.The ARSB gene was expressed during different development stages of B. tabaci Q biotype. The expression level was the highest in the egg stage and the lowest in the adult stage. The expression level of ARSB gene in the head and thorax was significantly higher than that in the abdomen, and the expression level of ARSB gene under TYLCV treatment for 72 h was significantly higher than that without TYLCV treatment. This study indicated that the expression level of ARSB varied at different developmental stages and in different tissues of B. tabaci Q biotype. The ARSB gene might play a potential role in the response of B. tabaci Q biotype to TYLCV.
In order to investigate whether the arylsulfatase B(ARSB) gene of Bemisia tabaci Q biotype can respond to Tomato yellow leaf curl virus(TYLCV) on tomatoes, the full-length cDNA of ARSB gene was cloned from B. tabaci Q biotype based on the genomic data; the sequence characteristics of the gene were analyzed with bioinformatics, and the expression levels of ARSB gene in different developmental stages, different tissues and under TYLCV treatment were detected in B. tabaci Q biotype by using quantitative real-time PCR. The results showed that the cDNA length of ARSB gene was 1 731 bp,encoding 576 amino acids, with a molecular weight of 64.89 kD. It had the conserved domain of ARSB.The ARSB gene was expressed during different development stages of B. tabaci Q biotype. The expression level was the highest in the egg stage and the lowest in the adult stage. The expression level of ARSB gene in the head and thorax was significantly higher than that in the abdomen, and the expression level of ARSB gene under TYLCV treatment for 72 h was significantly higher than that without TYLCV treatment. This study indicated that the expression level of ARSB varied at different developmental stages and in different tissues of B. tabaci Q biotype. The ARSB gene might play a potential role in the response of B. tabaci Q biotype to TYLCV.
引文
Bhattacharyya S,Look D,Tobacman JK.2007.Increased arylsulfatase B activity in cystic fibrosis cells following correction of CFTR.Clinica Chimica Acta,380(1/2):122-127
Brown JK,Coats SA,Bedford ID,Markham PG,Bird J,Frohlich DR.1995.Characterization and distribution of esterase electromorphs in the whitefly,Bemisia tabaci(Genn.)(Homoptera:Aleyrodidae).Biochemical Genetics,33(7/8):205-214
Brown JK,Czosnek H.2002.Whitefly transmission of plant viruses.Advances in Botanical Research,36:65-100
Byrne DN,Bellows TS Jr.1991.Whitefly biology.Annual Review of Entomology,36(1):431-457
Czosnek H,Ghanim M.2002.The circulative pathway of begomoviruses in the whitefly vector Bemisia tabaci:insights from studies with Tomato yellow leaf curl virus.Annals of Applied Biology,140(3):215-231
Czosnek H,Ghanim M.2012.Back to basics:are begomoviruses whitefly pathogens?Journal of Integrative Agriculture,11(2):225-234
de Barro PJ,Liu SS,Boykin LM,Dinsdale AB.2011.Bemisia tabaci:a statement of species status.Annual Review of Entomology,56:1-19
Dinsdale A,Cook L,Riginos C,Buckley YM,de Barro P.2010.Refined global analysis of Bemisia tabaci(Hemiptera:Sternorrhyncha:Aleyrodoidea:Aleyrodidae)mitochondrial cytochrome oxidase 1 to identify species level genetic boundaries.Annals of the Entomological Society of America,103(2):196-208
Dodgson KS,Rose FA.1970.Sulfoconjugation and sulfohydrolysis.Metabolic Conjugation and Metabolic Hydrolysis,1:239-325
Dzialoszyński LM,Fr?hlich A,Kuczyński J,Pydzik T.1967.The activity of arylsulfatase in some tumors of the genital organs in women.Archivum Immunologiae et Therapiae Experimentalis,15(1):97-99
Feferman L,Bhattacharyya S,Deaton R,Gann P,Guzman G,KajdacsyBalla A,Tobacman JK.2013.Arylsulfatase B(N-acetylgalactosamine-4-sulfatase):potential role as a biomarker in prostate cancer.Prostate Cancer and Prostatic Diseases,16(3):277-284
Gray S,Cilia M,Ghanim M.2014.Circulative,“nonpropagative”virus transmission:an orchestra of virus-,insect-,and plant-derived instruments.Advances in Virus Research,89:141-199
Hariton Shalev A,Sobol I,Ghanim M,Liu SS,Czosnek H.2016.The whitefly Bemisia tabaci knottin-1 gene is implicated in regulating the quantity of Tomato yellow leaf curl virus ingested and transmitted by the insect.Viruses,8(7):e205
Hasegawa DK,Chen W,Zheng Y,Kaur N,Wintermantel WM,Simmons AM,Fei Z,Ling KS.2018.Comparative transcriptome analysis reveals networks of genes activated in the whitefly,Bemisia tabaci when fed on tomato plants infected with Tomato yellow leaf curl virus.Virology,513:52-64
Hogenhout SA,Ammar ED,Whitfield AE,Redinbaugh MG.2008.Insect vector interactions with persistently transmitted viruses.Annual Review of Phytopathology,46:327-359
Hogenhout SA,Bos JIB.2011.Effector proteins that modulate plant-insect interactions.Current Opinion in Plant Biology,14(4):422-428
Hu J,de Barro P,Zhao H,Wang J,Nardi F,Liu SS.2011.An extensive field survey combined with a phylogenetic analysis reveals rapid and widespread invasion of two alien whiteflies in China.PLoSONE,6(1):e16061
Jiu M,Zhou XP,Liu SS.2006.Acquisition and transmission of two begomoviruses by the B and a non-B biotype of Bemisia tabaci from Zhejiang,China.Journal of Phytopathology,154(10):587-591
Jiu M,Zhou X P,Tong L,Xu J,Yang X,Wan FH,Liu SS.2007.Vectorvirus mutualism accelerates population increase of an invasive whitefly.PLoS ONE,2(1):e182
Kanakala S,Ghanim M.2016.Implication of the whitefly Bemisia tabaci cyclophilin B protein in the transmission of Tomato yellow leaf curl virus.Frontiers in Plant Science,7:1702
Kikal T,Smith JN.1959.Comparative detoxication 6:the metabolism of 6-amino-4 nitro-o-cresol and 4:6-dinitro-o-cresol in locusts.The Biochemical Journal,71(1):48-54
Klionsky DJ,Emr SD.2000.Autophagy as a regulated pathway of cellular degradation.Science,290(5497):1717-1721
Li MR,Wang R,Sheng Q.2014.Advances in research of arylsulfatase in tumro diagnosis and oncotherapy.Pharmaceutical Biotechnology,21(1):81-85(in Chinese)[李孟如,王蓉,盛清.2014.芳香基硫酸酯酶在肿瘤诊断和治疗中的研究进展.药物生物技术,21(1):81-85]
Li R,Xie W,Wang S,Wu Q,Yang N,Yang X,Pan H,Zhou X,Bai L,Xu B,et al.2013.Reference gene selection for qRT-PCR analysis in the sweetpotato whitefly,Bemisia tabaci(Hemiptera:Aleyrodidae).PLoS ONE,8(1):e53006
Luan JB,Li JM,Varela N,Wang YL,Li FF,Bao YY,Zhang CX,Liu SS,Wang XW.2011.Global analysis of the transcriptional response of whitefly to Tomato yellow leaf curl China virus reveals the relationship of coevolved adaptations.Journal of Virology,85(7):3330-3340
Mitsunaga-Nakatsubo K,Kusunoki S,Kawakami H,Akasaka K,Akimoto Y.2009.Cell-surface arylsulfatase A and B on sinusoidal endothelial cells,hepatocytes,and Kupffer cells in mammalian livers.Medical Molecular Morphology,42(2):63-69
Mugiira RB,Liu SS,Zhou X.2008.Tomato yellow leaf curl virus and Tomato leaf curl Taiwan virus invade south-east coast of China.Journal of Phytopathology,156(4):217-221
Pan LL,Chen QF,Zhao JJ,Guo T,Wang XW,Hariton-Shalev A,Czosnek H,Liu SS.2017.Clathrin-mediated endocytosis is involved in Tomato yellow leaf curl virus transport across the midgut barrier of its whitefly vector.Virology,502:152-159
Prabhu SV,Bhattacharyya S,Guzman-Hartman G,Macias V,KajdacsyBalla A,Tobacman JK.2011.Extra-lysosomal localization of arylsulfatase B in human colonic epithelium.Journal of Histochemistry&Cytochemistry,59(3):328-335
Ren SX,Qiu BL,Ge F,Zhang YJ,Du YZ,Chen XX,Guo JY,Lin KJ,Peng ZQ,Yao SL,et al.2011.Research progress of the monitoring,forecast and sustainable management of whitefly pests in China.Chinese Journal of Applied Entomology,48(1):7-15(in Chinese)[任顺祥,邱宝利,戈峰,张友军,杜予州,陈学新,郭建英,林克剑,彭正强,姚松林,等.2011.粉虱类害虫的监测预警与可持续治理技术透视.应用昆虫学报,48(1):7-15]
Robinson D,Smith JN,Williams RT.1953.Studies in detoxication.XLVI.β-glucuronidase and arylsulphatase in the crop fluid of locusts.The Biochemical Journal,53(1):125-129
Shatters RG Jr.,Powell CA,Boykin LM,Liansheng H,McKenzie CL.2009.Improved DNA barcoding method for Bemisia tabaci and related Aleyrodidae:development of universal and Bemisia tabaci biotype-specific mitochondrial cytochrome c oxidase I polymerase chain reaction primers.Journal of Economic Entomology,102(2):750-758
Su YL,He WB,Wang J,Li JM,Liu SS,Wang XW.2013.Selection of endogenous reference genes for gene expression analysis in the Mediterranean species of the Bemisia tabaci(Hemiptera:Aleyrodidae)complex.Journal of Economic Entomology,106(3):1446-1455
Wang K,Li LK.1981.Arylsulfatase and its isoenzyme.Foreign Medical Sciences:Clinical Biochemistry and Laboratory Book,(4):1-6(in Chinese)[王坤,李立群.1981.芳香基硫酸酯酶及其同工酶.国外医学:临床生物化学与检验学分册,(4):1-6]
Wang LL,Wang XR,Wei XM,Huang H,Wu JX,Chen XX,Liu SS,Wang XW.2016.The autophagy pathway participates in resistance to Tomato yellow leaf curl virus infection in whiteflies.Autophagy,12(9):1560-1574
Xie W,Chen CH,Yang ZH,Guo LT,Yang X,Wang D,Chen M,Huang JQ,Wen YN,Zeng Y,et al.2017.Genome sequencing of the sweetpotato whitefly Bemisia tabaci MED/Q.GigaScience,6(5):1-7
Xu J,de Barro PJ,Liu SS.2010.Reproductive incompatibility among genetic groups of Bemisia tabaci supports the proposition that the whitefly is a cryptic species complex.Bulletin of Entomological Research,100(3):359-366
Yang RSH,Pelliccia JG,Wilkinson CF.1973.Age-dependent arylsulphatase and sulphotransferase activities in the southern armyworm:a possible insect endocrine regulatory mechanism?Biochemical Journal,136(3):817-820
Yang RSH,Wilkinson CF.1972.Enzymic sulphation of p-nitrophenol and steroids by larval gut tissues of the southern armyworm(Prodenia eridania Cramer).The Biochemical Journal,130(2):487-493
Yorimitsu T,Klionsky DJ.2005.Autophagy:molecular machinery for self-eating.Cell Death&Differentiation,12(S2):1542-1552
Brown JK,Coats SA,Bedford ID,Markham PG,Bird J,Frohlich DR.1995.Characterization and distribution of esterase electromorphs in the whitefly,Bemisia tabaci(Genn.)(Homoptera:Aleyrodidae).Biochemical Genetics,33(7/8):205-214
Brown JK,Czosnek H.2002.Whitefly transmission of plant viruses.Advances in Botanical Research,36:65-100
Byrne DN,Bellows TS Jr.1991.Whitefly biology.Annual Review of Entomology,36(1):431-457
Czosnek H,Ghanim M.2002.The circulative pathway of begomoviruses in the whitefly vector Bemisia tabaci:insights from studies with Tomato yellow leaf curl virus.Annals of Applied Biology,140(3):215-231
Czosnek H,Ghanim M.2012.Back to basics:are begomoviruses whitefly pathogens?Journal of Integrative Agriculture,11(2):225-234
de Barro PJ,Liu SS,Boykin LM,Dinsdale AB.2011.Bemisia tabaci:a statement of species status.Annual Review of Entomology,56:1-19
Dinsdale A,Cook L,Riginos C,Buckley YM,de Barro P.2010.Refined global analysis of Bemisia tabaci(Hemiptera:Sternorrhyncha:Aleyrodoidea:Aleyrodidae)mitochondrial cytochrome oxidase 1 to identify species level genetic boundaries.Annals of the Entomological Society of America,103(2):196-208
Dodgson KS,Rose FA.1970.Sulfoconjugation and sulfohydrolysis.Metabolic Conjugation and Metabolic Hydrolysis,1:239-325
Dzialoszyński LM,Fr?hlich A,Kuczyński J,Pydzik T.1967.The activity of arylsulfatase in some tumors of the genital organs in women.Archivum Immunologiae et Therapiae Experimentalis,15(1):97-99
Feferman L,Bhattacharyya S,Deaton R,Gann P,Guzman G,KajdacsyBalla A,Tobacman JK.2013.Arylsulfatase B(N-acetylgalactosamine-4-sulfatase):potential role as a biomarker in prostate cancer.Prostate Cancer and Prostatic Diseases,16(3):277-284
Gray S,Cilia M,Ghanim M.2014.Circulative,“nonpropagative”virus transmission:an orchestra of virus-,insect-,and plant-derived instruments.Advances in Virus Research,89:141-199
Hariton Shalev A,Sobol I,Ghanim M,Liu SS,Czosnek H.2016.The whitefly Bemisia tabaci knottin-1 gene is implicated in regulating the quantity of Tomato yellow leaf curl virus ingested and transmitted by the insect.Viruses,8(7):e205
Hasegawa DK,Chen W,Zheng Y,Kaur N,Wintermantel WM,Simmons AM,Fei Z,Ling KS.2018.Comparative transcriptome analysis reveals networks of genes activated in the whitefly,Bemisia tabaci when fed on tomato plants infected with Tomato yellow leaf curl virus.Virology,513:52-64
Hogenhout SA,Ammar ED,Whitfield AE,Redinbaugh MG.2008.Insect vector interactions with persistently transmitted viruses.Annual Review of Phytopathology,46:327-359
Hogenhout SA,Bos JIB.2011.Effector proteins that modulate plant-insect interactions.Current Opinion in Plant Biology,14(4):422-428
Hu J,de Barro P,Zhao H,Wang J,Nardi F,Liu SS.2011.An extensive field survey combined with a phylogenetic analysis reveals rapid and widespread invasion of two alien whiteflies in China.PLoSONE,6(1):e16061
Jiu M,Zhou XP,Liu SS.2006.Acquisition and transmission of two begomoviruses by the B and a non-B biotype of Bemisia tabaci from Zhejiang,China.Journal of Phytopathology,154(10):587-591
Jiu M,Zhou X P,Tong L,Xu J,Yang X,Wan FH,Liu SS.2007.Vectorvirus mutualism accelerates population increase of an invasive whitefly.PLoS ONE,2(1):e182
Kanakala S,Ghanim M.2016.Implication of the whitefly Bemisia tabaci cyclophilin B protein in the transmission of Tomato yellow leaf curl virus.Frontiers in Plant Science,7:1702
Kikal T,Smith JN.1959.Comparative detoxication 6:the metabolism of 6-amino-4 nitro-o-cresol and 4:6-dinitro-o-cresol in locusts.The Biochemical Journal,71(1):48-54
Klionsky DJ,Emr SD.2000.Autophagy as a regulated pathway of cellular degradation.Science,290(5497):1717-1721
Li MR,Wang R,Sheng Q.2014.Advances in research of arylsulfatase in tumro diagnosis and oncotherapy.Pharmaceutical Biotechnology,21(1):81-85(in Chinese)[李孟如,王蓉,盛清.2014.芳香基硫酸酯酶在肿瘤诊断和治疗中的研究进展.药物生物技术,21(1):81-85]
Li R,Xie W,Wang S,Wu Q,Yang N,Yang X,Pan H,Zhou X,Bai L,Xu B,et al.2013.Reference gene selection for qRT-PCR analysis in the sweetpotato whitefly,Bemisia tabaci(Hemiptera:Aleyrodidae).PLoS ONE,8(1):e53006
Luan JB,Li JM,Varela N,Wang YL,Li FF,Bao YY,Zhang CX,Liu SS,Wang XW.2011.Global analysis of the transcriptional response of whitefly to Tomato yellow leaf curl China virus reveals the relationship of coevolved adaptations.Journal of Virology,85(7):3330-3340
Mitsunaga-Nakatsubo K,Kusunoki S,Kawakami H,Akasaka K,Akimoto Y.2009.Cell-surface arylsulfatase A and B on sinusoidal endothelial cells,hepatocytes,and Kupffer cells in mammalian livers.Medical Molecular Morphology,42(2):63-69
Mugiira RB,Liu SS,Zhou X.2008.Tomato yellow leaf curl virus and Tomato leaf curl Taiwan virus invade south-east coast of China.Journal of Phytopathology,156(4):217-221
Pan LL,Chen QF,Zhao JJ,Guo T,Wang XW,Hariton-Shalev A,Czosnek H,Liu SS.2017.Clathrin-mediated endocytosis is involved in Tomato yellow leaf curl virus transport across the midgut barrier of its whitefly vector.Virology,502:152-159
Prabhu SV,Bhattacharyya S,Guzman-Hartman G,Macias V,KajdacsyBalla A,Tobacman JK.2011.Extra-lysosomal localization of arylsulfatase B in human colonic epithelium.Journal of Histochemistry&Cytochemistry,59(3):328-335
Ren SX,Qiu BL,Ge F,Zhang YJ,Du YZ,Chen XX,Guo JY,Lin KJ,Peng ZQ,Yao SL,et al.2011.Research progress of the monitoring,forecast and sustainable management of whitefly pests in China.Chinese Journal of Applied Entomology,48(1):7-15(in Chinese)[任顺祥,邱宝利,戈峰,张友军,杜予州,陈学新,郭建英,林克剑,彭正强,姚松林,等.2011.粉虱类害虫的监测预警与可持续治理技术透视.应用昆虫学报,48(1):7-15]
Robinson D,Smith JN,Williams RT.1953.Studies in detoxication.XLVI.β-glucuronidase and arylsulphatase in the crop fluid of locusts.The Biochemical Journal,53(1):125-129
Shatters RG Jr.,Powell CA,Boykin LM,Liansheng H,McKenzie CL.2009.Improved DNA barcoding method for Bemisia tabaci and related Aleyrodidae:development of universal and Bemisia tabaci biotype-specific mitochondrial cytochrome c oxidase I polymerase chain reaction primers.Journal of Economic Entomology,102(2):750-758
Su YL,He WB,Wang J,Li JM,Liu SS,Wang XW.2013.Selection of endogenous reference genes for gene expression analysis in the Mediterranean species of the Bemisia tabaci(Hemiptera:Aleyrodidae)complex.Journal of Economic Entomology,106(3):1446-1455
Wang K,Li LK.1981.Arylsulfatase and its isoenzyme.Foreign Medical Sciences:Clinical Biochemistry and Laboratory Book,(4):1-6(in Chinese)[王坤,李立群.1981.芳香基硫酸酯酶及其同工酶.国外医学:临床生物化学与检验学分册,(4):1-6]
Wang LL,Wang XR,Wei XM,Huang H,Wu JX,Chen XX,Liu SS,Wang XW.2016.The autophagy pathway participates in resistance to Tomato yellow leaf curl virus infection in whiteflies.Autophagy,12(9):1560-1574
Xie W,Chen CH,Yang ZH,Guo LT,Yang X,Wang D,Chen M,Huang JQ,Wen YN,Zeng Y,et al.2017.Genome sequencing of the sweetpotato whitefly Bemisia tabaci MED/Q.GigaScience,6(5):1-7
Xu J,de Barro PJ,Liu SS.2010.Reproductive incompatibility among genetic groups of Bemisia tabaci supports the proposition that the whitefly is a cryptic species complex.Bulletin of Entomological Research,100(3):359-366
Yang RSH,Pelliccia JG,Wilkinson CF.1973.Age-dependent arylsulphatase and sulphotransferase activities in the southern armyworm:a possible insect endocrine regulatory mechanism?Biochemical Journal,136(3):817-820
Yang RSH,Wilkinson CF.1972.Enzymic sulphation of p-nitrophenol and steroids by larval gut tissues of the southern armyworm(Prodenia eridania Cramer).The Biochemical Journal,130(2):487-493
Yorimitsu T,Klionsky DJ.2005.Autophagy:molecular machinery for self-eating.Cell Death&Differentiation,12(S2):1542-1552