A protocol for the visualization and quantification of Dalbulus maidis (DeLong & Wolcott, 1923) (Hemiptera: Cicadellidae) endophytic eggs in maize leaf tissues

Abstract

This study aimed to develop a simple and efficient method for quantifying eggs of Dalbulus maidis (DeLong & Wolcott, 1923) (Hemiptera: Cicadellidae), the corn leafhopper. This insect exhibits endophytic oviposition, inserting eggs into maize leaf tissues, which hinders direct visualization and complicates fertility assessments and life-table studies. Traditional egg detection techniques often rely on labor-intensive clarification procedures, specialized reagents, or extended staining periods, limiting their practicality for large-scale experiments. Here, we present a rapid, low-cost protocol for the clarification and staining of maize tissues containing D. maidis eggs. The method enabled clear detection of endophytic eggs while preserving tissue integrity and allowed samples to be stored in the staining solution for up to three weeks prior to analysis. This protocol provides a practical and scalable approach for egg quantification in D. maidis and can be readily adapted to different plant developmental stages, tissue characteristics, or insect species exhibiting endophytic oviposition. Overall, this method facilitates efficient reproductive and ecological studies by simplifying egg visualization and enabling high-throughput sample processing.