Unveiling and Blocking the Interaction Between Tomato Spotted Wilt Virus and Its Insect Vector, Frankliniella Occidentalis


Book Description

Tomato spotted wilt virus (TSWV) is an economically important plant virus dependent on insects (thrips) for transmission to plant hosts. Like many animal-infecting viruses, TSWV replicates in the cells of its insect vector. The virus is an emergent disease threatening food and fiber crops worldwide. The aim of this work was to develop novel control strategies against TSWV through a better understanding of the virus-vector interaction. Previously, the TSWV GN protein was shown to be the viral attachment protein, a molecule mediating attachment of virus particles to the midgut epithelial cells of vector thrips. The specific goals of my research were to further examine the utility of disrupting the virus-vector interaction for effective virus control by exploiting GN properties, and to track the route of TSWV in thrips using confocal microscopy. To achieve these goals, I expressed soluble and insoluble forms of GN fused to green fluorescent protein (GFP) transiently and transgenically and examined their cellular localization in planta. GN::GFP recombinant protein localized to Golgi stacks throughout the cells as indicated by a punctate pattern or co-localization to a Golgi marker. In contrast, the soluble form of GN, GN-S::GFP, localized to the ER and apparently also to the cytoplasm. Virus acquisition and transmission assays with GN-S::GFP transgenic tomato plants demonstrated that transmission of TSWV by F. occidentalis was reduced by 35 to 100%. These results indicated that transgenic expression of GN-S in tomato plants may have the potential to prevent secondary spread of the virus. Novel features of the morphology of principal (PSGs) and tubular salivary glands (TSGs) of the insect vector F. occidentalis and of their infection with TSWV were described. The virus colonized different cell types and regions within the PSGs with variable intensity and distribution; and accumulated at the lumen of individual cells. The TSGs of F. occidentalis are proposed as a route for TSWV infection into the PSGs. The transgenic plants and the new knowledge of the virus vector interaction are promising tools to control TSWV and a model approach for the control of other vector-borne viruses.




Impact of Tomato Spotted Wilt Virus (family Bunyaviridae, Genus Tospovirus) on Western Flower Thrips (Frankliniella Occidentalis) Feeding Behaviors and Analysis of the Adult Salivary Gland Transcriptome


Book Description

The interactions between parasites and their vectors can cause profound changes in vector behavior, including changes that can enhance parasite transmission and alter trophic interactions. Parasite driven modification of vector feeding behavior is of broad adaptive significance, as vector borne parasite fitness relies on passage to a new host through vector feeding. We report for the first time, that vector infection by a plant virus alters feeding behavior. Here we show that infection with Tomato spotted wilt virus (TSWV), (family Bunyaviridae, genus Tospovirus), alters both the plant feeding behaviors and predatory behaviors of its omnivorous thrips vector, Frankliniella occidentalis (Pergande). Interestingly, the behavioral alterations reported herein are sexually dimorphic. TSWV infection altered the plant feeding behavior of male but not female thrips, and altered the predatory behavior of female thrips to a greater extent than male thrips. TSWV infection and cell-to-cell movement requires the inoculation of virus particles into a living, functional plant cell, therefore feeding behaviors that introduce virus particles into this type of environment are most likely to result in successful virus transmission. Male thrips infected with TSWV fed on plant material more than uninfected males, with the frequency of all feeding behaviors increasing by up to 3 fold. Importantly, infected males made almost 3 times more non-ingestion probes (probes in which they salivate, but leave cells largely undamaged), compared to uninfected males, thus increasing the probability of virus inoculation. F. occidentalis infected with TSWV also exhibited increased predatory behaviors and the presence and consumption of mite eggs as an alternative food source did not significantly alter vector plant feeding behaviors. Alteration to thrips predatory behavior is unlikely to be directly linked with increased virus transmission because increased predation did not affect plant feeding behaviors. Taken together these alterations to thrips feeding behaviors appear to be a compensatory response to alleviate detrimental effects of virus infection, as opposed to a direct manipulation of vector feeding behaviors to increase virus transmission. Furthermore, these finding shows an alternative pathway by which viruses can influence the structure of trophic interactions in food webs. Saliva is known to play a crucial role in insect feeding behavior and virus transmission. Currently, very little is known about the salivary glands and saliva of thrips. As a first step towards characterizing thrips salivary gland functions, we sequenced the transcriptome of the primary salivary glands of F. occidentalis using short read sequencing (Illumina) technology. We found 32,566 high quality contig sequences with an average size of 605 bp. A high percentage (57.29%) of contigs had no protein or nucleotide hits, but 13,275 contigs had significant BLAST hits (E-value (less than/equal to) 1.0E−6). We identified several genes that are likely to play a role in thrips feeding including: aldehyde dehydrogenases, metalloproteases, and glucose oxidase, which are likely to be involved in counteracting plant host defense responses; [beta] glucosidases and pectin lyases which are likely play a role in the extra-oral digestion of plant structural tissues; and [alpha] amylase, and [alpha] glucosidase while likely play a role in the extra-oral digestion of sugars. This is the first comprehensive study of a sialotranscriptome for any Thysanopteran species and provides a necessary tool to further our understanding of how thrips interact with their viruses and plant hosts.




Characterizing Global Gene Expression and Antiviral Response in Frankliniella Occidentalis Infected with Tomato Spotted Wilt Virus


Book Description

Frankliniella occidentalis, the western flower thrips, transmits the plant-pathogenic virus, Tomato spotted wilt virus (TSWV), through a circulative-propagative transmission strategy. The virus infects and replicates in the insect, traversing membrane barriers as it moves from the midgut to the salivary glands for subsequent inoculation of a plant host. Based on well-characterized virus-vector systems, many molecular interactions occur as the virus completes an infection cycle in the vector, and knowledge of transcriptome-wide response of thrips to TSWV has been limited. My research goals were to gain insight into i) the molecular responses that occur in thrips vectors of orthotospoviruses, ii) the role of antiviral defense in viruliferous thrips, and iii) plant transgenic-based strategies for studying thrips gene function and crop-pest control. To this end, my specific research objectives were to: 1) generate, assemble, and annotate a RNA-Seq-derived transcriptome for F. occidentalis using the thrips genome, and to quantify global gene expression in response to TSWV activity in larval, pre-pupal, and adult developmental stages, 2) conduct a time-course experiment to determine the effect(s) of challenging TSWV-exposed and non-exposed thrips with dsRNAs of F. occidentalis Dcr-2 or AGO2 by hemocoel injection, and 3) construct transgenic plants expressing a thrips-gene specific dsRNA hairpin to target a vital gene. My research has catalogued insect response to TSWV activity in thrips during development and provides candidate sequences for functional analysis of genes involved in insect development and defense. Successful silencing of the antiviral RNAi pathway in thrips revealed increased mortality and decreased offspring production in both virus-exposed and non-exposed insects. Arabidopsis plants were developed to express dsRNA of vacuolar ATP synthase (V-ATPase) and preliminary feeding bioassays to explore the effect of these transgenics on thrips fitness indicate a need for further description of thrips dsRNA uptake. In total, my research contributes new basic knowledge underpinning the complex and dynamic relationship between thrips vectors and the plant viruses they transmit.







Factors Controlling Virus-vector-host Plant Interactions


Book Description

Frankliniella occidentalis, tospovirus, Tomato spotted wilt virus, vector competence, haplodiploidy, intraspecific variation, predictive models, vector manipulation hypothesis, innate immunity. - Vektorkompetenz, Haplodiploidie, intraspezifische Variation, Vorhersagemodelle, angeborene Immunität




Management of Insect Pests to Agriculture


Book Description

Thanks to the application of new technologies such as whole-genome sequencing, analysis of transcriptome and proteome of insect pest to agriculture, great progress has been made in understanding the life style, reproduction, evolution and nuisance to crops caused by insect pests such as aphids, planthoppers, and whiteflies. We believe that time has come to summarize progress and to have a glance over the horizon. In this Book experts in the field discuss novel means to increase the different kinds of resistances of plants to better limit the effects of pest, to understand and disturb the hormonal regulation of embryogenesis, molting, metamorphosis and reproduction, to determine the function of insect genes in diverse processes such as metabolism, interaction with plants, virus transmission, development, and adaptation to a changing environment. The knowledge presented here is discussed with the aim of further improving control strategies of insect pestsman";mso-hansi-theme-font:minor-bidi;mso-bidi-theme-font:minor-bidi; mso-ansi-language:NL;mso-fareast-language:NL;mso-bidi-language:AR-SA">.