|
|
|
|
|
| A Fragment of Cadherin-Like Protein Enhances Bacillus thuringiensis Cry1B and Cry1C Toxicity to Spodoptera exigua (Lepidoptera: Noctuidae) |
| LU Qiong, ZHANG Yong-jun, CAO Guang-chun, ZHANG Li-li, LIANG Ge-mei, LU Yan-hui, WU Kong-ming, GAO Xi-wu , GUO Yu-yuan |
1.College of Agriculture and Biotechnology, China Agricultural University, Beijing 100193, P.R.China
2.State Key Laboratory for Biology of Plant Diseases and Insect Pests/Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100193, P.R.China |
|
|
|
摘要 Cry toxins produced by Bacillus thuringiensis (Bt) are effective biological insecticides against certain insect species. In this study, bioassay results indicated that Cry1B and Cry1C were toxic to Spodoptera exigua. We also identified a cadherin-like gene in S. exigua that could enhance the toxicity of Cry1B and Cry1C. The cadherin-like gene identified from the larvae midgut tissue was cloned by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). The full-length cDNA of the gene consisted of 5 220 bp encoding 1 740 amino acid with a predicted molecular mass of 196 kD. BLAST search analysis showed that the predicted amino acid sequence had a high sequence identity to the published sequences of cadherin-like proteins from other Lepidoptera insects. Spatial expression of the cadherin-like gene detected by qRT-PCR analysis revealed that the cadherin-like gene was mainly present in the gut of 4th instar larvae and during different life stages. The results suggested that the commercial development of this synergist has the potential to enhance Cry1B and Cry1C toxicity against Lepidoptera insects.
Abstract Cry toxins produced by Bacillus thuringiensis (Bt) are effective biological insecticides against certain insect species. In this study, bioassay results indicated that Cry1B and Cry1C were toxic to Spodoptera exigua. We also identified a cadherin-like gene in S. exigua that could enhance the toxicity of Cry1B and Cry1C. The cadherin-like gene identified from the larvae midgut tissue was cloned by reverse transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA ends (RACE). The full-length cDNA of the gene consisted of 5 220 bp encoding 1 740 amino acid with a predicted molecular mass of 196 kD. BLAST search analysis showed that the predicted amino acid sequence had a high sequence identity to the published sequences of cadherin-like proteins from other Lepidoptera insects. Spatial expression of the cadherin-like gene detected by qRT-PCR analysis revealed that the cadherin-like gene was mainly present in the gut of 4th instar larvae and during different life stages. The results suggested that the commercial development of this synergist has the potential to enhance Cry1B and Cry1C toxicity against Lepidoptera insects.
|
|
Received: 14 April 2011
Accepted:
|
| Fund: This work was supported by the National Basic Research Program of China (2012CB114104) and the National Natural Science Foundation of China (31071694 and 31171858). |
|
Corresponding Authors:
Correspondence GUO Yu-yuan, Tel/Fax: +86-10-62894786, E-mail: yuyuanguo@hotmail.com
E-mail: yuyuanguo@hotmail.com
|
| About author: LU Qiong, E-mail: luqiong19@sina.com |
Cite this article:
LU Qiong, ZHANG Yong-jun, CAO Guang-chun, ZHANG Li-li, LIANG Ge-mei, LU Yan-hui, WU Kong-ming, GAO Xi-wu , GUO Yu-yuan .
2012.
A Fragment of Cadherin-Like Protein Enhances Bacillus thuringiensis Cry1B and Cry1C Toxicity to Spodoptera exigua (Lepidoptera: Noctuidae). Journal of Integrative Agriculture, 12(4): 628-638.
|
| [1]Abdullah M A, Moussa S, Taylor M D, Adang M J. 2009. Manduca sexta (Lepidoptera: Sphingidae) cadherin fragments function as synergists for Cry1A and Cry1C Bacillus thuringiensis toxins against noctuid moths Helicoverpa zea, Agrotis ipsilon and Spodoptera exigua. Pest Management Science, 65, 1097-1103. [2]Abdul-Rauf M, Ellar D J. 1999. Toxicity and receptor binding properties of a Bacillus thuringiensis Cry1C toxin active against both lepidoptera and diptera. Journal of Invertebrate Pathology, 73, 52-58. [3]Aimanova K G, Zhuang M, Gill S S. 2006. Expression of Cry1Ac cadherin receptors in insect midgut and cell lines. Journal of Invertebrate Pathology, 92, 178-187. [4]Bradford M M. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry, 72, 248-254. [5]Bravo A, Gomez I, Conde J, Munoz-Garay C, Sanchez J, Miranda R, Zhuang M, Gill S S, Soberon M. 2004. Oligomerization triggers binding of a Bacillus thuringiensis Cry1Ab pore-forming toxin to aminopeptidase N receptor leading to insertion into membrane microdomains. Biochimica et Biophysica Acta, 1667, 38-46. [6]Broderick N A, Raffa K F, Handelsman J. 2006. Midgut bacteria required for Bacillus thuringiensis insecticidal activity. Proceedings of the National Academy of Sciences of the United States of America, 103, 15196-15199. [7]Carrièrea Y, Showalter A M, Fabrick J A, Sollome J, Ellers-Kirk C, Tabashnik B E. 2009. Cadherin gene expression and effects of Bt resistance on sperm transfer in pink bollworm. Journal of Invertebrate Pathology, 55, 1058-1064. [8]Chen J, Brown M R, Hua G, Adang M J. 2005. Comparison of the localization of Bacillus thuringiensis Cry1A delta-endotoxins and their binding proteins in larval midgut of tobacco hornworm, Manduca sexta. Cell Tissue Research, 321, 123-129. [9]Chen J, Hua G, Jurat-Fuentes J L, Abdullah M A, Adang M J. 2007. Synergism of Bacillus thuringiensis toxins by a fragment of a toxin-binding cadherin. Proceedings of the National Academy of Sciences of the United States of America, 104, 13901-13906. [10]Dror A, Haviva E, Menachem K, Noam R, Michal S, Baruch S, Aciah Z. 2009. The Bacillus thuringiensis deltaendotoxin Cry1C as a potential bioinsecticide in plants. Plant Science, 176, 315-324. [11]Dorsch J A, Candas M, Griko N B, Maaty W S A, Midboe E G, Vadlamudi R K, Bulla J L A. 2002. Cry1A toxins of Bacillus thuringiensis bind specifically to a region adjacent to the membrane-proximal extracellular domain of BT-R1 in Manduca sexta: involvement of a cadherin in the entomopathogenicity of Bacillus thuringiensis. Insect Biochemisry and Molecular Biology, 32, 1025-1036. [12]Fabrick J, Oppert C, Lorenzen M D, Morris K, Oppert B, Jurat-Fuents L. 2009. A novel Tenebrio Molitor cadherin is a functional receptor for Bacillus thuringiensis Cry3Aa toxin. The Journal of Biological Chemistry, 27, 18401-18410. [13]Flannagan R D, Yu C, Mathis J P, Meyer T E, Shi X, Siqueira H A A, Siegfried B D. 2005. Identification, cloning and expression of a Cry1Ab cadherin receptor from European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae). Insect Biochemisry and Molecular Biology, 35, 33-40. [14]Gahan L J, Gould F, Heckel D G. 2001. Identification of a gene associated with Bt resistance in Heliothis virescens. Science, 293, 857-860. [15]Gill S S, Cowles E A, Francis V. 1995. Identification, isolation, and cloning of a Bacillus thuringiensis Cry1Ac toxin-binding protein from the midgut of the Lepidopteran insect Heliothis virescens. Journal of Biological Chemistry, 270, 27277-27282. [16]Gilliland A, Chambers C E, Bone E J, Ellar D J. 2002. Role of Bacillus thuringiensis Cry1 ?-endotoxin binding in determining potency during Lepidopteran larval development. Applied and Environmental Microbiology, 68, 1509-1515. [17]Gómez I, Dean D H, Bravo A, Soberón M. 2003. Molecular basis for Bacillus thuringiensis Cry1Ab toxin specificity: two structural determinants in the Manduca sexta: Bt-R1 receptors interact with loops ?-8 and 2 in domain II of Cry1Ab toxin. ACS Chemical Biology, 42, 10482-10489. [18]Gómez I, Oltean D I, Gill S S, Bravo A, Soberón M. 2001. Mapping the epitope in Cadherin-like receptors involved in Bacillus thuringiensis Cry1A toxin interaction using phage display. Journal of Biological Chemistry, 276, 28906-28912. [19]Granados R R, Fu Y, Corsaro B, Hughes P R. 2001. Enhancement of Bacillus thuringiensis toxicity to lepidopterous species with the enhancin from Trichoplusia ni granulovirus. Biological Control, 20, 153-159. [20]Griffitts J S, Aroian R V. 2005. Many roads to resistance: how invertebrates adapt to Bt toxins. BioEssays, 27, 614-624. [21]Griko N, Candas M, Zhang X, Junker M. 2004. Selective antagonism to the cadherin BT-R1 interferes with calcium-induced adhesion of epithelial membrane vesicles. Biochemistry, 43, 1393-1400. [22]Hernández-Martínez P, Ferré J, Escriche B. 2008. Susceptibility of Spodoptera exigua to 9 toxins from Bacillus thringiensis. Journal of Invertebrate Pathology, 97, 245-250. [23]Herrero S, Gónzalez-Cabrera J, Tabashnik B E, Ferré J. 2001. Shared binding sites in Lepidoptera for Bacillus thuringiensis Cry1Ja and Cry1A toxins. Applied and Environmental Microbiology, 67, 5729-5734. [24]Hara H, Atsumi S, Yaoi K, Nakanishi K, Higurashi S, Miura N, Tabunoki H, Sato R. 2003. A cadherin-like protein functions as a receptor for Bacillus thuringiensis Cry1Aa and Cry1Ac toxins on midgut epithelial cells of Bombyx mori larvae. FEBS Letters, 538, 29-34. [25]Ibiza-Palacios M S, Ferré J, Higurashi S, Miyamoto K, Sato R, Escriche B. 2008. Selective inhibition of binding of Bacillus thuringiensis Cry1Ab toxin to cadherin-like and aminopeptidase proteins in brush-border membranes and dissociated epithelial cells from Bombyx mori. Biochemical Journal, 409, 215-221. [26]James C. 2010. Global Status of Commercialized Biotech/ GM Crops: 2010. ISAAA 41, Ithaca, New York, USA. Jurat-Fuentes J L, Adang M J. 2004. Characterization of a Cry1Ac receptor alkaline phosphatase in susceptible and resistant Heliothis virescens larvae. European Journal of Biochemistry, 271, 3127-3135. [27]Jurat-Fuentes J L, Adang M J. 2006. The Heliothis virescens cadherin protein expressed in Drosophila S2 cells functions as a receptor for Bacillus thuringiensis Cry1A but not Cry1Fa toxins. Biochemistry, 45, 9688-9695. [28]Knight P J, Knowles B H, Ellar D J. 1995. Molecular cloning of an insect aminopeptidase N that serves as a receptor for Bacillus thuringiensis Cry1A(c) toxin. Journal of Biological Chemistry, 270, 17765-17770. [29]Lee M K, Curtiss A, Alcantara E, Dean D H. 1996. Synergistic effect of the Bacillus thuringiensis toxins Cry1Aa and Cry1Ac on the gypsy moth, Lymantria dispar. Applied and Environmental Microbiology, 62, 583-596. [30]LeOra Software. 1987. POLO-PC: A User’s Manual for Probit or Logit Analysis. Berkeley, CA. Liang G M, Tan W J, Guo Y Y. 1999. Improvement technology of artificial feeding bollworm. Plant Protection, 25, 15-17. (in Chinese) [31]Liu C X, Wu K M, Wu Y D, Gao Y L, Ning C M, Oppert B. 2009. Reduction of Bacillus thuringiensis Cry1Ac toxicity against Helicoverpa armigera by a soluble toxin-binding cadherin fragment. Journal of Invertebrate Pathology, 55, 686-693. [32]Luo K, Banks D, Adang M J. 1998. Toxicity, binding, permeability analyses of four Bacillus thuringiensis Cry1 δ-endotoxins using brush border membrane vesicles of Spodoptera exigua and Spodoptera frugiperda. Applied and Environmental Microbiology, 99, 457-464. [33]Luttrell R G, Wan L, Knighten K. 1999. Variation in susceptibility of noctuid (Lepidoptera) larvae attacking cotton and soybean to purified endotoxin proteins and commercial formulations of Bacillus thuringiensis. Journal of Economic Entomology, 92, 21-32. [34]de Maagd R, Weemen-Hendriks M, Stiekema W, Bosch D. 2000. Bacillus thuringiensis delta-endotoxin Cry1C domain III can function as a specificity determinant for Spodoptera exigua in different, but not all, Cry1-Cry1C hybrids. Applied and Environmental Microbiology, 66, 1559-1563. [35]Moar W J, Pusztai-Carey M, Van Faassen H, Bosh D, Frutis R, Rang C, Luo K, Adang M J. 1995. Development of Bacillus thuringiensis Cry1C resistance by Spodoptera exigua (Hübner) (Lepidoptera: Noctuidae). Applied and Environmental Microbiology, 95, 2086-2092. [36]Morin S, Biggs R W, Sisterson M S, Shriver L, Ellers-Kirk C, Higginson D, Holley D, Gahan L J, Heckel D G, Carriére Y, et al. 2003. Three cadherin alleles associated with resistance to Bacillus thuringiensis in pink bollworm. Proceedings of the National Academy of Sciences of the United States of America, 100, 5004-5009. [37]Moulton J K, Pepper D A, Dennehy T J. 2000. Beet armyworm (Spodoptera exigua) resistance to spinosad. Pest Management Science, 56, 842-848. [38]Nagamatsu Y, Toda S, Koike T, Miyoshi Y, Shigematsu S, Kogure M. 1998a. Cloning, sequencing, and Expression of the Bombyx mori receptor for Bacillus thuringiensis insecticidal CryIA(a) toxin. Bioscience Biotechnology Biochemistry, 62, 727-734. [39]Nagamatsu Y, Toda S, Yamaguchi F, Ogo M, Kogure M, Nakamura M, Shibata Y, Katsumoto T. 1998b. Identification of Bombyx mori midgut receptor for Bacillus thuringiensis insecticidal CryIA(a) toxin. Bioscience Biotechnology Biochemistry, 62, 718-726. [40]Pacheco S, Gómez I, Gill S S, Bravo A, Soberón M. 2009. Enhancement of insecticidal activity of Bacillus thuringiensis Cry1A toxins by fragments of a toxinbinding cadherin correlates with oligomer formation. Peptides, 30, 533-588. [41]Pérez C, Fernandez L E, Sun J, Folch J L, Gill S S, Soberón M, Bravo A. 2005. Bacillus thuringiensis subsp. israelensis Cyt1Aa synergizes Cry11Aa toxin by functioning as a membrane-bound receptor. Proceedings of the National Academy of Sciences of the United States of America, 102, 18303-18308. [42]Pigott C R, Ellar D J. 2007. Role of receptors in Bacillus thuringiensis crystal toxin activity. Microbiology and Molecular Biology Reviews, 71, 255-281. [43]Regev A, Keller M, Strizhov N, Sneh B, Prudovsky E, Chet I, Ginzberg I, Koncz-Kalman Z, Koncz C, Schell J, et al. 1996. Synergistic activity of a Bacillus thuringiensis delta-endotoxin and a bacterial endochitinase against S p o d o p t e r a l i t t o r a l i s l a r v a e . Applied and Environmental Microbiology, 62, 3581-3586. [44]Sayed A, Nekl E R, Siqueira H A A, Wang H C, ffrench-Constant R H, Bagley M, Siegfried B D. 2007. A novel cadherin-like gene from western corn rootworm, Diabrotica vigifera (Coleoptera: Chrysomelidae), larval midgut tissue. Insect Biochemisry and Molecular Biology, 16, 591-600. [45]Smits P H, van Velden M C, van de Vrie M, Vlak J M. 1987. Feeding and dispersion of Spodoptera exigua larvae and its relevance for control with a nuclear polyhedrosis virus. Entomologia Experimentalis et Applicata, 43, 67-72. [46]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. [47]Tabashnik B E. 1992. Evalution of synergism among Bacillus thuringiensis toxins. Applied and Environmental Microbiology, 58, 3343-3346. [48]Tabashnik B E. 1994. Evolution of resistance to Bacillus thuringiensis. Annual Review of Entomology, 39, 47-79. [49]Tsuda Y, Nakatani F, Hashimoto K, Ikawa S, Matsuura C, Fukada T, Sugimoto K, Himeno M. 2003. Cytotoxic activity of Bacillus thuringiensis Cry proteins on mammalian cells transfected with cadherin-like Cry receptor gene of Bombyx mori (silkworm). Biochemical Journal, 369, 697-703. [50]Vadlamudi R K, Ji T H, Bulla Jr L A. 1993. A specific binding protein from Manduca sexta for the insecticidal toxin of Bacillus thuringiensis subsp. berliner. Journal of Biological Chemistry, 268, 12334-12340. [51]Vadlamudi R K, Weber E, Ji I, Ji T H, Bulla Jr L A. 1995. Cloning and expression of a receptor for an insecticidal toxin of Bacillus thuringiensis. Journal of Biological Chemistry, 270, 5490-5494. [52]Wang G, Wu K, Liang G, Guo Y. 2005. Gene cloning and expression of cadherin in midgut of Helicoverpa armigera and its Cry1A binding region. Science in China Series (C: Life Sciences), 48, 346-356. [53]Wu K, Lu Y, Feng H, Jiang Y, Zhao J. 2008. Suppression of cotton bollworm in multiple crops in China in areas with Bt toxin-containing cotton. Science, 321, 1676-1678. [54]Xie R, Zhuang M, Ross L S, Gómez I, Oltean D I, Bravo A, Soberon M, Gill S S. 2005. Single amino acid mutations in the cadherin receptor from Heliothis virescens affect its toxin binding ability to Cry1A toxins. The Journal of Biological Chemistry, 280, 8416-8425. [55]Zhang X, Candas M, Griko N B, Rose-Young L, Bulla Jr L A. 2005. Cytotoxicity of Bacillus thuringiensis Cry1Ab toxin depends on specific binding of the toxin to the cadherin receptor BT-R1 expressed in insect cells. Cell Death and Differentiation, 12, 1407-1416. [56]Zhang X, Candas M, Griko N B, Taussig R, Bulla Jr L A. 2006. A mechanism of cell death involving an adenylyl cyclase/PKA signaling pathway is induced by the Cry1Ab toxin of Bacillus thuringiensis. Proceedings of the National Academy of Sciences of the United States of America, 103, 9897-9902. [57]Zhang X, Griko N B, Corona S K, Bulla Jr L A. 2008. Enhanced exocytosis of the receptor BT-R1 induced by Cry1Ab toxin of Bacillus thuringiensis directly correlates to the execution of cell death. Comparative Biochemistry and Physiology (Part B), 149, 581-588. |
| No Suggested Reading articles found! |
|
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
