Development of an ELISA for evaluating the reproductive status of female brown planthopper, Nilaparvata lugens, by measuring vitellogenin and vitellin levels

Abstract

To evaluate the reproductive status of the female brown planthopper (BPH), Nilaparvata lugens (Stål) (Hemiptera: Delphacidae), an indirect sandwich enzyme-linked immunosorbent assay (ELISA) for monitoring vitellogenin (Vg) and vitellin (Vt) was developed by using monoclonal antibodies and polyclonal antiserum made specifically against BPH Vt. The ovarian development of BPH was divided into five stages according to ovariole development and morphological characteristics. Stages I–III, IV, and V represented the pre-oviposition, peak oviposition, and post-oviposition stages, respectively. Levels of Vt in the ovary and Vg/Vt in the whole female body during the five ovary stages appeared to relate well with the corresponding ovarian stages, suggesting that ovarian development can be evaluated by measuring ovarian Vt or whole body Vg/Vt in BPH. With this ELISA protocol, the reproductive status of macropterous BPH captured in rice fields during immigration, dwelling, and emigration was determined based on the levels of Vg/Vt in individual females. The females were mainly in stages I and II, as was confirmed by ovarian dissection. Therefore, this study presented an alternative method for evaluating the reproductive status of BPH in rice fields, which is more precise, convenient, and efficient than conventional techniques, such as dissection and classification of ovaries.