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pepper suppressor of the g2 allele of skp1 interacts with the receptor-like cytoplasmic kinase1 and type iii effector avrbst and promotes the hypersensitive cell death response in a phosphorylation-dependent manner.xanthomonas campestris pv vesicatoria type iii effector protein, avrbst, triggers hypersensitive cell death in pepper (capsicum annuum). here, we have identified the pepper sgt1 (for suppressor of the g2 allele of skp1) as a host interactor of avrbst and also the pepper pik1 (for receptor-like cytoplasmic kinase1). pik1 specifically phosphorylates sgt1 and avrbst in vitro. avrbst specifically binds to the chord-containing protein and sgt1 domain of sgt1, resulting in the inhibition of pik1-media ...201424686111
the tomato leucine-rich repeat receptor-like kinases slserk3a and slserk3b have overlapping functions in bacterial and nematode innate immunity.the somatic embryogenesis receptor kinase 3 (serk3)/brassinosteroid (br) insensitive 1-associated kinase 1 (bak1) is required for pattern-triggered immunity (pti) in arabidopsis thaliana and nicotiana benthamiana. tomato (solanum lycopersicum) has three slserk members. two of them exhibit particularly high levels of sequence similarity to atserk3 and, therefore, were named slserk3a and slserk3b. to characterize a role for slserk3a and slserk3b in defense, we suppressed each gene individually or ...201424675749
arabidopsis pectin methylesterase17 is co-expressed with and processed by sbt3.5, a subtilisin-like serine protease.in arabidopsis thaliana, the degree of methylesterification (dm) of homogalacturonans (hgs), the main pectic constituent of the cell wall, can be modified by pectin methylesterases (pmes). in all organisms, two types of protein structure have been reported for pmes: group 1 and group 2. in group 2 pmes, the active part (pme domain, pfam01095) is preceded by an n-terminal extension (pro part), which shows similarities to pme inhibitors (pmei domain, pfam04043). this pro part mediates retention of ...201424665109
genetic variability within the polish tomato torrado virus kra isolate caused by deletions in the 3'-untranslated region of genomic rna1.tomato torrado virus (totv) is in the genus torradovirus in the family secoviridae. totv contains a single-stranded, positive-sense, bipartite rna genome encapsidated in icosahedral particles. it is a serious tomato pathogen causing significant crop reductions. its occurrence has been reported from many countries worldwide. however, the state of knowledge of totv epidemiology, sequences and phylogeny is still rather poor. in this study we found that the polish totv isolates are characterized by ...201424657785
dige-abpp by click chemistry: pairwise comparison of serine hydrolase activities from the apoplast of infected plants.activity-based protein profiling (abpp) is a targeted functional proteomics method that displays the active proteome by using small molecule probes that react covalently with the active sites of protein classes. comparison of activity profiles from two different samples is not always easy, especially when using probes that generate too many signals. for accurate comparison of protein activities between two proteomes, we developed difference gel electrophoresis abpp (dige-abpp), which compares tw ...201424643562
phytophthora infestans rxlr effector pexrd2 interacts with host mapkkk ε to suppress plant immune signaling.mitogen-activated protein kinase cascades are key players in plant immune signaling pathways, transducing the perception of invading pathogens into effective defense responses. plant pathogenic oomycetes, such as the irish potato famine pathogen phytophthora infestans, deliver rxlr effector proteins to plant cells to modulate host immune signaling and promote colonization. our understanding of the molecular mechanisms by which these effectors act in plant cells is limited. here, we report that t ...201424632534
a diverse set of mirnas responsive to begomovirus-associated betasatellite in nicotiana benthamiana.roles of micrornas (mirnas) and short interfering rnas (sirnas) in biotic stress responses, e.g., viral infection, have been demonstrated in plants by many studies. tomato yellow leaf curl china virus (tylccnv) is a monopartite begomovirus that can systemically infect solanaceae plants, and induces leaf curling, yellowing and enation symptoms when co-inoculated with a betasatellite (tylccnb). the released genome sequence of nicotiana benthamiana provides an opportunity to identify mirnas and sir ...201424618068
the coat protein of alternanthera mosaic virus is the elicitor of a temperature-sensitive systemic necrosis in nicotiana benthamiana, and interacts with a host boron transporter protein.different isolates of alternanthera mosaic virus (altmv; potexvirus), including four infectious clones derived from altmv-sp, induce distinct systemic symptoms in nicotiana benthamiana. virus accumulation was enhanced at 15 °c compared to 25 °c; severe clone altmv 3-7 induced systemic necrosis (sn) and plant death at 15 °c. no interaction with potexvirus resistance gene rx was detected, although sn was ablated by silencing of sgt1, as for other cases of potexvirus-induced necrosis. substitution ...201424606704
tombusvirus-based vector systems to permit over-expression of genes or that serve as sensors of antiviral rna silencing in plants.a next generation tomato bushy stunt virus (tbsv) coat protein gene replacement vector system is described that can be applied by either rna inoculation or through agroinfiltration. a vector expressing gfp rapidly yields high levels of transient gene expression in inoculated leaves of various plant species, as illustrated for nicotiana benthamiana, cowpea, tomato, pepper, and lettuce. a start-codon mutation to down-regulate the dose of the p19 silencing suppressor reduces gfp accumulation, where ...201424606693
the p33 auxiliary replicase protein of cucumber necrosis virus targets peroxisomes and infection induces de novo peroxisome formation from the endoplasmic reticulum.tombusviruses replicate on pre-existing organelles such as peroxisomes or mitochondria, the membranes of which become extensively reorganized into multivesicular bodies (mvbs) during the infection process. cucumber necrosis virus (cnv) has previously been shown to replicate in association with peroxisomes in yeast. we show that cnv induces mvbs from peroxisomes in infected plants and that gfp-tagged p33 auxiliary replicase protein colocalizes with yfp(skl), a peroxisomal marker. most remarkably, ...201424606690
identification and functional analysis of mitogen-activated protein kinase kinase kinase (mapkkk) genes in canola (brassica napus l.).mitogen-activated protein kinase (mapk) signalling cascades, consisting of three types of reversibly phosphorylated kinases (mapkkk, mapkk, and mapk), are involved in important processes including plant immunity and hormone responses. the mapkkks comprise the largest family in the mapk cascades, yet only a few of these genes have been associated with physiological functions, even in the model plant arabidopsis thaliana. canola (brassica napus l.) is one of the most important oilseed crops in chi ...201424604738
the post-transcriptional regulator rsma/csra activates t3ss by stabilizing the 5' utr of hrpg, the master regulator of hrp/hrc genes, in xanthomonas.the rsma/csra family of the post-transcriptional regulators of bacteria is involved in the regulation of many cellular processes, including pathogenesis. in this study, we demonstrated that rsma not only is required for the full virulence of the phytopathogenic bacterium xanthomonas citri subsp. citri (xcc) but also contributes to triggering the hypersensitive response (hr) in non-host plants. deletion of rsma resulted in significantly reduced virulence in the host plant sweet orange and a delay ...201424586158
inactivation of the host lipin gene accelerates rna virus replication through viral exploitation of the expanded endoplasmic reticulum membrane.rna viruses take advantage of cellular resources, such as membranes and lipids, to assemble viral replicase complexes (vrcs) that drive viral replication. the host lipins (phosphatidate phosphatases) are particularly interesting because these proteins play key roles in cellular decisions about membrane biogenesis versus lipid storage. therefore, we examined the relationship between host lipins and tombusviruses, based on yeast model host. we show that deletion of pah1 (phosphatidic acid phosphoh ...201424586157
rpi-blb2-mediated late blight resistance in nicotiana benthamiana requires sgt1 and salicylic acid-mediated signaling but not rar1 or hsp90.the rpi-blb2 recognizes the presence of the phytophthora infestans avrblb2 and initiates effector-triggered immunity (eti). we performed gain-of-function and loss-of-function studies in nicotiana benthamiana to elucidate rpi-blb2-mediated resistance to p. infestans. rpi-blb2 triggered a hypersensitive response through sgt1-mediated, but not rar-mediated or hsp90-mediated, pathways. nbsgt1 was also required for basal and eti-mediated by rpi-blb2 in n. benthamiana. moreover, salicylic acid (sa) af ...201424582656
a procedure for the transient expression of genes by agroinfiltration above the permissive threshold to study temperature-sensitive processes in plant-pathogen interactions.localized expression of genes in plants from t-dnas delivered into plant cells by agrobacterium tumefaciens is an important tool in plant research. the technique, known as agroinfiltration, provides fast, efficient ways to transiently express or silence a desired gene without resorting to the time-consuming, challenging stable transformation of the host, the use of less efficient means of delivery, such as bombardment, or the use of viral vectors, which multiply and spread within the host causin ...201424581046
roles of different forms of lipopolysaccharides in ralstonia solanacearum pathogenesis.lipopolysaccharides (lps) are critical components for the fitness of most gram-negative bacteria. ralstonia solanacearum causes a deadly wilting disease in many crops; however, the pathogenic roles of different forms of lps and their pathways of biogenesis remain unknown. by screening for phage-resistant mutants of r. solanacearum pss4, whose genome sequence is unavailable, mutants with various types of structural defects in lps were isolated. pathogenesis assays of the mutants revealed that pro ...201424580105
molecular cloning and characterization of enhanced disease susceptibility 1 (eds1) from gossypium barbadense.arabidopsis enhanced disease susceptibility 1 (eds1) plays an important role in plant defense against biotrophic and necrotrophic pathogens. the necrotrophic pathogen verticillium dahliae infection of gossypium barbadense could lead to verticillium wilt which seriously reduces the cotton production. here, we cloned and characterized a g. barbadense homolog of eds1, designated as gbeds1. the full-length cdna of the gbeds1 gene was obtained by the technique of rapid-amplification of cdna ends. the ...201424570021
characterization of two genes for the biosynthesis of abietane-type diterpenes in rosemary (rosmarinus officinalis) glandular trichomes.rosemary (rosmarinus officinalis) produces the phenolic diterpenes carnosic acid and carnosol, which, in addition to their general antioxidant activities, have recently been suggested as potential ingredients for the prevention and treatment of neurodegenerative diseases. little is known about the biosynthesis of these diterpenes. here we show that the biosynthesis of phenolic diterpenes in rosemary predominantly takes place in the glandular trichomes of young leaves, and used this feature to id ...201424569175
a simple method for screening of plant nbs-lrr genes that confer a hypersensitive response to plant viruses and its application for screening candidate pepper genes against pepper mottle virus.plant nbs-lrr genes are abundant and have been increasingly cloned from plant genomes. in this study, a method based on agroinfiltration and virus inoculation was developed for the simple and inexpensive screening of candidate r genes that confer a hypersensitive response to plant viruses. the well-characterized resistance genes rx and n, which confer resistance to potato virus x (pvx) and tobamovirus, respectively, were used to optimize a transient expression assay for detection of hypersensiti ...201424552951
a modular plasmid assembly kit for multigene expression, gene silencing and silencing rescue in plants.the golden gate (gg) modular assembly approach offers a standardized, inexpensive and reliable way to ligate multiple dna fragments in a pre-defined order in a single-tube reaction. we developed a gg based toolkit for the flexible construction of binary plasmids for transgene expression in plants. starting from a common set of modules, such as promoters, protein tags and transcribed regions of interest, synthetic genes are assembled, which can be further combined to multigene constructs. as an e ...201424551083
suppression of rna silencing by a plant dna virus satellite requires a host calmodulin-like protein to repress rdr6 expression.in plants, rna silencing plays a key role in antiviral defense. to counteract host defense, plant viruses encode viral suppressors of rna silencing (vsrs) that target different effector molecules in the rna silencing pathway. evidence has shown that plants also encode endogenous suppressors of rna silencing (esrs) that function in proper regulation of rna silencing. the possibility that these cellular proteins can be subverted by viruses to thwart host defense is intriguing but has not been full ...201424516387
silencing of ds2 aminoacylase-like genes confirms basal resistance to phytophthora infestans in nicotiana benthamiana.nicotiana benthamiana is a potential host to several plant pathogens, and immature leaves of n. benthamiana are susceptible to phytophthora infestans. in contrast, mature leaves of n. benthamiana are weakly susceptible and show basal resistance to p. infestans. we screened a gene-silenced mature plant showing high resistance to p. infestans, designated as ds2 (disease suppression 2). the deduced amino acid sequence of cdna responsible for ds2 encoded a putative aminoacylase. growth of p. infesta ...201424514749
analysis of complementarity requirements for plant microrna targeting using a nicotiana benthamiana quantitative transient assay.micrornas (mirnas) guide rna-induced silencing complexes to target rnas based on mirna-target complementarity. using a dual-luciferase based sensor system in nicotiana benthamiana, we quantitatively assessed the relationship between mirna-target complementarity and silencing efficacy measured at both the rna and protein levels, using several conserved mirnas and their known target sites from arabidopsis thaliana. we found that naturally occurring sites have variable efficacies attributable to th ...201424510721
in nicotiana species, an artificial microrna corresponding to the virulence modulating region of potato spindle tuber viroid directs rna silencing of a soluble inorganic pyrophosphatase gene and the development of abnormal phenotypes.potato spindle tuber viroid (pstvd) is a small non-protein-coding rna pathogen that can induce disease symptoms in a variety of plant species. how pstvd induces disease symptoms is a long standing question. it has been suggested that pstvd-derived small rnas (srnas) could direct rna silencing of a targeted host gene(s) resulting in symptom development. to test this, we expressed pstvd sequences as artificial micrornas (amirnas) in nicotiana tabacum and nicotiana benthamiana. one amirna, amir46 t ...201424503090
the rice tal effector-dependent resistance protein xa10 triggers cell death and calcium depletion in the endoplasmic reticulum.the recognition between disease resistance (r) genes in plants and their cognate avirulence (avr) genes in pathogens can produce a hypersensitive response of localized programmed cell death. however, our knowledge of the early signaling events of the r gene-mediated hypersensitive response in plants remains limited. here, we report the cloning and characterization of xa10, a transcription activator-like (tal) effector-dependent r gene for resistance to bacterial blight in rice (oryza sativa). xa ...201424488961
the essential role of ngatha genes in style and stigma specification is widely conserved across eudicots.carpel development and evolution are central issues for plant biology. the conservation of genetic functions conferring carpel identity has been widely studied in higher plants. however, although genetic networks directing the development of characteristic features of angiosperm carpels such as stigma and style are increasingly known in arabidopsis thaliana, little information is available on the conservation and diversification of these networks in other species. here, we have studied the funct ...201424483275
transplastomic nicotiana benthamiana plants expressing multiple defence genes encoding protease inhibitors and chitinase display broad-spectrum resistance against insects, pathogens and abiotic stresses.plastid engineering provides several advantages for the next generation of transgenic technology, including the convenient use of transgene stacking and the generation of high expression levels of foreign proteins. with the goal of generating transplastomic plants with multiresistance against both phytopathogens and insects, a construct containing a monocistronic patterned gene stack was transformed into nicotiana benthamiana plastids harbouring sweet potato sporamin, taro cystatin and chitinase ...201424479648
lateral organ boundaries 1 is a disease susceptibility gene for citrus bacterial canker disease.citrus bacterial canker (cbc) disease occurs worldwide and incurs considerable costs both from control measures and yield losses. bacteria that cause cbc require one of six known type iii transcription activator-like (tal) effector genes for the characteristic pustule formation at the site of infection. here, we show that xanthomonas citri subspecies citri strain xcc306, with the type iii tal effector gene ptha4 or with the distinct yet biologically equivalent gene pthaw from strain xcca(w), ind ...201424474801
salicylic acid and jasmonic acid are essential for systemic resistance against tobacco mosaic virus in nicotiana benthamiana.systemic resistance is induced by pathogens and confers protection against a broad range of pathogens. recent studies have indicated that salicylic acid (sa) derivative methyl salicylate (mesa) serves as a long-distance phloem-mobile systemic resistance signal in tobacco, arabidopsis, and potato. however, other experiments indicate that jasmonic acid (ja) is a critical mobile signal. here, we present evidence suggesting both mesa and methyl jasmonate (meja) are essential for systemic resistance ...201424450774
dna-a of a highly pathogenic indian cassava mosaic virus isolated from jatropha curcas causes symptoms in nicotiana benthamiana.jatropha curcas mosaic disease (jcmd) is a newly emerging disease that has been reported in africa and india. here, we report the complete nucleotide sequence of a new indian cassava mosaic virus isolate (icmv-sg) from singapore. infection of icmv-sg showed more severe jcmd in jatropha curcas and nicotiana benthamiana than the other icmv isolates reported previously, though icmv-sg shares high sequence identity with the other icmv isolates. agroinfectious dna-a alone sufficiently induced systemi ...201424445901
agroinfiltration and pvx agroinfection in potato and nicotiana benthamiana.agroinfiltration and pvx agroinfection are two efficient transient expression assays for functional analysis of candidate genes in plants. the most commonly used agent for agroinfiltration is agrobacterium tumefaciens, a pathogen of many dicot plant species. this implies that agroinfiltration can be applied to many plant species. here, we present our protocols and expected results when applying these methods to the potato (solanum tuberosum), its related wild tuber-bearing solanum species (solan ...201424430891
molecular evidence for the occurrence of abutilon mosaic virus, a new world begomovirus in india.during an investigation in the year 2010, on the weed reservoir of begomovirus, abutilon pictum showing bright yellow mosaic symptoms was observed in udhagamandalam, tamil nadu, india. the complete bipartite genome of a begomovirus was cloned and sequenced which revealed association of abutilon mosaic virus (abmv). nicotiana benthamiana plants inoculated biolistically with the concatemers generated through rolling circle amplification of the cloned dnas were asymptomatic; however three out of ni ...201324426288
a nuclear factor y interacting protein of the gras family is required for nodule organogenesis, infection thread progression, and lateral root growth.a c subunit of the heterotrimeric nuclear factor y (nf-yc1) was shown to play a key role in nodule organogenesis and bacterial infection during the nitrogen fixing symbiosis established between common bean (phaseolus vulgaris) and rhizobium etli. to identify other proteins involved in this process, we used the yeast (saccharomyces cerevisiae) two-hybrid system to screen for nf-yc1-interacting proteins. one of the positive clones encodes a member of the phytochrome a signal transduction1 subfamil ...201424424321
deep sequencing-based transcriptome profiling reveals comprehensive insights into the responses of nicotiana benthamiana to beet necrotic yellow vein virus infections containing or lacking rna4.beet necrotic yellow vein virus (bnyvv), encodes either four or five plus-sense single stranded rnas and is the causal agent of sugar beet rhizomania disease, which is widely distributed in most regions of the world. bnyvv can also infect nicotiana benthamiana systemically, and causes severe curling and stunting symptoms in the presence of rna4 or mild symptoms in the absence of rna4.201424416380
the evolutionary conserved oil body associated protein obap1 participates in the regulation of oil body size.a transcriptomic approach has been used to identify genes predominantly expressed in maize (zea mays) scutellum during maturation. one of the identified genes is oil body associated protein1 (obap1), which is transcribed during seed maturation predominantly in the scutellum, and its expression decreases rapidly after germination. proteins similar to obap1 are present in all plants, including primitive plants and mosses, and in some fungi and bacteria. in plants, obap genes are divided in two sub ...201424406791
regulation of copper homeostasis and biotic interactions by microrna 398b in common bean.micrornas are recognized as important post-transcriptional regulators in plants. information about the roles of mirnas in common bean (phaseolus vulgaris l.), an agronomically important legume, is yet scant. the objective of this work was to functionally characterize the conserved mirna: mir398b and its target cu/zn superoxide dismutase 1 (csd1) in common bean. we experimentally validated a novel mir398 target: the stress up-regulated nodulin 19 (nod19). expression analysis of mir398b and target ...201424400089
novel type of adenylyl cyclase participates in tabtoxinine-β-lactam-induced cell death and occurrence of wildfire disease in nicotiana benthamiana.tabtoxinine-β-lactam (tβl), a non-specific bacterial toxin, is produced by pseudomonas syringae pv. tabaci, the causal agent of tobacco wildfire disease. tβl causes the plant cell death by the inhibiting glutamine synthetase, which leads to an abnormal accumulation of ammonium ions. to better understand the molecular mechanisms involved in tβl-induced cell death and necrotic wildfire lesions, we focused on adenylyl cyclase in nicotiana benthamiana. we isolated the gene designated as nbac (nicoti ...201424398509
two key arginine residues in the coat protein of bamboo mosaic virus differentially affect the accumulation of viral genomic and subgenomic rnas.the interactions between viral rnas and coat proteins (cps) are critical for the efficient completion of infection cycles of rna viruses. however, the specificity of the interactions between cps and genomic or subgenomic rnas remains poorly understood. in this study, bamboo mosaic virus (bamv) was used to analyse such interactions. using reversible formaldehyde cross-linking and mass spectrometry, two regions in cp, each containing a basic amino acid (r99 and r227, respectively), were identified ...201424393453
a simplified approach to construct infectious cdna clones of a tobamovirus in a binary vector.infectious cdna clones of rna viruses are important tools to study molecular processes such as replication and host-virus interactions. however, the cloning steps necessary for construction of cdnas of viral rna genomes in binary vectors are generally laborious. in this study, a simplified method of producing an agro-infectious pepper mild mottle virus (pmmov) clone is described in detail. initially, the complete genome of pmmov was amplified by a single-step rt-pcr, cloned, and subcloned into a ...201424388933
identification of fungus-responsive cis-acting element in the promoter of brassica juncea chitinase gene, bjchi1.chitinases are a group of pathogenesis-related proteins. the brassica juncea chitinase gene bjchi1 is highly inducible by pathogenic fungal infection, suggesting that the promoter of bjchi1 might contain specific cis-acting element responsive to fungal attack. to identify the fungus-responsive element in bjchi1 promoter (bjc-p), a series of binary plant transformation vectors were constructed by fusing the bjc-p or its deletion-derivatives to β-glucuronidase (gus) reporter gene. expression of th ...201424388530
host-specific adaptation of diverse betasatellites associated with distinct indian tomato-infecting begomoviruses.tomato leaf curl viruses cause major crop loss hindering tomato cultivation worldwide. the 'old world' begomoviruses are often associated with circular ssdna satellite molecules called betasatellites. in the present study, replication compatibility of five different betasatellites with three distinct indian tomato-infecting begomoviruses representing each of a monopartite, a mono-bipartite and a bipartite begomoviruses was studied. all the betasatellites could be trans-replicated by the begomovi ...201424384936
perception of the novel mamp emax from different xanthomonas species requires the arabidopsis receptor-like protein remax and the receptor kinase sobir.as part of their innate immune system plants carry a number of pattern recognition receptors (prrs) that can detect a broad range of microbe-associated molecular patterns (mamps). in a recently published article (1) we described a novel, proteinaceous mamp termed emax (enigmatic mamp of xanthomonas) that derives from xanthomonas and gets recognized by the receptor-like protein remax (receptor of emax) of arabidopsis thaliana. remax has no ortholog in nicotiana benthamiana and this species does n ...201324384530
accumulation of secreted antibodies in plant cell cultures varies according to the isotype, host species and culture conditions.nicotiana tabacum suspension cells have been widely used to produce monoclonal antibodies, but the yield of secreted antibodies is usually low probably because of proteolytic degradation. most iggs that have been expressed in suspension cells have been of the human igg1 isotype. in this study, we examined whether other isotypes displayed the same sensitivity to proteolytic degradation and whether the choice of plant host species mattered. human serum igg displayed different degradation profiles ...201424373507
the nia-pro protein of turnip mosaic virus improves growth and reproduction of the aphid vector, myzus persicae (green peach aphid).many plant viruses depend on aphids and other phloem-feeding insects for transmission within and among host plants. thus, viruses may promote their own transmission by manipulating plant physiology to attract aphids and increase aphid reproduction. consistent with this hypothesis, myzus persicae (green peach aphids) prefer to settle on nicotiana benthamiana infected with turnip mosaic virus (tumv) and fecundity on virus-infected n. benthamiana and arabidopsis thaliana (arabidopsis) is higher tha ...201424372679
mutational analysis of two highly conserved motifs in the silencing suppressor encoded by tomato spotted wilt virus (genus tospovirus, family bunyaviridae).tospoviruses cause serious economic losses to a wide range of field and horticultural crops on a global scale. the nss gene encoded by tospoviruses acts as a suppressor of host plant defense. we identified amino acid motifs that are conserved in all of the nss proteins of tospoviruses for which the sequence is known. using tomato spotted wilt virus (tswv) as a model, the role of these motifs in suppressor activity of nss was investigated. using site-directed point mutations in two conserved moti ...201424363189
characterization and expression of a gdsl-like lipase gene from brassica napus in nicotiana benthamiana.the gdsl esterase and lipase families play important roles in abiotic stress, pathogen defense, seed development and lipid metabolism. identifying the lipase activity of the putative gdsl lipase is the prerequisite for dissecting its function. according to the sequence similarity and the conserved domains, we cloned the brassica napus bnglip gene, which encodes a gdsl-like protein. we failed to identify the bnglip lipase activity in the bacterium and yeast expression systems. in this paper, we e ...201424363150
interference with jasmonic acid-regulated gene expression is a general property of viral suppressors of rna silencing but only partly explains virus-induced changes in plant-aphid interactions.the cucumber mosaic virus (cmv) 2b viral suppressor of rna silencing (vsr) inhibits host responses to jasmonic acid (ja), a chemical signal regulating resistance to insects. previous experiments with a cmv subgroup ia strain and its 2b gene deletion mutant suggested that vsrs might neutralize aphid (myzus persicae) resistance by inhibiting ja-regulated gene expression. to further investigate this, we examined ja-regulated gene expression and aphid performance in nicotiana benthamiana infected wi ...201424362960
expression and biological activity of the cystine knot bioinsecticide pa1b (pea albumin 1 subunit b).the pa1b (pea albumin 1, subunit b) peptide is an entomotoxin extract from legume seeds with lethal activity on several insect pests, such as mosquitoes, some aphids and cereal weevils. this 37 amino-acid cysteine-rich peptide has been, until now, obtained by biochemical purification or chemical synthesis. in this paper, we present our results for the transient production of the peptide in nicotiana benthamiana by agro-infiltration, with a yield of about 35 µg/g of fresh leaves and maximum produ ...201324349099
the gossypium hirsutum wrky gene ghwrky39-1 promotes pathogen infection defense responses and mediates salt stress tolerance in transgenic nicotiana benthamiana.our results indicate that overexpression of the ghwrky39 - 1 gene enhances resistance to pathogen infection and tolerance to high salt and oxidative stress in transgenic nicotiana benthamiana.201424337818
heat shock protein 70 is required for tabtoxinine-β-lactam-induced cell death in nicotiana benthamiana.tabtoxinine-β-lactam (tβl), a non-specific bacterial toxin, is produced by pseudomonas syringae pv. tabaci, the causal agent of tobacco wildfire disease. tβl causes death of plant cells through the inhibition of glutamine synthetase, which leads to an abnormal accumulation of ammonium ions and the characteristic necrotic wildfire lesions. to better understand the mechanisms involved in tβl-induced cell death, we studied its regulation in nicotiana benthamiana. tβl-induced lesions, similar to tho ...201424331433
high-level diterpene production by transient expression in nicotiana benthamiana.characterization of plant terpene synthases is typically done by production of recombinant enzymes in escherichia coli. this is often difficult due to solubility and codon usage issues. furthermore, plant terpene synthases which are targeted to the plastids, such as diterpene synthases, have to be shortened in a more or less empirical approach to improve expression. we report here an optimized agrobacterium-mediated transient expression assay in nicotiana benthamiana for plant diterpene synthase ...201324330621
methylation of translation elongation factor 1a by the mettl10-like see1 methyltransferase facilitates tombusvirus replication in yeast and plants.replication of tombusviruses and other plus-strand rna viruses depends on several host factors that are recruited into viral replicase complexes. previous studies have shown that eukaryotic translation elongation factor 1a (eef1a) is one of the resident host proteins in the highly purified tombusvirus replicase complex. in this paper, we show that methylation of eef1a by the mettl10-like see1p methyltransferase is required for tombusvirus and unrelated nodavirus rna replication in yeast model ho ...201424314635
the fusarium oxysporum effector six6 contributes to virulence and suppresses i-2-mediated cell death.plant pathogens secrete effectors to manipulate their host and facilitate colonization. fusarium oxysporum f. sp. lycopersici is the causal agent of fusarium wilt disease in tomato. upon infection, f. oxysporum f. sp. lycopersici secretes numerous small proteins into the xylem sap (six proteins). most six proteins are unique to f. oxysporum, but six6 is an exception; a homolog is also present in two colletotrichum spp. six6 expression was found to require living host cells and a knockout of six6 ...201424313955
pathogenesis-related protein 4b interacts with leucine-rich repeat protein 1 to suppress pr4b-triggered cell death and defense response in pepper.to control defense and cell-death signaling, plants contain an abundance of pathogen recognition receptors such as leucine-rich repeat (lrr) proteins. here we show that pepper (capsicum annuum) lrr1 interacts with the pepper pathogenesis-related (pr) protein 4b, pr4b, in yeast and in planta. pr4b is synthesized in the endoplasmic reticulum, interacts with lrr1 in the plasma membrane, and is secreted to the apoplast via the plasma membrane. binding of pr4b to lrr1 requires the chitin-binding doma ...201424304389
independent encapsidation of tomato golden mosaic virus a component dna in transgenic plants.tomato golden mosaic virus (tgmv), a member of the geminivirus group, has a genome consisting of two dna molecules designated the a and b components. both are required for infectivity in healthy plants, although the former has been shown to replicate independently in transgenic plants containing tandem direct repeats of the a genome component. in the studies presented here, petunia plants transgenic for either both components (a×b hybrids) or the a component alone were examined for the presence ...198724301310
novel secretory protein ss-caf1 of the plant-pathogenic fungus sclerotinia sclerotiorum is required for host penetration and normal sclerotial development.to decipher the mechanism of pathogenicity in sclerotinia sclerotiorum, a pathogenicity-defective mutant, sunf-mt6, was isolated from a t-dna insertional library. sunf-mt6 could not form compound appressorium and failed to induce lesions on leaves of rapeseed though it could produce more oxalic acid than the wild-type strain. however, it could enter into host tissues via wounds and cause typical necrotic lesions. furthermore, sunf-mt6 produced fewer but larger sclerotia than the wild-type strain ...201424299212
direct visualization of agrobacterium-delivered vire2 in recipient cells.agrobacterium tumefaciens is a natural genetic engineer widely used to deliver dna into various recipients, including plant, yeast and fungal cells. the bacterium can transfer single-stranded dna molecules (t-dnas) and bacterial virulence proteins, including vire2. however, neither the dna nor the protein molecules have ever been directly visualized after the delivery. in this report, we adopted a split-gfp approach: the small gfp fragment (gfp11) was inserted into vire2 at a permissive site to ...201424299048
virus-based microrna silencing in plants.micrornas (mirnas) play pivotal roles in various biological processes across kingdoms. many plant mirnas have been experimentally identified or predicted by bioinformatics mining of small rna databases. however, the functions of these mirnas remain largely unknown due to the lack of effective genetic tools. here, we report a virus-based microrna silencing (vbms) system that can be used for functional analysis of plant mirnas. vbms is performed through tobacco rattle virus-based expression of mir ...201424296072
effects and effectiveness of two rnai constructs for resistance to pepper golden mosaic virus in nicotiana benthamiana plants.tochlpv and pepgmv are begomoviruses that have adapted to a wide host range and are able to cause major diseases in agronomic crops. we analyzed the efficacy of induced resistance to pepgmv in nicotiana benthamiana plants with two constructs: one construct with homologous sequences derived from pepgmv, and the other construct with heterologous sequences derived from tochlpv. plants protected with the heterologous construct showed an efficacy to decrease the severity of symptoms of 45%, while pla ...201324287597
replication and packaging of turnip yellow mosaic virus rna containing flock house virus rna1 sequence.turnip yellow mosaic virus (tymv) is a spherical plant virus that has a single 6.3 kb positive strand rna as a genome. in this study, rna1 sequence of flock house virus (fhv) was inserted into the tymv genome to test whether tymv can accommodate and express another viral entity. in the resulting construct, designated ty-fhv, the fhv rna1 sequence was expressed as a tymv subgenomic rna. northern analysis of the nicotiana benthamiana leaves agroinfiltrated with the ty-fhv showed that both genomic ...201424286326
genome-wide expressional and functional analysis of calcium transport elements during abiotic stress and development in rice.ca²⁺ homeostasis is required to maintain a delicate balance of cytosolic ca²⁺ during normal and adverse growth conditions. various ca²⁺ transporters actively participate to maintain this delicate balance especially during abiotic stresses and developmental events in plants. in this study, we present a genome-wide account, detailing expression profiles, subcellular localization and functional analysis of rice ca²⁺ transport elements. exhaustive in silico data mining and analysis resulted in the i ...201424286292
gene coexpression analysis reveals complex metabolism of the monoterpene alcohol linalool in arabidopsis flowers.the cytochrome p450 family encompasses the largest family of enzymes in plant metabolism, and the functions of many of its members in arabidopsis thaliana are still unknown. gene coexpression analysis pointed to two p450s that were coexpressed with two monoterpene synthases in flowers and were thus predicted to be involved in monoterpenoid metabolism. we show that all four selected genes, the two terpene synthases (tps10 and tps14) and the two cytochrome p450s (cyp71b31 and cyp76c3), are simulta ...201324285789
a bromodomain-containing host protein mediates the nuclear importation of a satellite rna of cucumber mosaic virus.replication of the satellite rna (satrna) of cucumber mosaic virus is dependent on replicase proteins of helper virus (hv). however, we recently demonstrated that like with potato spindle tuber viroid (pstvd), a satrna associated with cucumber mosaic virus strain q (q-satrna) has the propensity to localize in the nucleus and generate multimers that subsequently serve as templates for hv-dependent replication. but the mechanism regulating the nuclear importation of q-satrna is unknown. here we sh ...201424284314
a dual gene-silencing vector system for monocot and dicot plants.plant virus-based gene-silencing vectors have been extensively and successfully used to elucidate functional genomics in plants. however, only limited virus-induced gene-silencing (vigs) vectors can be used in both monocot and dicot plants. here, we established a dual gene-silencing vector system based on bamboo mosaic virus (bamv) and its satellite rna (satbamv). both bamv and satbamv vectors could effectively silence endogenous genes in nicotiana benthamiana and brachypodium distachyon. the sa ...201424283212
xanthomonas euvesicatoria type iii effector xopq interacts with tomato and pepper 14-3-3 isoforms to suppress effector-triggered immunity.effector-triggered immunity (eti) to host-adapted pathogens is associated with rapid cell death at the infection site. the plant-pathogenic bacterium xanthomonas euvesicatoria (xcv) interferes with plant cellular processes by injecting effector proteins into host cells through the type iii secretion system. here, we show that the xcv effector xopq suppresses cell death induced by components of the eti-associated map kinase cascade mapkkkα mek2/sipk and by several r/avr gene pairs. inactivation o ...201424279912
investigations on the tobacco necrosis virus d p60 replicase protein.tobacco necrosis virus d (tnv-d), in the genus betanecrovirus (family tombusviridae), possesses a single-stranded, positive-sense rna genome containing six open reading frames (orfs). two 5'-proximal orfs (1 and 2) encode overlapping polypeptides of 22 and 82 kda (p22 and p82, respectively) which are both required for replication. the p22 auxiliary protein contains no replication motifs but the c-terminal region, downstream of a leaky stop codon, encodes a 60 kda polypeptide (p60) which contains ...201324278346
plant-based production of recombinant plasmodium surface protein pf38 and evaluation of its potential as a vaccine candidate.pf38 is a surface protein of the malarial parasite plasmodium falciparum. in this study, we produced and purified recombinant pf38 and a fusion protein composed of red fluorescent protein and pf38 (rfp-pf38) using a transient expression system in the plant nicotiana benthamiana. to our knowledge, this is the first description of the production of recombinant pf38. to verify the quality of the recombinant pf38, plasma from semi-immune african donors was used to confirm specific binding to pf38. e ...201324278216
a plant-produced pfs25 vlp malaria vaccine candidate induces persistent transmission blocking antibodies against plasmodium falciparum in immunized mice.malaria transmission blocking vaccines (tbvs) are considered an effective means to control and eventually eliminate malaria. the pfs25 protein, expressed predominantly on the surface of the sexual and sporogonic stages of plasmodium falciparum including gametes, zygotes and ookinetes, is one of the primary targets for tbv. it has been demonstrated that plants are an effective, highly scalable system for the production of recombinant proteins, including virus-like particles (vlps). we engineered ...201324260245
fungal endopolygalacturonases are recognized as microbe-associated molecular patterns by the arabidopsis receptor-like protein responsiveness to botrytis polygalacturonases1.plants perceive microbial invaders using pattern recognition receptors that recognize microbe-associated molecular patterns. in this study, we identified responsiveness to botrytis polygalacturonases1 (rbpg1), an arabidopsis (arabidopsis thaliana) leucine-rich repeat receptor-like protein, atrlp42, that recognizes fungal endopolygalacturonases (pgs) and acts as a novel microbe-associated molecular pattern receptor. rbpg1 recognizes several pgs from the plant pathogen botrytis cinerea as well as ...201324259685
identification of novel pepper genes involved in bax- or inf1-mediated cell death responses by high-throughput virus-induced gene silencing.hot pepper is one of the economically important crops in asia. a large number of gene sequences, including expressed sequence tag (est) and genomic sequences are publicly available. however, it is still a daunting task to determine gene function due to difficulties in genetic modification of a pepper plants. here, we show the application of the virus-induced gene silencing (vigs) repression for the study of 459 pepper ests selected as non-host pathogen-induced cell death responsive genes from pe ...201324256816
effect of mitochondria-targeted antioxidant skq1 on programmed cell death induced by viral proteins in tobacco plants.programmed cell death (pcd) is the main defense mechanism in plants to fight various pathogens including viruses. the best-studied example of virus-induced pcd in plants is tobacco mosaic virus (tmv)-elicited hypersensitive response in tobacco plants containing the n resistance gene. it was previously reported that the animal mitochondrial protein bcl-xl, which lacks a homolog in plants, effectively suppresses plant pcd induced by tmv p50 - the elicitor of hypersensitive response in nicotiana ta ...201324228922
the abscisic acid pathway has multifaceted effects on the accumulation of bamboo mosaic virus.accepted 29 october 2013. abscisic acid (aba) plays a key role in modulating plant responses to different biotic and abiotic stresses. however, the effect of aba on virus infection is not fully understood. here, we describe the effects of the aba pathway on the accumulation of bamboo mosaic virus (bamv) and cucumber mosaic virus (cmv) in two different hosts: arabidopsis thaliana and nicotiana benthamiana. we report that aba2 plays a critical role in the accumulation of bamv and cmv. mutants down ...201424224533
trv-gfp: a modified tobacco rattle virus vector for efficient and visualizable analysis of gene function.virus-induced gene silencing (vigs) is a useful tool for functional characterization of genes in plants. unfortunately, the efficiency of infection by tobacco rattle virus (trv) is relatively low for some non-solanaceae plants, which are economically important, such as rose (rosa sp.). here, to generate an easy traceable trv vector, a green fluorescent protein (gfp) gene was tagged to the 3' terminus of the coat protein gene in the original trv2 vector, and the silencing efficiency of the modifi ...201424218330
interaction between rice stripe virus disease-specific protein and host psbp enhances virus symptoms.rice stripe virus (rsv) causes severe diseases in oryza sativa (rice) in many eastern asian countries. disease-specific protein (sp) of rsv is a non-structural protein and its accumulation level in rice plant was shown to determine the severity of rsv symptoms. here, we present evidence that expression of rsv sp alone in rice or nicotiana benthamiana did not produce visible symptoms. expression of sp in these two plants, however, enhanced rsv- or potato virus x (pvx)-induced symptoms. through ye ...201424214893
coat protein mediated resistance to plum pox virus in nicotiana clevelandii and n. benthamiana.transgenic nicotiana benthamiana and n. clevelandii plants expressing the coat protein of plum pox virus under the control of the 35s promoter from cauliflower mosaic virus were engineered by agrobacterium tumefaciens mediated transformation. the phenomenon of virus resistance was observed at different levels when transgenic plants, expressing the coat protein and control plants were compared after challenge infection with plum pox virus. n. clevelandii coat protein transgenic plants circumvent ...199224213033
a putative rab-gtpase activation protein from nicotiana benthamiana is important for bamboo mosaic virus intercellular movement.the cdna-amplified fragment length polymorphism technique was applied to isolate the differentially expressed genes during bamboo mosaic virus (bamv) infection on nicotiana benthamiana plants. one of the upregulated genes was cloned and predicted to contain a tbc domain designated as nbrabgap1 (rab gtpase activation protein 1). no significant difference was observed in bamv accumulation in the nbrabgap1-knockdown and the control protoplasts. however, bamv accumulation was 50% and 2% in the inocu ...201324210126
rice black-streaked dwarf virus p10 induces membranous structures at the er and elicits the unfolded protein response in nicotiana benthamiana.endoplasmic reticular (er) membrane modifications play an important role in viral rna replication and virion assembly but little is known about the involvement of er-membrane remodeling in the infection cycle of fijiviruses in plant cells. the subcellular localization of rice black-streaked dwarf virus outer capsid p10 was therefore examined using live-cell imaging. p10 fused to egfp formed vesicular structures associated with er membranes in nicotiana benthamiana epidermal cells and in rice pro ...201324210107
protection against virus infection in tobacco plants expressing the coat protein of grapevine fanleaf nepovirus.grapevine fanleaf nepovirus (gflv) is responsible for the economically significant "court-noué" disease in vineyards. its genome is made up of two single-stranded rna molecules (rna1 and rna2) which direct the synthesis of polyproteins p1 and p2 respectively. a chimeric coat protein gene derived from the c-terminal part of p2 was constructed and subsequently introduced into a binary transformation vector. transgenic nicotiana benthamiana plants expressing the coat protein under the control of th ...199424193837
arabidopsis myosin xi sub-domains homologous to the yeast myo2p organelle inheritance sub-domain target subcellular structures in plant cells.myosin xi motor proteins transport plant organelles on the actin cytoskeleton. the arabidopsis gene family that encodes myosin xi has 13 members, 12 of which have sub-domains within the tail region that are homologous to well-characterized cargo-binding domains in the yeast myosin v myo2p. little is presently known about the cargo-binding domains of plant myosin xis. prior experiments in which most or all of the tail regions of myosin xis have been fused to yellow fluorescent protein (yfp) and t ...201324187546
v2 of tomato yellow leaf curl virus can suppress methylation-mediated transcriptional gene silencing in plants.tomato yellow leaf curl virus (tylcv) is a dna virus belonging to the genus begomovirus. tylcv replicates using double-stranded dna intermediates that can become the target of plant transcriptional gene silencing (tgs). here, we show that the v2 protein of tylcv can suppress tgs of a transcriptionally silenced green fluorescent protein (gfp) transgene in nicotiana benthamiana line 16-tgs. through bisulfite sequencing and chop-pcr, we demonstrated that the tylcv v2 can reverse gfp transgene silen ...201424187017
nonstructural protein p7-2 encoded by rice black-streaked dwarf virus interacts with skp1, a core subunit of scf ubiquitin ligase.rice black-streaked dwarf virus (rbsdv), a member of the genus fijivirus within the family reoviridae, causes severe damage to cereal crops in south east asia. the protein p7-2, encoded by the second open reading frame of segment s7, is conserved among most plant-infecting fijiviruses, but its function is still obscure.201324176102
detection of potato mop-top virus in soils and potato tubers using bait-plant bioassay, elisa and rt-pcr.the hilly region of northwest of pakistan is leading seed potato producing areas of the country. soil and plant samples were collected from the region and tested for pmtv using both conventional and molecular techniques. the bait plants exhibited pmtv-characteristic v-shaped yellow leaf markings in nicotiana debneyi plants grown in putative viruliferious soils from 20/26 locations. the results were confirmed by back inoculation of sap from both roots and leaves of bait plant on indicator hosts ( ...201424161813
efficient foreign gene expression in planta using a plantago asiatica mosaic virus-based vector achieved by the strong rna-silencing suppressor activity of tgbp1.plant virus expression vectors provide a powerful tool for basic research as well as for practical applications. here, we report the construction of an expression vector based on plantago asiatica mosaic virus (plamv), a member of the genus potexvirus. modification of a vector to enhance the expression of a foreign gene, combined with the use of the foot-and-mouth disease virus 2a peptide, allowed efficient expression of the foreign gene in two model plant species, arabidopsis thaliana and nicot ...201424154949
chloroplast phosphoglycerate kinase is involved in the targeting of bamboo mosaic virus to chloroplasts in nicotiana benthamiana plants.the bamboo mosaic virus (bamv) is a positive-sense, single-stranded rna virus. previously, we identified that the chloroplast phosphoglycerate kinase (chl-pgk) from nicotiana benthamiana is one of the viral rna binding proteins involved in the bamv infection cycle. because chl-pgk is transported to the chloroplast, we hypothesized that chl-pgk might be involved in viral rna localization in the chloroplasts. to test this hypothesis, we constructed two green fluorescent protein (gfp)-fused misloca ...201324154620
modification of turnip yellow mosaic virus coat protein and its effect on virion assembly.turnip yellow mosaic virus (tymv) is a positive strand rna virus. we have modified tymv coat protein (cp) by inserting a c-myc epitope peptide at the n- or c-terminus of the cp, and have examined its effect on assembly. we introduced the recombinant cp constructs into nicotiana benthamiana leaves by agroinfiltration. examination of the leaf extracts by agarose gel electrophoresis and western blot analysis showed that the cp modified at the n-terminus produced a band co-migrating with wild-type v ...201324148770
long-distance transport of gibberellic acid insensitive mrna in nicotiana benthamiana.the gibberellic acid (ga) signal is governed by the gai (gibberellic acid insensitive) repressor, which is characterized by a highly conserved n-terminal della domain. deletion of the della domain results in constitutive suppression of ga signaling. as the gai transcript is transportable in phloem elements, a δ-della gai (gai) transgenic stock plant can reduce the stature of a scion through transport of gai mrna from the stock. however, little is known about the characteristics of a scion on a g ...201324144190
differential expression of tomato spotted wilt virus-derived viral small rnas in infected commercial and experimental host plants.viral small rnas (vsirnas) in the infected host can be generated from viral double-stranded rna replicative intermediates, self-complementary regions of the viral genome or from the action of host rna-dependent rna polymerases on viral templates. the vsirna abundance and profile as well as the endogenous small rna population can vary between different hosts infected by the same virus influencing viral pathogenicity and host response. there are no reports on the analysis of vsirnas of tomato spot ...201324143182
expression of terpenoids 1, a glandular trichome-specific transcription factor from tomato that activates the terpene synthase 5 promoter.terpene biosynthesis in tomato glandular trichomes has been well studied, with most if not all terpene synthases (tpss) being identified. however, transcription factors (tfs) that regulate tpss have not yet been discovered from tomato. in order to unravel the transcriptional regulation of the solanum lycopersicum linalool synthase (slmts1, recently renamed sltps5) gene in glandular trichomes, we functionally dissected its promoter. a 207 bp fragment containing the minimal promoter and the 5'utr ...201424142382
genome-wide analysis of pectate-induced gene expression in botrytis cinerea: identification and functional analysis of putative d-galacturonate transporters.the fungal plant pathogen botrytis cinerea produces a spectrum of cell wall degrading enzymes for the decomposition of host cell wall polysaccharides and the consumption of the monosaccharides that are released. especially pectin is an abundant cell wall component, and the decomposition of pectin by b. cinerea has been extensively studied. an effective concerted action of the appropriate pectin depolymerising enzymes, monosaccharide transporters and catabolic enzymes is important for complete d- ...201424140151
an rxlr effector from phytophthora infestans prevents re-localisation of two plant nac transcription factors from the endoplasmic reticulum to the nucleus.the potato late blight pathogen phytophthora infestans secretes an array of effector proteins thought to act in its hosts by disarming defences and promoting pathogen colonisation. however, little is known about the host targets of these effectors and how they are manipulated by the pathogen. this work describes the identification of two putative membrane-associated nac transcription factors (tf) as the host targets of the rxlr effector pitg_03192 (pi03192). the effector interacts with nac targe ...201324130484
a β-glucuronosyltransferase from arabidopsis thaliana involved in biosynthesis of type ii arabinogalactan has a role in cell elongation during seedling growth.we have characterized a β-glucuronosyltransferase (atglcat14a) from arabidopsis thaliana that is involved in the biosynthesis of type ii arabinogalactan (ag). this enzyme belongs to the carbohydrate active enzyme database glycosyltransferase family 14 (gt14). the protein was localized to the golgi apparatus when transiently expressed in nicotiana benthamiana. the soluble catalytic domain expressed in pichia pastoris transferred glucuronic acid (glca) to β-1,6-galactooligosaccharides with degrees ...201324128328
molecular and biochemical analysis of the first ara6 homologue, a rab5 gtpase, from green algae.rab5 gtpases are important regulators of endosomal membrane traffic in yeast, plants, and animals. a specific subgroup of this family, the ara6 group, has been described in land plants including bryophytes, lycophytes, and flowering plants. here, we report on the isolation of an ara6 homologue in a green alga. caara6 (carabf1) from chara australis, a member of the characeae that is a close relative of land plants, encodes a polypeptide of 237 aa with a calculated molecular mass of 25.4 kda, whic ...201324127512
safety and immunogenicity of a plant-produced recombinant monomer hemagglutinin-based influenza vaccine derived from influenza a (h1n1)pdm09 virus: a phase 1 dose-escalation study in healthy adults.novel influenza viruses continue to pose a potential pandemic threat worldwide. in recent years, plants have been used to produce recombinant proteins, including subunit vaccines. a subunit influenza vaccine, hac1, based on recombinant hemagglutinin from the 2009 pandemic a/california/04/2009 (h1n1) strain of influenza virus, has been manufactured using a plant virus-based transient expression technology in nicotiana benthamiana plants and demonstrated to be immunogenic and safe in pre-clinical ...201424126211
the c2 protein of bhendi yellow vein mosaic virus plays an important role in symptom determination and virus replication.bhendi yellow vein mosaic virus (byvmv) that causes bhendi yellow vein mosaic disease is a monopartite begomovirus with an associated betasatellite. previous studies have shown that c2 protein of byvmv acts as a suppressor of post transcriptional gene silencing, activates transcription, localizes to nucleus, and interacts with karyopherin α. to probe the role of c2 in symptom determination and virus replication, the infectious clones of byvmv containing two stop codons in the c2 orf were created ...201424122068
two aspartate residues at the putative p10 subunit of a type ii metacaspase from nicotiana tabacum l. may contribute to the substrate-binding pocket.metacaspases are cysteine proteases present in plants, fungi, prokaryotes, and early branching eukaryotes, although a detailed description of their cellular function remains unclear. currently, three-dimensional (3d) structures are only available for two metacaspases: trypanosoma brucei (mca2) and saccharomyces cerevisiae (yca1). furthermore, metacaspases diverged from animal caspases of known structure, which limits straightforward homology-based interpretation of functional data. we report for ...201424121807
the production of recombinant cationic α-helical antimicrobial peptides in plant cells induces the formation of protein bodies derived from the endoplasmic reticulum.synthetic linear antimicrobial peptides with cationic α-helical structures, such as bp100, are valuable as novel therapeutics and preservatives. however, they tend to be toxic when expressed at high levels as recombinant peptides in plants, and they can be difficult to detect and isolate from complex plant tissues because they are strongly cationic and display low extinction coefficient and extremely limited immunogenicity. we therefore expressed bp100 with a c-terminal tag which preserved its a ...201424102775
the role of hydrogen peroxide and nitric oxide in the induction of plant-encoded rna-dependent rna polymerase 1 in the basal defense against tobacco mosaic virus.plant rna-dependent rna polymerase 1 (rdr1) is an important element of the rna silencing pathway in the plant defense against viruses. rdr1 expression can be elicited by viral infection and salicylic acid (sa), but the mechanisms of signaling during this process remains undefined. the involvement of hydrogen peroxide (h2o2) and nitric oxide (no) in rdr1 induction in the compatible interactions between tobacco mosaic tobamovirus (tmv) and nicotiana tabacum, nicotiana benthamiana, and arabidopsis ...201324098767
type i j-domain nbmip1 proteins are required for both tobacco mosaic virus infection and plant innate immunity.tm-2² is a coiled coil-nucleotide binding-leucine rich repeat resistance protein that confers durable extreme resistance against tomato mosaic virus (tomv) and tobacco mosaic virus (tmv) by recognizing the viral movement protein (mp). here we report that the nicotiana benthamiana j-domain mip1 proteins (nbmip1s) associate with tobamovirus mp, tm-2² and sgt1. silencing of nbmip1s reduced tmv movement and compromised tm-2²-mediated resistance against tmv and tomv. furthermore, silencing of nbmip1s ...201324098120
a strain-specific segment of the rna-dependent rna polymerase of grapevine fanleaf virus determines symptoms in nicotiana species.factors involved in symptom expression of viruses from the genus nepovirus in the family secoviridae such as grapevine fanleaf virus (gflv) are poorly characterized. to identify symptom determinants encoded by gflv, infectious cdna clones of rna1 and rna2 of strain ghu were developed and used alongside existing infectious cdna clones of strain f13 in a reverse genetics approach. in vitro transcripts of homologous combinations of rna1 and rna2 induced systemic infection in nicotiana benthamiana a ...201324088345
biological activity of the tzs gene of nopaline agrobacterium tumefaciens gv3101 in plant regeneration and genetic transformation.agrobacterium tumefaciens has been widely used in plant genetic transformation. hormone-encoding genes residing in the t-dna region have been removed, resulting in disarmed agrobacterium strains that are used in various transformation experiments. nopaline agrobacterium strains, however, carry another hormone gene, trans-zeatin synthesizing (tzs), that codes for trans-zeatin in the virulence region of the tumor-inducing plasmids. we investigated the activity and function of the tzs gene of a nop ...201324088018
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