One objective of this project is to determine how cold and heat acclimation and temperature-mediated gene expression influence Las transmission by Asian citrus psyllids (ACP). We previously reported that climatic conditions appear to influence the likelihood of Las transmission. Moreover, at some temperatures ACPs exhibit altered coloration indicating up-regulation of melanization genes. We continue to identify additional cryoptotective gene targets for quantification using qRT-PCR. Currently, qRT-PCR assays to quantify several heat shock protein and melanization genes expressed by psyllids under varying temperatures are ongoing. Understanding how cold and heat acclimation, and underlying temperature-mediated gene expression, influence transmission of Las should provide novel targets for managing transmission by ACP populations. We continue to develop isofemale lines of Wolbachia and Wolbachia-free psyllids derived from 10 distinct geographic locations in Florida. Currently, replicate isofemale lines from seven distinct regions of the state have been established. Additional collections will be conducted during the next month to establish additional psyllid lines. Phenotypic screening of isofemale lines is expected to begin in the next quarter. Recently, personnel have been hired by Handler to optimize rearing and egg collection protocols needed to support for germ-line transformation of ACP from these isofemale lines.