Comparative binding of Cry1Ab and Cry1F Bacillus thuringiensis toxins to brush border membrane proteins from Ostrinia nubilalis, Ostrinia furnacalis and Diatraea saccharalis (Lepidoptera: Crambidae) midgut tissue

Authors

Sek Yee Tan, University of Nebraska–Lincoln
Bonifacio F. Cayabyab, University of the Philippines, Los Baños
Edwin P. Alcantara, University of the Philippines, Los Baños
Fangneng Huang, Louisiana State University
Kanglai He, Chinese Academy of Agricultural Sciences
Kenneth Nickerson, University of Nebraska-LincolnFollow
Blair Siegfried, University of Nebraska--LincolnFollow

Document Type

Article

Date of this Version

2013

Citation

Published in Journal of Invertebrate Pathology 114:3 (2013), pp. 234–240. doi: 10.1016/j.jip.2013.08.007

Comments

Copyright © 2013 Elsevier. Used by permission.

Abstract

The European (Ostrinia nubilalis Hübner) and Asian corn borers (Ostrinia furnacalis Guenée) are closely related and display similar sensitivity to Cry1 toxins. In this study, we compared the binding patterns of Cry1Ab and Cry1F toxins between both Ostrinia spp., as well as the expression of putative cad­herin and aminopeptidase-N (APN)-like protein receptors. Additionally, cDNA sequences of these putative toxin receptors from both Ostrinia species were compared. Ligand blots for both species indicated a similar binding pattern for Cry1Ab with the strongest immunoreactive band at 260 kDa in both species. In addition, similar expression of the putative cadherin- and APN-like protein re­ceptors were observed at 260 and 135 kDa, respectively. A high degree of sim­ilarity (98% amino acid sequence identity) of cDNA sequences for both puta­tive receptor sequences was observed. The Cry1F ligand blot revealed that O. furnacalis and O. nubilalis BBMV exhibited slightly different binding patterns, with strong binding to putative proteins at 150 and 140 kDa, respectively. Both proteins appeared to also bind Cry1Ab, although the signal intensity was much reduced with Cry1Ab. O. furnacalis showed an additional but weaker band at 210 kDa relative to the 150 kDa band. Diatraea saccharalis (Fabricius), which was used as an outgroup species, exhibited different binding patterns than ei­ther Ostrinia species, with both Cry1Ab and Cry1F toxins binding to a 210 kDa protein. These results support the previous experiments indicating that O. nu­bilalis and O. furnacalis share similar patterns of susceptibility to Cry toxins.