Biochemical assays for identifying seeds of lodgepole pine and other conifers fed on by Leptoglossus occidentalis Heidemann (Hemiptera: Coreidae)

Abstract

Radiography is a valuable tool for assessing quality of conifer seeds, but it cannot differentiate between aborted seeds and seeds that have been emptied by western conifer seed bug (Leptoglossus occidentalis Heidemann) feeding. We tested three biochemical marker-based assays that were developed to identify L. occidentalis damage to seeds of Douglas-fir, Pseudostuga menziesii (Mirb.) Franco, for their use in lodgepole pine, Pinus contorta var. latifolia Engelmann. The three assays included measurement of storage protein reserve depletion, immunodetection of fragments of insoluble (crystalloid) storage proteins, and immunodetection of L. occidentalis salivary proteins. Aborted seeds contained significantly less soluble and insoluble protein than seeds that were fed on by L. occidentalis. Polyclonal antibodies raised against 11S globulin crystalloid proteins or L. occidentalis salivary gland extracts only immunoreacted with proteins in seeds exposed to L. occidentalis feeding. In a single-blind test, antibody raised against salivary-gland extracts correctly distinguished between undamaged full seeds, unfilled aborted seeds, and seeds fed on by L. occidentalis. Immunodetection of L. occidentalis salivary proteins was also performed on seeds of Abies amabilis Dougl. ex J. Forbes, Tsuga heterophylla (Raf.) Sarg., Picea sitchensis Bong (Carr.), Pinus ponderosa Lawson, and Pinus monticola Dougl. ex D. Don. For all species, immunoreactive polypeptides were only detected in seeds fed on by L. occidentalis. These biochemical marker-based techniques could help researchers and seed orchard managers estimate seed losses caused by L. occidentalis in commercial seed orchards and natural forest stands.Key words: Leptoglossus occidentalis, saliva, biochemical markers, polyclonal antibody, immunodetection, Pinus contorta.