Control of GWSS has focused on quarantine restrictions across California, as well as the application of chemical insecticides. The costs of PD alone are considerable, as GWSS endangers the Californian grape and wine industry that generates $57.6 billion annually 4, 5. It is a xylem-feeder with more than 100 host plants 1 and is an important vector of Xylella fastidiosa, which causes Pierce’s Disease (PD) in grapes and other pathologies its host plants 1– 3. GWSS is a polyphagous hemipteran pest that, while invasive to California, is native to the south-eastern United States and north-eastern Mexico. Here we describe deployment of this technology in Homalodisca vitripennis Germar (glassy-winged sharpshooter, GWSS). All that is required for direct mutagenesis is a high-quality draft genome, a means to introduce the Cas9 protein and single-guide RNAs (sgRNAs) to the germline, and the ability to perform genetic crosses. This will facilitate novel genetic control strategies.ĬRISPR/Cas9 technology provides the means to direct mutagenesis to a specific site in a genome its application to insect pests brings genetic control strategies to the forefront in insect species where genetic manipulations have either proven elusive or not been attempted. Our data show that GWSS can be easily developed as a genetic model system for the Hemiptera, enabling the study of traits that contribute to the success of invasive pests and vectors of plant pathogens. We used amplicon sequencing to examine the extent of off-target mutagenesis in adults arising from injected eggs, which was found to be negligible or non-existent.
Our analysis of wing phenotype revealed an unexpected discovery of the participation of pteridine pigments in wing and wing-vein coloration, indicating a role for these pigments beyond eye color. Through pair matings, we obtained 100% transmission of w and cn alleles to the G3 generation and also established that both genes are located on autosomes. We use a novel and simple approach of embryo microinjection in situ on the host plant and obtain high frequency mutagenesis, in excess of 55%, of the cinnabar and white eye pigmentation loci. We report the establishment of genetic analysis in the glassy-winged sharpshooter (GWSS), Homalodisca vitripennis, which is a significant leafhopper pest of agriculture in California. Published 21 March 2003.CRISPR/Cas9 technology enables the extension of genetic techniques into insect pests previously refractory to genetic analysis. In a field study that compared three biweekly Surround WP treatments to six weekly contact insecticide treatments, Surround WP performed as well as insecticides in reducing GWSS adult numbers and oviposition.Īccepted for publication 25 February 2003. This study showed that yellow was the most attractive color, followed by orange, and that white was among the least attractive colors, suggesting that Surround WP applications may make plants unrecognizable as hosts. Applications of Surround WP turn foliage white thus, we compared the attractiveness of white and other colors with yellow, which is extremely attractive to GWSS.
In caged field studies, we found that GWSS nymphs and adults were highly repelled by lemon trees treated with Surround WP. We studied how GWSS was affected by a new technology called particle film, Surround™ WP, that protects plants from insect feeding, oviposition, and infestation by coating the plant surfaces with a protective mineral barrier. Glassy-winged sharpshooter (GWSS) was recently introduced into California and poses a serious threat to the grape industry because it is a very effective vector of the bacterium that causes Pierce's disease.