Development of a monoclonal antibody to detect predation of the brown planthopper Nilaparvata lugens (Stål)

Abstract

A monoclonal antibody (McAb), 4B8, was developed to whole homogenates of the brown planthopper, Nilaparvata lugens (Stål), using hybridoma technology. It reacted with all the stages of BPH but did not cross-react with other insects and predator species tested with an indirect enzyme-linked immunosorbent assay (ELISA). Immunodiffusion revealed it to belong to the IgG₃ subclass. The ascites developed with 4B8 cell lines had high absorbance values until it was diluted over 6.5536 × 10⁷ times tested by indirect ELISA. It could recognize the BPH proteins after one female was ingested by Pardosa pseudoannulata for about 2.13 days at 25°C. The results of extended antigen detection period and the high specificity of the antibody indicated that 4B8 could be used to study interactions between planthoppers and their predators in the field. Application of the McAb to assess relative importance of four major spider species in subtropical rice ecosystem showed that P. pseudoannulata was more important than Ummeliata insecticeps, and those were more important than Tetragnatha sp. and Coleosoma octomaculatum.

Keywords:

• enzyme-linked immunosorbent assay (ELISA)
• hybridoma technology
• monoclonal antibodies (McAb)
• Nilaparvata lugens (Stål)

Acknowledgments

This study was undertaken with support from ‘Rice IPM Network’, ‘National Basic Research Priorities Program of China (TG2000016210)’ and National Natural Science Fundation (30370934). We wish to express our special thanks to Drs K. L. Heong, Cui Hu, Junhua He and Shusheng Liu for providing helpful comments on experimental design and an earlier draft of the manuscript. We also express our thanks to Jianxiang Wu for providing technical assistance.