Give and Take: Salmonella Enterica Alters Macrosteles Quadrilineatus Feeding Behaviors Resulting in Altered S. Enterica Populations and Distribution on Leaves

Hemipteran insects are ubiquitous inhabitants of the phyllosphere. Changes in microbial phyllosphere communities have recently been demonstrated following infestation by Macrosteles quadrilineatus (Aster Leafhopper). Although epiphytic Salmonella enterica populations naturally decline in the phyllosphere of plants, M. quadrilineatus infestation facilitated the growth of the bacterial pathogen populations. Here, we demonstrate that cellular damage by insect stylet penetration results in a localized beneficial niche on the leaf surface, leading to enhanced S. enterica populations. We measured S. enterica populations and colonization patterns on plants infested with Hemipterans with distinct feeding behaviors. M. quadrilineatus infestation resulted in higher solute leakage and significantly greater bacterial populations than plants absent of insects. Following immigration via contaminated irrigation water, the highest populations of S. enterica are naturally found on the tips of tomato leaflets. We discovered M. quadrilineatus feeding preference altered the natural distribution of S. enterica populations, and that the presence of S. enterica altered the distribution of probing attempts. These findings elucidate how cellular damage resulting from insect feeding drives changes in bacterial colonization of the phyllosphere.

Identifying Leafhopper Targets for Controlling Aster Yellows in Carrots and Celery

Aster yellows phytoplasma (Candidatus Phytoplasma asteris) is a multi-host plant pathogen and is transmitted by at least 24 leafhopper species. Pathogen management is complex and requires a thorough understanding of vector dynamics. In the American Midwest, aster yellows is of great concern for vegetable farmers who focus on controlling one vector, Macrosteles quadrilineatus—the aster leafhopper. However, vegetable-associated leafhopper communities can be diverse. To investigate whether additional species are important aster yellows vectors, we surveyed leafhopper communities at commercial celery and carrot farms in Michigan from 2018 to 2019 and conducted real-time PCR to determine infection status. Leafhoppers were collected within crop fields and field edges and identified with DNA barcoding. Overall, we collected 5049 leafhoppers, with the most abundant species being M. quadrilineatus (57%) and Empoasca fabae—the potato leafhopper (23%). Our results revealed the most abundant aster yellows vector in Michigan in both crops is M. quadrilineatus, but we also found that E. fabae may be a potential vector for this pathogen. While several taxa reside in and near these crops, we did not find strong evidence that they contribute to phytoplasma infection. These findings indicate that M. quadrilineatus should be the primary target for controlling this pathogen.

Genetic Variation Among Geographically Disparate Isolates of Aster Yellows Phytoplasma in the Contiguous United States

Aster Yellows phytoplasma (AYp; Candidatus Phytoplasma asteris) is associated with diseases of herbaceous plants, including ornamentals and important commercial vegetable and grain crops. The aster leafhopper (ALH; Macrosteles quadrilineatus Forbes) is the predominant vector of these bacteria, though other leafhopper species can acquire and transmit AYp. Potentially inoculative leafhoppers are reported to overwinter in the southern United States and migrate to northern latitudes in the spring. Examining the genetic similarities and differences in AYp associated with southern and northern populations of ALH may provide insight into the role that migrating ALH play in AYp disease development. To investigate similarities among geographically distinct populations of ALH and characterize the variation in AYp associated within these populations, we identified genetic variations in subgroup designation and the relative proportions of secreted AY-WB proteins from field-collected populations of AYp isolated from ALH from select locations in the southern (Arkansas, Kansas, Oklahoma, and Texas) and the northern United States (Wisconsin) in 2016, 2017, and 2018. Isolated phytoplasma were tested for variation of AYp genotypes, numbers of potentially inoculative (AYp-positive) ALH, and presence of specific AYp virulence (effector) genes. Geographically distinct populations of ALH collected in northern and southern regions were similar in CO1 genotype but carried different proportions of AYp genotypes. While similar AYp strains were detected in geographically distinct locations, the proportion of each genotype varied over time.

Evolution of host support for two ancient bacterial symbionts with differentially degraded genomes in a leafhopper host

Plant sap-feeding insects (Hemiptera) rely on bacterial symbionts for nutrition absent in their diets. These bacteria experience extreme genome reduction and require genetic resources from their hosts, particularly for basic cellular processes other than nutrition synthesis. The host-derived mechanisms that complete these processes have remained poorly understood. It is also unclear how hosts meet the distinct needs of multiple bacterial partners with differentially degraded genomes. To address these questions, we investigated the cell-specific gene-expression patterns in the symbiotic organs of the aster leafhopper (ALF), Macrosteles quadrilineatus (Cicadellidae). ALF harbors two intracellular symbionts that have two of the smallest known bacterial genomes: Nasuia (112 kb) and Sulcia (190 kb). Symbionts are segregated into distinct host cell types (bacteriocytes) and vary widely in their basic cellular capabilities. ALF differentially expresses thousands of genes between the bacteriocyte types to meet the functional needs of each symbiont, including the provisioning of metabolites and support of cellular processes. For example, the host highly expresses genes in the bacteriocytes that likely complement gene losses in nucleic acid synthesis, DNA repair mechanisms, transcription, and translation. Such genes are required to function in the bacterial cytosol. Many host genes comprising these support mechanisms are derived from the evolution of novel functional traits via horizontally transferred genes, reassigned mitochondrial support genes, and gene duplications with bacteriocyte-specific expression. Comparison across other hemipteran lineages reveals that hosts generally support the incomplete symbiont cellular processes, but the origins of these support mechanisms are generally specific to the host–symbiont system.

Influence ofprgHon the Persistence of Ingested Salmonella enterica in the Leafhopper Macrosteles quadrilineatus

ABSTRACTPhytophagous insects can encounterSalmonella entericaon contaminated plant surfaces and transmit externally adhered and internalized bacteria on and among leaves. Excretion of ingestedS. entericaby the leafhopperMacrosteles quadrilineatushas been previously reported; however, the sites of persistence of ingested bacteria remain undetermined. Fluorescence microscopy revealed the presence and persistence ofS. entericain various organs ofM. quadrilineatusfed an inoculated diet for 12 h and then moved to two consecutive noninoculated diets for a total of 48 h. IngestedS. entericawas predominantly observed in the filter chamber, midgut, and Malpighian tubules ofM. quadrilineatusdissected immediately after acquisition and at 24- and 48-h post-acquisition access periods (post-AAPs). Additionally, we examined the potential roles of theSalmonellapathogenicity island 1 (SPI-1) and SPI-2 type III secretion systems (T3SSs) in the persistence and excretion of ingestedS. enterica. In competition assays, aprgHmutant lacking a functional SPI-1 T3SS was recovered at significantly lower levels than the WT in insect homogenates at 24 h post-AAP, and complementation withprgHrestoredS. entericapersistence inM. quadrilineatus. Moreover, expression ofprgHinsideM. quadrilineatuswas observed up to 48 post-AAP. No differences were observed between the WT and anssaKmutant lacking a functional SPI-2 T3SS in insect homogenates or between the WT and either mutant in insect excretions. This study provides novel insight into the presence and persistence ofS. entericainsideM. quadrilineatusand demonstrates that the SPI-1 T3SS influences the persistence of the pathogen in the gut of a potential vector.

Interactions between Camelina sativa (Brassicaceae) and insect pests of canola

In an investigation of Camelina sativa (Linnaeus) Crantz (Brassicaceae) and five common insect pests of canola (Brassica napus Linnaeus) (Brassicaceae), little feeding damage to the plant was inflicted by crucifer-feeding specialist flea beetles (Phyllotreta Chevrolat species) (Coleoptera: Chrysomelidae), Delia Robineau-Desvoidy (Diptera: Anthomyiidae) root maggots, or diamondback moth (Plutella xylostella Linnaeus (Lepidoptera: Plutellidae)). In choice tests, diamondback moths laid fewer eggs on C. sativa than on B. napus leaves. Diamondback moth larvae consumed less C. sativa leaf tissue, and tended to have a longer developmental period on C. sativa. Larvae of the polyphagous bertha armyworm (Mamestra configurata Walker (Lepidoptera: Noctuidae)) had similar feeding levels on C. sativa and B. napus plants. However, there was a longer developmental period from larval to pupal stage and pupae weighed less when fed on C. sativa foliage, suggesting that C. sativa contains antibiosis factors against bertha armyworm. Two strains of aster yellows phytoplasma, 16SrI-A and 16SrI-B, were identified in C. sativa and in Macrosteles quadrilineatus (Forbes) (Hemiptera: Cicadellidae). Differences in incidence of aster yellows and abundance of M. quadrilineatus were observed among lines of C. sativa. The findings confirm that C. sativa is unlikely to support high populations of these insect pests on the Canadian prairies.