Production of Specific Polyclonal Antibody against the Larvae of Corallivorous Gastropod Drupella fragum for Prediction of Outbreaks

The term corallivorous gastropod refers to a group of snails that feeds on coral and inhabits coral communities worldwide. Outbreaks of these species cause serious damage to coral communities. There are various reasons behind the outbreaks; however, further clarifications are needed. It may be possible to predict outbreaks by measuring the number of floating larvae of corallivorous gastropods in seawater. Drupella fragum is the most damaging species in Japan, so we produced antibodies against D. fragum larvae in order to easily detect this species in the field. Antibody specificity analysis in aquarium-hatched corallivorous gastropods showed a higher specificity against D. fragum compared to D. cornus. A field study using the antibody showed that many D. fragum larvae were detected from June to November at all stations. The larvae at the Shirigai station were collected in June and July in large numbers compared to the other stations. Large groups of D. fragum were collected around the sampling point in Shirigai in September 2016. Our results imply that there is a possibility that outbreaks could be predicted using this antibody.

Novel Polyclonal Antibody Raised against Tetrodotoxin Using Its Haptenic Antigen Prepared from 4,9-anhydrotetrodotoxin Reacted with 1,2-Ethaneditiol and Further Reacted with Keyhole Limpet Hemocyanin

A novel polyclonal antibody against tetrodotoxin (TTX) was raised using its haptenic antigen, where 4,9-anhydroTTX was reacted with 1,2-ethanedithiol and this derivative was further reacted with keyhole limpet hemocyanin (KLH). This newly designed antigen (KLH-TTX) was inoculated into rabbits, resulting in the production of the specific polyclonal antibody, which reacted well with TTX and its analogs, 4-epiTTX, 11-oxoTTX and 5,6,11-trideoxyTTX, except for 4,9-anhydroTTX. The enzyme-linked immunosorbent assay (ELISA) system using this specific antibody was also developed in the present study. This newly developed polyclonal antibody with analytical procedures using direct one-step ELISA is useful to detect TTX and its analogs in toxic organisms and also disclose the mechanisms involved in their metabolic pathways and accumulation of TTX.

Characterization of a potential ripening regulator, SlNAC3, from Solanum lycopersicum

NAC (for NAM, ATAF1-2, and CUC2) proteins are one of the largest transcription factor families in plants. They have various functions and are closely related to developmental processes of fruits. Tomato (Solanum lycopersicum) is a model plant for studies of fruit growth patterns. In this study, the functional characteristics and action mechanisms of a new NAC-type transcription factor, SlNAC3 (SGN-U568609), were examined to determine its role in tomato development and ripening. The SlNAC3 protein was produced by prokaryotic expression and used to immunize New Zealand white rabbits to obtain a specific polyclonal antibody against SlNAC3. By co-immunoprecipitation and MALDI-TOF-MS assays, we showed that there was an interaction between the SlNAC3 protein and Polygalacturonase-2 (PG-2), which is related to the ripening and softening of fruit. Chromatin immunoprecipitation assays revealed the genome of the highly stress-tolerant Solanum pennellii chromosome 10 (sequence ID, HG975449.1), further demonstrating that SlNAC3 is a negative regulator of drought and salinity stress resistance in tomato, consistent with previous reports. These results indicate that SlNAC3 is not only involved in abiotic stress, but also plays a necessary role in mediating tomato ripening.