Expression in larvae fed on resistant or susceptible plants, and in larvae that are at different developmental stages
ABSTRACT: Genes expressed in the salivary glands and gut of Hessian fly (Mayetiola destructor) larvae are likely involved in interactions with host plants. Overall design: RNA samples were extracted from whole larvae. The microarray was used to determine the abundance of transcripts in larvae at different developmental stage or larvae that feed on resistant and susceptible host plants.
Project description:Genes expressed in the salivary glands and gut of Hessian fly (Mayetiola destructor) larvae are likely involved in interactions with host plants. RNA samples were extracted from whole larvae. The microarray was used to determine the abundance of transcripts in larvae at different developmental stage or larvae that feed on resistant and susceptible host plants.
Project description:The Hessian fly, Mayetiola destructor, is a destructive pest of wheat worldwide and mainly controlled by deploying resistant cultivars. In this study, we investigated the genes that were expressed differentially between larvae in resistant plants and those in susceptible plants through RNA sequencing on the Illumina platform. Informative genes were 11,832, 14,861, 15,708, and 15,071 for the comparisons between larvae in resistant versus susceptible plants for 0.5, 1, 3, and 5 days, respectively, after larvae had reached the feeding site. The transcript abundance corresponding to 5401, 6902, 8457, and 5202 of the informative genes exhibited significant differences (p ? 0.05) in the respective paired comparisons. Overall, genes involved in nutrient metabolism, RNA and protein synthesis exhibited lower transcript abundance in larvae from resistant plants, indicating that resistant plants inhibited nutrient metabolism and protein production in larvae. Interestingly, the numbers of cytochrome P450 genes with higher transcript abundance in larvae from resistant plants were comparable to, or higher than those with lower transcript abundance, indicating that toxic chemicals from resistant plants may have played important roles in Hessian fly larval death. Our study also identified several families of genes encoding secreted salivary gland proteins (SSGPs) that were expressed at early stage of 1(st) instar larvae and with more genes with higher transcript abundance in larvae from resistant plants. Those SSGPs are candidate effectors with important roles in plant manipulation.
Project description:Genes expressed in the salivary glands and gut of Hessian fly (Mayetiola destructor) larvae are likely involved in interactions with host plants. Overall design: Hessian fly larval tissues were derived from three day old larvae that were cultured on susceptible cultivar Newton. Each tissue sample contained 200 individuals dissected from the same stage of insects. The same type of dissected tissues were pooled and total RNA was extracted from development of microarray probes.
Project description:Genes expressed in the salivary glands and gut of Hessian fly (Mayetiola destructor) larvae are likely involved in interactions with host plants. Hessian fly larval tissues were derived from three day old larvae that were cultured on susceptible cultivar Newton. Each tissue sample contained 200 individuals dissected from the same stage of insects. The same type of dissected tissues were pooled and total RNA was extracted from development of microarray probes.
Project description:Hessian fly Mayetiola destructor is a notorious pest of wheat. Previous studies suggest that Hessian fly uses effector-based mechanisms to attack wheat plants during parasitism, but no direct evidence has been reported to support this postulation. Here, we produced recombinant proteins for five Family-1 candidate effectors and antibodies. Indirect immunostaining and western blots were carried out to examine the localization of Hessian fly Family-1 proteins in plant and insect tissues. Confocal images revealed that Family-1 putative effectors were exclusively produced in the basal region of larval salivary glands, which are directly linked to the mandibles' ducts for effector injection. The five Family-1 proteins were detected in infested host plants on western blots. Indirect immunostaining of sectioned host tissues around the feeding site revealed strikingly different localization patterns between resistant and susceptible plants. In susceptible plants, the Family-1 proteins penetrated from the feeding cell into deep tissues, indicative of movement between cells during nutritive cell formation. In contrast, the Hessian fly proteins were primarily limited to the initially attacked cells in resistant plants. The limitation of effectors' spread in resistant plants was likely due to wall strengthening and rapid hypersensitive cell death. Cell death was found in Nicotiana benthamiana in association with hypersensitive reaction triggered by the Family-1 effector SSGP-1A2. Our finding represents a significant progress in visualizing insect effectors in host tissues and mechanisms of plant resistance and susceptibility to gall midge pests.
Project description:AbstractBoth the wheat midge () (Géhin) (Diptera: Cecidomyiidae) and the Hessian fly () (Say) (Diptera: Cecidomyiidae) belong to a group of insects called gall midges (Diptera: Cecidomyiidae), and both are destructive pests of wheat. From Hessian fly larvae, a large number of genes have been identified to encode secreted salivary gland proteins (SSGPs), which are presumably critical for the insect to feed on and manipulate host plants. For comparison, we conducted an analysis on transcripts encoding SSGPs from the first instar larvae of the wheat midge. In total, 3,500 cDNA clones were sequenced, from which 1,301 high-quality sequences were obtained. Approximately 25% of the cDNAs with high-quality sequences encoded SSGPs. The SSGPs were grouped into 97 groups based on sequence homology. Among the SSGP-encoding transcripts, 206 encoded unique proteins with no sequence similarity to any known protein and 29 encoded proteins similar to known proteins including proteases, serpines, thioesterases, ankyrins, and ferritins. Most (~80%) SSGP-encoding genes appear under strong selection for mutations that generate amino acid changes within the coding region. Identification and characterization of SSGPs in wheat midge larvae provide a foundation for future work to reveal molecular mechanisms behind wheat midge–wheat interactions and the role of these putative effector proteins in insect virulence. Availability of the SSGP transcripts will also facilitate comparative analyses of insect effectors from related species.
Project description:Hessian fly (Mayetiola destructor), a member of the gall midge family, is one of the most destructive pests of wheat (Triticum aestivum) worldwide. Probing of wheat plants by the larvae results in either an incompatible (avirulent larvae, resistant plant) or a compatible (virulent larvae, susceptible plant) interaction. Virulent larvae induce the formation of a nutritive tissue, resembling the inside surface of a gall, in susceptible wheat. These nutritive cells are a rich source of proteins and sugars that sustain the developing virulent Hessian fly larvae. In addition, on susceptible wheat, larvae trigger a significant increase in levels of amino acids including proline and glutamic acid, which are precursors for the biosynthesis of ornithine and arginine that in turn enter the pathway for polyamine biosynthesis.Following Hessian fly larval attack, transcript abundance in susceptible wheat increased for several genes involved in polyamine biosynthesis, leading to higher levels of the free polyamines, putrescine, spermidine and spermine. A concurrent increase in polyamine levels occurred in the virulent larvae despite a decrease in abundance of Mdes-odc (ornithine decarboxylase) transcript encoding a key enzyme in insect putrescine biosynthesis. In contrast, resistant wheat and avirulent Hessian fly larvae did not exhibit significant changes in transcript abundance of genes involved in polyamine biosynthesis or in free polyamine levels.The major findings from this study are: (i) although polyamines contribute to defense in some plant-pathogen interactions, their production is induced in susceptible wheat during interactions with Hessian fly larvae without contributing to defense, and (ii) due to low abundance of transcripts encoding the rate-limiting ornithine decarboxylase enzyme in the larval polyamine pathway the source of polyamines found in virulent larvae is plausibly wheat-derived. The activation of the host polyamine biosynthesis pathway during compatible wheat-Hessian fly interactions is consistent with a model wherein the virulent larvae usurp the polyamine biosynthesis machinery of the susceptible plant to acquire nutrients required for their own growth and development.
Project description:Salivary secretions of neonate Hessian fly larvae initiate a two-way exchange of molecules with their wheat host. Changes in properties of the leaf surface allow larval effectors to enter the plant where they trigger plant processes leading to resistance and delivery of defence molecules, or susceptibility and delivery of nutrients. To increase understanding of the host plant's response, the timing and characteristics of the induced epidermal permeability were investigated. Resistant plant permeability was transient and limited in area, persisting just long enough to deliver defence molecules before gene expression and permeability reverted to pre-infestation levels. The abundance of transcripts for GDSL-motif lipase/hydrolase, thought to contribute to cuticle reorganization and increased permeability, followed the same temporal profile as permeability in resistant plants. In contrast, susceptible plants continued to increase in permeability over time until the entire crown of the plant became a nutrient sink. Permeability increased with higher infestation levels in susceptible but not in resistant plants. The ramifications of induced plant permeability on Hessian fly populations are discussed.
Project description:Evidence is emerging that some proteins secreted by gall-forming parasites of plants act as effectors responsible for systemic changes in the host plant, such as galling and nutrient tissue formation. A large number of secreted salivary gland proteins (SSGPs) that are the putative effectors responsible for the physiological changes elicited in susceptible seedling wheat by Hessian fly, Mayetiola destructor (Say), larvae have been documented. However, how the genes encoding these candidate effectors might respond under field conditions is unknown. The goal of this study was to use microarray analysis to investigate variation in SSGP transcript abundance amongst field collections from different geographical regions (southeastern USA, central USA, and the Middle East). Results revealed significant variation in SSGP transcript abundance amongst the field collections studied. The field collections separated into three distinct groups that corresponded to the wheat classes grown in the different geographical regions as well as to recently described Hessian fly populations. These data support previous reports correlating Hessian fly population structure with micropopulation differences owing to agro-ecosystem parameters such as cultivation of regionally adapted wheat varieties, deployment of resistance genes and variation in climatic conditions.
Project description:Carnivorous plants allocate more resources to carnivorous structures under nutrient-limited conditions, and relative investment can also be influenced by animals (infauna) that live in association with these plants and feed on their prey. We investigated these effects within a population of the pitcher plant Cephalotus follicularis containing varying densities of larvae of the fly Badisis ambulans. For plants with a relatively high proportion of adult pitchers, increasing larval density was associated with lower relative leaf allocation to new pitcher buds. For plants with relatively few adult pitchers, however, there was greater relative leaf allocation to pitcher buds with increasing larval density. In a field experiment, there was no significant effect of experimental larval presence or absence on the change in carbon-to-nitrogen (C/N) ratio of plants. Although the direction of the correlation between B. ambulans larvae and relative investment in carnivorous and non-carnivorous structures depends on the relative number of mature structures, whether the larvae enhance or reduce nutrient stress under different conditions remains unclear. The change in C/N was, however, less variable for pitchers that contained larvae, suggesting a stabilizing effect. Eighteen of 52 experimental pitchers were damaged by an unknown species, causing the pitcher fluid to drain. These pitchers were significantly more likely to survive if they contained larvae. These results suggest that the relationship between infauna and host varies with the initial resource status and environmental context of the host plant.