| production of camel-like antibodies in plants. | transgenic plants for the production of high-value recombinant complex and/or glycosylated proteins are a promising alternative for conventional systems, such as mammalian cells and bacteria. many groups use plants as production platform for antibodies and antibody fragments. here, we describe how bivalent camel-like antibodies can be produced in leaves and seeds. camel-like antibodies are fusions of the antigen-binding domain of heavy chain camel antibodies (vhh) with an fc fragment of choice. ... | 2012 | 22886260 |
| legume receptors perceive the rhizobial lipochitin oligosaccharide signal molecules by direct binding. | lipochitin oligosaccharides called nod factors function as primary rhizobial signal molecules triggering legumes to develop new plant organs: root nodules that host the bacteria as nitrogen-fixing bacteroids. here, we show that the lotus japonicus nod factor receptor 5 (nfr5) and nod factor receptor 1 (nfr1) bind nod factor directly at high-affinity binding sites. both receptor proteins were posttranslationally processed when expressed as fusion proteins and extracted from purified membrane frac ... | 2012 | 22859506 |
| a cytosolic acyltransferase contributes to triacylglycerol synthesis in sucrose-rescued arabidopsis seed oil catabolism mutants. | triacylglycerol (tag) levels and oil bodies persist in sucrose (suc)-rescued arabidopsis (arabidopsis thaliana) seedlings disrupted in seed oil catabolism. this study set out to establish if tag levels persist as a metabolically inert pool when downstream catabolism is disrupted, or if other mechanisms, such as fatty acid (fa) recycling into tag are operating. we show that tag composition changes significantly in suc-rescued seedlings compared with that found in dry seeds, with 18:2 and 18:3 acc ... | 2012 | 22760209 |
| functional analyses of agamous family members in nicotiana benthamiana clarify the evolution of early and late roles of c-function genes in eudicots. | the c-function, according to the abc model of floral organ identity, is required for stamen and carpel development and to provide floral meristem determinacy. members of the ag lineage of the large mads box gene family specify the c-function in a broadly conserved manner in angiosperms. in core eudicots, two sub-lineages co-exist, euag and ple, which have been extensively characterized in antirrhinum majus and arabidopsis thaliana, where strong sub-functionalization has led to highly divergent c ... | 2012 | 22563981 |
| very bright orange fluorescent plants: endoplasmic reticulum targeting of orange fluorescent proteins as visual reporters in transgenic plants. | the expression of fluorescent protein (fp) genes as real-time visual markers, both transiently and stably, has revolutionized plant biotechnology. a palette of colors of fps is now available for use, but the diversity has generally been underutilized in plant biotechnology. because of the green and far-red autofluorescent properties of many plant tissues and the fps themselves, red and orange fps (rfps, and ofps, respectfully) appear to be the colors with maximum utility in plant biotechnology. ... | 2012 | 22554231 |
| arabidopsis senescence-associated protein dmp1 is involved in membrane remodeling of the er and tonoplast. | arabidopsis dmp1 was discovered in a genome-wide screen for senescence-associated membrane proteins. dmp1 is a member of a novel plant-specific membrane protein family of unknown function. in rosette leaves dmp1 expression increases from very low background level several 100fold during senescence progression. | 2012 | 22530652 |
| the extreme n-terminal domain of a hordeivirus tgb1 movement protein mediates its localization to the nucleolus and interaction with fibrillarin. | the hordeiviral movement protein encoded by the first gene of the triple gene block (tgbp1) of poa semilatent virus (pslv), interacts with viral genomic rnas to form rnp particles which are considered to be a form of viral genome capable of cell-to-cell and long-distance transport in infected plants. the pslv tgbp1 contains a c-terminal ntpase/helicase domain (held) and an n-terminal extension region consisting of two structurally and functionally distinct domains: an extreme n-terminal domain ( ... | 2012 | 22349738 |
| the two endo-β-n-acetylglucosaminidase genes from arabidopsis thaliana encode cytoplasmic enzymes controlling free n-glycan levels. | endo-β-n-acetylglucosaminidases (engases) cleave n-glycans from proteins and/or peptides by hydrolyzing the o-glycosidic linkage between the two core-n-acetylglucosamine (glcnac) residues. although, two homologous genes potentially encoding engases have been identified in arabidopsis thaliana, their respective substrate specificity, their subcellular and their organ specific localization was hitherto unknown. in order to investigate the role of engases in this model plant species, we transiently ... | 2011 | 21796445 |
| labeling and enrichment of arabidopsis thaliana matrix metalloproteases using an active-site directed, marimastat-based photoreactive probe. | matrix metalloproteases (mmps) are secreted or membrane-bound zinc-containing proteases that play diverse roles in development and immunity in plants and in tissue remodeling in animals. we developed a photoreactive probe based on the mmp inhibitor marimastat, conjugated to a 4-azidotetrafluorobenzoyl moiety as photoreactive group and biotin as detection or sorting function. the probe labels at2-mmp, at4-mmp, at5-mmp, and likely other plant mmps in leaf extracts, as shown by transient at-mmp exp ... | 2012 | 21775155 |
| intron retention and 3'-utr analysis of arabidopsis dicer-like 2 transcripts. | arabidopsis thaliana dicer-like protein 2 (atdcl2) plays an essential role in the rna interference pathway. the function of atdcl2 and other dcls has been much studied but little has been done to characterize the dcls transcripts before they are translated into proteins. here, we investigated atdcl2 transcripts and showed that all 21 introns of atdcl2 except intron 9, 18, 20 and 21 could be retained although spliced sequences usually predominated. intron 10 was more frequently retained and trans ... | 2012 | 21698366 |
| salicylic acid and its location in response to biotic and abiotic stress. | salicylic acid (sa) is an important signal involved in the activation of plant defence responses against abiotic and biotic stress. sa may derive from the phenylpropanoid pathway or via isochorismate synthase as demonstrated in nicotiana benthamiana, tomato and arabidopsis thaliana. the phenylpropanoid pathway as well as isochorismate synthase are localized in the chloroplasts but it remains unknown if the end product sa is in the same organelle. we have studied the localization of sa in a. thal ... | 2011 | 21530511 |
| cell death mediated by the n-terminal domains of a unique and highly conserved class of nb-lrr protein. | plant genomes encode large numbers of nucleotide-binding, leucine-rich repeat (nb-lrr) proteins, many of which are active in pathogen detection and defense response induction. nb-lrr proteins fall into two broad classes: those with a toll and interleukin-1 receptor (tir) domain at their n-terminus and those with a coiled-coil (cc) domain at the n-terminus. within cc-nb-lrr-encoding genes, one basal clade is distinguished by having cc domains resembling the arabidopsis thaliana rpw8 protein, whic ... | 2011 | 21501087 |
| a putative arabidopsis thaliana glycosyltransferase, at4g01220, which is closely related to three plant cell wall-specific xylosyltransferases, is differentially expressed spatially and temporally. | plant cell wall polysaccharides are amongst the most complex, heterogeneous and abundant bio-molecules on earth. this makes the biosynthetic enzymes, namely the glycosyltransferases and polysaccharide synthases, important research targets in plant science and biotechnology. as an initial step to characterize at4g01220, a putative arabidopsis thaliana encoding glycosyltransferases in cazy gt-family-77 that is similar to three known xylosyltransferases involved in the biosynthesis of the pectic po ... | 2011 | 21421394 |
| metabolic engineering in nicotiana benthamiana reveals key enzyme functions in arabidopsis indole glucosinolate modification. | indole glucosinolates, derived from the amino acid trp, are plant secondary metabolites that mediate numerous biological interactions between cruciferous plants and their natural enemies, such as herbivorous insects, pathogens, and other pests. while the genes and enzymes involved in the arabidopsis thaliana core biosynthetic pathway, leading to indol-3-yl-methyl glucosinolate (i3m), have been identified and characterized, the genes and gene products responsible for modification reactions of the ... | 2011 | 21317374 |
| modulation of sulfur metabolism enables efficient glucosinolate engineering. | metabolic engineering in heterologous organisms is an attractive approach to achieve efficient production of valuable natural products. glucosinolates represent a good example of such compounds as they are thought to be the cancer-preventive agents in cruciferous plants. we have recently demonstrated that it is feasible to engineer benzylglucosinolate (bgls) in the non-cruciferous plant nicotiana benthamiana by transient expression of five genes from arabidopsis thaliana. in the same study, we s ... | 2011 | 21281472 |
| rpk2 is an essential receptor-like kinase that transmits the clv3 signal in arabidopsis. | the shoot apical meristem (sam) is the fundamental structure that is located at the growing tip and gives rise to all aerial parts of plant tissues and organs, such as leaves, stems and flowers. in arabidopsis thaliana, the clavata3 (clv3) pathway regulates the stem cell pool in the sam, in which a small peptide ligand derived from clv3 is perceived by two major receptor complexes, clv1 and clv2-coryne (crn)/suppressor of llp1 2 (sol2), to restrict wuschel (wus) expression. in this study, we use ... | 2010 | 20978082 |
| multiple regulatory elements in the arabidopsis nia1 promoter act synergistically to form a nitrate enhancer. | to accommodate fluctuating nutrient levels in the soil, plants modulate their metabolism and root development via signaling mechanisms that rapidly reprogram the plant transcriptome. in the case of nitrate, over 1,000 genes are induced or repressed within minutes of nitrate exposure. to identify cis-regulatory elements that mediate these responses, an enhancer screen was performed in transgenic arabidopsis (arabidopsis thaliana) plants. a 1.8-kb promoter fragment from the nitrate reductase gene ... | 2010 | 20668061 |
| methods to study pamp-triggered immunity using tomato and nicotiana benthamiana. | understanding the molecular basis of plant responses to pathogen-associated molecular patterns (pamps) is an active area of research in the field of plant-microbe interactions. a growing number of plant genes involved in various steps of pamp-triggered immunity (pti) pathways and microbial factors involved in the elicitation or suppression of pti have been identified. these studies have largely relied on arabidopsis thaliana and, therefore, most of the pti assays have been developed and optimize ... | 2010 | 20615110 |
| plastidial thioredoxin z interacts with two fructokinase-like proteins in a thiol-dependent manner: evidence for an essential role in chloroplast development in arabidopsis and nicotiana benthamiana. | here, we characterize a plastidial thioredoxin (trx) isoform from arabidopsis thaliana that defines a previously unknown branch of plastidial trxs lying between x- and y-type trxs and thus was named trx z. an arabidopsis knockout mutant of trx z had a severe albino phenotype and was inhibited in chloroplast development. quantitative real-time rt-pcr analysis of the mutant suggested that the expressions of genes that depend on a plastid-encoded rna polymerase (pep) were specifically decreased. si ... | 2010 | 20511297 |
| arabidopsis thaliana pattern recognition receptors for bacterial elongation factor tu and flagellin can be combined to form functional chimeric receptors. | the receptor kinase efr of arabidopsis thaliana detects the microbe-associated molecular pattern elf18, a peptide that represents the n terminus of bacterial elongation factor tu. here, we tested subdomains of efr for their importance in receptor function. transient expression of tagged versions of efr and efr lacking its cytoplasmic domain in leaves of nicotiana benthamiana resulted in functional binding sites for elf18. no binding of ligand was found with the ectodomain lacking the transmembra ... | 2010 | 20410299 |
| interfamily transfer of a plant pattern-recognition receptor confers broad-spectrum bacterial resistance. | plant diseases cause massive losses in agriculture. increasing the natural defenses of plants may reduce the impact of phytopathogens on agricultural productivity. pattern-recognition receptors (prrs) detect microbes by recognizing conserved pathogen-associated molecular patterns (pamps). although the overall importance of pamp-triggered immunity for plant defense is established, it has not been used to confer disease resistance in crops. we report that activity of a prr is retained after its tr ... | 2010 | 20231819 |
| multiple receptor complexes assembled for transmitting clv3 signaling in arabidopsis. | in arabidopsis, the feedback regulatory loop between clavata3 (clv3) signaling pathway and transcription factor, wuschel (wus) plays a significant role in shoot apical meristems (sam) maintenance. previously, clv1/clv2 heterodimers were supposed to perceive and transmit clv3 signaling. recent genetic analysis isolated a novel receptor kinase, coryne (crn), which was found to be involved in the clv3 pathway. therefore, new hypothesis was put forward that crn probably acts with clv2 to transmit cl ... | 2010 | 20220313 |
| a host rna helicase-like protein, atrh8, interacts with the potyviral genome-linked protein, vpg, associates with the virus accumulation complex, and is essential for infection. | the viral genome-linked protein, vpg, of potyviruses is a multifunctional protein involved in viral genome translation and replication. previous studies have shown that both eukaryotic translation initiation factor 4e (eif4e) and eif4g or their respective isoforms from the eif4f complex, which modulates the initiation of protein translation, selectively interact with vpg and are required for potyvirus infection. here, we report the identification of two dead-box rna helicase-like proteins, ppddx ... | 2010 | 19880609 |
| defining the determinants for dual targeting of amino acyl-trna synthetases to mitochondria and chloroplasts. | most of the organellar amino acyl-trna synthetases (aarss) are dually targeted to both mitochondria and chloroplasts using dual targeting peptides (dtps). we have investigated the targeting properties and domain structure of dtps of seven aarss by studying the in vitro and in vivo import of n-terminal deleted constructs of dtps fused to green fluorescent protein. the deletion constructs were designed based on prediction programs, targetp and predotar, as well as logoplots derived from organellar ... | 2009 | 19733576 |
| melandrium yellow fleck bromovirus infects arabidopsis thaliana and has genomic rna sequence characteristics that are unique among bromoviruses. | melandrium yellow fleck bromovirus (myfv) systemically infected arabidopsis thaliana, although the susceptibility of several a. thaliana accessions to myfv differed from their susceptibility to the other two bromoviruses infecting a. thaliana. we constructed full-length cdna clones of myfv genomic rnas 1, 2, and 3 and determined their complete nucleotide sequences. similar to broad bean mottle bromovirus, (1) the 5'-terminal nucleotide of the myfv genomic rnas was adenine, and (2) the "d-arm" wa ... | 2009 | 19597694 |
| hlmyb3, a putative regulatory factor in hop (humulus lupulus l.), shows diverse biological effects in heterologous transgenotes. | a hop-specific cdna library from glandular tissue-enriched hop cones was screened for myb transcription factors. cdna encoding for r2r3 myb, designated hlmyb3, was cloned and characterized. according to the amino acid (aa) sequence, hlmyb3 shows the highest homology to ghmyb5 from cotton and is unrelated to the previously characterized hlmyb1 from the hop. southern blot analyses indicated that hlmyb3 is a unique gene, which was detected in various humulus lupulus cultivars, but not in humulus ja ... | 2007 | 17708645 |
| spatio-temporal accumulation of micrornas is highly coordinated in developing plant tissues. | micrornas (mirnas) are an abundant class of small, endogenous non-protein-coding rnas, approximately 21 nucleotides in length, that modulate the expression of animal and plant target genes at the post-transcriptional level. recent work has shown that mirna-based gene regulation plays a crucial role in pathways involved in plant growth and development. however, knowledge about the timing and spatial regulation of plant mirna expression is still limited. here we used in situ analysis to demonstrat ... | 2006 | 16824182 |
| a novel thioredoxin h is secreted in nicotiana alata and reduces s-rnase in vitro. | thioredoxins type h are classified into three subgroups. the subgroup ii includes thioredoxins containing an n-terminal extension, the role of which is still unclear. although thioredoxin secretion has been observed in animal cells, there is no evidence suggesting that any thioredoxin h is secreted in plants. in this study, we report that a thioredoxin h, subgroup ii, from nicotiana alata (natrxh) is secreted into the extracellular matrix of the stylar transmitting tract tissue. fractionation st ... | 2006 | 16354655 |
| rapid one-step protein purification from plant material using the eight-amino acid strepii epitope. | beyond the rewards of plant genome analysis and gene identification, characterisation of protein activities, post-translational modifications and protein complex composition remains a challenge for plant biologists. ideally, methods should allow rapid isolation of proteins from plant material achieving a high degree of purity. we tested three purification strategies based on the eight-amino acid strepii, six-amino acid his(6) and 181-amino acid tandem affinity purification (tap) affinity tags fo ... | 2004 | 15604670 |
| analysis of thermal stress-mediated pstvd variation and biolistic inoculation of progeny of viroid "thermomutants" to tomato and brassica species. | thermal stress of pstvd-infected nicotiana benthamiana led to appearance of a broad pstvd sequence distribution, where most of mutations accumulated in the left half of the viroid's secondary structure including the "pathogenicity" domain. a similar effect had been reported for hop latent viroid [virology 287 (2001) 349]. the pool of viroid "thermomutants" progenies was transcribed into cdna and used for biolistic inoculation of raphanus sativa, where the pstvd infection was detectable by revers ... | 2004 | 15165815 |
| a chemical-regulated inducible rnai system in plants. | constitutive expression of an intron-containing self-complementary 'hairpin' rna (ihprna) has recently been shown to efficiently silence target genes in transgenic plants. however, this technique cannot be applied to genes whose silencing may block plant regeneration or result in embryo lethality. to obviate these potential problems, we have used a chemical-inducible cre/loxp (clx) recombination system to trigger the expression of an intron-containing inverted-repeat rna (rnai) in plants. a deta ... | 2003 | 12713544 |
| nuclear localization of the plant protein ser/thr phosphatase pp7. | recently we identified novel plant ser/thr phosphatases, termed pp7, which belong to the ppp family and have no known close homologs in other kingdoms. we now addressed the intracellular location of arabidopsis thaliana pp7 using gfp fusions and confocal laser scanning microscopy. pp7. gfp fusion was expressed transiently or stably in nicotiana benthamiana. pp7. gfp was found to be a predominantly nuclear protein. effects of cytoskeleton-disrupting drugs indicate that cytoskeleton may be require ... | 2001 | 11703093 |
| nonsense and missense translational suppression in plant cells mediated by trna(lys). | a nuclear trna(lys) gene from arabidopsis thaliana was cloned and mutated so as to express trnas with altered anticodons which bind to a uag nonsense (amber) codon and to the arg (agg), asn (aac,aat), gln (cag) or glu (gag) codons. concomitantly, a codon in the firefly luciferase gene for a functionally important lys was altered to an amber codon, or to arg, asn, gln, glu, thr and trp codons, so as to construct reporter genes reliant upon incorporation of lys. the altered trna(lys) and luciferas ... | 1998 | 9484471 |
| identification of low ca(2+) stress-induced embryo apoptosis response genes in arachis hypogaea by ssh-associated library lift (sshall). | calcium is a universal signal in the regulation of wide aspects in biology, but few are known about the function of calcium in the control of early embryo development. ca(2+) deficiency in soil induces early embryo abortion in peanut, producing empty pods, which is a general problem; however, the underlying mechanism remains unclear. in this study, embryo abortion was characterized to be caused by apoptosis marked with cell wall degradation. using a method of ssh cdna libraries associated with l ... | 2016 | 26079063 |
| techno-economic analysis of horseradish peroxidase production using a transient expression system in nicotiana benthamiana. | despite the advantages of plant-based transient expression systems relative to microbial or mammalian cell systems, the commercial production of recombinant proteins using plants has not yet been achieved to any significant extent. one of the challenges has been the lack of published data on the costs of manufacture for products other than biopharmaceuticals. in this study, we report on the techno-economic analysis of the production of a standard commercial enzyme, namely, horseradish peroxidase ... | 2015 | 25344434 |
| transient production of artemisinin in nicotiana benthamiana is boosted by a specific lipid transfer protein from a. annua. | our lack of full understanding of transport and sequestration of the heterologous products currently limit metabolic engineering in plants for the production of high value terpenes. for instance, although all genes of the artemisinin/arteannuin b (an/ab) biosynthesis pathway (an-pw) from artemisia annua have been identified, ectopic expression of these genes in nicotiana benthamiana yielded mostly glycosylated pathway intermediates and only very little free (dihydro)artemisinic acid [(dh)aa]. he ... | 2016 | 27421621 |
| the metabolite chemotype of nicotiana benthamiana transiently expressing artemisinin biosynthetic pathway genes is a function of cyp71av1 type and relative gene dosage. | artemisia annua, which produces the anti-malaria compound artemisinin, occurs as high-artemisinin production (hap) and low-artemisinin production (lap) chemotypes. understanding the basis of the difference between these chemotypes would assist breeding and optimising artemisinin biosynthesis. here we present a systematic comparison of artemisinin biosynthesis genes that may be involved in determining the chemotype (cyp71av1, dbr2 and aldh1). these genes were isolated from the two chemotypes and ... | 2013 | 23638869 |
| the conformation of a plasma membrane-localized somatic embryogenesis receptor kinase complex is altered by a potato aphid-derived effector. | somatic embryogenesis receptor kinases (serks) are transmembrane receptors involved in plant immunity. tomato (solanum lycopersicum) carries three serk members. one of these, slserk1, is required for mi-1.2-mediated resistance to potato aphids (macrosiphum euphorbiae). mi-1.2 encodes a coiled-coil nucleotide-binding leucine-rich repeat protein that in addition to potato aphids confers resistance to two additional phloem-feeding insects and to root-knot nematodes (meloidogyne spp.). how slserk1 p ... | 2016 | 27208261 |
| identification and genome organization of saponin pathway genes from a wild crucifer, and their use for transient production of saponins in nicotiana benthamiana. | the ability to evolve novel metabolites has been instrumental for the defence of plants against antagonists. a few species in the barbarea genus are the only crucifers known to produce saponins, some of which make plants resistant to specialist herbivores, like plutella xylostella, the diamondback moth. genetic mapping in barbarea vulgaris revealed that genes for saponin biosynthesis are not clustered but are located in different linkage groups. using co-location with quantitative trait loci (qt ... | 2015 | 26333142 |
| rna1-independent replication and gfp expression from tomato marchitez virus isolate m cloned cdna. | tomato marchitez virus (tomarv; synonymous with tomato apex necrosis virus) is a positive-strand rna virus in the genus torradovirus within the family secoviridae. tomarv is an emergent whitefly-transmitted virus that causes important diseases in tomato (solanum lycopersicum) in mexico. here, the genome sequence of the tomarv isolate m (tomarv-m) was determined. we engineered full-length cdna clones of the tomarv-m genomic rna (rna1 and rna2), separately, into a binary vector. coinfiltration of ... | 2016 | 26756828 |
| diverse population of a new bipartite begomovirus infecting tomato crops in uruguay. | several isolates of a novel begomovirus were characterized from tomato samples collected in northern uruguay exhibiting disease symptoms associated with bemisia tabaci infestations. analysis of full-length sequences of dna-a and dna-b components revealed the presence of a new begomovirus with the typical genome organization of a new world begomovirus, for which the name tomato rugose yellow leaf curl virus (torylcv) is proposed. a high degree of nucleotide sequence diversity was found for both c ... | 2012 | 22383056 |
| a mutation in the lettuce infectious yellows virus minor coat protein disrupts whitefly transmission but not in planta systemic movement. | the lettuce infectious yellows virus (liyv) rna 2 mutant p1-5b was previously isolated from bemisia tabaci-transmitted virus maintained in chenopodium murale plants. p1-5b rna 2 contains a single-nucleotide deletion in the minor coat protein (cpm) open reading frame (orf) that is predicted to result in a frameshift and premature termination of the protein. using the recently developed agroinoculation system for liyv, we tested rna 2 containing the p1-5b cpm mutant genotype (agro-pr6-5b) in nicot ... | 2010 | 20861267 |
| bean dwarf mosaic virus: a model system for the study of viral movement. | bean dwarf mosaic virus-[colombia:1987] (bdmv-[co:87]) is a single-stranded plant dna virus, a member of the genus begomovirus of the family geminiviridae. | 2010 | 20618704 |
| agroinoculation of the crinivirus, lettuce infectious yellows virus, for systemic plant infection. | lettuce infectious yellows virus (liyv) is phloem-limited, non-mechanically transmissible, and is transmitted to plants only by bemisia tabaci. here, we developed agroinoculation to deliver liyv to plants thereby obviating the need for b. tabaci. agroinfiltration of rna 1 containing a green fluorescent protein gene into nicotiana benthamiana leaves resulted in subliminal infections, as judged by green fluorescence. agroinfiltration of liyv wild-type rna 1 and 2 constructs resulted in systemic in ... | 2009 | 19632699 |
| the infectivity of dimeric potato yellow mosaic geminivirus clones in different hosts. | head-to-tail dimeric clones of both dna a and dna b of potato yellow mosaic geminivirus (pymv) were constructed. these constructs were infectious when inoculated onto nicotiana benthamiana plants either as dna or by agroinoculation and were also infectious for tomato plants by agroinoculation. the dimers were not infectious for potato plants following inoculation by either method. symptom induction required both dna a and dna b but agroinoculation with dna a alone resulted in virus spread in 30% ... | 1994 | 7931179 |
| molecular cloning and characterization of the promoter for the multiple stress-inducible gene bjchi1 from brassica juncea. | we have previously isolated a brassica juncea cdna encoding a novel chitinase bjchi1 with two chitin-binding domains (zhao and chye in plant mol biol 40:1009-1018, 1999). the expression of bjchi1 was highly inducible by methyl jasmonate (meja) treatment, wounding, caterpillar feeding, and pathogenic fungal infection. these observations suggest that the promoter of bjchi1 gene might contain specific cis-acting elements for stress responses. here, we report the cloning and characterization of the ... | 2009 | 19277702 |
| methods to quantify pamp-triggered oxidative burst, map kinase phosphorylation, gene expression, and lignification in brassicas. | quantitative disease resistance (qdr) based on pamp-triggered immunity (pti) could be durable and effective against many pathogens (broad spectrum). development of methods to evaluate pti responses in crops could therefore accelerate breeding for durable qdr. most pti-studies involved model plants such as arabidopsis and nicotiana benthamiana or cell cultures, and cannot be directly applied to diverse germplasm of crop plants.we developed methods to measure pti in brassica crop species (lloyd et ... | 2017 | 28220438 |
| functional characterization of nac55 transcription factor from oilseed rape (brassica napus l.) as a novel transcriptional activator modulating reactive oxygen species accumulation and cell death. | nac transcription factors (tfs) are plant-specific and play important roles in development, responses to biotic and abiotic cues and hormone signaling. so far, only a few nac genes have been reported to regulate cell death. in this study, we identified and characterized a nac55 gene isolated from oilseed rape (brassica napus l.). bnanac55 responds to multiple stresses, including cold, heat, abscisic acid (aba), jasmonic acid (ja) and a necrotrophic fungal pathogen sclerotinia sclerotiorum. bnana ... | 2016 | 27312204 |
| the receptor-like kinase sobir1 interacts with brassica napus lepr3 and is required for leptosphaeria maculans avrlm1-triggered immunity. | the fungus leptosphaeria maculans (l. maculans) is the causal agent of blackleg disease of canola/oilseed rape (brassica napus) worldwide. we previously reported cloning of the b. napus blackleg resistance gene, lepr3, which encodes a receptor-like protein. lepr3 triggers localized cell death upon recognition of its cognate avr protein, avrlm1. here, we exploited the nicotiana benthamiana model plant to investigate the recognition mechanism of avrlm1 by lepr3. co-expression of the lepr3/avrlm1 g ... | 2015 | 26579176 |
| crystal structure of the effector avrlm4-7 of leptosphaeria maculans reveals insights into its translocation into plant cells and recognition by resistance proteins. | the avirulence gene avrlm4-7 of leptosphaeria maculans, the causal agent of stem canker in brassica napus (oilseed rape), confers a dual specificity of recognition by two resistance genes (rlm4 and rlm7) and is strongly involved in fungal fitness. in order to elucidate the biological function of avrlm4-7 and understand the specificity of recognition by rlm4 and rlm7, the avrlm4-7 protein was produced in pichia pastoris and its crystal structure was determined. it revealed the presence of four di ... | 2015 | 26082394 |
| enhanced seed oil content by overexpressing genes related to triacylglyceride synthesis. | oilseed rape (brassica napus) is one of the most important oilseed crops globally. to meet increasing demand for oil-based products, the ability to enhance desirable oil content in the seed is required. this study assessed the capability of five genes in the triacylglyceride (tag) synthesis pathway to enhance oil content. the genes bngpdh, bngpat, bndgat, scgpdh and sclpaat were overexpressed separately in a tobacco (nicotiana benthamiana) model system, and simultaneously by pyramiding in b. nap ... | 2015 | 25523093 |
| oxidation and phosphorylation of map kinase 4 cause protein aggregation. | mitogen-activated protein kinase (mpk) cascades are highly conserved signaling pathways that respond to environmental cues. arabidopsis mpk4 has been identified as a stress-responsive protein kinase. here we demonstrate that brassica napus mpk4 (bnmpk4) is activated by hydrogen peroxide (h2o2) and phytohormone abscisic acid (aba). transient expression of a constitutively active bnmpk4 causes h2o2 production and cell death in nicotiana benthamiana leaves. however, little is known about how h2o2 c ... | 2015 | 25433264 |
| role of a gdsl lipase-like protein as sinapine esterase in brassicaceae. | the seeds of most members of the brassicaceae accumulate high amounts of sinapine (sinapoylcholine) that is rapidly hydrolyzed during early stages of seed germination. one of three isoforms of sinapine esterase activity (bnsce3) has been isolated from brassica napus seedlings and subjected to trypsin digestion and spectrometric sequencing. the peptide sequences were used to isolate bnsce3 cdna, which was shown to contain an open reading frame of 1170 bp encoding a protein of 389 amino acids, inc ... | 2008 | 18036206 |
| pigeonpea sterility mosaic virus: a legume-infecting emaravirus from south asia. | pigeonpea sterility mosaic virus (ppsmv), a species of the genus emaravirus, is the causal agent of sterility mosaic disease (smd) of pigeonpea [cajanus cajan (l.) millsp]. this disease, dubbed the 'green plague', as the infected plants remain in the vegetative state without flower production, has been reported from india and a few other south-east asian countries. smd is estimated to result in an annual yield loss of over us$300 million in india alone. the aetiology of smd, which remained a mys ... | 2015 | 25640756 |
| a novel mite-transmitted virus with a divided rna genome closely associated with pigeonpea sterility mosaic disease. | abstract the agent of sterility mosaic, a disease that is a major constraint on pigeonpea (cajanus cajan) production in the indian subcontinent, is transmitted by the eriophyid mite, aceria cajani. this agent has remained elusive for decades despite intensive efforts but we report the isolation of highly flexuous filamentous virus-like particles (vlps) of 3 to 10 nm in width and of undefined lengths from sterility mosaic disease (smd)-affected pigeonpea plants. purified vlp preparations from vir ... | 2003 | 18944159 |
| superoxide serves as a putative signal molecule for plant cell division: overexpression of carlk1 promotes the plant cell cycle via accumulation of o2(-) and decrease in h2 o2. | reactive oxygen species (ros) exert both positive and negative effects on plant growth and development and therefore receive a great deal of attention in current research. a hot pepper, capsicum annuum receptor-like kinase 1 (carlk1) was ectopically expressed in nicotiana tabacum by-2 cell and nicotiana benthamiana plants. this ectopic expression of carlk1 enhanced cell division and proliferation in both heterologous systems. apparently, carlk1 is involved in controlling the cell cycle, possibly ... | 2017 | 27528370 |
| a common plant cell-wall protein hyprp1 has dual roles as a positive regulator of cell death and a negative regulator of basal defense against pathogens. | although hybrid proline-rich proteins (hyprps) are ubiquitous in plants, little is known about their roles other than as cell-wall structural proteins. we identified the gene hyprp1 in capsicum annuum and nicotiana benthamiana, which encodes a protein containing proline-rich domain and eight-cysteine motif (8cm) that is constitutively expressed in various organs, mostly in the root, but is down-regulated upon inoculation with either incompatible or compatible pathogens. ectopic expression of hyp ... | 2012 | 22023393 |
| 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 ... | 2013 | 23001197 |
| "self" and "non-self" in the control of phytoalexin biosynthesis: plant phospholipases a2 with alkaloid-specific molecular fingerprints. | the overproduction of specialized metabolites requires plants to manage the inherent burdens, including the risk of self-intoxication. we present a control mechanism that stops the expression of phytoalexin biosynthetic enzymes by blocking the antecedent signal transduction cascade. cultured cells of eschscholzia californica (papaveraceae) and catharanthus roseus (apocynaceae) overproduce benzophenanthridine alkaloids and monoterpenoid indole alkaloids, respectively, in response to microbial eli ... | 2015 | 25670767 |
| inhibitors of plant invertases do not affect the structurally related enzymes of fructan metabolism. | plant fructan active enzymes (fazys), including the enzymes involved in inulin metabolism, namely sucrose:sucrose 1-fructosyltransferase (1-sst; ec 2.4.1.99), fructan:fructan 1-fructosyltransferase (1-fft; ec 2.4.1.100) and fructan 1-exohydrolase (1-feh; ec 3.2.1.153), are evolutionarily related to acid invertases (ais), that is, plant cell wall invertase (cwi) and vacuolar invertase (vi). acid invertases are post-translationally controlled by proteinaceous inhibitors. whether fazys are subject ... | 2009 | 19037899 |
| metabolic engineering of medium-chain fatty acid biosynthesis in nicotiana benthamiana plant leaf lipids. | various research groups are investigating the production of oil in non-seed biomass such as leaves. recently, high levels of oil accumulation have been achieved in plant biomass using a combination of biotechnological approaches which also resulted in significant changes to the fatty acid composition of the leaf oil. in this study, we were interested to determine whether medium-chain fatty acids (mcfa) could be accumulated in leaf oil. mcfa are an ideal feedstock for biodiesel and a range of ole ... | 2015 | 25852716 |
| cloning, molecular characterization, and expression analysis of a nucleoporin gene (rgnup98-96) from rehmannia glutinosa. | nucleoporin 98 (nup98) and nucleoporin 96 (nup96) are essential components of the nuclear pore complex (npc) in eukaryote cells. however, there is a lack of available information about complete rehmannia glutinosa nup98-96 (rgnup98-96) sequences. here, the full-length cdna sequence of rgnup96-98 was isolated from r. glutinosa using rapid amplification of cdna ends (race) technology, based on a cloned cdna sequence (genbank accession no. jz483329). the identified rgnup98-96 was 3476 bp, and it en ... | 2015 | 26505455 |
| plant expression systems for early stage discovery and development of lead therapeutic antibodies. | antibodies for human clinical applications are predominantly produced in mammalian expression systems, with chinese hamster ovary (cho) cells being the gold standard. cho cells are ideal for the manufacturing of the igg class of antibodies, but not for the production of complex antibodies like secretory igas (sigas) and igms, which remain unavailable for clinical use. in contrast, plant seeds and leaves hold the promise to produce sigas, igms and similar complex antibody formats to scalable amou ... | 2015 | 27472860 |
| functional characterization of csbglu12, a β-glucosidase from crocus sativus, provides insights into its role in abiotic stress through accumulation of antioxidant flavonols. | glycosylation and deglycosylation are impressive mechanisms that allow plants to regulate the biological activity of an array of secondary metabolites. although glycosylation improves solubility and renders the metabolites suitable for transport and sequestration, deglycosylation activates them to carry out biological functions. herein, we report the functional characterization of csbglu12, a β-glucosidase from crocus sativus. csbglu12 has a characteristic glucoside hydrolase 1 family (α/β)8 tri ... | 2017 | 28154174 |
| genome-wide analysis reveals dynamic changes in expression of micrornas during vascular cambium development in chinese fir, cunninghamia lanceolata. | micrornas (mirnas) are small noncoding regulatory rnas that play key roles in the process of plant development. to date, extensive studies of mirnas have been performed in a few model plants, but few efforts have focused on small rnas (srnas) in conifers because of the lack of reference sequences for their enormous genomes. in this study, solexa sequencing of three srna libraries obtained from dormant, reactivating, and active vascular cambium in chinese fir (cunninghamia lanceolata) using tangen ... | 2015 | 25795740 |
| identification of the subgenomic promoter of the coat protein gene of cucumber fruit mottle mosaic virus and development of a heterologous expression vector. | heterologous gene expression using plant virus vectors enables research on host-virus interactions and the production of useful proteins, but the host range of plant viruses limits the practical applications of such vectors. here, we aimed to develop a viral vector based on cucumber fruit mottle mosaic virus (cfmmv), a member of the genus tobamovirus, whose members infect cucurbits. the subgenomic promoter (sgp) in the coat protein (cp) gene, which was used to drive heterologous expression, was ... | 2016 | 26976138 |
| effect of c-terminal deletions in the movement protein of cowpea chlorotic mottle virus on cell-to-cell and long-distance movement. | in order to elucidate the function of the c-terminal region of cowpea chlorotic mottle bromovirus (ccmv) movement protein (mp) in cell-to-cell movement, a set of deletions ranging from 10 to 80 amino acids (deltamp10, deltamp20, deltamp33, deltamp43, deltamp60 and deltamp80) was engineered into the mp gene encoded by the biologically active clone c3/deltacp-egfp, a variant of ccmv rna3 that contained wild-type (wt) mp and the enhanced green fluorescent protein (egfp) gene in place of the coat pr ... | 1999 | 10374951 |
| infectious transcripts of an asymptomatic panicovirus identified from a metagenomic survey. | a new panicovirus was identified from a metagenomic survey of plant viruses in the tallgrass prairie preserve (tpp), osage county, oklahoma, usa. thin paspalum asymptomatic virus (tpav) was isolated from paspalum setaceum var. muehlenbergii. the complete 4195 nucleotide sequence encodes six proteins of 44, 109, 8, 6.8, 26, and 15kda, whose genes are similar in size and location to those of panicum mosaic virus (pmv) and cocksfoot mild mosaic virus (cmmv). infectious transcripts made from cdnas o ... | 2013 | 23774065 |
| identification of a new tospovirus causing necrotic ringspot on tomato in china. | emerging tospoviruses cause significant yield losses and quality reduction in vegetables, ornamentals, and legumes throughout the world. so far, eight tospoviruses were reported in china. tomato fruits displaying necrotic and concentric ringspot symptoms were found in guizhou province of southwest china. | 2014 | 25465801 |
| engineering of sialylated mucin-type o-glycosylation in plants. | proper n- and o-glycosylation of recombinant proteins is important for their biological function. although the n-glycan processing pathway of different expression hosts has been successfully modified in the past, comparatively little attention has been paid to the generation of customized o-linked glycans. plants are attractive hosts for engineering of o-glycosylation steps, as they contain no endogenous glycosyltransferases that perform mammalian-type ser/thr glycosylation and could interfere w ... | 2012 | 22948156 |
| comparative transcriptome analysis of three oil palm fruit and seed tissues that differ in oil content and fatty acid composition. | oil palm (elaeis guineensis) produces two oils of major economic importance, commonly referred to as palm oil and palm kernel oil, extracted from the mesocarp and the endosperm, respectively. while lauric acid predominates in endosperm oil, the major fatty acids (fas) of mesocarp oil are palmitic and oleic acids. the oil palm embryo also stores oil, which contains a significant proportion of linoleic acid. in addition, the three tissues display high variation for oil content at maturity. to gain ... | 2013 | 23735505 |
| functional analysis of the naturally recombinant dna-a of the bipartite begomovirus tomato chlorotic mottle virus. | all geminiviruses found in brazil belong to the begomovirus genus with a bipartite genome that is split between two genomic components, dna-a and dna-b. the dna-a of the bipartite begomovirus tocmov-[mg-bt] (tomato chlorotic mottle virus), however, possesses as a peculiar characteristic the capacity to systemically infect nicotiana benthamiana. here we further characterize this variant dna-a and show that it also infects solanum lycopersicum and other host plants, in the absence of dna-b. the to ... | 2007 | 17367887 |
| a plant-produced bacteriophage tailspike protein for the control of salmonella. | the receptor binding domain of the tailspike protein gp9 from the p22 bacteriophage was recently shown to reduce salmonella colonization in the chicken gut. in this study, we transiently expressed the receptor binding domain of the gp9 tailspike protein in nicotiana benthamiana, and targeted it to the endoplasmic reticulum (er) or to the chloroplasts. gp9 was also fused to either an elastin-like polypeptide (elp) or hydrophobin i tag, which were previously described to improve accumulation level ... | 2015 | 26779243 |
| plant-produced recombinant influenza vaccine based on virus-like hbc particles carrying an extracellular domain of m2 protein. | conventional influenza vaccines are based on a virus obtained in chicken embryos or its components. the high variability of the surface proteins of influenza virus, hemagglutinin and neuraminidase, requires strain-specific vaccines matching the antigenic specificity of newly emerging virus strains to be developed. a recombinant vaccine based on a highly conservative influenza virus protein m2 fused to a nanosized carrier particle can be an attractive alternative to traditional vaccines. we have ... | 2012 | 22339631 |
| heterologous expression of moss light-harvesting complex stress-related 1 (lhcsr1), the chlorophyll a-xanthophyll pigment-protein complex catalyzing non-photochemical quenching, in nicotiana sp. | oxygenic photosynthetic organisms evolved mechanisms for thermal dissipation of energy absorbed in excess to prevent formation of reactive oxygen species. the major and fastest component, called non-photochemical quenching, occurs within the photosystem ii antenna system by the action of two essential light-harvesting complex (lhc)-like proteins, photosystem ii subunit s (psbs) in plants and light-harvesting complex stress-related (lhcsr) in green algae and diatoms. in the evolutionary intermedi ... | 2015 | 26260788 |
| 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 ... | 2015 | 22989991 |
| genome-wide analysis and expression profiling of the phospholipase c gene family in soybean (glycine max). | phosphatidylinositol-specific phospholipase c (pi-plc) hydrolyses phosphatidylinositol-4,5-bisphosphate to produce diacylglycerol and inositol 1,4,5-trisphosphate. it plays an important role in plant development and abiotic stress responses. however, systematic analysis and expression profiling of the phospholipase c (plc) gene family in soybean have not been reported. in this study, 12 putative plc genes were identified in the soybean genome. soybean plcs were found on chromosomes 2, 11, 14 and ... | 2015 | 26421918 |
| identification of photoperiod-regulated gene in soybean and functional analysis in nicotiana benthamiana. | soybean (glycine max) is a short-day crop and the photoperiod is a crucial factor regulating its flowering time. to investigate the molecular mechanism controlling the flowering time by photoperiod in soybean, cdna-amplified fragment length polymorphism (cdna-aflp) was used to identify photoperiod-regulated genes in leaves of soybean growing under short-day length, neutral photoperiod and long-day length. a total of 36 transcript-derived fragments (tdfs) were identified to be regulated by photop ... | 2014 | 24840822 |
| 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 ... | 2013 | 23483290 |
| molecular cloning and characterization of enhanced disease susceptibility 1 (eds1) from gossypium barbadense. | arabidopsis enhanced disease susceptibility 1 (eds1) plays an important role in plant defense against biotrophic and necrotrophic pathogens. the necrotrophic pathogen verticillium dahliae infection of gossypium barbadense could lead to verticillium wilt which seriously reduces the cotton production. here, we cloned and characterized a g. barbadense homolog of eds1, designated as gbeds1. the full-length cdna of the gbeds1 gene was obtained by the technique of rapid-amplification of cdna ends. the ... | 2014 | 24570021 |
| genome-wide identification of bahd acyltransferases and in vivo characterization of hqt-like enzymes involved in caffeoylquinic acid synthesis in globe artichoke. | globe artichoke (cynara cardunculus l. var. scolymus) is a rich source of compounds promoting human health (phytonutrients), among them caffeoylquinic acids (cqas), mainly represented by chlorogenic acid (cga), and dicaffeoylquinic acids (dicqas). the enzymes involved in their biosynthesis belong to the large family of bahd acyltransferases. following a survey of the globe artichoke genome, we identified 69 bahd proteins carrying the catalytic site (hxxxd). their phylogenetic analysis together w ... | 2016 | 27721818 |
| accumulation of cynaropicrin in globe artichoke and localization of enzymes involved in its biosynthesis. | globe artichoke (cynara cardunculus var. scolymus) belongs to the asteraceae family, in which one of the most biologically significant class of secondary metabolites are sesquiterpene lactones (stls). in globe artichoke the principal stl is the cynaropicrin, which contributes to approximately 80% of its characteristic bitter taste. cynaropicrin content was assessed in globe artichoke tissues and was observed to accumulate in leaves of different developmental stages. in the receptacle, a progress ... | 2015 | 26398797 |
| the cotton wrky gene ghwrky41 positively regulates salt and drought stress tolerance in transgenic nicotiana benthamiana. | wrky transcription factors constitute a very large family of proteins in plants and participate in modulating plant biological processes, such as growth, development and stress responses. however, the exact roles of wrky proteins are unclear, particularly in non-model plants. in this study, gossypium hirsutum wrky41 (ghwrky41) was isolated and transformed into nicotiana benthamiana. our results showed that overexpression of ghwrky41 enhanced the drought and salt stress tolerance of transgenic ni ... | 2015 | 26562293 |
| overexpression of ghwrky27a reduces tolerance to drought stress and resistance to rhizoctonia solani infection in transgenic nicotiana benthamiana. | wrky proteins constitute transcriptional regulators involved in various biological processes, especially in coping with diverse biotic and abiotic stresses. however, in contrast to other well-characterized wrky groups, the functions of group iii wrky transcription factors are poorly understood in the economically important crop cotton (gossypium hirsutum). in this study, a group iii wrky gene from cotton, ghwrky27a, was isolated and characterized. our data indicated that ghwrky27a localized to t ... | 2015 | 26483697 |
| engineering the anthocyanin regulatory complex of strawberry (fragaria vesca). | the woodland strawberry, fragaria vesca is a model fruit for a number of rosaceous crops. we have engineered altered concentrations of anthocyanin in f. vesca, to determine the impact on plant growth and fruit quality. anthocyanin concentrations were significantly increased by over-expression or decreased by knock-down of the r2r3 myb activator, myb10. in contrast, a potential bhlh partner for myb10 (bhlh33) did not affect the anthocyanin pathway when knocked down using rnai constructs. metaboli ... | 2014 | 25477896 |
| the cotton mitogen-activated protein kinase kinase 3 functions in drought tolerance by regulating stomatal responses and root growth. | mitogen-activated protein kinase (mapk) cascades play critical roles in signal transduction processes in eukaryotes. the mapk kinases (mapkks) that link mapkk kinases (mapkkks) and mapks are key components of mapk cascades. however, the intricate regulatory mechanisms that control mapkks under drought stress conditions are not fully understood, especially in cotton (gossypium hirsutum) here, we isolated and characterized the cotton group b mapkk gene ghmkk3 overexpressing ghmkk3 in nicotiana ben ... | 2016 | 27335349 |
| the cotton wrky transcription factor ghwrky17 functions in drought and salt stress in transgenic nicotiana benthamiana through aba signaling and the modulation of reactive oxygen species production. | drought and high salinity are two major environmental factors that significantly limit the productivity of agricultural crops worldwide. wrky transcription factors play essential roles in the adaptation of plants to abiotic stresses. however, wrky genes involved in drought and salt tolerance in cotton (gossypium hirsutum) are largely unknown. here, a group iid wrky gene, ghwrky17, was isolated and characterized. ghwrky17 was found to be induced after exposure to drought, salt, h2o2 and aba. the ... | 2014 | 25261532 |
| cloning and functional analysis of the promoter of an ascorbate oxidase gene from gossypium hirsutum. | apoplastic ascorbate oxidase (ao) plays significant roles in plant cell growth. however, the mechanism of underlying the transcriptional regulation of ao in gossypium hirsutum remains unclear. here, we obtained a 1,920-bp promoter sequence from the gossypium hirsutum ascorbate oxidase (ghao1) gene, and this ghao1 promoter included a number of known cis-elements. promoter activity analysis in overexpressing pghao1::gfp-gus tobacco (nicotiana benthamiana) showed that the ghao1 promoter exhibited h ... | 2016 | 27597995 |