Application of droplet digital PCR in detection of seed-transmitted pathogen Acidovorax citrulli

Bacterial fruit blotch caused by Acidovorax citrulli is a serious threat to cucurbit industry worldwide.The pathogen is seedtransmitted,so seed detection to prevent distribution of contaminated seed is crucial in disease management.In this study,we adapted a quantitative real-time PCR(qPCR)assay to droplet digital PCR(ddPCR)format for A.citrulli detection by optimizing reaction conditions.The performance of ddPCR in detecting A.citrulli pure culture,DNA,infested watermelon/melon seed and commercial seed samples were compared with multiplex PCR,qPCR,and dilution plating method.The lowest concentrations detected(LCD)by ddPCR reached up to 2 fg DNA,and 102 CFU mL–1 bacterial cells,which were ten times more sensitive than those of the qPCR.When testing artificially infested watermelon and melon seed,0.1%infestation level was detectable using ddPCR and dilution plating method.The 26 positive samples were identified in 201 commercial seed samples through ddPCR,which was the highest positive number among all the methods.High detection sensitivity achieved by ddPCR demonstrated a promising technique for improving seed-transmitted pathogen detection threshold in the future.

The effects of seed detectability and seed traits on hoarding preference of two rodent species

Seed traits play an important role in affecting seed preference and hoarding behaviors of small rodents.Despite greatly affected by seed traits,seed detectability of competitors represents pilfering risks and may also modify seed hoarding preference of animals.However,whether seed traits and seed detectability show consistent effects on seed hoarding preference of animals remain largely unknown.Here,we explored how seed traits and seed detectability correlate with seed hoarding preference of Leopoldamys edwardsi and Apodemus chevrieri in a subtropical forest.Despite the effects of seed coat thickness and caloric value on hoarding preference of L.edwardsi,we detected no significant effects of other seed traits on hording preference of the 2 rodent species.There was no correlation between larder-hoarding preference and inter-or intra-specific seed detectability of L.edwardsi;however,seed detectability of L.edwardsi was negatively correlated with its own scatter-hoarding preference.Although scatter-hoarding preference of A.chevrieri was not correlated with inter-and intra-specific seed detectability,larder-hoarding preference of A.chevrieri was positively correlated with intra-specific seed detectability.Our study may provide evidence that intra-specific seed detectability rather than seed traits and inter-specific pilfering risks play an important role in modifying seed hoarding preference of rodents.