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Published on May 8, 2008; 10.1104/pp.108.116145

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Received January 10, 2008
Accepted May 5, 2008

Functional characterization of HFR-1, a high mannose N-glycan-specific wheat lectin induced by Hessian fly larvae

Subhashree Subramanyam , David F. Smith , James C. Clemens , Mary A.Webb , Nagesh Sardesai , and Christie E. Williams *

Department of Agronomy, Purdue University, West Lafayette, IN 47907; Protein-Carbohydrate Interaction Core H, Emory University School of Medicine, Atlanta, GA 30322; Department of Biochemistry, Purdue University, West Lafayette, IN 47907; Department of Botany and Plant Pathology, Purdue University, West Lafayette, IN 47907; Department of Biological Sciences, Purdue University, West Lafayette, IN 47907; USDA-ARS Crop Production and Pest Control Research Unit, West Lafayette, IN 47907; Department of Entomology, Purdue University, West Lafayette, IN 47907

* Corresponding author; email: christie.williams{at}ars.usda.gov.

We previously cloned and characterized a novel jacalin-like lectin gene from wheat (Triticum aestivum L.) plants that responds to infestation by Hessian fly [Mayetiola destructor (Say)] larvae, a major dipteran pest of this crop. The infested resistant plants accumulated higher levels of Hfr-1 (Hessian fly-responsive gene 1) transcripts compared to uninfested or susceptible plants. Here we characterize the soluble and active recombinant His6-HFR1 protein isolated from Escherichia coli. Functional characterization of the protein using hemagglutination assays revealed lectin activity. Glycan microarray-binding assays indicated strong affinity of His6-HFR1 to Man{alpha}1-6(Man{alpha}1-3)Man trisaccharide structures. Resistant wheat plants accumulated high levels of HFR-1 at the larval feeding sites as revealed by immunodetection, but the avirulent larvae were deterred from feeding and consumed only small amounts of the lectin. Behavioral studies revealed that avirulent Hessian fly larvae on resistant plants exhibited prolonged searching and writhing behaviors as they unsuccessfully attempted to establish feeding sites. During His6-HFR1 feeding bioassays, Drosophila melanogaster larvae experienced significant delays in growth and pupation, while percent mortality increased with progressively higher concentrations of His6-HFR1 in the diet. Thus, HFR-1 is an anti-nutrient to dipteran larvae and may play a significant role in deterring Hessian fly larvae from feeding on resistant wheat plants.






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