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prevention and treatment of clostridium perfringens epsilon toxin intoxication in mice with a neutralizing monoclonal antibody (c4d7) produced in nicotiana benthamiana.epsilon toxin (etx), produced by clostridium perfringens types b and d, is among the most lethal toxins known. etx is a potential bioterrorism threat that was listed as a category b agent by the u.s. centers for disease control until 2012 and it still remains a toxin of interest for several government agencies. we produced a monoclonal antibody (mab) against etx (etx mab c4d7) in nicotiana benthamiana and characterized its preventive and therapeutic efficacy in mice. the etx preparation used was ...201424950050
effectiveness of gene silencing induced by viral vectors based on citrus leaf blotch virus is different in nicotiana benthamiana and citrus plants.virus induced gene silencing (vigs) is an effective technology for gene function analysis in plants. we assessed the vigs effectiveness in nicotiana benthamiana and citrus plants of different citrus leaf blotch virus (clbv)-based vectors, using insets of the phytoene desaturase (pds) gene. while in n. benthamiana the silencing phenotype was induced only by the construct carrying a 58-nt pds hairpin, in citrus plants all the constructs induced the silencing phenotype. differences in the generatio ...201425010281
interaction between the transcription factor attify4b and begomovirus al2 protein impacts pathogenicity.the begomovirus al2 protein is a transcriptional activator, a silencing suppressor, and inhibitor of basal defense. al2 forms a complex at the cp promoter, through interaction with a plant-specific dna-binding protein, arabidopsis peapod2 (also known as tify4b). attify4b has three domains (ppd, tify and cct_2) conserved between homologs from different plant species. we confirmed that the al2 protein from tomato golden mosaic virus and cabbage leaf curl virus interacts with tify4b from arabidopsi ...201425011501
overexpression and self-assembly of virus-like particles in nicotiana benthamiana by a single-vector dna replicon system.based on recent developments, virus-like particles (vlps) are considered to be perfect candidates as nanoplatforms for applications in materials science and medicine. to succeed, mass production of vlps and self-assembly into a correct form in plant systems are key factors. here, we report expression of synthesized coat proteins of the three viruses, brome mosaic virus, cucumber mosaic virus, and maize rayado fino virus, in nicotiana benthamiana and production of self-assembled vlps by transient ...201424965559
recent evolution of a novel begomovirus causing tomato leaf curl disease in the al-batinah region of oman.for last two decades, begomoviruses (family geminiviridae) have been a major constraint for tomato production in oman, particularly in the al-batinah region, the major agricultural area of oman. farms in the al-batinah region were surveyed during january-march and november-december in 2012 and january-february in 2013. leaf samples of tomato plants showing typical leaf curl disease symptoms were collected and analyzed for begomoviruses. out of fifteen begomovirus clones sequenced, seven were sho ...201424052149
induction and maintenance of dna methylation in plant promoter sequences by apple latent spherical virus-induced transcriptional gene silencing.apple latent spherical virus (alsv) is an efficient virus-induced gene silencing vector in functional genomics analyses of a broad range of plant species. here, an agrobacterium-mediated inoculation (agroinoculation) system was developed for the alsv vector, and virus-induced transcriptional gene silencing (vitgs) is described in plants infected with the alsv vector. the cdnas of alsv rna1 and rna2 were inserted between the cauliflower mosaic virus 35s promoter and the nos-t sequences in a binar ...201425426109
molecular and biological characterization of an isolate of cucumber mosaic virus from glycine soja by generating its infectious full-genome cdna clones.molecular and biological characteristics of an isolate of cucumber mosaic virus (cmv) from glycine soja (wild soybean), named as cmv-209, was examined in this study. comparison of nucleotide sequences and phylogenetic analyses of cmv-209 with the other cmv strains revealed that cmv-209 belonged to cmv subgroup i. however, cmv-209 showed some genetic distance from the cmv strains assigned to subgroup ia or subgroup ib. infectious full-genome cdna clones of cmv-209 were generated under the control ...201425288998
association of the p6 protein of cauliflower mosaic virus with plasmodesmata and plasmodesmal proteins.the p6 protein of cauliflower mosaic virus (camv) is responsible for the formation of inclusion bodies (ibs), which are the sites for viral gene expression, replication, and virion assembly. moreover, recent evidence indicates that ectopically expressed p6 inclusion-like bodies (i-lbs) move in association with actin microfilaments. because camv virions accumulate preferentially in p6 ibs, we hypothesized that p6 ibs have a role in delivering camv virions to the plasmodesmata. we have determined ...201425239023
cucumber pti1-l is a cytoplasmic protein kinase involved in defense responses and salt tolerance.homologs of the cytoplasmic protein kinase pti1 are found in diverse plant species. a clear role of pti1 in plant defense response has not been established. we identified a pti1 homolog in cucumber (cspti1-l). cspti1-l expression was induced when cucumber plants were challenged with the fungal pathogen sphaerotheca fuliginea or with salt treatment. cspti1-l expression in cucumber leaves also was induced by methyl jasmonate, salicylic acid, and abscisic acid. cspti1-l exhibited autophosphorylatio ...201424877673
the arabidopsis lecrk-vi.2 associates with the pattern-recognition receptor fls2 and primes nicotiana benthamiana pattern-triggered immunity.pattern-triggered immunity (pti) is broad spectrum and manipulation of pti is believed to represent an attractive way to engineer plants with broad-spectrum disease resistance. pti is activated upon perception of microbe-associated molecular patterns (mamps) by pattern-recognition receptors (prrs). we have recently demonstrated that the l-type lectin receptor kinase-vi.2 (lecrk-vi.2) positively regulates arabidopsis thaliana pti. here we show through in vitro pull-down, bimolecular fluorescence ...201424844677
the endo-arabinanase bcara1 is a novel host-specific virulence factor of the necrotic fungal phytopathogen botrytis cinerea.the plant cell wall is one of the first physical interfaces encountered by plant pathogens and consists of polysaccharides, of which arabinan is an important constituent. during infection, the necrotrophic plant pathogen botrytis cinerea secretes a cocktail of plant cell-wall-degrading enzymes, including endo-arabinanase activity, which carries out the breakdown of arabinan. the roles of arabinan and endo-arabinanases during microbial infection were thus far elusive. in this study, the gene bcar ...201424725206
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
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
divergence of host range and biological properties between natural isolate and full-length infectious cdna clone of the beet mild yellowing virus 2itb.plant infection by poleroviruses is restricted to phloem tissues, preventing any classical leaf rub inoculation with viral rna or virions. efficient virus inoculation to plants is achieved by viruliferous aphids that acquire the virus by feeding on infected plants. the use of promoter-driven infectious cdna is an alternative means to infect plants and allows reverse genetic studies to be performed. using beet mild yellowing virus isolate 2itb (bmyv-2itb), we produced a full-length infectious cdn ...201423855287
infection of beet necrotic yellow vein virus with rna4-encoded p31 specifically up-regulates pathogenesis-related protein 10 in nicotiana benthamiana.beet necrotic yellow vein virus (bnyvv) is the infectious agent of sugar beet rhizomania, which consists of four or five plus-sense rnas. rna4 of bnyvv is not essential for virus propagation in nicotiana benthamiana but has a major effect on symptom expression. early reports showed that rna4-encoded p31 was associated with severe symptoms, such as curling and dwarfing, in n. benthamiana.201424961274
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
proof by synthesis of tobacco mosaic virus.synthetic biology is a discipline that includes making life forms artificially from chemicals. here, a dna molecule was enzymatically synthesized in vitro from dna templates made from oligonucleotides representing the text of the first tobacco mosaic virus (tmv) sequence elucidated in 1982. no infectious dna molecule of that seminal reference sequence exists, so the goal was to synthesize it and then build viral chimeras.201424887356
positive and negative roles for soybean mpk6 in regulating defense responses.it has been well established that mpk6 is a positive regulator of defense responses in model plants such as arabidopsis and tobacco. however, the functional importance of soybean mpk6 in disease resistance has not been investigated. here, we showed that silencing of gmmpk6 in soybean using virus-induced gene silencing mediated by bean pod mottle virus (bpmv) caused stunted growth and spontaneous cell death on the leaves, a typical phenotype of activated defense responses. consistent with this ph ...201424762222
phosphorylation of coat protein by protein kinase ck2 regulates cell-to-cell movement of bamboo mosaic virus through modulating rna binding.in this study, we investigated the fine regulation of cell-to-cell movement of bamboo mosaic virus (bamv). we report that the coat protein (cp) of bamv is phosphorylated in planta at position serine 241 (s241), in a process involving nicotiana benthamiana casein kinase 2α (nbck2α). bamv cp and nbck2α colocalize at the plasmodesmata, suggesting that phosphorylation of bamv may be involved in its movement. s241 was mutated to examine the effects of temporal and spatial dysregulation of phosphoryla ...201425025779
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 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
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
the movement protein (nsm) of tomato spotted wilt virus is the avirulence determinant in the tomato sw-5 gene-based resistance.the avirulence determinant triggering the resistance conferred by the tomato gene sw-5 against tomato spotted wilt virus (tswv) is still unresolved. sequence comparison showed two substitutions (c118y and t120n) in the movement protein nsm present only in tswv resistance-breaking (rb) isolates. in this work, transient expression of nsm of three tswv isolates [rb1 (t120n), rb2 (c118y) and non-resistance-breaking (nrb)] in nicotiana benthamiana expressing sw-5 showed a hypersensitive response (hr) ...201424690181
selective interaction between chloroplast β-atpase and tgb1l88 retards severe symptoms caused by alternanthera mosaic virus infection.the multifunctional triple gene block protein 1 (tgb1) of the potexvirus alternanthera mosaic virus (altmv) has been reported to have silencing suppressor, cell-to-cell movement, and helicase functions. yeast two hybrid screening using an arabidopsis thaliana cdna library with tgb1 as bait, and co-purification with tgb1 inclusion bodies identified several host proteins which interact with altmv tgb1. host protein interactions with tgb1 were confirmed by biomolecular fluorescence complementation, ...201425288986
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
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
pollen transmission of asparagus virus 2 (av-2) may facilitate mixed infection by two av-2 isolates in asparagus plants.asparagus virus 2 (av-2) is a member of the genus ilarvirus and thought to induce the asparagus decline syndrome. av-2 is known to be transmitted by seed, and the possibility of pollen transmission was proposed 25 years ago but not verified. in av-2 sequence analyses, we have unexpectedly found mixed infection by two distinct av-2 isolates in two asparagus plants. because mixed infections by two related viruses are normally prevented by cross protection, we suspected that pollen transmission of ...201425116643
expression of aspergillus nidulans phy gene in nicotiana benthamiana produces active phytase with broad specificities.a full-length phytase gene (phy) of aspergillus nidulans was amplified from the cdna library by polymerase chain reaction (pcr), and it was introduced into a bacterial expression vector, pet-28a. the recombinant protein (rphy-e, 56 kda) was overexpressed in the insoluble fraction of escherichia coli culture, purified by ni-nta resin under denaturing conditions and injected into rats as an immunogen. to express a. nidulans phytase in a plant, the full-length of phy was cloned into a plant express ...201425192284
production and characterization of in planta transiently produced polygalacturanase from aspergillus niger and its fusions with hydrophobin or elp tags.pectinases play an important role in plant cell wall deconstruction and have potential in diverse industries such as food, wine, animal feed, textile, paper, fuel, and others. the demand for such enzymes is increasing exponentially, as are the efforts to improve their production and to implement their use in several industrial processes. the goal of this study was to examine the potential of producing polygalacturonase i from aspergillus niger in plants and to investigate the effects of subcellu ...201424970673
agrobacterium tumefaciens mediated transient expression of plant cell wall-degrading enzymes in detached sunflower leaves.for biofuel applications, synthetic endoglucanase e1 and xylanase (xyn10a) derived from acidothermus cellulolyticus were transiently expressed in detached whole sunflower (helianthus annuus l.) leaves using vacuum infiltration. three different expression systems were tested, including the constitutive camv 35s-driven, cmvar (cucumber mosaic virus advanced replicating), and trbo (tobacco mosaic virus rna-based overexpression vector) systems. for 6-day leaf incubations, codon-optimized e1 and xyla ...201425180328
successful agrobacterium mediated transformation of thielaviopsis basicola by optimizing multiple conditions.thielaviopsis basicola is a hemibiotrophic root pathogen causing black root rot in a wide range of economically important crops. our initial attempts to transform t. basicola using standard agrobacterium tumefaciens-mediated transformation (atmt) protocols were unsuccessful. successful transformation required the addition of v8 juice (to induce germination of t. basicola chlamydospores) and higher concentrations of acetosyringone in the co-cultivation medium, and of chlamydospores/endoconidia, a ...201425110130
a golden gate modular cloning toolbox for plants.plant synthetic biology requires robust and efficient methods for assembling multigene constructs. golden gate cloning provides a precision module-based cloning technique for facile assembly of multiple genes in one construct. we present here a versatile resource for plant biologists comprising a set of cloning vectors and 96 standardized parts to enable golden gate construction of multigene constructs for plant transformation. parts include promoters, untranslated sequences, reporters, antigeni ...201424933124
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
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
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
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
plant cell death caused by fungal, bacterial, and viral elicitors: protective effect of mitochondria-targeted quinones.chitosan (partially deacetylated chitin), a component of fungal cell walls, caused epidermal cell (ec) death in the leaves of pea (pisum sativum l.) and tobacco nicotiana tabacum or nicotiana benthamiana detected by destruction of cell nuclei. the mitochondria-targeted quinone skq1 prevented the destruction of ec nuclei induced by chitosan. chitosan increased and skq1 suppressed the activity of protein kinases in n. benthamiana and p. sativum and eliminated the effect of chitosan. chitosan induc ...201425716725
the mir9863 family regulates distinct mla alleles in barley to attenuate nlr receptor-triggered disease resistance and cell-death signaling.barley (hordeum vulgare l.) mla alleles encode coiled-coil (cc), nucleotide binding, leucine-rich repeat (nb-lrr) receptors that trigger isolate-specific immune responses against the powdery mildew fungus, blumeria graminis f. sp. hordei (bgh). how mla or nb-lrr genes in grass species are regulated at post-transcriptional level is not clear. the microrna family, mir9863, comprises four members that differentially regulate distinct mla alleles in barley. we show that mir9863 members guide the cle ...201425502438
pepper heat shock protein 70a interacts with the type iii effector avrbst and triggers plant cell death and immunity.heat shock proteins (hsps) function as molecular chaperones and are essential for the maintenance and/or restoration of protein homeostasis. the genus xanthomonas type iii effector protein avrbst induces hypersensitive cell death in pepper (capsicum annuum). here, we report the identification of the pepper cahsp70a as an avrbst-interacting protein. bimolecular fluorescence complementation and coimmunoprecipitation assays confirm the specific interaction between cahsp70a and avrbst in planta. the ...201425491184
the tomato ubiquitin-conjugating enzyme variant suv, but not sluev1c and sluev1d regulates fen-mediated programmed cell death in nicotiana benthamiana.the unconventional, lysine-63-linked ubiquitination has been shown to play a central role in regulating human and animal innate and adaptive immunity. by contrast, the role and mechanism of k63-linked ubiquitination in plant biology remain largely unexplored. the tomato (solanum lycopersicum) fni3 ubiquitin-conjugating enzyme and its co-factor, suv ubiquitin e2 variant (uev) were shown recently to catalyze k63-linked ubiquitination and are essential for protein fen and other resistance protein-m ...201425482775
tomato sr/camta transcription factors slsr1 and slsr3l negatively regulate disease resistance response and slsr1l positively modulates drought stress tolerance.the sr/camta proteins represent a small family of transcription activators that play important roles in plant responses to biotic and abiotic stresses. seven slsr/camta genes were identified in tomato as tomato counterparts of sr/camta; however, the involvement of slsrs/camtas in biotic and abiotic stress responses is not clear. in this study, we performed functional analysis of the slsr/camta family for their possible functions in defense response against pathogens and tolerance to drought stre ...201425348703
micrornas suppress nb domain genes in tomato that confer resistance to fusarium oxysporum.micrornas (mirnas) suppress the transcriptional and post-transcriptional expression of genes in plants. several mirna families target genes encoding nucleotide-binding site-leucine-rich repeat (nb-lrr) plant innate immune receptors. the fungus fusarium oxysporum f. sp. lycopersici causes vascular wilt disease in tomato. we explored a role for mirnas in tomato defense against f. oxysporum using comparative mirna profiling of susceptible (moneymaker) and resistant (motelle) tomato cultivars. slmir ...201425330340
pepper mitochondrial formate dehydrogenase1 regulates cell death and defense responses against bacterial pathogens.formate dehydrogenase (fdh; ec 1.2.1.2) is an nad-dependent enzyme that catalyzes the oxidation of formate to carbon dioxide. here, we report the identification and characterization of pepper (capsicum annuum) mitochondrial fdh1 as a positive regulator of cell death and defense responses. transient expression of fdh1 caused hypersensitive response (hr)-like cell death in pepper and nicotiana benthamiana leaves. the d-isomer -: specific 2-hydroxyacid dehydrogenase signatures of fdh1 were required ...201425237129
na⁺/h⁺ exchanger 1 participates in tobacco disease defence against phytophthora parasitica var. nicotianae by affecting vacuolar ph and priming the antioxidative system.despite the importance of nhx1 (na(+)/h(+) exchanger 1) in plant salt tolerance, little is known about its other functions. in this study, intriguingly, it was found that nhx1 participated in plant disease defence against phytophthora parasitica var. nicotianae (ppn) in nicotiana benthamiana. nbnhx1 was originally isolated from n. benthamiana, and characterized. the subcellular localization of nbnhx1 with its c-terminus fused with green fluorescent protein indicated that nbnhx1 localized primari ...201425170102
involvement of potato (solanum tuberosum l.) mkk6 in response to potato virus y.mitogen-activated protein kinase (mapk) cascades have crucial roles in the regulation of plant development and in plant responses to stress. plant recognition of pathogen-associated molecular patterns or pathogen-derived effector proteins has been shown to trigger activation of several mapks. this then controls defence responses, including synthesis and/or signalling of defence hormones and activation of defence related genes. the mapk cascade genes are highly complex and interconnected, and thu ...201425111695
induction of innate immune responses by flagellin from the intracellular bacterium, 'candidatus liberibacter solanacearum'.'candidatus liberibacter solanacearum' (lso) is a phloem-limited alphaproteobacterium associated with the devastating zebra chip disease of potato (solanum tuberosum). like other members of liberibacter, lso-zc1 encodes a flagellin domain-containing protein (fla lso ) with a conserved 22 amino-acid peptide (flg22 lso ). to understand the innate immune responses triggered by this unculturable intracellular bacterium, we studied the pathogen-associated molecular patterns (pamps) that triggered imm ...201425091183
role of intron-mediated enhancement on accumulation of an arabidopsis nb-lrr class r-protein that confers resistance to cucumber mosaic virus.the accumulation of rcy1 protein, which is encoded by resistance to cmv(y) (rcy1), a cc-nb-lrr class r-gene, is tightly correlated with the strength of the resistance to a yellow strain of cucumber mosaic virus [cmv(y)] in arabidopsis thaliana. in order to enhance resistance to cmv by overexpression of rcy1, a. thaliana was transformed with intron-less rcy1 cdna construct under the control of strong camv35s promoter. remarkably, a relative amount of rcy1 protein accumulation in the transformants ...201424915153
agrobacterium-derived cytokinin influences plastid morphology and starch accumulation in nicotiana benthamiana during transient assays.agrobacterium tumefaciens-based transient assays have become a common tool for answering questions related to protein localization and gene expression in a cellular context. the use of these assays assumes that the transiently transformed cells are observed under relatively authentic physiological conditions and maintain 'normal' sub-cellular behaviour. although this premise is widely accepted, the question of whether cellular organization and organelle morphology is altered in agrobacterium-inf ...201424886417
functionally redundant rxlr effectors from phytophthora infestans act at different steps to suppress early flg22-triggered immunity.genome sequences of several economically important phytopathogenic oomycetes have revealed the presence of large families of so-called rxlr effectors. functional screens have identified rxlr effector repertoires that either compromise or induce plant defense responses. however, limited information is available about the molecular mechanisms underlying the modes of action of these effectors in planta. the perception of highly conserved pathogen- or microbe-associated molecular patterns (pamps/mam ...201424763622
aly proteins participate in multifaceted nep1mo-triggered responses in nicotiana benthamiana and arabidopsis thaliana.previously, it was found that nep1mo (a nep1-like protein from magnaporthe oryzae) could trigger a variety of plant responses, including stomatal closure, hypersensitive cell death (hcd), and defence-related gene expression, in nicotiana benthamiana. in this study, it was found that nep1mo-induced cell death could be inhibited by the virus-induced gene silencing of nbaly916 in n. benthamiana. nbaly916-silenced plants showed impaired nep1mo-induced stomatal closure, decreased nep1mo-induced produ ...201424723400
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
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
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
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
protein fragment bimolecular fluorescence complementation analyses for the in vivo study of protein-protein interactions and cellular protein complex localizations.the analyses of protein-protein interactions are crucial for understanding cellular processes including signal transduction, protein trafficking, and movement. protein fragment complementation assays are based on the reconstitution of protein function when non-active protein fragments are brought together by interacting proteins that were genetically fused to these protein fragments. bimolecular fluorescence complementation (bifc) relies on the reconstitution of fluorescent proteins and enables ...201424057390
domain dissection of avrrxo1 for suppressor, avirulence and cytotoxicity functions.avrrxo1, a type iii effector from xanthomonas oryzae pv. oryzicola (xoc) which causes bacterial leaf streak (bls) in rice, can be recognised by non-host resistance protein rxo1. it triggers a hypersensitive response (hr) in maize. little is known regarding the virulence function of avrrxo1. in this study, we determined that avrrxo1 is able to suppress the hr caused by the non-host resistance recognition of xanthomonas oryzae pv. oryzae (xoo) by nicotiana benthamiana. it is toxic, inducing cell d ...201425437277
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 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
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
a peroxisomal long-chain acyl-coa synthetase from glycine max involved in lipid degradation.seed storage oil, in the form of triacylglycerol (tag), is degraded to provide carbon and energy during germination and early seedling growth by the fatty acid β-oxidation in the peroxisome. although the pathways for lipid degradation have been uncovered, understanding of the exact involved enzymes in soybean is still limited. long-chain acyl-coa synthetase (acsl) is a critical enzyme that activates free fatty acid released from tag to form the fatty acyl-coa. recent studies have shown the impor ...201424992019
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
specific argonautes selectively bind small rnas derived from potato spindle tuber viroid and attenuate viroid accumulation in vivo.the identification of viroid-derived small rnas (vd-srnas) of 21 to 24 nucleotides (nt) in plants infected by viroids (infectious non-protein-coding rnas of just 250 to 400 nt) supports their targeting by dicer-like enzymes, the first host rna-silencing barrier. however, whether viroids, like rna viruses, are also targeted by the rna-induced silencing complex (risc) remains controversial. at the risc core is one argonaute (ago) protein that, guided by endogenous or viral srnas, targets complemen ...201425100851
β-carboxysomal proteins assemble into highly organized structures in nicotiana chloroplasts.the photosynthetic efficiency of c3 plants suffers from the reaction of ribulose 1,5-bisphosphate carboxylase/oxygenase (rubisco) with o2 instead of co2 , leading to the costly process of photorespiration. increasing the concentration of co2 around rubisco is a strategy used by photosynthetic prokaryotes such as cyanobacteria for more efficient incorporation of inorganic carbon. engineering the cyanobacterial co2 -concentrating mechanism, the carboxysome, into chloroplasts is an approach to enha ...201424810513
characterization of necrosis-inducing nlp proteins in phytophthora capsici.effector proteins function not only as toxins to induce plant cell death, but also enable pathogens to suppress or evade plant defense responses. nlp-like proteins are considered to be effector proteins, and they have been isolated from bacteria, fungi, and oomycete plant pathogens. there is increasing evidence that nlps have the ability to induce cell death and ethylene accumulation in plants.201424886309
structural and functional characterization of an anti-west nile virus monoclonal antibody and its single-chain variant produced in glycoengineered plants.previously, our group engineered a plant-derived monoclonal antibody (mab pe16) that efficiently treated west nile virus (wnv) infection in mice. in this study, we developed a pe16 variant consisting of a single-chain variable fragment (scfv) fused to the heavy chain constant domains (ch) of human igg (pe16scfv-ch). pe16 and pe16scfv-ch were expressed and assembled efficiently in nicotiana benthamiana ∆xf plants, a glycosylation mutant lacking plant-specific n-glycan residues. glycan analysis re ...201424975464
a novel plant-produced pfs25 fusion subunit vaccine induces long-lasting transmission blocking antibody responses.malaria transmission blocking vaccines (tbv) directed against proteins expressed on sexual stages of plasmodium falciparum in the mosquito midgut are considered an effective means to reduce malaria transmission. antibodies induced by tbv block sporogonic development in the mosquito, and thus transmission to the next human host. the pfs25 protein, expressed on the surface of gametes, zygotes and ookinetes, is one of the primary targets for tbv development. using a plant virus-based transient expr ...201425483525
the rxl motif of the african cassava mosaic virus rep protein is necessary for rereplication of yeast dna and viral infection in plants.geminiviruses, single-stranded dna plant viruses, encode a replication-initiator protein (rep) that is indispensable for virus replication. a potential cyclin interaction motif (rxl) in the sequence of african cassava mosaic virus rep may be an alternative link to cell cycle controls to the known interaction with plant homologs of retinoblastoma protein (prbr). mutation of this motif abrogated rereplication in fission yeast induced by expression of wildtype rep suggesting that rep interacts via ...201424999043
characterization of the cryptic av3 promoter of ageratum yellow vein virus in prokaryotic and eukaryotic systems.a cryptic prokaryotic promoter, designated av3 promoter, has been previously identified in certain begomovirus genus, including ageratum yellow vein virus isolate nt (ayvv-nt). in this study, we demonstrated that the core nucleotides in the putative -10 and -35 boxes are necessary but not sufficient for promoter activity in escherichia coli, and showed that ayvv-nt av3 promoter could specifically interact with single-stranded dna-binding protein and sigma 70 of e. coli involved in transcription. ...201425268755
molecular chaperons and co-chaperons, hsp90, rar1, and sgt1 negatively regulate bacterial wilt disease caused by ralstonia solanacearum in nicotiana benthamiana.ralstonia solanacearum is the causal agent of bacterial wilt disease. to better understand the molecular mechanisms involved in interaction between nicotiana benthamiana and r. solanacearum, we focused on hsp90, rar1 and sgt1. appearances of wilt symptom were significantly suppressed in hsp90, rar1 and sgt1-silenced plants compared with control plants. in rar1-silenced plants, population of r. solanacearum increased in a similar manner to control plants. in contrast, multiplication of r. solanac ...201425482800
the nb-lrr proteins rga4 and rga5 interact functionally and physically to confer disease resistance.plant resistance proteins of the class of nucleotide-binding and leucine-rich repeat domain proteins (nb-lrrs) are immune sensors which recognize pathogen-derived molecules termed avirulence (avr) proteins. we show that rga4 and rga5, two nb-lrrs from rice, interact functionally and physically to mediate resistance to the fungal pathogen magnaporthe oryzae and accomplish different functions in avr recognition. rga4 triggers an avr-independent cell death that is repressed in the presence of rga5 ...201425024433
tomato slmkk2 and slmkk4 contribute to disease resistance against botrytis cinerea.mitogen-activated protein kinase (mapk) cascades are highly conserved signaling modules that mediate the transduction of extracellular stimuli via receptors/sensors into intracellular responses and play key roles in plant immunity against pathogen attack. however, the function of tomato mapk kinases, slmkks, in resistance against botrytis cinerea remains unclear yet.201424930014
sec14 phospholipid transfer protein is involved in lipid signaling-mediated plant immune responses in nicotiana benthamiana.we previously identified a gene related to the sec14-gene phospholipid transfer protein superfamily that is induced in nicotiana benthamiana (nbsec14) in response to infection with ralstonia solanacearum. we here report that nbsec14 plays a role in plant immune responses via phospholipid-turnover. nbsec14-silencing compromised expression of defense-related pr-4 and accumulation of jasmonic acid (ja) and its derivative ja-ile. transient expression of nbsec14 induced pr-4 gene expression. activiti ...201424845602
ghwrky40, a multiple stress-responsive cotton wrky gene, plays an important role in the wounding response and enhances susceptibility to ralstonia solanacearum infection in transgenic nicotiana benthamiana.wrky transcription factors form one of the largest transcription factor families and function as important components in the complex signaling processes that occur during plant stress responses. however, relative to the research progress in model plants, far less information is available on the function of wrky proteins in cotton. in the present study, we identified the ghwrky40 gene in cotton (gossypium hirsutum) and determined that the ghwrky40 protein is targeted to the nucleus and is a stres ...201424747610
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
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
suppression of ds1 phosphatidic acid phosphatase confirms resistance to ralstonia solanacearum in nicotiana benthamiana.nicotianabenthamiana is susceptible to ralstonia solanacearum. to analyze molecular mechanisms for disease susceptibility, we screened a gene-silenced plant showing resistance to r. solanacearum, designated as ds1 (disease suppression 1). the deduced amino acid sequence of ds1 cdna encoded a phosphatidic acid phosphatase (pap) 2. ds1 expression was induced by infection with a virulent strain of r. solanacearum in an hrp-gene-dependent manner. ds1 rescued growth defects of the temperature-sensiti ...201324073238
cotton ghmkk1 induces the tolerance of salt and drought stress, and mediates defence responses to pathogen infection in transgenic nicotiana benthamiana.mitogen-activated protein kinase kinases (mapkk) mediate a variety of stress responses in plants. so far little is known on the functional role of mapkks in cotton. in the present study, gossypium hirsutum mkk1 (ghmkk1) function was investigated. ghmkk1 protein may activate its specific targets in both the nucleus and cytoplasm. treatments with salt, drought, and h2o2 induced the expression of ghmkk1 and increased the activity of ghmkk1, while overexpression of ghmkk1 in nicotiana benthamiana en ...201323844212
the role of tir-nbs and tir-x proteins in plant basal defense responses.toll/interleukin receptor (tir) domain-containing proteins encoded in the arabidopsis (arabidopsis thaliana) genome include the tir-nucleotide binding site (tn) and tir-unknown site/domain (tx) families. we investigated the function of these proteins. transient overexpression of five tx and tn genes in tobacco (nicotiana benthamiana) induced chlorosis. this induced chlorosis was dependent on enhanced disease resistance1, a dependency conserved in both tobacco and arabidopsis. stable overexpressi ...201323735504
a remarkable synergistic effect at the transcriptomic level in peach fruits doubly infected by prunus necrotic ringspot virus and peach latent mosaic viroid.microarray profiling is a powerful technique to investigate expression changes of large amounts of genes in response to specific environmental conditions. the majority of the studies investigating gene expression changes in virus-infected plants are limited to interactions between a virus and a model host plant, which usually is arabidopsis thaliana or nicotiana benthamiana. in the present work, we performed microarray profiling to explore changes in the expression profile of field-grown prunus ...201323710752
genome-wide identification and functional analyses of calmodulin genes in solanaceous species.calmodulin (cam) is a major calcium sensor in all eukaryotes. it binds calcium and modulates the activity of a wide range of downstream proteins in response to calcium signals. however, little is known about the cam gene family in solanaceous species, including the economically important species, tomato (solanum lycopersicum), and the gene silencing model plant, nicotiana benthamiana. moreover, the potential function of cam in plant disease resistance remains largely unclear.201323621884
discovering host genes involved in the infection by the tomato yellow leaf curl virus complex and in the establishment of resistance to the virus using tobacco rattle virus-based post transcriptional gene silencing.the development of high-throughput technologies allows for evaluating gene expression at the whole-genome level. together with proteomic and metabolomic studies, these analyses have resulted in the identification of plant genes whose function or expression is altered as a consequence of pathogen attacks. members of the tomato yellow leaf curl virus (tylcv) complex are among the most important pathogens impairing production of agricultural crops worldwide. to understand how these geminiviruses su ...201323524390
breaking restricted taxonomic functionality by dual resistance genes.nb-lrr-type disease resistance (r) genes have been used in traditional breeding programs for crop protection. however, functional transfer of nb-lrr-type r genes to plants in taxonomically distinct families to establish pathogen resistance has not been successful. here we demonstrate that a pair of arabidopsis (brassicaceae) nb-lrr-type r genes, rps4 and rrs1, properly function in two other brassicaceae, brassica rapa and b. napus, but also in two solanaceae, nicotiana benthamiana and tomato (so ...201323518587
interfamily transfer of dual nb-lrr genes confers resistance to multiple pathogens.a major class of disease resistance (r) genes which encode nucleotide binding and leucine rich repeat (nb-lrr) proteins have been used in traditional breeding programs for crop protection. however, it has been difficult to functionally transfer nb-lrr-type r genes in taxonomically distinct families. here we demonstrate that a pair of arabidopsis (brassicaceae) nb-lrr-type r genes, rps4 and rrs1, properly function in two other brassicaceae, brassica rapa and brassica napus, but also in two solana ...201323437080
phosphorylation of hopq1, a type iii effector from pseudomonas syringae, creates a binding site for host 14-3-3 proteins.hopq1 (for hrp outer protein q), a type iii effector secreted by pseudomonas syringae pv phaseolicola, is widely conserved among diverse genera of plant bacteria. it promotes the development of halo blight in common bean (phaseolus vulgaris). however, when this same effector is injected into nicotiana benthamiana cells, it is recognized by the immune system and prevents infection. although the ability to synthesize hopq1 determines host specificity, the role it plays inside plant cells remains u ...201323396834
interaction of medicago truncatula lysin motif receptor-like kinases, nfp and lyk3, produced in nicotiana benthamiana induces defence-like responses.receptor(-like) kinases with lysin motif (lysm) domains in their extracellular region play crucial roles during plant interactions with microorganisms; e.g. arabidopsis thaliana cerk1 activates innate immunity upon perception of fungal chitin/chitooligosaccharides, whereas medicago truncatula nfp and lyk3 mediate signalling upon perception of bacterial lipo-chitooligosaccharides, termed nod factors, during the establishment of mutualism with nitrogen-fixing rhizobia. however, little is still kno ...201323750228
rnase j participates in a pentatricopeptide repeat protein-mediated 5' end maturation of chloroplast mrnas.nucleus-encoded ribonucleases and rna-binding proteins influence chloroplast gene expression through their roles in rna maturation and stability. one mechanism for mrna 5' end maturation posits that sequence-specific pentatricopeptide repeat (ppr) proteins define termini by blocking the 5'→3' exonucleolytic activity of ribonuclease j (rnase j). to test this hypothesis in vivo, virus-induced gene silencing was used to reduce the expression of three ppr proteins and rnase j, both individually and ...201323921629
allelic variation in two distinct pseudomonas syringae flagellin epitopes modulates the strength of plant immune responses but not bacterial motility.the bacterial flagellin (flic) epitopes flg22 and flgii-28 are microbe-associated molecular patterns (mamps). although flg22 is recognized by many plant species via the pattern recognition receptor fls2, neither the flgii-28 receptor nor the extent of flgii-28 recognition by different plant families is known. here, we tested the significance of flgii-28 as a mamp and the importance of allelic diversity in flg22 and flgii-28 in plant-pathogen interactions using purified peptides and a pseudomonas ...201323865782
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
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
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
the tomato fni3 lysine-63-specific ubiquitin-conjugating enzyme and suv ubiquitin e2 variant positively regulate plant immunity.the activation of an immune response in tomato (solanum lycopersicum) against pseudomonas syringae relies on the recognition of e3 ligase-deficient forms of avrptob by the host protein kinase, fen. to investigate the mechanisms by which fen-mediated immunity is regulated, we characterize in this study a fen-interacting protein, fni3, and its cofactor, s. lycoperiscum uev (suv). fni3 encodes a homolog of the ubc13-type ubiquitin-conjugating enzyme that catalyzes exclusively lys-63-linked ubiquiti ...201324076975
jasmonate zim-domain (jaz) protein regulates host and nonhost pathogen-induced cell death in tomato and nicotiana benthamiana.the nonhost-specific phytotoxin coronatine (cor) produced by several pathovars of pseudomonas syringae functions as a jasmonic acid-isoleucine (ja-ile) mimic and contributes to disease development by suppressing plant defense responses and inducing reactive oxygen species in chloroplast. it has been shown that the f-box protein coronatine insensitive 1 (coi1) is the receptor for cor and ja-ile. jasmonate zim domain (jaz) proteins act as negative regulators for ja signaling in arabidopsis. howeve ...201324086622
vigs-mediated forward genetics screening for identification of genes involved in nonhost resistance.nonhost disease resistance of plants against bacterial pathogens is controlled by complex defense pathways. understanding this mechanism is important for developing durable disease-resistant plants against wide range of pathogens. virus-induced gene silencing (vigs)-based forward genetics screening is a useful approach for identification of plant defense genes imparting nonhost resistance. tobacco rattle virus (trv)-based vigs vector is the most efficient vigs vector to date and has been efficie ...201323995956
the tomato calcium sensor cbl10 and its interacting protein kinase cipk6 define a signaling pathway in plant immunity.ca(2+) signaling is an early and necessary event in plant immunity. the tomato (solanum lycopersicum) kinase pto triggers localized programmed cell death (pcd) upon recognition of pseudomonas syringae effectors avrpto or avrptob. in a virus-induced gene silencing screen in nicotiana benthamiana, we independently identified two components of a ca(2+)-signaling system, cbl10 (for calcineurin b-like protein) and cipk6 (for calcineurin b-like interacting protein kinase), as their silencing inhibited ...201323903322
the receptor-like protein remax of arabidopsis detects the microbe-associated molecular pattern emax from xanthomonas.as part of their immune system, plants have pattern recognition receptors (prrs) that can detect a broad range of microbe-associated molecular patterns (mamps). here, we identified a prr of arabidopsis thaliana with specificity for the bacterial mamp emax from xanthomonads. response to emax seems to be restricted to the brassicaceae family and also varied among different accessions of arabidopsis. in crosses between sensitive accessions and the insensitive accession shakhdara, emax perception ma ...201323898033
pepper arginine decarboxylase is required for polyamine and γ-aminobutyric acid signaling in cell death and defense response.the xanthomonas campestris pv vesicatoria (xcv) effector avrbst induces a hypersensitive cell death in pepper (capsicum annuum). however, the molecular mechanisms underlying avrbst-triggered cell death are not fully understood. here, we identified pepper arginine decarboxylase (caadc1) as an avrbst-interacting protein, which is early and strongly induced in incompatible pepper-xcv interactions. bimolecular fluorescence complementation and coimmunoprecipitation assays showed that the caadc1-avrbs ...201323784462
members of the xb3 family from diverse plant species induce programmed cell death in nicotiana benthamiana.programmed cell death has been associated with plant immunity and senescence. the receptor kinase xa21 confers resistance to bacterial blight disease of rice (oryza sativa) caused by xanthomonas oryzae pv. oryzae (xoo). here we show that the xa21 binding protein 3 (xb3) is capable of inducing cell death when overexpressed in nicotiana benthamiana. xb3 is a ring finger-containing e3 ubiquitin ligase that has been positively implicated in xa21-mediated resistance. mutation abolishing the xb3 e3 ac ...201323717500
drought stress acclimation imparts tolerance to sclerotinia sclerotiorum and pseudomonas syringae in nicotiana benthamiana.acclimation of plants with an abiotic stress can impart tolerance to some biotic stresses. such a priming response has not been widely studied. in particular, little is known about enhanced defense capacity of drought stress acclimated plants to fungal and bacterial pathogens. here we show that prior drought acclimation in nicotiana benthamiana plants imparts tolerance to necrotrophic fungus, sclerotinia sclerotiorum, and also to hemi-biotrophic bacterial pathogen, pseudomonas syringae pv. tabac ...201323644883
oxylipin biosynthesis genes positively regulate programmed cell death during compatible infections with the synergistic pair potato virus x-potato virus y and tomato spotted wilt virus.one of the most severe symptoms caused by compatible plant-virus interactions is systemic necrosis, which shares common attributes with the hypersensitive response to incompatible pathogens. although several studies have identified viral symptom determinants responsible for systemic necrosis, mechanistic models of how they contribute to necrosis in infected plants remain scarce. here, we examined the involvement of different branches of the oxylipin biosynthesis pathway in the systemic necrosis ...201323487466
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