Polycalin is involved in the action mechanism of Cry2Aa toxin in Helicoverpa armigera(Hübner)
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摘要
Receptor proteins on the brush border membrane of the insect midgut epithelium are involved in the mode of action of insecticidal Cry proteins from Bacillus thuringiensis(Bt). Polycalin has been identified as a binding protein of the Bt Cry1 Ac toxin in several Lepidoptera including Helicoverpa armigera, but its role in the action mechanism of Cry2 Aa is still unclear. In this study, we investigated the binding characteristics of polycalin from the midgut of H. armigera with Cry2 Aa and its role in the toxicity of Cry2 Aa. The results demonstrated that heterologously expressed H. armigera polycalin peptide could bind with Cry2 Aa with high affinity(K_d=32 nmol L~(–1)). The toxicity of Cry2 Aa decreased by 27% after H. armigera larvae ingested polycalin antisera. These results suggested that polycalin could be a potential functional receptor for Cry2 Aa, and it plays an important role in the susceptibility of H. armigera to Cry2 Aa.
Receptor proteins on the brush border membrane of the insect midgut epithelium are involved in the mode of action of insecticidal Cry proteins from Bacillus thuringiensis(Bt). Polycalin has been identified as a binding protein of the Bt Cry1 Ac toxin in several Lepidoptera including Helicoverpa armigera, but its role in the action mechanism of Cry2 Aa is still unclear. In this study, we investigated the binding characteristics of polycalin from the midgut of H. armigera with Cry2 Aa and its role in the toxicity of Cry2 Aa. The results demonstrated that heterologously expressed H. armigera polycalin peptide could bind with Cry2 Aa with high affinity(K_d=32 nmol L~(–1)). The toxicity of Cry2 Aa decreased by 27% after H. armigera larvae ingested polycalin antisera. These results suggested that polycalin could be a potential functional receptor for Cry2 Aa, and it plays an important role in the susceptibility of H. armigera to Cry2 Aa.
Receptor proteins on the brush border membrane of the insect midgut epithelium are involved in the mode of action of insecticidal Cry proteins from Bacillus thuringiensis(Bt). Polycalin has been identified as a binding protein of the Bt Cry1 Ac toxin in several Lepidoptera including Helicoverpa armigera, but its role in the action mechanism of Cry2 Aa is still unclear. In this study, we investigated the binding characteristics of polycalin from the midgut of H. armigera with Cry2 Aa and its role in the toxicity of Cry2 Aa. The results demonstrated that heterologously expressed H. armigera polycalin peptide could bind with Cry2 Aa with high affinity(K_d=32 nmol L~(–1)). The toxicity of Cry2 Aa decreased by 27% after H. armigera larvae ingested polycalin antisera. These results suggested that polycalin could be a potential functional receptor for Cry2 Aa, and it plays an important role in the susceptibility of H. armigera to Cry2 Aa.
引文
Adang M J,Crickmore N,Jurat-Fuentes J L.2014.Diversity of Bacillus thuringiensis crystal toxins and mechanism of action.In:Dhadialla T S,Gill S S,eds.,Advances in Insect Physiology.Academic Press,New York.pp.39-87.
Angelucci C,Barrettwilt G A,Hunt D F,Akhurst R J,East P D,Gordon K H,Campbell P M.2008.Diversity of aminopeptidases,derived from four lepidopteran gene duplications,and polycalins expressed in the midgut of Helicoverpa armigera:Identification of proteins binding theδ-endotoxin,Cry1Ac of Bacillus thuringiensis.Insect Biochemistry and Molecular Biology,38,685-696.
Aronson A I,Beckman W,Dunn P.1986.Bacillus thuringiensis and related insect pathogens.FEMS Microbiology Reviews,50,1-24.
Baxter S W,Badenes-Pérez F R,Morrison A,Vogel H,Crickmore N,Kain W,Wang P,Heckel D G,Jiggins CD.2011.Parallel evolution of Bacillus thuringiensis toxin resistance in Lepidoptera.Genetics,189,675-679.
Bradford M M.1976.A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding.Analytical Biochemistry,72,248-254.
Brévault T,Heuberger S,Zhang M,Ellerskirk C,Ni X,Masson L,Li X,Tabashnik B E,Carrière Y.2013.Potential shortfall of pyramided transgenic cottonfor insect resistance management.Proceedings of the National Academy of Sciences of the United States of America,110,5806-5811.
Caccia S,Hernández-Rodríguez C S,Mahon R J,Downes S,James W,Bautsoens M N,Van Rie J,FerréJ.2010.Binding site alteration is responsible for field-isolated resistance to Bacillus thuringiensis Cry2A insecticidal proteins in two Helicoverpa species.PLoS ONE,5,e9975.
Carrière Y,Crickmore N,Tabashnik B E.2015.Optimizing pyramided transgenic Bt crops for sustainable pest management.Nature Biotechnology,33,161-168.
Chen W B,Liu C X,Xiao Y T,Zhang D D,Zhang Y D,Li X C,Tabashnik B E,Wu K M.2015.A toxin-binding alkaline phosphatase fragment synergizes Bt toxin Cry1Ac against susceptible and resistant Helicoverpa armigera.PLoSONE,10,e0126288.
FerréJ,Rie J V.2002.Biochemistry and genetics of insect resistance to Bacillus thuringiensis insecticidal crystal proteins.Annual Review of Entomology,47,501-533.
Gahan L J,Gould F,Heckel D G.2001.Identification of a gene associated with Bt resistance in Heliothis virescens.Science,293,857-860.
Gahan L J,Pauchet Y,Vogel H,Heckel D G.2010.An ABCtransporter mutation is correlated with insect resistance to Bacillus thuringiensis Cry1Ac toxin.PLoS Genetics,6,e1001248.
Griffitts J S,Haslam S M,Yang T,Garczynski S F,Mulloy B,Morris H,Cremer P S,Dell A,Adang M J,Aroian R V.2005.Glycolipids as receptors for Bacillus thuringiensis crystal toxin.Science,37,922-925.
Guo Z,Kang S,Chen D,Wu Q,Wang S,Xie W,Zhu X,Baxter S,Zhou X,Jurat-Fuentes J,Zhang Y.2015.MAPK signaling pathway alters expression of midgut ALP and ABCC genes and causes resistance to Bacillus thuringiensis Cry1Ac toxin in diamondback moth.PLo S Genetics,11,e1005124.
Hernandez-Rodriguez C S,Van Vliet A,Bautsoens N,Van Rie J,Ferre J.2008.Specific binding of Bacillus thuringiensis Cry2A insecticidal proteins to a common site in the midgut of Helicoverpa species.Applied and Environmental Microbiology,74,7654-7659.
Hossain D M,Shitomi Y,Moriyama K,Higuchi M,Hayakawa T,Mitsui T,Sato R,Hori H.2004.Characterization of a novel plasma membrane protein,expressed in the midgut epithelia of Bombyx mori,that binds to Cry1A toxins.Applied and Environmental Microbiology,70,4604-4612.
Hossain D M,Shitomi Y,Nanjo Y,Takano D,Nishiumi T,Hayakawa T,Mitsui T,Sato R,Hori H.2005.Localization of a novel 252-kDa plasma membrane protein that binds Cryl A toxins in the midgut epithelia of Bombyx mori.Applied Entomology and Zoology,40,125-135.
Jakka S R,Gong L,Hasler J,Banerjee R,Sheets J J,Narva K,Blanco C A,Jurat-Fuentes J L.2016.Field-evolved mode 1 resistance of the fall armyworm to transgenic Cry1Fa-expressing corn associated with reduced Cry1Fa toxin binding and midgut alkaline phosphatase expression.Applied and Environmental Microbiology,82,1023-1034.
Jurat-Fuentes J L,Karumbaiah L,Jakka S R,Ning C,Liu C,Wu K,Jackson J,Gould F,Blanco C,Portilla M,Perera O,Adang M.2011.Reduced levels of membrane-bound alkaline phosphatase are common to lepidopteran strains resistant to Cry toxins from Bacillus thuringiensis.PLoSONE,6,e17606.
Liang G M,Tan W J,Guo Y Y.1999.An improvement in the technique of artificial rearing cotton bollworm.Plant Protection,25,15-17.(in Chinese)
Liu C Y,Gao Y,Ning C,Wu K M,Oppert B,Guo Y Y.2010.Antisera-mediated in vivo reduction of Cry1Ac toxicity in Helicoverpa armigera.Journal of Insect Physiology,56,718-724.
Ma W H,Zhang Z,Peng C H,Wang X,Li F,Lin Y.2012.Correction:Exploring the midgut transcriptome and brush border membrane vesicle proteome of the rice stem borer,Chilo suppressalis(Walker).PLoS ONE,7,e38151.
Mauchamp B,Royer C,Garel A,Jalabert A,Da Rocha M,Grenier A M,Labas V,Vinh J,Mita K,Kadono K,Chavancy G.2006.Polycalin(chlorophyllid a binding protein):Anovel,very large fluorescent lipocalin from the midgut of the domestic silkworm Bombyx mori L.Insect Biochemistry and Molecular Biology,36,623-633.
Nagamatsu Y,Toda S,Yamaguchi F,Ogo M,Kogure M,Nakamura M,Shibata Y,Katsumoto T.1998.Identification of Bombyx mori midgut receptor for Bacillus thuringiensis insecticidal Cry1A(a)toxin.Journal of the Agricultural Chemical Society of Japan,62,718-726.
Pandian G N,Ishikawa T,Togashi M,Shitomi Y,Haginoya K,Yamamoto S,Nishiumi T,Hori H.2008.Bombyx mori midgut membrane protein p252,which binds to Bacillus thuringiensis Cry1A,is a chlorophyllide-binding protein,and the resulting complex has antimicrobial activity.Applied and Environmental Microbiology,74,1324-1331.
Pandian G N,Ishikawa T,Vaijayanthi T,Hossain D M,Yamamoto S,Nishiumi T,Angsuthanasombat C,Haginoya K,Mitsui T,Hori H.2010.Formation of macromolecule complex with Bacillus thuringiensis Cry1A toxins and chlorophyllide binding 252-kDa lipocalin-like protein locating on Bombyx mori midgut membrane.The Journal of Membrane Biology,237,125-136.
Pardo-López L,Soberon M,Bravo A.2013.Bacillus thuringiensis insecticidal three-domain Cry toxins:Mode of action,insect resistance and consequences for crop protection.FEMS Microbiology Reviews,37,3-22.
Pigott C R,Ellar D J.2007.Role of receptors in Bacillus thuringiensis crystal toxin activity.Microbiology and Molecular Biology Reviews,71,255-281.
Qiu L,Zhang B,Liu L,Ma W,Wang X,Lei C,Chen L Z.2017.Proteomic analysis of Cry2Aa-binding proteins and their receptor function in Spodoptera exigua.Scientific Reports,7,40222.
Sivakumar S,Rajagopal R,Venkatesh G R,Srivastava A,Bhatnagar R K.2007.Knockdown of aminopeptidase-Nfrom Helicoverpa armigera larvae and in transfected Sf21cells by RNA interference reveals its functional interaction with Bacillus thuringiensis insecticidal protein Cry1Ac.Journal of Biological Chemistry,282,7312-7319.
Tabashnik B E.2015.ABCs of insect resistance to Bt.PLoSGenetics,11,e1005646.
Tabashnik B E,Brévault T,Carrière Y.2013.Insect resistance to Bt crops:Lessons from the first billion acres.Nature Biotechnology,31,510-521.
Tabashnik B E,Gassmann A J,Crowder D W,Carriere Y.2008.Insect resistance to Bt crops:Evidence versus theory.Nature Biotechnology,26,199-202.
Tabashnik B E,Unnithan G C,Masson L,Crowder D W,Li X,Carrière Y.2009.Asymmetrical cross-resistance between Bacillus thuringiensis toxins Cry1Ac and Cry2Ab in pink bollworm.Proceedings of the National Academy of Sciences of the United States of America,106,11889-11894.
Tanaka S,Miyamoto K,Noda H,Jurat-Fuentes J L,Yoshizawa Y,Endo H,Sato R.2013.The ATP-binding cassette transporter subfamily C member 2 in Bombyx mori larvae is a functional receptor for Cry toxins from Bacillus thuringiensis.FEBS Journal,280,1782-1794.
Wang Y N,Zhong F,Wei J Z,Xie B T,Zhang W N,Chen L Z,Liang G M.2015.Cloning and expression analysis of the polycalin gene in the cotton bollworm Helicoverpa armigera.Chinese Journal of Applied Entomology,52,549-556.(in Chinese)
Wei J,Guo Y,Liang G,Wu K,Zhang J,Tabashnik B E,Li X.2015.Cross-resistance and interactions between Bt toxins Cry1Ac and Cry2Ab against the cotton bollworm.Scientific Reports,5,7714.
Welch K L,Unnithan G C,Degain B A,Wei J Z,Zhang J,Li X C,Tabashnik B E,Carrière Y.2015.Cross-resistance to toxins used in pyramided Bt crops and resistance to Bt sprays in Helicoverpa zea.Journal of Invertebrate Pathology,132,149-156.
Xiao Y T,Zhang T,Liu C X,Heckel D,Li X C,Tabashnik BE,Wu K M.2014.Mis-splicing of the ABCC2 gene linked with Bt toxin resistance in Helicoverpa armigera.Scientific Reports,4,6184.
Xie R,Zhuang M,Ross L S,Gomez I,Oltean D I,Bravo A,SoberonM,Gill S S.2005.Single amino acid mutations in the cadherin receptor from Heliothis virescens affect its toxin binding ability to Cry1A toxins.Journal of Biological Chemistry,280,8416-8425.
Xu X,Wu Y.2008.Disruption of Ha_BtR alters binding of Bacillus thuringiensisδ-endotoxin Cry1Ac to midgut BBMVs of Helicoverpa armigera.Journal of Invertebrate Pathology,97,27-32.
Xu X J,Yu L Y,Wu Y D.2005.Disruption of a cadherin gene associated with resistance to Cry1Acδ-endotoxin of Bacillus thuringiensis in Helicoverpa armigera.Applied and Environmental Microbiology,71,948-954.
Yang Y,Zhu Y,Ottea J,Husseneder C,Leonard B R,Abel C,Luttrell R,Huang F N.2011.Down regulation of a gene for cadherin,but not alkaline phosphatase,associated with Cry1Ab resistance in the sugarcane borer Diatraea saccharalis.PLoS ONE,6,e25783.
Zhang S,Cheng H,Gao Y,Wang G,Liang G M,Wu K M.2009.Mutation of an aminopeptidase N gene is associated with Helicoverpa armigera resistance to Bacillus thuringiensis Cry1Ac toxin.Insect Biochemistry and Molecular Biology,39,421-429.
Zhang T,Coates B S,Wang Y,Wang Y,Bai S,Wang Z,He K.2017.Down-regulation of aminopeptidase N and ABCtransporter subfamily G transcripts in Cry1Ab and Cry1Ac resistant Asian corn borer,Ostrinia furnacalis(Lepidoptera:Crambidae).International Journal of Biological Sciences,13,835-851.
Zhao M,Yuan X D,Wei J Z,Zhang W N,Wang B J,Myint K,Liang G M.2017.Functional roles of cadherin,aminopeptidase-Nand alkaline phosphatase from Helicoverpa armigera(Hübner)in the action mechanism of Bacillus thuringiensis Cry2Aa.Scientific Reports,7,46555
Zhou Z S,Wang Z Y,Liu Y X,Liang G M,Shu C G,Song F P,Zhou X P,Bravo A,Soberon M,Zhang J.2016.Identification of ABCC2 as a binding protein of Cry1Ac on brush border membrane vesicles from Helicoverpa armigera by an improved pull-down assay.Microbiology Open,5,659-669.
Angelucci C,Barrettwilt G A,Hunt D F,Akhurst R J,East P D,Gordon K H,Campbell P M.2008.Diversity of aminopeptidases,derived from four lepidopteran gene duplications,and polycalins expressed in the midgut of Helicoverpa armigera:Identification of proteins binding theδ-endotoxin,Cry1Ac of Bacillus thuringiensis.Insect Biochemistry and Molecular Biology,38,685-696.
Aronson A I,Beckman W,Dunn P.1986.Bacillus thuringiensis and related insect pathogens.FEMS Microbiology Reviews,50,1-24.
Baxter S W,Badenes-Pérez F R,Morrison A,Vogel H,Crickmore N,Kain W,Wang P,Heckel D G,Jiggins CD.2011.Parallel evolution of Bacillus thuringiensis toxin resistance in Lepidoptera.Genetics,189,675-679.
Bradford M M.1976.A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding.Analytical Biochemistry,72,248-254.
Brévault T,Heuberger S,Zhang M,Ellerskirk C,Ni X,Masson L,Li X,Tabashnik B E,Carrière Y.2013.Potential shortfall of pyramided transgenic cottonfor insect resistance management.Proceedings of the National Academy of Sciences of the United States of America,110,5806-5811.
Caccia S,Hernández-Rodríguez C S,Mahon R J,Downes S,James W,Bautsoens M N,Van Rie J,FerréJ.2010.Binding site alteration is responsible for field-isolated resistance to Bacillus thuringiensis Cry2A insecticidal proteins in two Helicoverpa species.PLoS ONE,5,e9975.
Carrière Y,Crickmore N,Tabashnik B E.2015.Optimizing pyramided transgenic Bt crops for sustainable pest management.Nature Biotechnology,33,161-168.
Chen W B,Liu C X,Xiao Y T,Zhang D D,Zhang Y D,Li X C,Tabashnik B E,Wu K M.2015.A toxin-binding alkaline phosphatase fragment synergizes Bt toxin Cry1Ac against susceptible and resistant Helicoverpa armigera.PLoSONE,10,e0126288.
FerréJ,Rie J V.2002.Biochemistry and genetics of insect resistance to Bacillus thuringiensis insecticidal crystal proteins.Annual Review of Entomology,47,501-533.
Gahan L J,Gould F,Heckel D G.2001.Identification of a gene associated with Bt resistance in Heliothis virescens.Science,293,857-860.
Gahan L J,Pauchet Y,Vogel H,Heckel D G.2010.An ABCtransporter mutation is correlated with insect resistance to Bacillus thuringiensis Cry1Ac toxin.PLoS Genetics,6,e1001248.
Griffitts J S,Haslam S M,Yang T,Garczynski S F,Mulloy B,Morris H,Cremer P S,Dell A,Adang M J,Aroian R V.2005.Glycolipids as receptors for Bacillus thuringiensis crystal toxin.Science,37,922-925.
Guo Z,Kang S,Chen D,Wu Q,Wang S,Xie W,Zhu X,Baxter S,Zhou X,Jurat-Fuentes J,Zhang Y.2015.MAPK signaling pathway alters expression of midgut ALP and ABCC genes and causes resistance to Bacillus thuringiensis Cry1Ac toxin in diamondback moth.PLo S Genetics,11,e1005124.
Hernandez-Rodriguez C S,Van Vliet A,Bautsoens N,Van Rie J,Ferre J.2008.Specific binding of Bacillus thuringiensis Cry2A insecticidal proteins to a common site in the midgut of Helicoverpa species.Applied and Environmental Microbiology,74,7654-7659.
Hossain D M,Shitomi Y,Moriyama K,Higuchi M,Hayakawa T,Mitsui T,Sato R,Hori H.2004.Characterization of a novel plasma membrane protein,expressed in the midgut epithelia of Bombyx mori,that binds to Cry1A toxins.Applied and Environmental Microbiology,70,4604-4612.
Hossain D M,Shitomi Y,Nanjo Y,Takano D,Nishiumi T,Hayakawa T,Mitsui T,Sato R,Hori H.2005.Localization of a novel 252-kDa plasma membrane protein that binds Cryl A toxins in the midgut epithelia of Bombyx mori.Applied Entomology and Zoology,40,125-135.
Jakka S R,Gong L,Hasler J,Banerjee R,Sheets J J,Narva K,Blanco C A,Jurat-Fuentes J L.2016.Field-evolved mode 1 resistance of the fall armyworm to transgenic Cry1Fa-expressing corn associated with reduced Cry1Fa toxin binding and midgut alkaline phosphatase expression.Applied and Environmental Microbiology,82,1023-1034.
Jurat-Fuentes J L,Karumbaiah L,Jakka S R,Ning C,Liu C,Wu K,Jackson J,Gould F,Blanco C,Portilla M,Perera O,Adang M.2011.Reduced levels of membrane-bound alkaline phosphatase are common to lepidopteran strains resistant to Cry toxins from Bacillus thuringiensis.PLoSONE,6,e17606.
Liang G M,Tan W J,Guo Y Y.1999.An improvement in the technique of artificial rearing cotton bollworm.Plant Protection,25,15-17.(in Chinese)
Liu C Y,Gao Y,Ning C,Wu K M,Oppert B,Guo Y Y.2010.Antisera-mediated in vivo reduction of Cry1Ac toxicity in Helicoverpa armigera.Journal of Insect Physiology,56,718-724.
Ma W H,Zhang Z,Peng C H,Wang X,Li F,Lin Y.2012.Correction:Exploring the midgut transcriptome and brush border membrane vesicle proteome of the rice stem borer,Chilo suppressalis(Walker).PLoS ONE,7,e38151.
Mauchamp B,Royer C,Garel A,Jalabert A,Da Rocha M,Grenier A M,Labas V,Vinh J,Mita K,Kadono K,Chavancy G.2006.Polycalin(chlorophyllid a binding protein):Anovel,very large fluorescent lipocalin from the midgut of the domestic silkworm Bombyx mori L.Insect Biochemistry and Molecular Biology,36,623-633.
Nagamatsu Y,Toda S,Yamaguchi F,Ogo M,Kogure M,Nakamura M,Shibata Y,Katsumoto T.1998.Identification of Bombyx mori midgut receptor for Bacillus thuringiensis insecticidal Cry1A(a)toxin.Journal of the Agricultural Chemical Society of Japan,62,718-726.
Pandian G N,Ishikawa T,Togashi M,Shitomi Y,Haginoya K,Yamamoto S,Nishiumi T,Hori H.2008.Bombyx mori midgut membrane protein p252,which binds to Bacillus thuringiensis Cry1A,is a chlorophyllide-binding protein,and the resulting complex has antimicrobial activity.Applied and Environmental Microbiology,74,1324-1331.
Pandian G N,Ishikawa T,Vaijayanthi T,Hossain D M,Yamamoto S,Nishiumi T,Angsuthanasombat C,Haginoya K,Mitsui T,Hori H.2010.Formation of macromolecule complex with Bacillus thuringiensis Cry1A toxins and chlorophyllide binding 252-kDa lipocalin-like protein locating on Bombyx mori midgut membrane.The Journal of Membrane Biology,237,125-136.
Pardo-López L,Soberon M,Bravo A.2013.Bacillus thuringiensis insecticidal three-domain Cry toxins:Mode of action,insect resistance and consequences for crop protection.FEMS Microbiology Reviews,37,3-22.
Pigott C R,Ellar D J.2007.Role of receptors in Bacillus thuringiensis crystal toxin activity.Microbiology and Molecular Biology Reviews,71,255-281.
Qiu L,Zhang B,Liu L,Ma W,Wang X,Lei C,Chen L Z.2017.Proteomic analysis of Cry2Aa-binding proteins and their receptor function in Spodoptera exigua.Scientific Reports,7,40222.
Sivakumar S,Rajagopal R,Venkatesh G R,Srivastava A,Bhatnagar R K.2007.Knockdown of aminopeptidase-Nfrom Helicoverpa armigera larvae and in transfected Sf21cells by RNA interference reveals its functional interaction with Bacillus thuringiensis insecticidal protein Cry1Ac.Journal of Biological Chemistry,282,7312-7319.
Tabashnik B E.2015.ABCs of insect resistance to Bt.PLoSGenetics,11,e1005646.
Tabashnik B E,Brévault T,Carrière Y.2013.Insect resistance to Bt crops:Lessons from the first billion acres.Nature Biotechnology,31,510-521.
Tabashnik B E,Gassmann A J,Crowder D W,Carriere Y.2008.Insect resistance to Bt crops:Evidence versus theory.Nature Biotechnology,26,199-202.
Tabashnik B E,Unnithan G C,Masson L,Crowder D W,Li X,Carrière Y.2009.Asymmetrical cross-resistance between Bacillus thuringiensis toxins Cry1Ac and Cry2Ab in pink bollworm.Proceedings of the National Academy of Sciences of the United States of America,106,11889-11894.
Tanaka S,Miyamoto K,Noda H,Jurat-Fuentes J L,Yoshizawa Y,Endo H,Sato R.2013.The ATP-binding cassette transporter subfamily C member 2 in Bombyx mori larvae is a functional receptor for Cry toxins from Bacillus thuringiensis.FEBS Journal,280,1782-1794.
Wang Y N,Zhong F,Wei J Z,Xie B T,Zhang W N,Chen L Z,Liang G M.2015.Cloning and expression analysis of the polycalin gene in the cotton bollworm Helicoverpa armigera.Chinese Journal of Applied Entomology,52,549-556.(in Chinese)
Wei J,Guo Y,Liang G,Wu K,Zhang J,Tabashnik B E,Li X.2015.Cross-resistance and interactions between Bt toxins Cry1Ac and Cry2Ab against the cotton bollworm.Scientific Reports,5,7714.
Welch K L,Unnithan G C,Degain B A,Wei J Z,Zhang J,Li X C,Tabashnik B E,Carrière Y.2015.Cross-resistance to toxins used in pyramided Bt crops and resistance to Bt sprays in Helicoverpa zea.Journal of Invertebrate Pathology,132,149-156.
Xiao Y T,Zhang T,Liu C X,Heckel D,Li X C,Tabashnik BE,Wu K M.2014.Mis-splicing of the ABCC2 gene linked with Bt toxin resistance in Helicoverpa armigera.Scientific Reports,4,6184.
Xie R,Zhuang M,Ross L S,Gomez I,Oltean D I,Bravo A,SoberonM,Gill S S.2005.Single amino acid mutations in the cadherin receptor from Heliothis virescens affect its toxin binding ability to Cry1A toxins.Journal of Biological Chemistry,280,8416-8425.
Xu X,Wu Y.2008.Disruption of Ha_BtR alters binding of Bacillus thuringiensisδ-endotoxin Cry1Ac to midgut BBMVs of Helicoverpa armigera.Journal of Invertebrate Pathology,97,27-32.
Xu X J,Yu L Y,Wu Y D.2005.Disruption of a cadherin gene associated with resistance to Cry1Acδ-endotoxin of Bacillus thuringiensis in Helicoverpa armigera.Applied and Environmental Microbiology,71,948-954.
Yang Y,Zhu Y,Ottea J,Husseneder C,Leonard B R,Abel C,Luttrell R,Huang F N.2011.Down regulation of a gene for cadherin,but not alkaline phosphatase,associated with Cry1Ab resistance in the sugarcane borer Diatraea saccharalis.PLoS ONE,6,e25783.
Zhang S,Cheng H,Gao Y,Wang G,Liang G M,Wu K M.2009.Mutation of an aminopeptidase N gene is associated with Helicoverpa armigera resistance to Bacillus thuringiensis Cry1Ac toxin.Insect Biochemistry and Molecular Biology,39,421-429.
Zhang T,Coates B S,Wang Y,Wang Y,Bai S,Wang Z,He K.2017.Down-regulation of aminopeptidase N and ABCtransporter subfamily G transcripts in Cry1Ab and Cry1Ac resistant Asian corn borer,Ostrinia furnacalis(Lepidoptera:Crambidae).International Journal of Biological Sciences,13,835-851.
Zhao M,Yuan X D,Wei J Z,Zhang W N,Wang B J,Myint K,Liang G M.2017.Functional roles of cadherin,aminopeptidase-Nand alkaline phosphatase from Helicoverpa armigera(Hübner)in the action mechanism of Bacillus thuringiensis Cry2Aa.Scientific Reports,7,46555
Zhou Z S,Wang Z Y,Liu Y X,Liang G M,Shu C G,Song F P,Zhou X P,Bravo A,Soberon M,Zhang J.2016.Identification of ABCC2 as a binding protein of Cry1Ac on brush border membrane vesicles from Helicoverpa armigera by an improved pull-down assay.Microbiology Open,5,659-669.