Plant Sciences & Plant Pathology
Permanent URI for this communityhttps://scholarworks.montana.edu/handle/1/12
The Department of Plant Sciences and Plant Pathology is part of the College of Agriculture at Montana State University in Bozeman. An exciting feature of this department is the diversity of programs in Plant Biology, Crop Science, Plant Pathology, Horticulture, Mycology, Plant Genetics and Entomology. The department offers BS, MS, and Ph.D. degree program
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Item Honey Bee and Bumble Bee Antiviral Defense(2018-08) McMenamin, Alexander J.; Daughenbaugh, Katie F.; Parekh, Fenali; Pizzorno, Marie C.; Flenniken, Michelle L.Bees are important plant pollinators in both natural and agricultural ecosystems. Managed and wild bees have experienced high average annual colony losses, population declines, and local extinctions in many geographic regions. Multiple factors, including virus infections, impact bee health and longevity. The majority of bee-infecting viruses are positive-sense single-stranded RNA viruses. Bee-infecting viruses often cause asymptomatic infections but may also cause paralysis, deformity or death. The severity of infection is governed by bee host immune responses and influenced by additional biotic and abiotic factors. Herein, we highlight studies that have contributed to the current understanding of antiviral defense in bees, including the Western honey bee (Apis mellifera), the Eastern honey bee (Apis cerana) and bumble bee species (Bombus spp.). Bee antiviral defense mechanisms include RNA interference (RNAi), endocytosis, melanization, encapsulation, autophagy and conserved immune pathways including Jak/STAT (Janus kinase/signal transducer and activator of transcription), JNK (c-Jun N-terminal kinase), MAPK (mitogen-activated protein kinases) and the NF-κB mediated Toll and Imd (immune deficiency) pathways. Studies in Dipteran insects, including the model organism Drosophila melanogaster and pathogen-transmitting mosquitos, provide the framework for understanding bee antiviral defense. However, there are notable differences such as the more prominent role of a non-sequence specific, dsRNA-triggered, virus limiting response in honey bees and bumble bees. This virus-limiting response in bees is akin to pathways in a range of organisms including other invertebrates (i.e., oysters, shrimp and sand flies), as well as the mammalian interferon response. Current and future research aimed at elucidating bee antiviral defense mechanisms may lead to development of strategies that mitigate bee losses, while expanding our understanding of insect antiviral defense and the potential evolutionary relationship between sociality and immune function.Item Antiviral Defense in Invertebrates(2018-08) Flenniken, Michelle L.Invertebrate organisms include vectors of human viruses (mosquitoes, sand flies), model organisms (fruit fly), insect pollinators (honey bees and bumble bees), plant virus vectors (aphids), and commercially valuable aquatic species (oysters and shrimp) that play important roles in shaping ecosystems throughout the world. Like all organisms, invertebrates are infected by viruses and have, in turn, evolved strategies to limit virus infection. There are some fundamental similarities in host defense mechanisms, including the host recognition of non-self, pathogen-associated molecular patterns (e.g., viral dsRNA) that in turn stimulate the activation of host proteins, and expression of genes required to restrict virus replication, as well as unique aspects of specific host–virus interactions that are a result of co-evolution. Invertebrate antiviral defense mechanisms include canonical immune signaling cascades (e.g., Jak/STAT, Toll, Imd), heat shock responses, apoptosis, and dsRNA-triggered responses including the sequence-specific RNA interference mechanism and a less well characterized, non-sequence-specific dsRNA mediated response.Item Melanoma and Lymphocyte Cell Specific Targeting Incorporated into a Heat Shock Protein Cage Architecture(2006) Flenniken, Michelle L.; Willits, Deborah A.; Harmsen, Ann L.; Liepold, Lars O.; Harmsen, Allen G.; Young, Mark J.; Douglas, TrevorProtein cages, including viral capsids, ferritins, and heat shock proteins (Hsps), can serve as nanocontainers for biomedical applications. They are genetically and chemically malleable platforms, with potential as therapeutic and imaging agent delivery systems. Here, both genetic and chemical strategies were used to impart cell-specific targeting to the Hsp cage from Methanococcus jannaschii. A tumor vasculature targeting peptide was incorporated onto the exterior surface of the Hsp cage. This protein cage bound to αvβ3 integrin-expressing cells. Cellular tropism was also imparted by conjugating anti-CD4 antibodies to the exterior of Hsp cages. These Ab-Hsp cage conjugates specifically bound to CD4+ cells. Protein cages have the potential to simultaneously incorporate multiple functionalities, including cell-specific targeting, imaging, and therapeutic agent delivery. We demonstrate the simultaneous incorporation of two functionalities, imaging and cell-specific targeting, onto the Hsp protein cage.Item Microbe manufacturers of semiconductors(2004-11) Flenniken, Michelle L.; Allen, Mark; Douglas, TrevorSynthesis of cadmium sulfide (CdS) semiconductor nanoparticles within a prokaryotic organism is reported for the first time by Sweeney et al. [1]. This paper demonstrates the utility of microorganisms to perform chemistries outside the scope of their “normal” metabolism and offers an environmentally benign synthesis of CdS nanoparticles.Item Biodistribution studies of protein cage nanoparticles demonstrate broad tissue distribution and rapid clearance in vivo(2007-12) Kaiser, Coleen R.; Flenniken, Michelle L.; Gillitzer, Eric; Harmsen, Ann L.; Harmsen, Allen G.; Jutila, Mark A.; Douglas, Trevor; Young, Mark J.Protein cage nanoparticles have the potential to serve as multifunctional cell targeted, imaging and therapeutic platforms for broad applications in medicine. However, before they find applications in medicine, their biocompatibility in vivo needs to be demonstrated. We provide here baseline biodistribution information of two different spherical protein cage nanoplatforms, the 28 nm viral Cowpea chlorotic mottle virus (CCMV) and the 12 nm heat shock protein (Hsp) cage. In naïve and immunized mice both nanoplatforms show similar broad distribution and movement throughout most tissues and organs, rapid excretion, the absence of long term persistence within mice tissue and organs, and no overt toxicity after a single injection. These results suggest that protein cage based nanoparticles may serve as safe, biocompatible, nanoplatforms for applications in medicine.Item Temporal Analysis of the Honey Bee Microbiome Reveals Four Novel Viruses and Seasonal Prevalence of Known Viruses, Nosema and Crithidia(2011-06) Runckel, Charles; Flenniken, Michelle L.; Engel, Juan C.; Ruby, J. Graham; Ganem, Donald; Andino, Raul; DeRisi, Joseph L.Honey bees (Apis mellifera) play a critical role in global food production as pollinators of numerous crops. Recently, honey bee populations in the United States, Canada, and Europe have suffered an unexplained increase in annual losses due to a phenomenon known as Colony Collapse Disorder (CCD). Epidemiological analysis of CCD is confounded by a relative dearth of bee pathogen field studies. To identify what constitutes an abnormal pathophysiological condition in a honey bee colony, it is critical to have characterized the spectrum of exogenous infectious agents in healthy hives over time. We conducted a prospective study of a large scale migratory bee keeping operation using high-frequency sampling paired with comprehensive molecular detection methods, including a custom microarray, qPCR, and ultra deep sequencing. We established seasonal incidence and abundance of known viruses, Nosema sp., Crithidia mellificae, and bacteria. Ultra deep sequence analysis further identified four novel RNA viruses, two of which were the most abundant observed components of the honey bee microbiome (∼1011 viruses per honey bee). Our results demonstrate episodic viral incidence and distinct pathogen patterns between summer and winter time-points. Peak infection of common honey bee viruses and Nosema occurred in the summer, whereas levels of the trypanosomatid Crithidia mellificae and Lake Sinai virus 2, a novel virus, peaked in January.Item Non-specific dsRNA-Mediated Innate Immune Response in the Honey Bee(2013-10) Flenniken, Michelle L.; Andino, RaulHoney bees are essential pollinators of numerous agricultural crops. Since 2006, honey bee populations have suffered considerable annual losses that are partially attributed to Colony Collapse Disorder (CCD). CCD is an unexplained phenomenon that correlates with elevated incidence of pathogens, including RNA viruses. Honey bees are eusocial insects that live in colonies of genetically related individuals that work in concert to gather and store nutrients. Their social organization provides numerous benefits, but also facilitates pathogen transmission between individuals. To investigate honey bee antiviral defense mechanisms, we developed an RNA virus infection model and discovered that administration of dsRNA, regardless of sequence, reduced virus infection. Our results suggest that dsRNA, a viral pathogen associated molecular pattern (PAMP), triggers an antiviral response that controls virus infection in honey bees.Item A draft genome of the honey bee trypanosomatid parasite Crithidia mellificae(2014-04) Runckel, Charles; DeRisi, Joseph; Flenniken, Michelle L.Since 2006, honey bee colonies in North America and Europe have experienced increased annual mortality. These losses correlate with increased pathogen incidence and abundance, though no single etiologic agent has been identified. Crithidia mellificae is a unicellular eukaryotic honey bee parasite that has been associated with colony losses in the USA and Belgium. C. mellificae is a member of the family Trypanosomatidae, which primarily includes other insect-infecting species (e.g., the bumble bee pathogen Crithidia bombi), as well as species that infect both invertebrate and vertebrate hosts including human pathogens (e.g.,Trypanosoma cruzi, T. brucei, and Leishmania spp.). To better characterize C. mellificae, we sequenced the genome and transcriptome of strain SF, which was isolated and cultured in 2010. The 32 megabase draft genome, presented herein, shares a high degree of conservation with the related species Leishmania major. We estimate that C. mellificae encodes over 8,300 genes, the majority of which are orthologs of genes encoded by L. major and other Leishmania or Trypanosoma species. Genes unique to C. mellificae, including those of possible bacterial origin, were annotated based on function and include genes putatively involved in carbohydrate metabolism. This draft genome will facilitate additional investigations of the impact of C. mellificae infection on honey bee health and provide insight into the evolution of this unique family.Item Honey Bee Infecting “Plant Virus” with Implications on Honey Bee Colony Health(2014-02) Flenniken, Michelle L.Honey bees are eusocial insects that are commercially managed to provide pollination services for agricultural crops. Recent increased losses of honey bee colonies (averaging 32% annually since 2006) are associated with the incidence and abundance of pathogens. In their study in mBio, J. L. Li et al. [mBio 5(1):e00898-13, 2014, doi:10.1128/mBio.00898-13] share their discovery that a plant virus, tobacco ring spot virus (TRSV), replicates in honey bees and that the prevalence of this virus was high in weak colonies. Their findings increase our understanding of the role of viruses in honey bee colony losses and underscore the importance of surveying for new and/or emerging viruses in honey bees. Furthermore, their findings will pique the interest of virologists and biologists across all disciplines. The discovery that a plant virus (TRSV) replicates, spreads, and negatively affects the health of an insect host will lead to additional studies on the mechanisms of host-specific adaptation and the role of cross-kingdom infections in the transmission of this virus.Item Antiviral Defense Mechanisms in Honey Bees(2015-08) Brutscher, Laura M.; Daughenbaugh, Katie F.; Flenniken, Michelle L.Honey bees are significant pollinators of agricultural crops and other important plant species. High annual losses of honey bee colonies in North America and in some parts of Europe have profound ecological and economic implications. Colony losses have been attributed to multiple factors including RNA viruses, thus understanding bee antiviral defense mechanisms may result in the development of strategies that mitigate colony losses. Honey bee antiviral defense mechanisms include RNA-interference, pathogen-associated molecular pattern (PAMP) triggered signal transduction cascades, and reactive oxygen species generation. However, the relative importance of these and other pathways is largely uncharacterized. Herein we review the current understanding of honey bee antiviral defense mechanisms and suggest important avenues for future investigation.
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