Publications

ralstonia solanacearum type iii effector ripay is a glutathione-degrading enzyme that is activated by plant cytosolic thioredoxins and suppresses plant immunity.	the plant pathogen ralstonia solanacearum uses a large repertoire of type iii effector proteins to succeed in infection. to clarify the function of effector proteins in host eukaryote cells, we expressed effectors in yeast cells and identified seven effector proteins that interfere with yeast growth. one of the effector proteins, ripay, was found to share homology with the chac family proteins that function as γ-glutamyl cyclotransferases, which degrade glutathione (gsh), a tripeptide that plays ...	2016	27073091
isolation and characterization of scglud2, a new sugarcane beta-1,3-glucanase d family gene induced by sporisorium scitamineum, aba, h2o2, nacl, and cdcl2 stresses.	beta-1,3-glucanases (ec 3.2.1.39), commonly known as pathogenesis-related (pr) proteins, play an important role not only in plant defense against fungal pathogens but also in plant physiological and developmental processes. however, only a limited number of sugarcane beta-1,3-glucanase genes have been isolated. in the present study, we identified and characterized a new beta-1,3-glucanase gene scglud2 (genbank acc no. kf664181) from sugarcane. an x8 domain was present at the c terminal region of ...	2016	27642288
a cysteine-rich protein kinase associates with a membrane immune complex and is required for cell death.	membrane localized proteins perceive and respond to biotic and abiotic stresses. we performed quantitative proteomics on plasma membrane-enriched samples from arabidopsis treated with bacterial flagellin. we identified multiple receptor-like protein kinases (rlks) changing in abundance, including cysteine-rich receptor-like kinases (crks) that are upregulated upon perception of flagellin. crks possess extracellular cysteine-rich domains and comprise a gene family consisting of 46 members in arab ...	2016	27852951
overexpression of nictaba-like lectin genes from glycine max confers tolerance toward pseudomonas syringae infection, aphid infestation and salt stress in transgenic arabidopsis plants.	plants have evolved a sophisticated immune system that allows them to recognize invading pathogens by specialized receptors. carbohydrate-binding proteins or lectins are part of this immune system and especially the lectins that reside in the nucleocytoplasmic compartment are known to be implicated in biotic and abiotic stress responses. the class of nictaba-like lectins (nll) groups all proteins with homology to the tobacco (nicotiana tabacum) lectin, known as a stress-inducible lectin. here we ...	2016	27826309
detecting n-myristoylation and s-acylation of host and pathogen proteins in plants using click chemistry.	the plant plasma membrane is a key battleground in the war between plants and their pathogens. plants detect the presence of pathogens at the plasma membrane using sensor proteins, many of which are targeted to this lipophilic locale by way of fatty acid modifications. pathogens secrete effector proteins into the plant cell to suppress the plant's defense mechanisms. these effectors are able to access and interfere with the surveillance machinery at the plant plasma membrane by hijacking the hos ...	2016	27493678
alternative splicing of rice wrky62 and wrky76 transcription factor genes in pathogen defense.	the wrky family of transcription factors (tfs) functions as transcriptional activators or repressors in various signaling pathways. in this study, we discovered that oswrky62 and oswrky76, two genes of the wrky iia subfamily, undergo constitutive and inducible alternative splicing. the full-length oswrky62.1 and oswrky76.1 proteins formed homocomplexes and heterocomplexes, and the heterocomplex dominates in the nuclei when analyzed in nicotiana benthamiana leaves. transgenic overexpression of os ...	2016	27208272
nbcspr underlies age-dependent immune responses to bacterial cold shock protein in nicotiana benthamiana.	plants use receptor kinases (rks) and receptor-like proteins (rlps) as pattern recognition receptors (prrs) to sense pathogen-associated molecular patterns (pamps) that are typical of whole classes of microbes. after ligand perception, many leucine-rich repeat (lrr)-containing prrs interact with the lrr-rk bri1-associated kinase 1 (bak1). bak1 is thus expected to interact with unknown prrs. here, we used bak1 as molecular bait to identify a previously unknown lrr-rlp required for the recognition ...	2016	26944079
cloning and expression analysis of cinnamoyl-coa reductase (ccr) genes in sorghum.	cinnamoyl-coa reductase (ccr) is the first enzyme in the monolignol-specific branch of the lignin biosynthetic pathway. in this research, three sorghum ccr genes including sbccr1, sbccr2-1 and sbccr2-2 were cloned and characterized. analyses of the structure and phylogeny of the three ccr genes showed evolutionary conservation of the functional domains and divergence of function. transient expression assays in nicotiana benthamiana leaves demonstrated that the three ccr proteins were localized i ...	2016	27231650
isolation and characterization of pepper genes interacting with the cmv-p1 helicase domain.	cucumber mosaic virus (cmv) is a destructive pathogen affecting capsicum annuum (pepper) production. the pepper cmr1 gene confers resistance to most cmv strains, but is overcome by cmv-p1 in a process dependent on the cmv-p1 rna1 helicase domain (p1 helicase). here, to identify host factors involved in cmv-p1 infection in pepper, a yeast two-hybrid library derived from a c. annuum 'bukang' cdna library was screened, producing a total of 76 potential clones interacting with the p1 helicase. beta- ...	2016	26751216
avrrxo1 is a bifunctional type iii secreted effector and toxin-antitoxin system component with homologs in diverse environmental contexts.	toxin-antitoxin (ta) systems are ubiquitous bacterial systems that may function in genome maintenance and metabolic stress management, but are also thought to play a role in virulence by helping pathogens survive stress. we previously demonstrated that the xanthomonas oryzae pv. oryzicola protein avrrxo1 is a type iii-secreted virulence factor that has structural similarities to the zeta family of ta toxins, and is toxic to plants and bacteria in the absence of its predicted chaperone arc1. in t ...	2016	27391081
apoplastic recognition of multiple candidate effectors from the wheat pathogen zymoseptoria tritici in the nonhost plant nicotiana benthamiana.	the fungus zymoseptoria tritici is a strictly apoplastic, host-specific pathogen of wheat leaves and causal agent of septoria tritici blotch (stb) disease. all other plants are considered nonhosts, but the mechanism of nonhost resistance (nhr) to z. tritici has not been addressed previously. we sought to develop nicotiana benthamiana as a system to study nhr against z. tritici. fluorescence microscopy and quantitative reverse transcription polymerase chain reactions were used to establish the in ...	2016	27696417
environmental and genetic factors associated with solanesol accumulation in potato leaves.	solanesol is a high value 45-carbon, unsaturated, all-trans-nonaprenol isoprenoid. recently solanesol has received particular attention because of its utility, both in its own right and as a precursor in the production of numerous compounds used in the treatment of disease states. solanesol is found mainly in solanaceous crops such as potato, tomato, tobacco and pepper where it accumulates in the foliage. there is considerable potential to explore the extraction of solanesol from these sources a ...	2016	27610114
a rice gene homologous to arabidopsis agd2-like defense1 participates in disease resistance response against infection with magnaporthe oryzae.	ald1 (aberrant growth and death2 [agd2]-like defense1) is one of the key defense regulators in arabidopsis thaliana and nicotiana benthamiana. in these model plants, ald1 is responsible for triggering basal defense response and systemic resistance against bacterial infection. as well ald1 is involved in the production of pipecolic acid and an unidentified compound(s) for systemic resistance and priming syndrome, respectively. these previous studies proposed that ald1 is a potential candidate for ...	2016	27493611
role of brassinosteroid signaling in modulating tobacco mosaic virus resistance in nicotiana benthamiana.	plant steroid hormones, brassinosteroids (brs), play essential roles in plant growth, development and stress responses. however, mechanisms by which brs interfere with plant resistance to virus remain largely unclear. in this study, we used pharmacological and genetic approaches in combination with infection experiments to investigate the role of brs in plant defense against tobacco mosaic virus (tmv) in nicotiana benthamiana. exogenous applied brs enhanced plant resistance to virus infection, w ...	2016	26838475
an effector of the irish potato famine pathogen antagonizes a host autophagy cargo receptor.	plants use autophagy to safeguard against infectious diseases. however, how plant pathogens interfere with autophagy-related processes is unknown. here, we show that pexrd54, an effector from the irish potato famine pathogen phytophthora infestans, binds host autophagy protein atg8cl to stimulate autophagosome formation. pexrd54 depletes the autophagy cargo receptor joka2 out of atg8cl complexes and interferes with joka2's positive effect on pathogen defense. thus, a plant pathogen effector has ...	2016	26765567
regulation of the cobalt/nickel efflux operon dmerf in agrobacterium tumefaciens and a link between the iron-sensing regulator rira and cobalt/nickel resistance.	the agrobacterium tumefaciens c58 genome harbors an operon containing the dmer (atu0890) and dmef (atu0891) genes, which encode a transcriptional regulatory protein belonging to the rcnr/csor family and a metal efflux protein belonging to the cation diffusion facilitator (cdf) family, respectively. the dmerf operon is specifically induced by cobalt and nickel, with cobalt being the more potent inducer. promoter-lacz transcriptional fusion, an electrophoretic mobility shift assay, and dnase i foo ...	2016	27235438
role of viral rna and co-opted cellular escrt-i and escrt-iii factors in formation of tombusvirus spherules harboring the tombusvirus replicase.	plus-stranded rna viruses induce membrane deformations in infected cells in order to build viral replication complexes (vrcs). tomato bushy stunt virus (tbsv) co-opts cellular escrt (endosomal sorting complexes required for transport) proteins to induce the formation of vesicle (spherule)-like structures in the peroxisomal membrane with tight openings toward the cytosol. in this study, using a yeast (saccharomyces cerevisiae) vps23δ bro1δ double-deletion mutant, we showed that the vps23p escrt-i ...	2016	26792735
comparative analysis of rnai-based methods to down-regulate expression of two genes expressed at different levels in myzus persicae.	with the increasing availability of aphid genomic data, it is necessary to develop robust functional validation methods to evaluate the role of specific aphid genes. this work represents the first study in which five different techniques, all based on rna interference and on oral acquisition of double-stranded rna (dsrna), were developed to silence two genes, aly and eph, potentially involved in polerovirus transmission by aphids. efficient silencing of only eph transcripts, which are less abund ...	2016	27869783
production of functionally active and immunogenic non-glycosylated protective antigen from bacillus anthracis in nicotiana benthamiana by co-expression with peptide-n-glycosidase f (pngase f) of flavobacterium meningosepticum.	bacillus anthracis has long been considered a potential biological warfare agent, and therefore, there is a need for a safe, low-cost and highly efficient anthrax vaccine with demonstrated long-term stability for mass vaccination in case of an emergency. many efforts have been made towards developing an anthrax vaccine based on recombinant protective antigen (rpa) of b. anthracis, a key component of the anthrax toxin, produced using different expression systems. plants represent a promising reco ...	2016	27101370
the potato aphid salivary effector me47 is a glutathione-s-transferase involved in modifying plant responses to aphid infestation.	polyphagous aphid pests cause considerable economic damage to crop plants, primarily through the depletion of photoassimilates and transfer of viruses. the potato aphid (macrosiphum euphorbiae) is a notable pest of solanaceous crops, however, the molecular mechanisms that underpin the ability to colonize these hosts are unknown. it has recently been demonstrated that like other aphid species, m. euphorbiae injects a battery of salivary proteins into host plants during feeding. it is speculated t ...	2016	27536306
the ectopic expression of carop1 modulates the response of tobacco plants to ralstonia solanacearum and aphids.	in plants, rho-related gtpases (rops) are versatile molecular switches that regulate various biological processes, although their exact roles are not fully understood. herein, we provide evidence that the ectopic expression of a rop derived from capsicum annuum, designated carop1, in tobacco plants modulates the response of these plants to ralstonia solanacearum or aphid attack. the deduced amino acid sequence of carop1 harbors a conserved rho domain and is highly homologous to rops of other pla ...	2016	27551287
bjmyb1, a transcription factor implicated in plant defence through activating bjchi1 chitinase expression by binding to a w-box-like element.	we previously identified the w-box-like-4 (wbl-4) element (gtagtgactcat), one of six wbl elements in the bjc-p promoter of the unusual chitinase gene bjchi1 from brassica juncea, as the core element responsive to fungal infection. here, we report the isolation and characterization of the cognate transcription factor interacting with the wbl-4 element. using wbl-4 as a target, we performed yeast one-hybrid screening of a b. juncea cdna library and isolated an r2r3-myb transcription factor designa ...	2016	27353280
overexpression of cotton ghmpk11 decreases disease resistance through the gibberellin signaling pathway in transgenic nicotiana benthamiana.	many changes in development, growth, hormone activity and environmental stimuli responses are mediated by mitogen-activated protein kinase (mapk) cascades. however, in plants, studies on mapks have mainly focused on mpk3, mpk4 and mpk6. here, a novel group b mapk gene, ghmpk11, was isolated from cotton (gossypium hirsutum l.) and characterized. both promoter and expression pattern analyses revealed that ghmpk11 is involved in defense responses and signaling pathways. ghmpk11 overexpression in ni ...	2016	27242882
wrky transcription factors phosphorylated by mapk regulate a plant immune nadph oxidase in nicotiana benthamiana.	pathogen attack sequentially confers pattern-triggered immunity (pti) and effector-triggered immunity (eti) after sensing of pathogen patterns and effectors by plant immune receptors, respectively. reactive oxygen species (ros) play pivotal roles in pti and eti as signaling molecules. nicotiana benthamiana rbohb, an nadph oxidase, is responsible for both the transient pti ros burst and the robust eti ros burst. here, we show that rbohb transactivation mediated by mapk contributes to r3a/avr3a-tr ...	2015	26373453
inter-organ defense networking: leaf whitefly sucking elicits plant immunity to crown gall disease caused by agrobacterium tumefaciens.	plants have elaborate defensive machinery to protect against numerous pathogens and insects. plant hormones function as modulators of defensive mechanisms to maintain plant resistance to natural enemies. our recent study suggests that salicylic acid (sa) is the primary phytohormone regulating plant responses to agrobacterium tumefaciens infection. tobacco (nicotiana benthamiana domin.) immune responses against agrobacterium-mediated crown gall disease were activated by exposure to the sucking in ...	2015	26357873
ghwrky68 reduces resistance to salt and drought in transgenic nicotiana benthamiana.	the wrky transcription factors modulate numerous physiological processes, including plant growth, development and responses to various environmental stresses. currently, our understanding of the functions of the majority of the wrky family members and their possible roles in signalling crosstalk is limited. in particular, very few wrkys have been identified and characterised from an economically important crop, cotton. in this study, we characterised a novel group iic wrky gene, ghwrky68, which ...	2015	25793865
the tyrosine aminomutase tam1 is required for β-tyrosine biosynthesis in rice.	non-protein amino acids, often isomers of the standard 20 protein amino acids, have defense-related functions in many plant species. a targeted search for jasmonate-induced metabolites in cultivated rice (oryza sativa) identified (r)-β-tyrosine, an isomer of the common amino acid (s)-α-tyrosine in the seeds, leaves, roots, and root exudates of the nipponbare cultivar. assays with 119 diverse cultivars showed a distinct presence/absence polymorphism, with β-tyrosine being most prevalent in temper ...	2015	25901084
pebl1, a novel protein elicitor from brevibacillus laterosporus strain a60, activates defense responses and systemic resistance in nicotiana benthamiana.	we report the identification, characterization, and gene cloning of a novel protein elicitor (pebl1) secreted from brevibacillus laterosporus strain a60. through a purification process consisting of ion-exchange chromatography and high-performance liquid chromatography (hplc), we isolated a protein that was identified by electrospray ionization quadrupole time of flight tandem mass spectrometry (esi-q-tof-ms-ms). the 351-bp pebl1 gene produces a 12,833-da protein with 116 amino acids that contai ...	2015	25662975
morphogenesis of endoplasmic reticulum membrane-invaginated vesicles during beet black scorch virus infection: role of auxiliary replication protein and new implications of three-dimensional architecture.	all well-characterized positive-strand rna viruses[(+)rna viruses] induce the formation of host membrane-bound viral replication complexes (vrcs), yet the underlying mechanism and machinery for vrc formation remain elusive. we report here the biogenesis and topology of the beet black scorch virus (bbsv) replication complex. distinct cytopathological changes typical of endoplasmic reticulum (er) aggregation and vesiculation were observed in bbsv-infected nicotiana benthamiana cells. immunogold la ...	2015	25833056
leishmania tarentolae: an alternative approach to the production of monoclonal antibodies to treat emerging viral infections.	monoclonal antibody therapy has an important role to play as a post-exposure prophylactic and therapeutic for the treatment of viral infections, including emerging infections. for example, several patients of the present ebola virus outbreak in west africa were treated with zmapp, a cocktail of three monoclonal antibodies which are expressed in nicotiana benthamiana.	2015	26191408
improving the large scale purification of the hiv microbicide, griffithsin.	griffithsin is a broad spectrum antiviral lectin that inhibits viral entry and maturation processes through binding clusters of oligomannose glycans on viral envelope glycoproteins. an efficient, scaleable manufacturing process for griffithsin active pharmaceutical ingredient (api) is essential for particularly cost-sensitive products such as griffithsin -based topical microbicides for hiv-1 prevention in resource poor settings. our previously published purification method used ceramic filtratio ...	2015	25887919
plant-made vaccines against west nile virus are potent, safe, and economically feasible.	the threat of west nile virus (wnv) epidemics with increasingly severe neuroinvasive infections demands the development and licensing of effective vaccines. to date, vaccine candidates based on inactivated, live-attenuated, or chimeric virus, and viral dna and wnv protein subunits have been developed. some have been approved for veterinary use or are under clinical investigation, yet no vaccine has been licensed for human use. reaching the milestone of a commercialized human vaccine, however, ma ...	2015	25676782
a geminivirus betasatellite damages the structural and functional integrity of chloroplasts leading to symptom formation and inhibition of photosynthesis.	geminivirus infection often causes severe vein clearing symptoms in hosts. recently a betasatellite has emerged as a key regulator of symptom induction. to understand the host-betasatellite interactions in the process of symptom development, a systematic study was carried out involving symptoms induced by a betasatellite associated with radish leaf curl disease (ralcb) in nicotiana benthamiana. it has been found that βc1 protein localized to chloroplasts of host cells, and ralcb lacking βc1, whi ...	2015	26113193
comparative genomic analysis of multiple strains of two unusual plant pathogens: pseudomonas corrugata and pseudomonas mediterranea.	the non-fluorescent pseudomonads, pseudomonas corrugata (pcor) and p. mediterranea (pmed), are closely related species that cause pith necrosis, a disease of tomato that causes severe crop losses. however, they also show strong antagonistic effects against economically important pathogens, demonstrating their potential for utilization as biological control agents. in addition, their metabolic versatility makes them attractive for the production of commercial biomolecules and bioremediation. an e ...	2015	26300874
a library of synthetic transcription activator-like effector-activated promoters for coordinated orthogonal gene expression in plants.	a library of synthetic promoters containing the binding site of a single designer transcription activator-like effector (dtale) was constructed. the promoters contain a constant sequence, consisting of an 18-base long dtale-binding site and a tata box, flanked by degenerate sequences of 49 bases downstream and 19 bases upstream. forty-three of these promoters were sequenced and tested in transient assays in nicotiana benthamiana using a gus reporter gene. the strength of expression of the promot ...	2015	25846505
temporal and spatial resolution of activated plant defense responses in leaves of nicotiana benthamiana infected with dickeya dadantii.	the necrotrophic bacteria dickeya dadantii is the causal agent of soft-rot disease in a broad range of hosts. the model plant nicotiana benthamiana, commonly used as experimental host for a very broad range of plant pathogens, is susceptible to infection by d. dadantii. the inoculation with d. dadantii at high dose seems to overcome the plant defense capacity, inducing maceration and death of the tissue, although restricted to the infiltrated area. by contrast, the output of the defense response ...	2015	26779238
an oomycete crn effector reprograms expression of plant hsp genes by targeting their promoters.	oomycete pathogens produce a large number of crn effectors to manipulate plant immune responses and promote infection. however, their functional mechanisms are largely unknown. here, we identified a phytophthora sojae crn effector pscrn108 which contains a putative dna-binding helix-hairpin-helix (hhh) motif and acts in the plant cell nucleus. silencing of the pscrn108 gene reduced p. sojae virulence to soybean, while expression of the gene in nicotiana benthamiana and arabidopsis thaliana enhan ...	2015	26714171
a phytophthora sojae glycoside hydrolase 12 protein is a major virulence factor during soybean infection and is recognized as a pamp.	we identified a glycoside hydrolase family 12 (gh12) protein, xeg1, produced by the soybean pathogen phytophthora sojae that exhibits xyloglucanase and β-glucanase activity. it acts as an important virulence factor during p. sojae infection but also acts as a pathogen-associated molecular pattern (pamp) in soybean (glycine max) and solanaceous species, where it can trigger defense responses including cell death. gh12 proteins occur widely across microbial taxa, and many of these gh12 proteins in ...	2015	26163574
functional dissection of a plant argonaute.	rna guided ribonuclease complexes play central role in rna interference. members of the evolutionarily conserved argonaute protein family form the catalytic cores of these complexes. unlike a number of other plant argonautes, the role of ago2 has been obscure until recently. newer data, however, have indicated its involvement in various biotic and abiotic stress responses. despite its suggested importance, there is no detailed characterization of this protein to date. here we report cloning and ...	2015	26673719
high levels of cyclic-di-gmp in plant-associated pseudomonas correlate with evasion of plant immunity.	the plant innate immune system employs plasma membrane-localized receptors that specifically perceive pathogen/microbe-associated molecular patterns (pamps/mamps). this induces a defence response called pattern-triggered immunity (pti) to fend off pathogen attack. commensal bacteria are also exposed to potential immune recognition and must employ strategies to evade and/or suppress pti to successfully colonize the plant. during plant infection, the flagellum has an ambiguous role, acting as both ...	2015	26202381
turnip vein clearing virus movement protein nuclear activity: do tobamovirus movement proteins play a role in immune response suppression?	plant viruses' cell-to-cell movement requires the function of virally encoded movement proteins (mps). the tobamovirus, tobacco mosaic virus (tmv) has served as the model virus to study the activities of single mps. however, since tmv does not infect the model plant arabidopsis thaliana i have used a related tobamovirus, turnip vein-clearing virus (tvcv). i recently showed that, despite belonging to the same genus, the behavior of the 2 viruses mps differ significantly during infection. most not ...	2015	26237173
tomato slrbohb, a member of the nadph oxidase family, is required for disease resistance against botrytis cinerea and tolerance to drought stress.	nadph oxidases (also known as respiratory burst oxidase homologs, rbohs) are key enzymes that catalyze the generation of reactive oxygen species (ros) in plants. in the present study, eight slrboh genes were identified in tomato and their possible involvement in resistance to botrytis cinerea and drought tolerance was examined. expression of slrbohs was induced by b. cinerea and pseudomonas syringae pv. tomato but displayed distinct patterns. virus-induced gene silencing based silencing of slrbo ...	2015	26157450
phosphatidic acid produced by phospholipase d promotes rna replication of a plant rna virus.	eukaryotic positive-strand rna [(+)rna] viruses are intracellular obligate parasites replicate using the membrane-bound replicase complexes that contain multiple viral and host components. to replicate, (+)rna viruses exploit host resources and modify host metabolism and membrane organization. phospholipase d (pld) is a phosphatidylcholine- and phosphatidylethanolamine-hydrolyzing enzyme that catalyzes the production of phosphatidic acid (pa), a lipid second messenger that modulates diverse intr ...	2015	26020241
cytoplastic glyceraldehyde-3-phosphate dehydrogenases interact with atg3 to negatively regulate autophagy and immunity in nicotiana benthamiana.	autophagy as a conserved catabolic pathway can respond to reactive oxygen species (ros) and plays an important role in degrading oxidized proteins in plants under various stress conditions. however, how ros regulates autophagy in response to oxidative stresses is largely unknown. here, we show that autophagy-related protein 3 (atg3) interacts with the cytosolic glyceraldehyde-3-phosphate dehydrogenases (gapcs) to regulate autophagy in nicotiana benthamiana plants. we found that oxidative stress ...	2015	25829441
the n-terminal domain of the tomato immune protein prf contains multiple homotypic and pto kinase interaction sites.	resistance to pseudomonas syringae bacteria in tomato (solanum lycopersicum) is conferred by the prf recognition complex, composed of the nucleotide-binding leucine-rich repeats protein prf and the protein kinase pto. the complex is activated by recognition of the p. syringae effectors avrpto and avrptob. the n-terminal domain is responsible for prf homodimerization, which brings two pto kinases into close proximity and holds them in inactive conformation in the absence of either effector. negat ...	2015	25792750
plant ribosomal proteins, rpl12 and rpl19, play a role in nonhost disease resistance against bacterial pathogens.	characterizing the molecular mechanism involved in nonhost disease resistance is important to understand the adaptations of plant-pathogen interactions. in this study, virus-induced gene silencing (vigs)-based forward genetics screen was utilized to identify genes involved in nonhost resistance in nicotiana benthamiana. genes encoding ribosomal proteins, rpl12 and rpl19, were identified in the screening. these genes when silenced in n. benthamiana caused a delay in nonhost bacteria induced hyper ...	2015	26779226
tomato sl3-mmp, a member of the matrix metalloproteinase family, is required for disease resistance against botrytis cinerea and pseudomonas syringae pv. tomato dc3000.	matrix metalloproteinases (mmps) are a family of zinc-dependent endopeptidases. mmps have been characterized in detail in mammals and shown to play key roles in many physiological and pathological processes. although mmps in some plant species have been identified, the function of mmps in biotic stress responses remains elusive.	2015	26070456
five xanthomonas type iii effectors suppress cell death induced by components of immunity-associated map kinase cascades.	mitogen-activated protein kinase (mapk) cascades play a fundamental role in signaling of plant immunity and mediate elicitation of cell death. xanthomonas spp. manipulate plant signaling by using a type iii secretion system to deliver effector proteins into host cells. we examined the ability of 33 xanthomonas effectors to inhibit cell death induced by overexpression of components of mapk cascades in nicotiana benthamiana plants. five effectors inhibited cell death induced by overexpression of m ...	2015	26237448
the xanthomonas effector xopj triggers a conditional hypersensitive response upon treatment of n. benthamiana leaves with salicylic acid.	xopj is a xanthomonas type iii effector protein that promotes bacterial virulence on susceptible pepper plants through the inhibition of the host cell proteasome and a resultant suppression of salicylic acid (sa) - dependent defense responses. we show here that nicotiana benthamiana leaves transiently expressing xopj display hypersensitive response (hr) -like symptoms when exogenously treated with sa. this apparent avirulence function of xopj was further dependent on effector myristoylation as w ...	2015	26284106
the alternative respiratory pathway is involved in brassinosteroid-induced environmental stress tolerance in nicotiana benthamiana.	brassinosteroids (brs), plant steroid hormones, play essential roles in modulating cell elongation, vascular differentiation, senescence, and stress responses. however, the mechanisms by which brs regulate plant mitochondria and resistance to abiotic stress remain largely unclear. mitochondrial alternative oxidase (aox) is involved in the plant response to a variety of environmental stresses. in this report, the role of aox in br-induced tolerance against cold, polyethylene glycol (peg), and hig ...	2015	26175355
role of proline and pyrroline-5-carboxylate metabolism in plant defense against invading pathogens.	pyrroline-5-carboxylate (p5c) is an intermediate product of both proline biosynthesis and catabolism. recent evidences indicate that proline-p5c metabolism is tightly regulated in plants, especially during pathogen infection and abiotic stress. however, role of p5c and its metabolism in plants has not yet been fully understood. studies indicate that p5c synthesized in mitochondria has a role in both resistance (r)-gene-mediated and non-host resistance against invading pathogens. proline dehydrog ...	2015	26217357
tight regulation of plant immune responses by combining promoter and suicide exon elements.	effector-triggered immunity (eti) is activated when plant disease resistance (r) proteins recognize the presence of pathogen effector proteins delivered into host cells. the eti response generally encompasses a defensive 'hypersensitive response' (hr) that involves programmed cell death at the site of pathogen recognition. while many r protein and effector protein pairs are known to trigger hr, other components of the eti signaling pathway remain elusive. effector genes regulated by inducible pr ...	2015	26138488
a terpenoid phytoalexin plays a role in basal defense of nicotiana benthamiana against potato virus x.	terpenoid phytoalexins function as defense compound against a broad spectrum of pathogens and pests in the plant kingdom. however, the role of phytoalexin in antiviral defense is still elusive. in this study, we identified the biosynthesis pathway of a sesquiterpenoid phytoalexin, capsidiol 3-acetate as an antiviral response against rna virus potato virus x (pvx) in nicotiana benthamiana. nbtps1 and nbeah genes were found strongly induced by pvx-infection. enzymatic activity and genetic evidence ...	2015	25993114
growth and stress response in arabidopsis thaliana, nicotiana benthamiana, glycine max, solanum tuberosum and brassica napus cultivated under polychromatic leds.	the use of light emitting diodes (leds) brings several key advantages over existing illumination technologies for indoor plant cultivation. among these are that leds have predicted lifetimes from 50-100.000 hours without significant drops in efficiency and energy consumption is much lower compared to traditional fluorescent tubes. recent advances allow leds to be used with customized wavelengths for plant growth. however, most of these led growth systems use mixtures of chips emitting in several ...	2015	27408615
pepper aldehyde dehydrogenase caaldh1 interacts with xanthomonas effector avrbst and promotes effector-triggered cell death and defence responses.	xanthomonas type iii effector avrbst induces hypersensitive cell death and defence responses in pepper (capsicum annuum) and nicotiana benthamiana. little is known about the host factors that interact with avrbst. here, we identified pepper aldehyde dehydrogenase 1 (caaldh1) as an avrbst-interacting protein. bimolecular fluorescence complementation and co-immunoprecipitation assays confirmed the interaction between caaldh1 and avrbst in planta. caaldh1:smgfp fluorescence was detected in the cyto ...	2015	25873668
phylogenomics of xanthomonas field strains infecting pepper and tomato reveals diversity in effector repertoires and identifies determinants of host specificity.	bacterial spot disease of pepper and tomato is caused by four distinct xanthomonas species and is a severely limiting factor on fruit yield in these crops. the genetic diversity and the type iii effector repertoires of a large sampling of field strains for this disease have yet to be explored on a genomic scale, limiting our understanding of pathogen evolution in an agricultural setting. genomes of 67 xanthomonas euvesicatoria (xe), xanthomonas perforans (xp), and xanthomonas gardneri (xg) strai ...	2015	26089818
functional analysis of plant defense suppression and activation by the xanthomonas core type iii effector xopx.	many phytopathogenic type iii secretion effector proteins (t3es) have been shown to target and suppress plant immune signaling but perturbation of the plant immune system by t3es can also elicit a plant response. xopx is a "core" xanthomonas t3e that contributes to growth and symptom development during xanthomonas euvesicatoria infection of tomato but its functional role is undefined. we tested the effect of xopx on several aspects of plant immune signaling. xopx promoted ethylene production and ...	2015	25338145
genome-wide characterization of mirnas involved in n gene-mediated immunity in response to tobacco mosaic virus in nicotiana benthamiana.	micrornas (mirnas) are a class of endogenous small rnas (srnas) that play pivotal roles in plant development, abiotic stress response, and pathogen response. mirnas have been extensively studied in plants, but rarely in nicotiana benthamiana, despite its wide use in plant virology studies, particularly for studying n protein-tobacco mosaic virus (tmv) interactions. we report an efficient method using high-throughput sequencing and bioinformatics to identify genome-wide mirnas in n. benthamiana. ...	2015	25673968
maize homologs of hydroxycinnamoyltransferase, a key enzyme in lignin biosynthesis, bind the nucleotide binding leucine-rich repeat rp1 proteins to modulate the defense response.	in plants, most disease resistance genes encode nucleotide binding leu-rich repeat (nlr) proteins that trigger a rapid localized cell death called a hypersensitive response (hr) upon pathogen recognition. the maize (zea mays) nlr protein rp1-d21 derives from an intragenic recombination between two nlrs, rp1-d and rp1-dp2, and confers an autoactive hr in the absence of pathogen infection. from a previous quantitative trait loci and genome-wide association study, we identified a single-nucleotide ...	2015	26373661
a new cyanogenic metabolite in arabidopsis required for inducible pathogen defence.	thousands of putative biosynthetic genes in arabidopsis thaliana have no known function, which suggests that there are numerous molecules contributing to plant fitness that have not yet been discovered. prime among these uncharacterized genes are cytochromes p450 upregulated in response to pathogens. here we start with a single pathogen-induced p450 (ref. 5), cyp82c2, and use a combination of untargeted metabolomics and coexpression analysis to uncover the complete biosynthetic pathway to 4-hydr ...	2015	26352477
the activation of phytophthora effector avr3b by plant cyclophilin is required for the nudix hydrolase activity of avr3b.	plant pathogens secrete an arsenal of effector proteins to impair host immunity. some effectors possess enzymatic activities that can modify their host targets. previously, we demonstrated that a phytophthora sojae rxlr effector avr3b acts as a nudix hydrolase when expressed in planta; and this enzymatic activity is required for full virulence of p. sojae strain p6497 in soybean (glycine max). interestingly, recombinant avr3b produced by e. coli does not have the hydrolase activity unless it was ...	2015	26317500
xanthomonas campestris cell-cell signalling molecule dsf (diffusible signal factor) elicits innate immunity in plants and is suppressed by the exopolysaccharide xanthan.	several secreted and surface-associated conserved microbial molecules are recognized by the host to mount the defence response. one such evolutionarily well-conserved bacterial process is the production of cell-cell signalling molecules which regulate production of multiple virulence functions by a process known as quorum sensing. here it is shown that a bacterial fatty acid cell-cell signalling molecule, dsf (diffusible signal factor), elicits innate immunity in plants. the dsf family of signal ...	2015	26248667
form follows function in geminiviral minichromosome architecture.	a comprehensive survey on the viral minichromosomes of the begomoviruses abutilon mosaic virus, tomato yellow leaf curl sardinia virus, african cassava mosaic virus, indian cassava mosaic virus (family geminiviridae) during the course of infections in nicotiana benthamiana is summarized. using optimized one-dimensional and two-dimensional gel systems combined with blot hybridization and a standardized evaluation, discrete and heterogeneous virus-specific signals with different dna forms were com ...	2015	25445344
ageratum enation virus-a begomovirus of weeds with the potential to infect crops.	samples of two ageratum conyzoides, one sonchus oleraceus and one turnip (brassica rapa var. rapa) exhibiting virus-like symptoms were collected from pakistan and nepal. full-length begomovirus clones were obtained from the four plant samples and betasatellite clones from three of these. the begomovirus sequences were shown to be isolates of ageratum enation virus (aev) with greater than 89.1% nucleotide sequence identity to the 26 aev sequences available in the databases. the three betasatellit ...	2015	25674770
gibson assembly: an easy way to clone potyviral full-length infectious cdna clones expressing an ectopic vpg.	approaches to simplify and accelerate the construction of full-length infectious cdna clones for plant potyviruses have been described, based on cloning strategies involving in vitro ligation or homologous recombination in yeast. in the present study, we developed a faster and more efficient in vitro recombination system using gibson assembly (ga), to engineer a lettuce mosaic virus (lmv) infectious clone expressing an ectopic mcherry-tagged vpg (viral protein genome-linked) for in planta subcel ...	2015	26070311
co-infection with two strains of brome mosaic bromovirus reveals common rna recombination sites in different hosts.	we have previously reported intra-segmental crossovers in brome mosaic virus (bmv) rnas. in this work, we studied the homologous recombination of bmv rna in three different hosts: barley (hordeum vulgare), chenopodium quinoa, and nicotiana benthamiana that were co-infected with two strains of bmv: russian (r) and fescue (f). our work aimed at (1) establishing the frequency of recombination, (2) mapping the recombination hot spots, and (3) addressing host effects. the f and r nucleotide sequences ...	2015	27774290
application of a scalable plant transient gene expression platform for malaria vaccine development.	despite decades of intensive research efforts there is currently no vaccine that provides sustained sterile immunity against malaria. in this context, a large number of targets from the different stages of the plasmodium falciparum life cycle have been evaluated as vaccine candidates. none of these candidates has fulfilled expectations, and as long as we lack a single target that induces strain-transcending protective immune responses, combining key antigens from different life cycle stages seem ...	2015	26779197
plant translation elongation factor 1bβ facilitates potato virus x (pvx) infection and interacts with pvx triple gene block protein 1.	the eukaryotic translation elongation factor 1 (eef1) has two components: the g-protein eef1a and the nucleotide exchange factor eef1b. in plants, eef1b is itself composed of a structural protein (eef1bγ) and two nucleotide exchange subunits (eef1bα and eef1bβ). to test the effects of elongation factors on virus infection, we isolated eef1a and eef1b genes from pepper (capsicum annuum) and suppressed their homologs in nicotiana benthamiana using virus-induced gene silencing (vigs). the accumulat ...	2015	26020533
pseudomonas syringae pv. tomato dc3000 type iii secretion effector polymutants reveal an interplay between hopad1 and avrptob.	the bacterial pathogen pseudomonas syringae pv. tomato dc3000 suppresses the two-tiered plant innate immune system by injecting a complex repertoire of type iii secretion effector (t3e) proteins. beyond redundancy and interplay, individual t3es may interact with multiple immunity-associated proteins, rendering their analysis challenging. we constructed a pst dc3000 polymutant lacking all 36 t3es and restored individual t3es or their mutants to explore the interplay among t3es. the weakly express ...	2015	26067603
purification of monoclonal antibody against ebola gp1 protein expressed in nicotiana benthamiana.	monoclonal antibodies (mabs) are one of the fastest growing drug molecules targeting the treatment of diseases ranging from arthritis, immune disorders, and infectious diseases to cancer. due to its unique application principle, antibodies are commonly produced in large quantities. plants, such as nicotiana benthamiana, offer a unique production platform for bio-therapeutics due to their ability to produce large amounts of biomolecules in a relatively quick manner. however, purification of a tar ...	2015	25746758
a population genetics perspective on geminivirus infection.	the selective accumulation of both dna components of a bipartite geminivirus, abutilon mosaic virus, was recorded during early systemic infection of nicotiana benthamiana plants. purified nuclei were diagnosed for viral dna using hybridization specific for dna a or dna b to detect these individual genome components either alone or both simultaneously by dual-color staining. although this virus needs both components for symptomatic infection, dna a alone was transported to upper leaves, where it ...	2015	26378173
the agrobacterium tumefaciens ti plasmid virulence gene vire2 reduces sri lankan cassava mosaic virus infection in transgenic nicotiana benthamiana plants.	cassava mosaic disease is a major constraint to cassava cultivation worldwide. in india, the disease is caused by indian cassava mosaic virus (icmv) and sri lankan cassava mosaic virus (slcmv). the agrobacterium ti plasmid virulence gene vire2, encoding a nuclear-localized, single-stranded dna binding protein, was introduced into nicotiana benthamiana to develop tolerance against slcmv. leaf discs of transgenic n. benthamiana plants, harboring the vire2 gene, complemented a vire2 mutation in a. ...	2015	26008704
snare-rnai results in higher terpene emission from ectopically expressed caryophyllene synthase in nicotiana benthamiana.	plants produce numerous terpenes and much effort has been dedicated to the identification and characterization of the terpene biosynthetic genes. however, little is known about how terpenes are transported within the cell and from the cell into the apoplast. to investigate a putative role of vesicle fusion in this process, we used agrobacterium tumefaciens-mediated transient coexpression in nicotiana benthamiana of an mtvamp721e-rnai construct (vi) with either a caryophyllene synthase or a linal ...	2015	25598143
tobacco against ebola virus disease.	the ebola virus disease (evd), formerly known as a hemorrhagic fever and discovered in 1976, is dangerous, highly infectious disease with very high mortality. there are no licensed therapeutics against evd, although a range of medicines and therapies are currently being evaluated. during the 2014 ebola outbreak, an experimental drug named zmapp was administered on an emergency basis to seven patients of which five were recovered. currently, since february 2015, zmapp is tested in clinical trials ...	2015	26946569
an improved cucumber mosaic virus-based vector for efficient decoying of plant micrornas.	we previously devised a cucumber mosaic virus (cmv)-based vector system carrying microrna target mimic sequences for analysis of microrna function in arabidopsis thaliana. we describe an improved version in which target mimic cloning is achieved by annealing two partly-overlapping complementary dna oligonucleotides for insertion into an infectious clone of cmv rna3 (ls strain) fused to the cauliflower mosaic virus-derived 35s promoter. ls-cmv variants carrying mimic sequences were generated by c ...	2015	26278008
the protein disulfide isomerase 1 of phytophthora parasitica (pppdi1) is associated with the haustoria-like structures and contributes to plant infection.	protein disulfide isomerase (pdi) is a ubiquitous and multifunction enzyme belonging to the thioredoxin (trx) superfamily, which can reduce, oxidize, and catalyze dithiol-disulfide exchange reactions. other than performing housekeeping functions in helping to maintain proteins in a more stable conformation, there is some evidence to indicate that pdi is involved in pathogen infection processes. in a high-throughput screening for necrosis-inducing factors by agrobacterium tumefaciens-mediated tra ...	2015	26347756
development of an agroinoculation system for full-length and gfp-tagged cdna clones of cucumber green mottle mosaic virus.	the complete 6243-nucleotide sequence of a cucumber green mottle mosaic virus (cgmmv) isolate from bottle gourd in zhejiang province, china, was determined. a full-length cdna clone of this isolate was constructed by inserting the cdna between the 35s promoter and the ribozyme in the binary plasmid pcb301-ch. a suspension of an agrobacterium tumefaciens eha105 clone carrying this construct was highly infectious in nicotiana benthamiana and bottle gourd. another infectious clone containing the gr ...	2015	26323263
lotus japonicus clathrin heavy chain1 is associated with rho-like gtpase rop6 and involved in nodule formation.	mechanisms underlying nodulation factor signaling downstream of the nodulation factor receptors (nfrs) have not been fully characterized. in this study, clathrin heavy chain1 (chc1) was shown to interact with the rho-like gtpase rop6, an interaction partner of nfr5 in lotus japonicus. the chc1 gene was found to be expressed constitutively in all plant tissues and induced in mesorhizobium loti-infected root hairs and nodule primordia. when expressed in leaves of nicotiana benthamiana, chc1 and ro ...	2015	25717037
phosphorylation of beet black scorch virus coat protein by pka is required for assembly and stability of virus particles.	plant virus coat proteins (cps) play a fundamental role in protection of genomic rnas, virion assembly, and viral movement. although phosphorylation of several cps during virus infection have been reported, little information is available about cp phosphorylation of the spherical rna plant viruses. here, we demonstrate that the cp of beet black scorch virus (bbsv), a member of the genus necrovirus, can be phosphorylated at threonine-41 (t41) by camp-dependent protein kinase (pka)-like kinase in ...	2015	26108567
analysis of geminivirus al2 and l2 proteins reveals a novel al2 silencing suppressor activity.	both posttranscriptional and transcriptional gene silencing (ptgs and tgs, respectively) participate in defense against the dna-containing geminiviruses. as a countermeasure, members of the genus begomovirus (e.g., cabbage leaf curl virus) encode an al2 protein that is both a transcriptional activator and a silencing suppressor. the related l2 protein of beet curly top virus (genus curtovirus) lacks transcription activation activity. previous studies showed that both al2 and l2 suppress silencin ...	2015	25552721
phosphorylation of tgb1 by protein kinase ck2 promotes barley stripe mosaic virus movement in monocots and dicots.	the barley stripe mosaic virus (bsmv) triple gene block 1 (tgb1) protein is required for virus cell-to-cell movement. however, little information is available about how these activities are regulated by post-translational modifications. in this study, we showed that the bsmv xinjiang strain tgb1 (xjtgb1) is phosphorylated in vivo and in vitro by protein kinase ck2 from barley and nicotiana benthamiana. liquid chromatography tandem mass spectrometry analysis and in vitro phosphorylation assays de ...	2015	25998907
viral elements and host cellular proteins in intercellular movement of bamboo mosaic virus.	as a member of the genus potexvirus, bamboo mosaic virus (bamv) also belongs to the plant viruses that encode triple gene block proteins (tgbps) for intercellular movement within the host plants. recent studies of the movement mechanisms of bamv have revealed similarities and differences between bamv and other potexviruses. this review focuses on the general aspects of viral and host elements involved in bamv movement, the interactions among these elements, and the possible pathways for intra- a ...	2015	25951346
mild and severe cereal yellow dwarf viruses differ in silencing suppressor efficiency of the p0 protein.	viral pathogenicity has often been correlated to the expression of the viral encoded-rna silencing suppressor protein (ssp). the silencing suppressor activity of the p0 protein encoded by cereal yellow dwarf virus-rpv (cydv-rpv) and -rps (cydv-rps), two poleroviruses differing in their symptomatology was investigated. cydv-rpv displays milder symptoms in oat and wheat whereas cydv-rps is responsible for more severe disease. we showed that both p0 proteins (p0(cy-rpv) and p0(cy-rps)) were able to ...	2015	26116275
characterization and comparative analysis of promoters from three plant pararetroviruses associated with dahlia (dahlia variabilis).	two distinct caulimoviruses, dahlia mosaic virus (dmv) and dahlia common mosaic virus (dcmv), and an endogenous plant pararetroviral sequence (dveprs, formerly known as dmv-d10) were reported from dahlia (dahlia spp). promoter elements from these dahlia-associated pararetroviruses were identified and characterized. the tata box, the caat box, the transcription start site, the polyadenylation signal, and regulation factors, characteristic of caulimovirus promoters, were present in each of these p ...	2015	25947569
aboveground insect infestation attenuates belowground agrobacterium-mediated genetic transformation.	agrobacterium tumefaciens causes crown gall disease. although agrobacterium can be popularly used for genetic engineering, the influence of aboveground insect infestation on agrobacterium induced gall formation has not been investigated. nicotiana benthamiana leaves were exposed to a sucking insect (whitefly) infestation and benzothiadiazole (bth) for 7 d, and these exposed plants were inoculated with a tumorigenic agrobacterium strain. we evaluated, both in planta and in vitro, how whitefly inf ...	2015	25676198
insights into the polerovirus-plant interactome revealed by coimmunoprecipitation and mass spectrometry.	identification of host proteins interacting with the aphidborne potato leafroll virus (plrv) from the genus polerovirus, family luteoviridae, is a critical step toward understanding how plrv and related viruses infect plants. however, the tight spatial distribution of plrv to phloem tissues poses challenges. a polyclonal antibody raised against purified plrv virions was used to coimmunoprecipitate virus-host protein complexes from nicotiana benthamiana tissue inoculated with an infectious plrv c ...	2015	25496593
characterization of the promoter of grapevine vein clearing virus.	grapevine vein clearing virus (gvcv) is a recently discovered dna virus in grapevine that is closely associated with the grapevine vein clearing syndrome observed in vineyards in missouri and surrounding states. the genome sequence of gvcv indicates that it belongs to the genus badnavirus in the family caulimoviridae. to identify the gvcv promoter, we cloned portions of the gvcv large intergenic region in front of a gfp gene present in an agrobacterium tumefaciens binary vector. gfp expression w ...	2015	25281563
new tools for ntd vaccines: a case study of quality control assays for product development of the human hookworm vaccine na-apr-1m74.	na-apr-1(m74) is an aspartic protease that is rendered enzymatically inactive by site-directed mutagenesis and is a candidate antigen component in the human hookworm vaccine. the mutant protease exerts vaccine efficacy by inducing antibodies that neutralize the enzymatic activity of wild type enzyme (na-apr-1wt) in the gut of the hookworm, thereby depriving the worm of its ability to digest its blood meal. previously, canines immunized with na-apr-1(m74) and challenged with ancylostoma caninum w ...	2015	26018444
roles of agrobacterium tumefaciens c58 znta and zntb and the transcriptional regulator zntr in controlling cd2+/zn2+/co2+ resistance and the peroxide stress response.	the putative zinc exporters znta (a p1b-type atpase) and zntb (2-tm-gxn family) in agrobacterium tumefaciens were characterized. the expression of the znta gene is inducible by cdcl2, zncl2 and cocl2, of which cdcl2 is the most potent inducer, whereas zntb is constitutively expressed. the metal-induced expression of znta is controlled by the merr-like regulator zntr. the znta and zntr mutants were highly sensitive to cdcl2 and zncl2, and cocl2 sensitivity was demonstrated to a lesser extent. by ...	2015	26296876
leaf proteome rebalancing in nicotiana benthamiana for upstream enrichment of a transiently expressed recombinant protein.	a key factor influencing the yield of biopharmaceuticals in plants is the ratio of recombinant to host proteins in crude extracts. postextraction procedures have been devised to enrich recombinant proteins before purification. here, we assessed the potential of methyl jasmonate (meja) as a generic trigger of recombinant protein enrichment in nicotiana benthamiana leaves before harvesting. previous studies have reported a significant rebalancing of the leaf proteome via the jasmonate signalling p ...	2015	26286859
characterization, expression patterns and functional analysis of the mapk and mapkk genes in watermelon (citrullus lanatus).	mitogen-activated protein kinase (mapk) cascades, which consist of three functionally associated protein kinases, namely mekks, mkks and mpks, are universal signaling modules in all eukaryotes and have been shown to play critical roles in many physiological and biochemical processes in plants. however, little or nothing is known about the mpk and mkk families in watermelon.	2015	26700161
establishing a crispr-cas-like immune system conferring dna virus resistance in plants.	crispr-cas (clustered, regularly interspaced short palindromic repeats-crispr-associated proteins) is an adaptive immune system in many archaea and bacteria that cleaves foreign dna on the basis of sequence complementarity. here, using the geminivirus, beet severe curly top virus (bsctv), transient assays performed in nicotiana benthamiana demonstrate that the sgrna-cas9 constructs inhibit virus accumulation and introduce mutations at the target sequences. further, transgenic arabidopsis and n. ...	2015	27251395
development of a single-replicon minibyv vector for co-expression of heterologous proteins.	in planta production of recombinant proteins, including vaccine antigens and monoclonal antibodies, continues gaining acceptance. with the broadening range of target proteins, the need for vectors with higher performance is increasing. here, we have developed a single-replicon vector based on beet yellows virus (byv) that enables co-delivery of two target genes into the same host cell, resulting in transient expression of each target. this byv vector maintained genetic stability during systemic ...	2015	25280556
blocking single-stranded transferred dna conversion to double-stranded intermediates by overexpression of yeast dna replication factor a.	agrobacterium tumefaciens delivers its single-stranded transferred dna (t-strand) into the host cell nucleus, where it can be converted into double-stranded molecules. various studies have revealed that double-stranded transfer dna (t-dna) intermediates can serve as substrates by as yet uncharacterized integration machinery. nevertheless, the possibility that t-strands are themselves substrates for integration cannot be ruled out. we attempted to block the conversion of t-strands into double-str ...	2015	25424309
construction of infectious clones of tomato torrado virus and their delivery by agroinfiltration.	the first biologically active infectious clones of tomato torrado virus (totv) were generated and delivered into nicotiana benthamiana and solanum lycopersicum plants via agrobacterium tumefaciens. the engineered constructs consisted of pcr-amplified complementary dnas derived from the totv rna1 and rna2 components, individually inserted into an engineered pgreen binary vector between the camv 35s promoter and nopaline synthase terminator. these constructs were introduced into the plant hosts by ...	2015	25416854
nitric oxide is involved in brassinosteroid-induced alternative respiratory pathway in nicotiana benthamiana seedlings' response to salt stress.	recent studies reported that brassinosteroids (brs) can induce plant tolerance to different environmental stresses via the nitric oxide (no) signaling pathway. previous reports have indicated that alternative oxidase (aox) plays an important role in plants under various stresses. the mechanisms governing how no is involved as a signal molecule which connects br with aox in regulating stress tolerance are still unknown. recently, we found that nicotiana benthamiana seedlings which were pretreated ...	2015	26419322
complete genome sequence of nine isolates of canna yellow streak virus reveals its relationship to the sugarcane mosaic virus (scmv) subgroup of potyviruses.	complete genome sequences were obtained from nine isolates of canna yellow streak virus (caysv). caysv belongs to the sugarcane mosaic virus (scmv) subgroup of potyviruses with johnsongrass mosaic virus (jgmv) as its closest relative. multiple sequence alignments showed a pattern of amino acid substitutions in the cp sequences, which enabled us to relate these isolates to south east asian or european isolates. biological characterization of caysv identified nicotiana benthamiana, chenopodium qui ...	2015	25567205
elicitin-like proteins oli-d1 and oli-d2 from pythium oligandrum trigger hypersensitive response in nicotiana benthamiana and induce resistance against botrytis cinerea in tomato.	the biocontrol agent pythium oligandrum and its elicitin-like proteins oligandrins have been shown to induce disease resistance in a range of plants. in the present study, the ability of two oligandrins, oli-d1 and oli-d2, to induce an immune response and the possible molecular mechanism regulating the defence responses in nicotiana benthamiana and tomato were investigated. infiltration of recombinant oli-d1 and oli-d2 proteins induced a typical immune response in n. benthamiana including the in ...	2015	25047132
establishment of an agrobacterium-mediated inoculation system for cucumber green mottle mosaic virus.	the infectious full-length cdna clones of cucumber green mottle mosaic virus (cgmmv) isolates kw and kom, which were isolated from watermelon and oriental melon, respectively, were constructed under the control of the cauliflower mosaic virus 35s promoter. we successfully inoculated nicotiana benthamiana with the cloned cgmmv isolates kw and kom by agrobacterium-mediated infiltration. virulence and symptomatic characteristics of the cloned cgmmv isolates kw and kom were tested on several indicat ...	2015	26674677
overexpression of a phytophthora cytoplasmic crn effector confers resistance to disease, salinity and drought in nicotiana benthamiana.	the crinkler (crn) effector family is produced by oomycete pathogens and may manipulate host physiological and biochemical events inside host cells. here, pscrn161 was identified from phytophthora sojae based on its broad and strong cell death suppression activities. the effector protein contains two predicted nuclear localization signals and localized to nuclei of plant cells, indicating that it may target plant nuclei to modify host cell physiology and function. the chimeric gene gfp:pscrn161 ...	2015	26546319