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raba members act in distinct steps of subcellular trafficking of the flagellin sensing2 receptor.cell surface proteins play critical roles in the perception of environmental stimuli at the plasma membrane (pm) and ensuing signal transduction. intracellular localization of such proteins must be strictly regulated, which requires elaborate integration of exocytic and endocytic trafficking pathways. subcellular localization of arabidopsis thaliana flagellin sensing2 (fls2), a receptor that recognizes bacterial flagellin, also depends on membrane trafficking. however, our understanding about th ...201323532067
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
beet yellows virus replicase and replicative compartments: parallels with other rna viruses.in eukaryotic virus systems, infection leads to induction of membranous compartments in which replication occurs. virus-encoded subunits of the replication complex mediate its interaction with membranes. as replication platforms, rna viruses use the cytoplasmic surfaces of different membrane compartments, e.g., endoplasmic reticulum (er), golgi, endo/lysosomes, mitochondria, chloroplasts, and peroxisomes. closterovirus infections are accompanied by formation of multivesicular complexes from cell ...201323508802
the endophytic lifestyle of escherichia coli o157:h7: quantification and internal localization in roots.the foodborne pathogen escherichia coli o157:h7 is increasingly associated with fresh produce (fruit and vegetables). bacterial colonization of fresh produce plants can occur to high levels on the external tissue but bacteria have also been detected within plant tissue. however, questions remain about the extent of internalization, its molecular basis, and internal location of the bacteria. we have determined the extent of internalization of e. coli o157:h7 in live spinach and lettuce plants and ...201323506361
biological and biochemical characterization of hiv-1 gag/dgp41 virus-like particles expressed in nicotiana benthamiana.the transmembrane hiv-1 envelope protein gp41 has been shown to play critical roles in the viral mucosal transmission and infection of cd4⁺ cells. gag is a structural protein configuring the enveloped viral particles and has been suggested to constitute a target of the cellular immunity that may control viral load. we hypothesized that hiv enveloped virus-like particles (vlps) consisting of gag and a deconstructed form of gp41 comprising the membrane proximal external, transmembrane and cytoplas ...201323506331
targeting of envelope domain iii protein of denv type 2 to dec-205 receptor elicits neutralizing antibodies in mice.dengue virus (denv) is the causal agent of severe disease and, in some cases, mortality in humans, but no licensed vaccines against dengue are available. an effective vaccine against dengue requires long-term humoral and cellular immune responses. several viral proteins have been the subjects of intense research, especially the envelope (e) protein, aimed at developing a vaccine. domain iii of the envelope protein (ediii) has been identified as a potential candidate because it is involved in bin ...201323499580
characterization of the complete genome of a novel citrivirus infecting actinidia chinensis.a ssrna virus from kiwifruit (actinidia spp.) was identified as a member of the family betaflexiviridae. it was mechanically transmitted to the herbaceous indicators nicotiana benthamiana, n. clevelandii, n. glutinosa and n. occidentalis. the complete genome was comprised of three orfs and a 3'poly (a) tail. phylogenetic analysis of the entire genome indicated it was a novel member of the genus citrivirus (family betaflexiviridae). the complete nucleotide sequence differed from that of citrus le ...201323494225
translation elongation factor 1b (eef1b) is an essential host factor for tobacco mosaic virus infection in plants.identifying host factors provides an important clue to understand virus infection. we selected 10 host factor candidate genes and each gene was silenced in nicotiana benthamiana (n. benthamiana) to investigate their roles in virus infection. the resulting plants were infected with tobacco mosaic virus (tmv). the accumulation of viral coat protein and the spread of virus were greatly reduced in the plants that eukaryotic translation elongation factor 1a (eef1a) or 1b (eef1b) was silenced. these r ...201323490052
differential requirements for tombusvirus coat protein and p19 in plants following leaf versus root inoculation.traditional virus inoculation of plants involves mechanical rubbing of leaves, whereas in nature viruses like tomato bushy stunt virus (tbsv) are often infected via the roots. a method was adapted to compare leaf versus root inoculation of nicotiana benthamiana and tomato with transcripts of wild-type tbsv (wttbsv), a capsid (tcp) replacement construct expressing gfp (t-gfp), or mutants not expressing the silencing suppressor p19 (tbsvδp19). in leaves, t-gfp remained restricted to the cells imme ...201323490050
interaction of the microtubule-associated host protein hip2 with viral helper component proteinase is important in infection with potato virus a.microtubules (mt) outline and maintain the overall shape of cells and can reorganize cellular membranes to serve as sites of rna virus replication. here, we provide data on involvement of an mt-associated protein in infection of plants with a potyvirus, potato virus a (pva), representing the largest family of plant-infecting rna viruses. our results showed that helper-component proteinase (hcpro)-interacting protein 2 (hip2) of potato (solanum tuberosum) is an mt-associated protein similar to ar ...201323489059
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
gmnfya3, a target gene of mir169, is a positive regulator of plant tolerance to drought stress.nuclear factor y (nf-y) is a heterotrimeric transcription factor composed of nf-ya, nf-yb and nf-yc proteins. in this study, we identified and characterized a gene, gmnfya3, which encodes the nf-ya subunit of the nf-y complex in soybeans (glycine max l.). real time rt-pcr analysis indicated that gmnfya3 was induced by abscisic acid (aba) and abiotic stresses, such as polyethylene glycol, nacl and cold. subcellular localization analysis suggested that gmnfya3 may activate its specific targets in ...201323483290
intracellular and extracellular phosphatidylinositol 3-phosphate produced by phytophthora species is important for infection.rxlr effectors produced by phytophthora pathogens have been proposed to bind to phosphatidylinositol 3-phosphate (ptdins(3)p) to mediate their translocation into host cells and/or to increase their stability in planta. since the levels of ptdins(3)p in plants are low, we examined whether phytophthora species may produce ptdins(3)p to promote infection. we observed that ptdins(3)p-specific gfp biosensors could bind to p. parasitica and p. sojae hyphae during infection of nicotiana benthamiana lea ...201323475996
interactions between the p6 and p5-1 proteins of southern rice black-streaked dwarf fijivirus in yeast and plant cells.southern rice black-streaked dwarf virus (srbsdv) is a recently described member of the genus fijivirus, family reoviridae. the roles of the proteins encoded by the srbsdv genome have rarely been studied. in a yeast two-hybrid (yth) assay in which srbsdv p6, a putatively multifunctional protein, was used as bait and an srbsdv cdna library was used as prey, there was a strong interaction between the p6 and p5-1 proteins. the interaction was confirmed by bimolecular fluorescence complement (bifc) ...201323474918
scion on a stock producing sirnas of potato spindle tuber viroid (pstvd) attenuates accumulation of the viroid.plants can attenuate the replication of plant viruses and viroids by rna silencing induced by virus and viroid infection. in higher plants, silencing signals such as small interfering rnas (sirnas) produced by rna silencing can be transported systemically through phloem, so it is anticipated that antiviral sirna signals produced in a stock would have the potential to attenuate propagation of viruses or viroids in the scion. to test whether this is indeed the case, we prepared transgenic tobacco ...201323469061
pest protection conferred by a beta vulgaris serine proteinase inhibitor gene.proteinase inhibitors provide a means of engineering plant resistance to insect pests. a beta vulgaris serine proteinase inhibitor gene (bvsti) was fused to the constitutive camv35s promoter for over-expression in nicotiana benthamiana plants to study its effect on lepidopteran insect pests. independently derived bvsti transgenic tobacco t2 homozygous progeny were shown to have relatively high bvsti gene transcript levels. bvsti-specific polyclonal antibodies cross-reacted with the expected 30 k ...201323468963
molecular characterization of the interaction between the n-terminal region of potato virus x (pvx) coat protein (cp) and nicotiana benthamiana pvx cp-interacting protein, nbpcip1.using yeast two-hybrid assays and a nicotiana benthamiana cdna library, we previously identified an n. benthamiana protein, nbpcip1, that interacts with potato virus x (pvx) coat protein (cp). we also previously determined that nbpcip1 enhances pvx replication in plants. to determine the domains and/or amino acid residues required for pvx cp and nbpcip1 interaction, here we used yeast two-hybrid and β-galactosidase filter assays to test the effects of deletion and site-directed mutations on the ...201323463175
tgbp3 triggers the unfolded protein response and skp1-dependent programmed cell death.the potato virus x (pvx) triple gene block protein 3 (tgbp3), an 8-kda membrane binding protein, aids virus movement and induces the unfolded protein response (upr) during pvx infection. tgbp3 was expressed from the tobacco mosaic virus (tmv) genome (tmv-p3), and we noted the up-regulation of skp1 and several endoplasmic reticulum (er)-resident chaperones, including the er luminal binding protein (bip), protein disulphide isomerase (pdi), calreticulin (crt) and calmodulin (cam). local lesions we ...201323458484
deployment of the burkholderia glumae type iii secretion system as an efficient tool for translocating pathogen effectors to monocot cells.genome sequences of plant fungal pathogens have enabled the identification of effectors that cooperatively modulate the cellular environment for successful fungal growth and suppress host defense. identification and characterization of novel effector proteins are crucial for understanding pathogen virulence and host-plant defense mechanisms. previous reports indicate that the pseudomonas syringae pv. tomato dc3000 type iii secretion system (t3ss) can be used to study how non-bacterial effectors ...201323451734
construction of an infectious cdna clone of radish mosaic virus, a crucifer-infecting comovirus.radish mosaic virus (ramv) is a crucifer-infecting comovirus that has been detected worldwide. here, we report the successful construction of a full-length infectious cdna clone of ramv. the full-length cdna clones corresponding to rna1 and rna2 of a japanese isolate of ramv were cloned into the pbluescript plasmid or the binary vector pcambia1301 downstream of the cauliflower mosaic virus 35s promoter. mechanical inoculation or agroinoculation of nicotiana benthamiana with these vectors resulte ...201323447094
jasmonate controls leaf growth by repressing cell proliferation and the onset of endoreduplication while maintaining a potential stand-by mode.phytohormones regulate plant growth from cell division to organ development. jasmonates (jas) are signaling molecules that have been implicated in stress-induced responses. however, they have also been shown to inhibit plant growth, but the mechanisms are not well understood. the effects of methyl jasmonate (meja) on leaf growth regulation were investigated in arabidopsis (arabidopsis thaliana) mutants altered in ja synthesis and perception, allene oxide synthase and coi1-16b (for coronatine ins ...201323439917
analysis of the solvent accessibility of cysteine residues on maize rayado fino virus virus-like particles produced in nicotiana benthamiana plants and cross-linking of peptides to vlps.mimicking and exploiting virus properties and physicochemical and physical characteristics holds promise to provide solutions to some of the world's most pressing challenges. the sheer range and types of viruses coupled with their intriguing properties potentially give endless opportunities for applications in virus-based technologies. viruses have the ability to self- assemble into particles with discrete shape and size, specificity of symmetry, polyvalence, and stable properties under a wide r ...201323439009
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
a new set of reversibly photoswitchable fluorescent proteins for use in transgenic plants.fluorescent reporter proteins that allow repeated switching between a fluorescent and a non-fluorescent state in response to specific wavelengths of light are novel tools for monitoring of protein trafficking and super-resolution fluorescence microscopy in living organisms. here, we describe variants of the reversibly photoswitchable fluorescent proteins rsfastlime, bsdronpa, and padron that have been codon-optimized for the use in transgenic arabidopsis plants. the synthetic proteins, designate ...201323434876
a coat-independent superinfection exclusion rapidly imposed in nicotiana benthamiana cells by tobacco mosaic virus is not prevented by depletion of the movement protein.new evidence is emerging which indicates that population variants in plant virus infections are not uniformly distributed along the plant, but structured in a mosaic-like pattern due to limitation to the superinfection imposed by resident viral clones. the mechanisms that prevent the infection of a challenge virus into a previously infected cell, a phenomenon known as superinfection exclusion (se) or homologous interference, are only partially understood. by taking advantage of a deconstructed t ...201323417583
mungbean yellow mosaic virus (mymv) ac4 suppresses post-transcriptional gene silencing and an ac4 hairpin rna gene reduces mymv dna accumulation in transgenic tobacco.mungbean yellow mosaic virus (mymv) is a legume-infecting geminivirus that causes yellow mosaic disease in blackgram, mungbean, soybean, frenchbean and mothbean. ac4/c4, which is nested completely within the rep gene, is less conserved among geminiviruses. much less is known about its role in viral pathogenesis other than its known role in the suppression of host-mediated gene silencing. transient expression of mymv ac4 by agroinfiltration suppressed post-transcriptional gene silencing in nicoti ...201323417222
establishment of an agroinoculation system for broad bean wilt virus 2.we determined the complete nucleotide sequence of a broad bean wilt virus 2 (bbwv-2) isolate from gentian in japan. the full-length rna1 and rna2 sequences, excluding poly(a) tails, were 5955 and 3600 nucleotides long, respectively. analysis indicated that, in contrast to other bbwv-2 isolates, the 5' end of both rna1 and rna2 starts with a guu sequence. we successfully inoculated nicotiana benthamiana with bbwv-2 by infiltrating a mixed suspension of two agrobacterium tumefaciens clones carryin ...201323404460
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
immunogenicity and protective efficacy of candidate universal influenza a nanovaccines produced in plants by tobacco mosaic virus-based vectors.a new approach for super-expression of the influenza virus epitope m2e in plants has been developed on the basis of a recombinant tobacco mosaic virus (tmv, strain u1) genome designed for agrobacterium-mediated delivery into the plant cell nucleus. the tmv coat protein (cp) served as a carrier and three versions of the m2e sequence were inserted into the surface loop between amino acid residues 155 and 156. cysteine residues in the heterologous peptide were thought likely to impede efficient ass ...201323394564
citrus tristeza virus p23: determinants for nucleolar localization and their influence on suppression of rna silencing and pathogenesis.citrus tristeza virus (ctv) encodes a singular protein (p23, 209 amino acids) with multiple functions, including rna silencing suppression (rss). confocal laser-scanning microscopy of green fluorescent protein (gfp)-p23 agroexpressed in nicotiana benthamiana revealed its accumulation in the nucleolus, cajal bodies, and plasmodesmata. to dissect the nucleolar localization signal (nols) typically associated with basic motifs, seven truncated and 10 point-mutated versions of p23 were assayed. delet ...201323387469
insights into alternanthera mosaic virus tgb3 functions: interactions with nicotiana benthamiana psbo correlate with chloroplast vesiculation and veinal necrosis caused by tgb3 over-expression.alternanthera mosaic virus (altmv) triple gene block 3 (tgb3) protein is involved in viral movement. altmv tgb3 subcellular localization was previously shown to be distinct from that of potato virus x (pvx) tgb3, and a chloroplast binding domain identified; veinal necrosis and chloroplast vesiculation were observed in nicotiana benthamiana when altmv tgb3 was over-expressed from pvx. plants with over-expressed tgb3 showed more lethal damage under dark conditions than under light. yeast-two-hybri ...201323386854
virus-induced gene silencing using artificial mirnas in nicotiana benthamiana.virus-induced gene silencing using artificial micrornas (mir vigs) is a newly developed technique for plant reverse genetic studies. traditional virus-induced gene silencing (vigs) assays introduce a large gene fragment, which is expressed and then converted into small rnas by the endogenous sirna-based gene silencing machinery of the plant host. by contrast, mir vigs uses well-designed mirnas to induce rna-mediated silencing of the target gene. using a single artificial mirna can provide greate ...201323386298
agroinoculation of beet necrotic yellow vein virus cdna clones results in plant systemic infection and efficient polymyxa betae transmission.agroinoculation is a quick and easy method for the infection of plants with viruses. this method involves the infiltration of tissue with a suspension of agrobacterium tumefaciens carrying binary plasmids harbouring full-length cdna copies of viral genome components. when transferred into host cells, transcription of the cdna produces rna copies of the viral genome that initiate infection. we produced full-length cdna corresponding to beet necrotic yellow vein virus (bnyvv) rnas and derived repl ...201323384276
production, characterization, and antigen specificity of recombinant 62-71-3, a candidate monoclonal antibody for rabies prophylaxis in humans.rabies kills many people throughout the developing world every year. the murine monoclonal antibody (mab) 62-71-3 was recently identified for its potential application in rabies postexposure prophylaxis (pep). the purpose here was to establish a plant-based production system for a chimeric mouse-human version of mab 62-71-3, to characterize the recombinant antibody and investigate at a molecular level its interaction with rabies virus glycoprotein. chimeric 62-71-3 was successfully expressed in ...201323371065
sce1, the sumo-conjugating enzyme in plants that interacts with nib, the rna-dependent rna polymerase of turnip mosaic virus, is required for viral infection.sumoylation, which is catalyzed by small ubiquitin-like modifier (sumo) enzymes, is a transient, reversible posttranslational protein modification that regulates diverse cellular processes. potyviruses, the largest group of known plant viruses, comprise many agriculturally important viruses, such as turnip mosaic virus (tumv). the potyviral genome encodes 11 mature proteins. to investigate if sumoylation plays a role in potyvirus infection, a yeast two-hybrid screen was performed to examine poss ...201323365455
identification of a movement protein of mirafiori lettuce big-vein ophiovirus.mirafiori lettuce big-vein virus (milbvv) is a member of the genus ophiovirus, which is a segmented negative-stranded rna virus. in microprojectile bombardment experiments to identify a movement protein (mp) gene of ophioviruses that can trans-complement intercellular movement of an mp-deficient heterologous virus, a plasmid containing an infectious clone of a tomato mosaic virus (tomv) derivative expressing the gfp was co-bombarded with plasmids containing one of three genes from milbvv rnas 1, ...201323364193
modification of tobacco rattle virus rna1 to serve as a vigs vector reveals that the 29k movement protein is an rna silencing suppressor of the virus.tobacco rattle virus (trv) has a bipartite, positive-sense single-stranded rna genome and is widely used for virus-induced gene silencing (vigs) in plants. rna1 of trv that lacks the gene for the cysteine-rich 16k silencing-suppression protein infects plants systemically in the absence of rna2. here, we attempted to engineer rna1 for use as a vigs vector by inserting heterologous gene fragments to replace 16k. the rna1 vector systemically silenced the phytoene desaturase (pds) gene, although les ...201323360458
tsw gene-based resistance is triggered by a functional rna silencing suppressor protein of the tomato spotted wilt virus.as a result of contradictory reports, the avirulence (avr) determinant that triggers tsw gene-based resistance in capsicum annuum against the tomato spotted wilt virus (tswv) is still unresolved. here, the n and nss genes of resistance-inducing (ri) and resistance-breaking (rb) isolates were cloned and transiently expressed in resistant capsicum plants to determine the identity of the avr protein. it was shown that the nss(ri) protein triggered a hypersensitive response (hr) in tsw-containing ca ...201323360130
production of a chimeric allergen derived from the major allergen group 1 of house dust mite species in nicotiana benthamiana.plants are widely accepted as a general platform for the large-scale production of recombinant proteins, which has been demonstrated by the successful expression of various exogenous proteins. using plants as a bioreactor for mass production of target proteins for vaccines is thought to show the most potential. this study explores whether a chimeric allergen r8, derived from the major allergen group 1 of house dust mites species (dermatophagoides farinae and dermatophagoides pteronyssinus), is e ...201323354320
a ubiquitin carboxyl extension protein secreted from a plant-parasitic nematode globodera rostochiensis is cleaved in planta to promote plant parasitism.nematode effector proteins originating from esophageal gland cells play central roles in suppressing plant defenses and in formation of the plant feeding cells that are required for growth and development of cyst nematodes. a gene (grubcep12) encoding a unique ubiquitin carboxyl extension protein (ubcep) that consists of a signal peptide for secretion, a mono-ubiquitin domain, and a 12 amino acid carboxyl extension protein (cep12) domain was cloned from the potato cyst nematode globodera rostoch ...201323346875
sgt1 interacts with the prf resistance protein and is required for prf accumulation and prf-mediated defense signaling.the highly conserved eukaryotic co-chaperone sgt1 (suppressor of the g2 allele of skp1) is an important signaling component of plant defense responses and positively regulates disease resistance conferred by many resistance (r) proteins. in this study, we investigated the contribution of sgt1 in the prf-mediated defense responses in both nicotiana benthamiana and tomato (solanum lycopersicum). sgt1 was demonstrated to interact with prf in plant cells by co-immunoprecipitation. the requirement of ...201323333384
a plant-produced protective antigen vaccine confers protection in rabbits against a lethal aerosolized challenge with bacillus anthracis ames spores.the potential use of bacillus anthracis as a bioterrorism weapon threatens the security of populations globally, requiring the immediate availability of safe, efficient and easily delivered anthrax vaccine for mass vaccination. extensive research efforts have been directed toward the development of recombinant subunit vaccines based on protective antigen (pa), the principal virulence factor of b. anthracis. among the emerging technologies for the production of these vaccine antigens is our launc ...201323324615
crystal structure of arabidopsis thaliana dawdle forkhead-associated domain reveals a conserved phospho-threonine recognition cleft for dicer-like 1 binding.dawdle (ddl) is a microrna processing protein essential for the development of arabidopsis. ddl contains a putative nuclear localization signal at its amino-terminus and forkhead-associated (fha) domain at the carboxyl-terminus. here, we report the crystal structure of the fha domain of arabidopsis dawdle, determined by multiple-wavelength anomalous dispersion method at 1.7-å resolution. ddl fha structure displays a seven-stranded β-sandwich architecture that contains a unique structural motif c ...201323313986
infection of tomato leaf curl new delhi virus (tolcndv), a bipartite begomovirus with betasatellites, results in enhanced level of helper virus components and antagonistic interaction between dna b and betasatellites.tomato leaf curl new delhi virus (tolcndv) (geminiviridae) is an important pathogen that severely affects tomato production. an extensive survey was carried out during 2003-2010 to study the diversity of begomoviruses found in tomato, potato, and cucurbits that showed symptoms of leaf puckering, distortion, curling, vein clearing, and yellow mosaic in various fields in different regions of india. ten begomovirus isolates were cloned from infected samples and identified as belonging to the specie ...201323306645
gene-for-gene relationship in the host-pathogen system malus × robusta 5-erwinia amylovora.fire blight is a destructive bacterial disease caused by erwinia amylovora affecting plants in the family rosaceae, including apple. host resistance to fire blight is present mainly in accessions of malus spp. and is thought to be quantitative in this pathosystem. in this study we analyzed the importance of the e. amylovora effector avrrpt2(ea) , a homolog of pseudomonas syringae avrrpt2, for resistance of malus × robusta 5 (mr5). the deletion mutant e. amylovora ea1189δavrrpt2(ea) was able to o ...201323301854
advanced engineering of lipid metabolism in nicotiana benthamiana using a draft genome and the v2 viral silencing-suppressor protein.the transient leaf assay in nicotiana benthamiana is widely used in plant sciences, with one application being the rapid assembly of complex multigene pathways that produce new fatty acid profiles. this rapid and facile assay would be further improved if it were possible to simultaneously overexpress transgenes while accurately silencing endogenes. here, we report a draft genome resource for n. benthamiana spanning over 75% of the 3.1 gb haploid genome. this resource revealed a two-member nbfad2 ...201223300750
inhibition of the host proteasome facilitates papaya ringspot virus accumulation and proteosomal catalytic activity is modulated by viral factor hcpro.the ubiquitin/26s proteasome system plays an essential role not only in maintaining protein turnover, but also in regulating many other plant responses, including plant-pathogen interactions. previous studies highlighted different roles of the 20s proteasome in plant defense during virus infection, either indirectly through viral suppressor-mediated degradation of argonaute proteins, affecting the rna interference pathway, or directly through modulation of the proteolytic and rnase activity of t ...201223300704
effects of the virus satellite gene βc1 on host plant defense signaling and volatile emission.tomato yellow leaf curl china virus spreads together with its invasive vector, the silverleaf whitefly b biotype, which exhibits higher growth rates on infected plants. previous studies indicate that the virus satellite gene βc1 accounts for the visible symptoms of infection and inhibits the constitutive expression of jasmonic acid (ja)--a phytohormone involved in plant defense against whiteflies--and of some ja-regulated genes. here we present new details of the effects of on plant signaling an ...201323299332
a plant-produced h1n1 trimeric hemagglutinin protects mice from a lethal influenza virus challenge.the increased worldwide awareness of seasonal and pandemic influenza, including pandemic h1n1 virus, has stimulated interest in the development of economic platforms for rapid, large-scale production of safe and effective subunit vaccines. in recent years, plants have demonstrated their utility as such a platform and have been used to produce vaccine antigens against various infectious diseases. previously, we have produced in our transient plant expression system a recombinant monomeric hemaggl ...201323296194
bis-aryl methanone compound is a candidate of nitric oxide producing elicitor and induces resistance in nicotiana benthamiana against phytophthora infestans.nitric oxide (no) is important in some physiological responses of plants and plays a crucial role in the regulation of both defense responses and inducing resistance to fungal pathogens. nubs-4190, a new bis-aryl-methanone compound elicited no production and defense responses in nicotiana benthamiana against phytophthora infestans. nubs-4190 induced resistance in n. benthamiana to p. infestans, without association of reactive oxygen generation and hypersensitive cell death. callose induction was ...201323291305
a large and functionally diverse family of fad2 genes in safflower (carthamus tinctorius l.).the application and nutritional value of vegetable oil is highly dependent on its fatty acid composition, especially the relative proportion of its two major fatty acids, i.e oleic acid and linoleic acid. microsomal oleoyl phosphatidylcholine desaturase encoded by fad2 gene is known to introduce a double bond at the δ12 position of an oleic acid on phosphatidylcholine and convert it to linoleic acid. the known plant fad2 enzymes are encoded by small gene families consisting of 1-4 members. in ad ...201323289946
overexpression of a soybean nuclear localized type-iii dnaj domain-containing hsp40 reveals its roles in cell death and disease resistance.heat-shock proteins such as hsp70 and hsp90 are important molecular chaperones that play critical roles in biotic and abiotic stress responses; however, the involvement of their co-chaperones in stress biology remains largely uninvestigated. in a screen for candidate genes stimulating cell death in glycine max (soybean), we transiently overexpressed full-length cdnas of soybean genes that are highly induced during soybean rust infection in nicotiana benthamiana leaves. overexpression of a type-i ...201323289813
high level resistance against rhizomania disease by simultaneously integrating two distinct defense mechanisms.with the aim of achieving durable resistance against rhizomania disease of sugar beet, the employment of different sources of resistance to beet necrotic yellow vein virus was pursued. to this purpose, nicotiana benthamiana transgenic plants that simultaneously produce dsrna originating from a conserved region of the bnyvv replicase gene and the hrpz(psph) protein in a secreted form (sp/hrpz(psph)) were produced. the integration and expression of both transgenes as well as proper production of t ...201223284692
gmrin4 protein family members function nonredundantly in soybean race-specific resistance against pseudomonas syringae.the pseudomonas syringae effector avrb interacts with four related soybean (glycine max) proteins (gmrin4a-d), three (gmrin4b, c, d) of which also interact with the cognate resistance (r) protein, rpg1-b. here, we investigated the specific requirements for the gmrin4 proteins in r-mediated resistance and examined the mechanism of rpg1-b activation. using virus-induced gene silencing, we show that only gmrin4a and b are required for rpg1-b-mediated resistance. in planta binding assays show that g ...201323278373
the secretory pathway and the actomyosin motility system are required for plasmodesmatal localization of the p7-1 of rice black-streaked dwarf virus.rice black-streaked dwarf virus (rbsdv), a plant-infecting reovirus (genus fijivirus), generally induces virus-containing tubules in infected cells. the nonstructural protein p7-1, encoded by the first open reading frame of segment 7, is involved in forming the structural matrix of these tubules. in experiments to investigate the subcellular localization of p7-1 in nicotiana benthamiana epidermal cells, fluorescence of p7-1:egfp was observed in the nucleus, cytoplasm and cell periphery, and in p ...201223271163
the potato mop-top virus tgb2 protein and viral rna associate with chloroplasts and viral infection induces inclusions in the plastids.the potato mop-top virus (pmtv) triple gene block 2 (tgb2) movement proteins fused to monomeric red fluorescent protein (mrfp-tgb2) was expressed under the control of the pmtv subgenomic promoter from a pmtv vector. the subcellular localizations and interactions of mrfp-tgb2 were investigated using confocal imaging [confocal laser-scanning microscope, (clsm)] and biochemical analysis. the results revealed associations with membranes of the endoplasmic reticulum (er), mobile granules, small round ...201223269927
comparative genomic and transcriptomic analyses reveal the hemibiotrophic stage shift of colletotrichum fungi.hemibiotrophic fungal plant pathogens represent a group of agronomically significant disease-causing agents that grow first on living tissue and then cause host death in later, necrotrophic growth. among these, colletotrichum spp. are devastating pathogens of many crops. identifying expanded classes of genes in the genomes of phytopathogenic colletotrichum, especially those associated with specific stages of hemibiotrophy, can provide insights on how these pathogens infect a large number of host ...201323252678
production of a de-novo designed antimicrobial peptide in nicotiana benthamiana.antimicrobial peptides are important defense compounds of higher organisms that can be used as therapeutic agents against bacterial and/or viral infections. we designed several antimicrobial peptides containing hydrophobic and positively charged clusters that are active against plant and human pathogens. especially peptide sp1-1 is highly active with a mic value of 0.1 μg/ml against xanthomonas vesicatoria, pseudomonas corrugata and pseudomonas syringae pv syringae. however, for commercial appli ...201323242916
role of rice stripe virus nsvc4 in cell-to-cell movement and symptom development in nicotiana benthamiana.our previous work has demonstrated that the nsvc4 protein of rice stripe virus (rsv) functions as a cell-to-cell movement protein. however, the mechanisms whereby rsv traffics through plasmodesmata (pd) are unknown. here we provide evidence that the nsvc4 moves on the actin filament and endoplasmic reticulum network, but not microtubules, to reach cell wall pd. disruption of cytoskeleton using different inhibitors altered nsvc4 localization to pd, thus impeding rsv infection of nicotiana bentham ...201223233857
functional analysis of potato genes involved in quantitative resistance to phytophthora infestans.the most significant threat to potato production worldwide is the late blight disease, which is caused by the oomycete pathogen phytophthora infestans. based on previous cdna microarrays and cdna-amplified fragment length polymorphism analysis, 63 candidate genes that are expected to contribute to developing a durable resistance to late blight were selected for further functional analysis. we performed virus-induced gene silencing (vigs) to these candidate genes on both nicotiana benthamiana and ...201323224656
rapid and efficient gene delivery into plant cells using designed peptide carriers.to develop a new easy and quick gene delivery system for any types of plants, we prepared ionic complexes of plasmid dna with designed peptide carriers, each of which combined a cell-penetrating peptide (bp100 or tat(2)) with a polycation (nona-arginine or a copolymer of histidine and lysine). the present system via the designed peptides demonstrated rapid and efficient transient transfections into intact leaf cells of nicotiana benthamiana and arabidopsis thaliana without protoplast preparation ...201323215041
the magnaporthe oryzae effector avrpiz-t targets the ring e3 ubiquitin ligase apip6 to suppress pathogen-associated molecular pattern-triggered immunity in rice.although the functions of a few effector proteins produced by bacterial and oomycete plant pathogens have been elucidated in recent years, information for the vast majority of pathogen effectors is still lacking, particularly for those of plant-pathogenic fungi. here, we show that the avirulence effector avrpiz-t from the rice blast fungus magnaporthe oryzae preferentially accumulates in the specialized structure called the biotrophic interfacial complex and is then translocated into rice (oryza ...201223204406
safety and immunogenicity of a plant-produced recombinant hemagglutinin-based influenza vaccine (hai-05) derived from a/indonesia/05/2005 (h5n1) influenza virus: a phase 1 randomized, double-blind, placebo-controlled, dose-escalation study in healthy adults.recently, we have reported [1,2] on a subunit influenza vaccine candidate based on the recombinant hemagglutinin protein from the a/indonesia/05/2005 (h5n1) strain of influenza virus, produced it using 'launch vector'-based transient expression technology in nicotiana benthamiana, and demonstrated its immunogenicity in pre-clinical studies. here, we present the results of a first-in-human, phase 1 randomized, double-blind, placebo-controlled study designed to investigate safety, reactogenicity a ...201223202523
in planta expression or delivery of potato aphid macrosiphum euphorbiae effectors me10 and me23 enhances aphid fecundity.the interactions between aphids and their host plants seem to be analogous to those of plant-microbial pathogens. unlike microbial pathogen effectors, little is known about aphid effectors and their ability to interfere with host immunity. to date, only three functional aphid effectors have been reported. to identify potato aphid (macrosiphum euphorbiae) effectors, we developed a salivary gland transcriptome using illumina technology. we generated 85 million illumina reads from salivary glands a ...201323194342
tombusvirus y-shaped translational enhancer forms a complex with eif4f and can be functionally replaced by heterologous translational enhancers.certain plus-strand rna plant viruses that are uncapped and nonpolyadenylated rely on rna elements in their 3' untranslated region, termed 3'-cap-independent translational enhancers (3'cites), for efficient translation of their proteins. here, we have investigated the properties of the y-shaped class of 3'cite present in the tombusvirus carnation italian ringspot virus (cirv). while some types of 3'cite have been found to function through recruitment of translation initiation factors to the vira ...201323192876
the gef1 proton-chloride exchanger affects tombusvirus replication via regulation of copper metabolism in yeast.replication of plus-strand rna viruses [(+)rna viruses] is performed by viral replicases, whose function is affected by many cellular factors in infected cells. in this paper, we demonstrate a surprising role for gef1p proton-chloride exchanger in replication of tomato bushy stunt virus (tbsv) model (+)rna virus. a genetic approach revealed that gef1p, which is the only proton-chloride exchanger in saccharomyces cerevisiae, is required for tbsv replication in the yeast model host. we also show t ...201223192874
viral nanoparticles for in vivo tumor imaging.the use of nanomaterials has the potential to revolutionize materials science and medicine. currently, a number of different nanoparticles are being investigated for applications in imaging and therapy. viral nanoparticles (vnps) derived from plants can be regarded as self-assembled bionanomaterials with defined sizes and shapes. plant viruses under investigation in the steinmetz lab include icosahedral particles formed by cowpea mosaic virus (cpmv) and brome mosaic virus (bmv), both of which ar ...201223183850
protein-protein and protein-membrane associations in the lignin pathway.supramolecular organization of enzymes is proposed to orchestrate metabolic complexity and help channel intermediates in different pathways. phenylpropanoid metabolism has to direct up to 30% of the carbon fixed by plants to the biosynthesis of lignin precursors. effective coupling of the enzymes in the pathway thus seems to be required. subcellular localization, mobility, protein-protein, and protein-membrane interactions of four consecutive enzymes around the main branch point leading to ligni ...201223175744
[effects of seven rna silencing suppressors on heterologous expression of green fluorescence protein expression mediated by a plant virus-based system in nicotiana benthamiana].to test the effects of 7 virus-encoded rna silencing suppressors (rsss) for enhancement of a plant virus-based vector system-mediated heterologous expression of green fluorescence protein (gfp) in nicotiana benthamiana.201223174571
the cbl-interacting protein kinase cipk26 is a novel interactor of arabidopsis nadph oxidase atrbohf that negatively modulates its ros-producing activity in a heterologous expression system.the plant nadph oxidases, known as respiratory burst oxidase homologues (rbohs), play an indispensable role in a wide array of cellular and developmental processes. arabidopsis thaliana rbohf (atrbohf)-mediated production of reactive oxygen species (ros) is involved in biotic and abiotic stress responses. because of the toxicity of excess amount of ros, the ros-producing activity of rbohs is speculated to be negatively regulated. however, its mechanism is mostly unknown to date. here, we report ...201323162070
high-level transient expression of er-targeted human interleukin 6 in nicotiana benthamiana.tobacco plants can be used to express recombinant proteins that cannot be produced in a soluble and active form using traditional platforms such as escherichia coli. we therefore expressed the human glycoprotein interleukin 6 (il6) in two commercial tobacco cultivars (nicotiana tabacum cv. virginia and cv. geudertheimer) as well as the model host n. benthamiana to compare different transformation strategies (stable vs. transient expression) and subcellular targeting (apoplast, endoplasmic reticu ...201223152824
the rubisco small subunit is involved in tobamovirus movement and tm-2²-mediated extreme resistance.the multifunctional movement protein (mp) of tomato mosaic tobamovirus (tomv) is involved in viral cell-to-cell movement, symptom development, and resistance gene recognition. however, it remains to be elucidated how tomv mp plays such diverse roles in plants. here, we show that tomv mp interacts with the rubisco small subunit (rbcs) of nicotiana benthamiana in vitro and in vivo. in susceptible n. benthamiana plants, silencing of nbrbcs enabled tomv to induce necrosis in inoculated leaves, thus ...201323148080
pseudomonas syringae pv. tomato dc3000 cmal (pspto4723), a duf1330 family member, is needed to produce l-allo-isoleucine, a precursor for the phytotoxin coronatine.pseudomonas syringae pv. tomato dc3000 produces the phytotoxin coronatine, a major determinant of the leaf chlorosis associated with dc3000 pathogenesis. the dc3000 pspto4723 (cmal) gene is located in a genomic region encoding type iii effectors; however, it promotes chlorosis in the model plant nicotiana benthamiana in a manner independent of type iii secretion. coronatine is produced by the ligation of two moieties, coronafacic acid (cfa) and coronamic acid (cma), which are produced by biosynt ...201323144243
a plant derived multifunctional tool for nanobiotechnology based on tomato bushy stunt virus.structure, size, physicochemical properties and production strategies make many plant viruses ideal protein based nanoscaffolds, nanocontainers and nano-building blocks expected to deliver a multitude of applications in different fields such as biomedicine, pharmaceutical chemistry, separation science, catalytic chemistry, crop pest control and biomaterials science. functionalization of viral nanoparticles through modification by design of their external and internal surfaces is essential to ful ...201323108557
specificity of ocimum basilicum geraniol synthase modified by its expression in different heterologous systems.numerous aromatic plant species produce high levels of monoterpenols, using geranyl diphosphate (gpp) as a precursor. sweet basil (ocimum basilicum) geraniol synthase (ges) was used to evaluate the monoterpenol profiles arising from heterologous expressions in various plant models. grapevine (vitis vinifera) calli were transformed using agrobacterium tumefasciens and the plants were regenerated. thale cress (arabidopsis thaliana) was transformed using the floral dip method. tobacco (nicotiana be ...201323108028
consequences of flagellin export through the type iii secretion system of pseudomonas syringae reveal a major difference in the innate immune systems of mammals and the model plant nicotiana benthamiana.bacterial flagellin is perceived as a microbe (or pathogen)-associated molecular pattern (mamp or pamp) by the extracellular pattern recognition receptors, fls2 and tlr5, of plants and mammals respectively. flagellin accidently translocated into mammalian cells by pathogen type iii secretion systems (t3sss) is recognized by nucleotide-binding leucine-rich repeat receptor nlrc4 as a pattern of pathogenesis and induces a death-associated immune response. the non-pathogen pseudomonas fluorescens pf ...201323107228
transient gene expression in epidermal cells of plant leaves by biolistic dna delivery.transient gene expression is a useful approach for studying the functions of gene products. in the case of plants, agrobacterium infiltration is a method of choice for transient introduction of genes for many species. however, this technique does not work efficiently in some species, such as arabidopsis thaliana. moreover, the infection of agrobacterium is known to induce dynamic changes in gene expression patterns in the host plants, possibly affecting the function and localization of the prote ...201323104330
dicer-like 4 but not dicer-like 2 may have a positive effect on potato spindle tuber viroid accumulation in nicotiana benthamiana. 201323100483
construction and biological activities of the first infectious cdna clones of the genus foveavirus.grapevine rupestris stem pitting-associated virus (grspav, genus foveavirus, family betaflexiviridae) is one of the most prevalent viruses in grapevines and is associated with three distinct diseases: rupestris stem pitting, vein necrosis and syrah decline. little is known about the biology and pathological properties of grspav. in this work, we engineered a full-length infectious cdna clone for grspav and a gfp-tagged variant, both under the transcriptional control of cauliflower mosaic virus 3 ...201323099206
influence of host chloroplast proteins on tobacco mosaic virus accumulation and intercellular movement.tobacco mosaic virus (tmv) forms dense cytoplasmic bodies containing replication-associated proteins (virus replication complexes [vrcs]) upon infection. to identify host proteins that interact with individual viral components of vrcs or vrcs in toto, we isolated viral replicase- and vrc-enriched fractions from tmv-infected nicotiana tabacum plants. two host proteins in enriched fractions, atp-synthase γ-subunit (atpc) and rubisco activase (rca) were identified by matrix-assisted laser-desorptio ...201323096159
the pepper extracellular xyloglucan-specific endo-β-1,4-glucanase inhibitor protein gene, caxegip1, is required for plant cell death and defense responses.plants produce various proteinaceous inhibitors to protect themselves against microbial pathogen attack. a xyloglucan-specific endo-β-1,4-glucanase inhibitor1 gene, caxegip1, was isolated and functionally characterized in pepper (capsicum annuum) plants. caxegip1 was rapidly and strongly induced in pepper leaves infected with avirulent xanthomonas campestris pv vesicatoria, and purified caxegip1 protein significantly inhibited the hydrolytic activity of the glycoside hydrolase74 family xylogluca ...201223093361
cauliflower mosaic virus protein p6 inhibits signaling responses to salicylic acid and regulates innate immunity.cauliflower mosaic virus (camv) encodes a multifunctional protein p6 that is required for translation of the 35s rna and also acts as a suppressor of rna silencing. here we demonstrate that p6 additionally acts as a pathogenicity effector of an unique and novel type, modifying npr1 (a key regulator of salicylic acid (sa)- and jasmonic acid (ja)-dependent signaling) and inhibiting sa-dependent defence responses we find that that transgene-mediated expression of p6 in arabidopsis and transient exp ...201223071821
delayed treatment of ebola virus infection with plant-derived monoclonal antibodies provides protection in rhesus macaques.filovirus infections can cause a severe and often fatal disease in humans and nonhuman primates, including great apes. here, three anti-ebola virus mouse/human chimeric mabs (c13c6, h-13f6, and c6d8) were produced in chinese hamster ovary and in whole plant (nicotiana benthamiana) cells. in pilot experiments testing a mixture of the three mabs (mb-003), we found that mb-003 produced in both manufacturing systems protected rhesus macaques from lethal challenge when administered 1 h postinfection. ...201223071322
agrobacterium may delay plant nonhomologous end-joining dna repair via xrcc4 to favor t-dna integration.agrobacterium tumefaciens is a soilborne pathogen that causes crown gall disease in many dicotyledonous plants by transfer of a portion of its tumor-inducing plasmid (t-dna) into the plant genome. several plant factors that play a role in agrobacterium attachment to plant cells and transport of t-dna to the nucleus have been identified, but the t-dna integration step during transformation is poorly understood and has been proposed to occur via nonhomologous end-joining (nhej)-mediated double-str ...201223064322
dynamic localization of two tobamovirus orf6 proteins involves distinct organellar compartments.orf6 is a small gene that overlaps the movement and coat protein genes of subgroup 1a tobamoviruses. the orf6 protein of tomato mosaic virus (tomv) strain l (l-orf6), interacts in vitro with eukaryotic elongation factor 1α, and mutation of the orf6 gene of tobacco mosaic virus (tmv) strain u1 (u1-orf6) reduces the pathogenicity in vivo of tmv, whereas expression of this gene from two other viruses, tobacco rattle virus (trv) and potato virus x (pvx), increases their pathogenicity. in this work, ...201323052393
identification and characterization of in planta-expressed secreted effector proteins from magnaporthe oryzae that induce cell death in rice.interactions between rice and magnaporthe oryzae involve the recognition of cellular components and the exchange of complex molecular signals from both partners. how these interactions occur in rice cells is still elusive. we employed robust-long serial analysis of gene expression, massively parallel signature sequencing, and sequencing by synthesis to examine transcriptome profiles of infected rice leaves. a total of 6,413 in planta-expressed fungal genes, including 851 genes encoding predicted ...201323035914
aphid protein effectors promote aphid colonization in a plant species-specific manner.microbial pathogens and pests produce effectors to modulate host processes. aphids are phloem-feeding insects, which introduce effectors via saliva into plant cells. however, it is not known if aphid effectors have adapted to modulate processes in specific plant species. myzus persicae is a polyphagous insect that colonizes arabidopsis thaliana and nicotiana benthamiana, while the pea aphid acyrthosiphon pisum specializes on colonizing plant species of the family fabaceae. we found that m. persi ...201323035913
validation of reference genes for gene expression studies in virus-infected nicotiana benthamiana using quantitative real-time pcr.nicotiana benthamiana is the most widely-used experimental host in plant virology. the recent release of the draft genome sequence for n. benthamiana consolidates its role as a model for plant-pathogen interactions. quantitative real-time pcr (qpcr) is commonly employed for quantitative gene expression analysis. for valid qpcr analysis, accurate normalisation of gene expression against an appropriate internal control is required. yet there has been little systematic investigation of reference ge ...201223029521
the destructive citrus pathogen, 'candidatus liberibacter asiaticus' encodes a functional flagellin characteristic of a pathogen-associated molecular pattern.huanglongbing (hlb) is presently the most devastating citrus disease worldwide. as an intracellular plant pathogen and insect symbiont, the hlb bacterium, 'candidatus liberibacter asiaticus' (las), retains the entire flagellum-encoding gene cluster in its significantly reduced genome. las encodes a flagellin and hook-associated protein (fla) of 452 amino acids that contains a conserved 22 amino acid domain (flg22) at positions 29 to 50 in the n-terminus. the phenotypic alteration in motility of ...201223029520
population diversity of rice stripe virus-derived sirnas in three different hosts and rnai-based antiviral immunity in laodelphgax striatellus.small rna-mediated gene silencing plays evolutionarily conserved roles in gene regulation and defense against invasive nucleic acids. virus-derived small interfering rnas (vsirnas) are one of the key elements involved in rna silencing-based antiviral activities in plant and insect. vsirnas produced after viruses infecting hosts from a single kingdom (i.e., plant or animal) are well described. in contrast, vsirnas derived from viruses capable of infecting both plants and their insect vectors have ...201223029445
the benyvirus rna silencing suppressor is essential for long-distance movement, requires both zinc-finger and nols basic residues but not a nucleolar localization for its silencing-suppression activity.the rna silencing-suppression properties of beet necrotic yellow vein virus (bnyvv) and beet soil-borne mosaic virus (bsbmv) cysteine-rich p14 proteins have been investigated. suppression of rna silencing activities were made evident using viral infection of silenced nicotiana benthamiana 16c, n. benthamiana agroinfiltrated with green fluorescent protein (gfp), and gf-fg hairpin triggers supplemented with viral suppressor of rna silencing (vsr) constructs or using complementation of a silencing- ...201323013437
roles of the different components of magnesium chelatase in abscisic acid signal transduction.the h subunit of mg-chelatase (chlh) was shown to regulate abscisic acid (aba) signaling and the i subunit (chli) was also reported to modulate aba signaling in guard cells. however, it remains essentially unknown whether and how the mg-chelatase-catalyzed mg-protoporphyrin ix-production differs from aba signaling. using a newly-developed surface plasmon resonance system, we showed that aba binds to chlh, but not to the other mg-chelatase components/subunits chli, chld (d subunit) and gun4. a ne ...201223011401
an enzyme activity capable of endotransglycosylation of heteroxylan polysaccharides is present in plant primary cell walls.heteroxylans in the plant cell wall have been proposed to have a role analogous to that of xyloglucans or heteromannans, forming growth-restraining networks by interlocking cellulose microfibrils. a xylan endotransglycosylase has been identified that can transglycosylate heteroxylan polysaccharides in the presence of xylan-derived oligosaccharides. high activity was detected in ripe fruit of papaya (carica papaya), but activity was also found in a range of other fruits, imbibed seeds and rapidly ...201323001197
characterization of in vivo functions of nicotiana benthamiana rabe1.we characterized the gene expression, subcellular localization, and in vivo functions of a nicotiana benthamiana small gtpase belonging to the rabe family, designated nbrabe1. the nbrabe1 promoter drove strong β-glucuronidase reporter expression in young tissues containing actively dividing cells and in stomata guard cells. gfp fusion proteins of nbrabe1 and its dominant-negative and constitutively active mutants were all localized to the golgi apparatus and the plasma membrane but showed differ ...201323001196
bipartite and tripartite cucumber mosaic virus-based vectors for producing the acidothermus cellulolyticus endo-1,4-β-glucanase and other proteins in non-transgenic plants.using plant viruses to produce desirable proteins in plants allows for using non-transgenic plant hosts and if necessary, the ability to make rapid changes in the virus construct for increased or modified protein product yields. the objective of this work was the development of advanced cmv-based protein production systems to produce acidothermus cellulolyticus endo-1, 4-β-glucanase (e1) in non-transgenic plants.201222999234
engineering of glucosinolate biosynthesis: candidate gene identification and validation.the diverse biological roles of glucosinolates as plant defense metabolites and anticancer compounds have spurred a strong interest in their biosynthetic pathways. since the completion of the arabidopsis genome, functional genomics approaches have enabled significant progress on the elucidation of glucosinolate biosynthesis, although in planta validation of candidate gene function often is hampered by time-consuming generation of knockout and overexpression lines in arabidopsis. to better exploi ...201222999179
disruption of sphingolipid biosynthesis in nicotiana benthamiana activates salicylic acid-dependent responses and compromises resistance to alternaria alternata f. sp. lycopersici.sphingolipids play an important role in signal transduction pathways that regulate physiological functions and stress responses in eukaryotes. in plants, recent evidence suggests that their metabolic precursors, the long-chain bases (lcbs) act as bioactive molecules in the immune response. interestingly, the virulence of two unrelated necrotrophic fungi, fusarium verticillioides and alternaria alternata, which are pathogens of maize and tomato plants, respectively, depends on the production of s ...201322990908
challenges in electrochemical pre-purification of recombinant proteins from green plant tissues: sgfp produced in tobacco leaves.the use of recombinant proteins has increased greatly in recent years, as have the number of techniques and materials used for their production and purification. the principal advantage of using plants as bioreactors is the cost of the recombinant protein production, which is about 1000-fold lower as in the case of using cho cells commonly applied in industry today. among the different types of "green" bioreactors being studied today, there is a general consensus among scientists that production ...201522989991
distinct regions of the pseudomonas syringae coiled-coil effector avrrps4 are required for activation of immunity.gram-negative phytopathogenic bacteria translocate effector proteins into plant cells to subvert host defenses. these effectors can be recognized by plant nucleotide-binding-leucine-rich repeat immune receptors, triggering defense responses that restrict pathogen growth. avrrps4, an effector protein from pseudomonas syringae pv. pisi, triggers rps4-dependent immunity in resistant accessions of arabidopsis. to better understand the molecular basis of avrrps4-triggered immunity, we determined the ...201222988101
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