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isolation and functional characterization of a novel seed-specific promoter region from peanut.the importance of using tissue-specific promoters in the genetic transformation of plants has been emphasized increasingly. here, we report the isolation of a novel seed-specific promoter region from peanut and its validation in arabidopsis and tobacco seeds. the reported promoter region referred to as groundnut seed promoter (gsp) confers seed-specific expression in heterologous systems, which include putative promoter regions of the peanut (arachis hypogaea l.) gene 8a4r19g1. this region was i ...201424078220
production and secretion of a heterologous protein by turnip hairy roots with superiority over tobacco hairy roots.a fully contained and efficient heterologous protein production system was designed using brassica rapa rapa (turnip) hairy roots. two expression cassettes containing a cauliflower mosaic virus (camv) 35s promoter with a duplicated enhancer region, an arabidopsis thaliana sequence encoding a signal peptide and the camv polyadenylation signal were constructed. one cassette was used to express the green fluorescent protein (gfp)-encoding gene in hairy roots grown in flasks. a stable and fast-growi ...201424078130
overexpression of artemisinic aldehyde δ11 (13) reductase gene-enhanced artemisinin and its relative metabolite biosynthesis in transgenic artemisia annua l.artemisinic aldehyde δ11 (13) reductase (dbr2) is the checkpoint enzyme catalyzing artemisinic aldehyde to form dihydroartemisinic aldehyde directly involved in artemisinin biosynthetic pathway. in the present study, dbr2 was employed to engineer the biosynthetic pathway of artemisinin in transgenic plants of artemisia annua l. seven independent transgenic plants of a. annua with dbr2 overexpression driven by the cauliflower mosaic virus 35s promoter were obtained by agrobacterium-mediated genet ...201525040292
symptoms induced by transgenic expression of p23 from citrus tristeza virus in phloem-associated cells of mexican lime mimic virus infection without the aberrations accompanying constitutive expression.citrus tristeza virus (ctv) is phloem restricted in natural citrus hosts. the 23-kda protein (p23) encoded by the virus is an rna silencing suppressor and a pathogenicity determinant. the expression of p23, or its n-terminal 157-amino-acid fragment comprising the zinc finger and flanking basic motifs, driven by the constitutive 35s promoter of cauliflower mosaic virus, induces ctv-like symptoms and other aberrations in transgenic citrus. to better define the role of p23 in ctv pathogenesis, we c ...201525171669
transgenic arabidopsis thaliana plants expressing a β-1,3-glucanase from sweet sorghum (sorghum bicolor l.) show reduced callose deposition and increased tolerance to aluminium toxicity.seventy-one cultivars of sweet sorghum (sorghum bicolor l.) were screened for aluminium (al) tolerance by measuring relative root growth (rrg). two contrasting cultivars, roma (al tolerant) and potchetstrm (al sensitive), were selected to study shorter term responses to al stress. potchetstrm had higher callose synthase activity, lower β-1,3-glucanase activity and more callose deposition in the root apices during al treatment compared with roma. we monitored the expression of 12 genes involved i ...201525311645
characterization of a small constitutive promoter from arabidopsis translationally controlled tumor protein (attctp) gene for plant transformation.a plant-derived 0.3 kb constitutive promoter was obtained from attctp expression analysis, and successfully applied to the expression of a selectable marker gene for production of transgenic creeping bentgrass plants. the isolation and use of an efficient promoter is essential to develop a vector system for efficient genetic transformation of plants, and constitutive promoters are particularly useful for the expression of selectable marker genes. in this study, we characterized a small size of t ...201525410250
enhanced accumulation of carotenoids in sweetpotato plants overexpressing ibor-ins gene in purple-fleshed sweetpotato cultivar.sweetpotato [ipomoea batatas (l.) lam] is an important root crop that produces low molecular weight antioxidants such as carotenoids and anthocyanin. the sweetpotato orange (ibor) protein is involved in the accumulation of carotenoids. to increase the levels of carotenoids in the storage roots of sweetpotato, we generated transgenic sweetpotato plants overexpressing ibor-ins under the control of the cauliflower mosaic virus (camv) 35s promoter in an anthocyanin-rich purple-fleshed cultivar (refe ...201525438140
development of gateway binary vector series with four different selection markers for the liverwort marchantia polymorpha.we previously reported agrobacterium-mediated transformation methods for the liverwort marchantia polymorpha using the hygromycin phosphotransferase gene as a marker for selection with hygromycin. in this study, we developed three additional markers for m. polymorpha transformation: the gentamicin 3'-acetyltransferase gene for selection with gentamicin; a mutated acetolactate synthase gene for selection with chlorsulfuron; and the neomycin phosphotransferase ii gene for selection with g418. base ...201526406247
overexpression of plastid transketolase in tobacco results in a thiamine auxotrophic phenotype.to investigate the effect of increased plastid transketolase on photosynthetic capacity and growth, tobacco (nicotiana tabacum) plants with increased levels of transketolase protein were produced. this was achieved using a cassette composed of a full-length arabidopsis thaliana transketolase cdna under the control of the cauliflower mosaic virus 35s promoter. the results revealed a major and unexpected effect of plastid transketolase overexpression as the transgenic tobacco plants exhibited a sl ...201525670766
effect of orientation of transcription of a gene in an inverted transferred dna repeat on transcriptional gene silencing in rice transgenics-a case study.we studied transgene silencing in two transgenic rice plants, osm25 and cot-osm4, which harboured two different types of right border (rb)-centered inverted transferred dna (t-dna) repeats (irs). the t-dna in osm25 has three genes gus, osm and hph, all under the transcriptional control of the cauliflower mosaic virus 35s promoter (p35s). the gus gene, which is proximal to the rb, is in a convergent orientation of transcription in the ir. osm25 displayed silencing of all three transgenes. nuclear ...201525648206
methyl-cpg-binding domain protein mbd7 is required for active dna demethylation in arabidopsis.although researchers have established that dna methylation and active demethylation are dynamically regulated in plant cells, the molecular mechanism for the regulation of active dna demethylation is not well understood. by using an arabidopsis (arabidopsis thaliana) line expressing the promoter responsive to dehydration 29a:luciferase (prord29a:luc) and promoter cauliflower mosaic virus 35s:neomycin phosphotransferase ii (pro35s:nptii) transgenes, we isolated an mbd7 (for methyl-cpg-binding dom ...201525593350
impact of cis-acting elements' frequency in transcription activity in dicot and monocot plants.the production of new cultivars via recombinant dna technology is important in applied agriculture. promoters play fundamental roles in successful transformation and gene expression. fragments of the upstream regulatory region of the movement protein gene of the tomato yellow leaf curl virus (tylcv; two fragments) and watermelon chlorotic stunt virus (wmcsv, two fragments) and one fragment of the coat protein putative promoter of tylcv (cpty-pro) were isolated to assess their abilities to drive ...201528324408
migration of plant viruses: time correlations with the agriculture history and human immigration.in this review, i made the phylodynamic comparisons of three plant viruses, turnip mosaic virus (tumv), cauliflower mosaic virus (camv) and cucumber mosaic virus (cmv), using the genomic sequences of a large numbers of isolates collected worldwide. we analyzed these genomic nucleotide sequences, in combination with published sequences, to estimate the timescale and rate of evolution of the individual genes of tumv, camv and cmv. the main hosts of the viruses are brassicaceae crops. we also compa ...201527760921
mutations within a 35 amino acid region of p6 influence self-association, inclusion body formation, and caulimovirus infectivity.cauliflower mosaic virus gene vi product (p6) is an essential protein that forms cytoplasmic, inclusion bodies (ibs). p6 contains four regions involved in self-association, termed d1-d4. d3 binds to d1, along with d4 and contains a spacer region (termed d3b) between two rna-binding domains. here we show d3b binds full-length p6 along with d1 and d4. full-length p6s harboring single amino acid substitutions within d3b showed reduced binding to both d1 and d4. full-length p6s containing d3b mutati ...201525506670
isolation and characterization of an ubiquitin extension protein gene (jcuep) promoter from jatropha curcas.the jcuep promoter is active constitutively in the bio-fuel plant jatropha curcas , and is an alternative to the widely used camv35s promoter for driving constitutive overexpression of transgenes in jatropha. well-characterized promoters are required for transgenic breeding of jatropha curcas, a biofuel feedstock with great potential for production of bio-diesel and bio-jet fuel. in this study, an ubiquitin extension protein gene from jatropha, designated jcuep, was identified to be ubiquitously ...201525502690
development of three full-length infectious cdna clones of distinct brassica yellows virus genotypes for agrobacterium-mediated inoculation.brassica yellows virus is a newly identified species in the genus of polerovirus within the family luteoviridae. brassica yellows virus (bryv) is prevalently distributed throughout mainland china and south korea, is an important virus infecting cruciferous crops. based on six bryv genomic sequences of isolates from oilseed rape, rutabaga, radish, and cabbage, three genotypes, bryv-a, bryv-b, and bryv-c, exist, which mainly differ in the 5' terminal half of the genome. bryv is an aphid-transmitte ...201525499296
priming with a double-stranded dna virus alters brassica rapa seed architecture and facilitates a defense response.abiotic and biotic stresses alter genome stability and physiology of plants. under some stressful situations, a state of stress tolerance can be passed on to the offspring rendering them more suitable to stressful events than their parents. in plants, the exploration of transgenerational response has remained exclusive to model species, such as arabidopsis thaliana. here, we expand transgenerational research to include brassica rapa, a close relative to economically important plant canola (brass ...201525498335
water-soluble chlorophyll-binding proteins from arabidopsis thaliana and raphanus sativus target the endoplasmic reticulum body.non-photosynthetic chlorophyll (chl) proteins called water-soluble chl-binding proteins are distributed in brassicaceae plants. brassica oleracea wscp (bowscp) and lepidium virginicum wscp (lvwscp) are highly expressed in leaves and stems, while arabidopsis thaliana wscp (atwscp) and raphanus sativus wscp (rshwscp) are highly transcribed in floral organs. bowscp and lvwscp exist in the endoplasmic reticulum (er) body. however, the subcellular localization of atwscp and rshwscp is still unclear. ...201526289422
dna extraction techniques compared for accurate detection of genetically modified organisms (gmos) in maize food and feed products.in this paper, dna extraction methods have been evaluated to detect the presence of genetically modified organisms (gmos) in maize food and feed products commercialised in turkey. all the extraction methods tested performed well for the majority of maize foods and feed products analysed. however, the highest dna content was achieved by the wizard, genespin or the ctab method, all of which produced optimal dna yield and purity for different maize food and feed products. the samples were then scre ...201526243938
retraction for moissiard and voinnet, rna silencing of host transcripts by cauliflower mosaic virus requires coordinated action of the four arabidopsis dicer-like proteins. 201526216957
targeting helicase-dependent amplification products with an electrochemical genosensor for reliable and sensitive screening of genetically modified organisms.cultivation of genetically modified organisms (gmos) and their use in food and feed is constantly expanding; thus, the question of informing consumers about their presence in food has proven of significant interest. the development of sensitive, rapid, robust, and reliable methods for the detection of gmos is crucial for proper food labeling. in response, we have experimentally characterized the helicase-dependent isothermal amplification (hda) and sequence-specific detection of a transgene from ...201526198403
cauliflower mosaic virus transcriptome reveals a complex alternative splicing pattern.the plant pararetrovirus cauliflower mosaic virus (camv) uses alternative splicing to generate several isoforms from its polycistronic pregenomic 35s rna. this pro-cess has been shown to be essential for infectivity. previous works have identified four splice donor sites and a single splice acceptor site in the 35s rna 5' region and suggested that the main role of camv splicing is to downregulate expression of open reading frames (orfs) i and ii. in this study, we show that alternative splicing ...201526162084
complete genome sequence and construction of infectious full-length cdna clones of tobacco ringspot nepovirus, a viral pathogen causing bud blight in soybean.tobacco ringspot virus (trsv, genus nepovirus), causes severe diseases in soybean and tobacco plants. trsv-induced bud blight disease significantly reduced both the yield and quality of soybeans. the function of the encoded viral gene product involved in trsv infection was unclear due to the limitation of reverse genetics studies on the viral genome. here, we represent the successful construction of infectious full-length cdna clones of trsv genome (rna1 and rna2). the cdnas of trsv rna1 and rna ...201526159876
rna based viral silencing suppression in plant pararetroviruses.the 35s promoter of cauliflower mosaic virus and that of other plant pararetroviruses gives rise to an rna, which is both a pre-genome and a polycistronic mrna. the 600 nucleotide long very structured leader of this rna is also transcribed separately. the resulting 8s rna is then converted to a double strand giving rise to a huge set of sirnas, which suppress silencing. in this mini-review i discuss how this versatile stretch of 600 nts constitutes a masterpiece of evolution.201526113850
truncated ubiquitin 5' regulatory region from erianthus arundinaceus drives enhanced transgene expression in heterologous systems.characterization of novel plant gene promoters underpins the development of transgenic crop plants. here, we report a novel 5' regulatory sequence (eriubi d7) of the ubiquitin gene from erianthus arundinaceus, a wild relative of sugarcane resistant to many biotic and abiotic stresses. a 3.2-kb regulatory sequence of ubiquitin gene was isolated through random amplification of genomic ends technique and characterized in rice, tobacco, and sugarcane. in silico analysis revealed that the regulatory ...201525985900
altered fruit and seed development of transgenic rapeseed (brassica napus) over-expressing microrna394.fruit and seed development in plants is a complex biological process mainly involved in input and biosynthesis of many storage compounds such as proteins and oils. although the basic biochemical pathways for production of the storage metabolites in plants are well characterized, their regulatory mechanisms are not fully understood. in this study, we functionally identified rapeseed (brassica napus) mir394 with its target gene brassica napus leaf curling responsiveness (bnlcr) to dissect a role o ...201525978066
traealdh7b1-5a, encoding aldehyde dehydrogenase 7 in wheat, confers improved drought tolerance in arabidopsis.traealdh7b1 - 5a , encoding aldehyde dehydrogenase 7 in wheat, conferred significant drought tolerance to arabidopsis , supported by molecular biological and physiological experiments. drought stress significantly affects wheat yields. aldehyde dehydrogenase (aldh) is a family of enzymes catalyzing the irreversible conversion of aldehydes into acids to decrease the damage caused by abiotic stresses. however, no wheat aldh member has been functionally characterized to date. here, we obtained a di ...201525893867
functional characterization of a strong bi-directional constitutive plant promoter isolated from cotton leaf curl burewala virus.cotton leaf curl burewala virus (clcubuv), belonging to the genus begomovirus, possesses single-stranded monopartite dna genome. the bidirectional promoters representing rep and coat protein (cp) genes of clcubuv were characterized and their efficacy was assayed. rep and cp promoters of clcubuv and 35s promoter of cauliflower mosaic virus (camv) were fused with β-glucuronidase (gus) and green fluorescent protein (gfp) reporter genes. gus activity in individual plant cells driven by rep, cp and 3 ...201525799504
platinum anniversary: virus and lichen alga together more than 70 years.trebouxia aggregata (archibald) gärtner (phylum chlorophyta, family trebouxiaceae), a lichen symbiotic alga, has been identified as host of the well-known herbaceous plant virus cauliflower mosaic virus (camv, family caulimoviridae). the alga had been isolated from xanthoria parietina more than 70 years ago and has been maintained in a collection since that time. the camv detected in this collection entry has now been completely sequenced. the virus from t. aggregata is mechanically transmissibl ...201525789995
[evaluation of salt tolerance of transgenic tobacco plants bearing with p5cs1 gene of arabidopsis thaliana].arabidopsis thaliana delta1-pyrroline-5-carhoxylate synthase 1 gene (p5cs1) cdna was cloned under the control of the potent constitutive 35s rna promoter of the cauliflower mosaic virus and transferred into genome of tobacco cv. petit havana sr-1 (nicotiana tabacum l.) plants. it is shown that the constitutive level of proline in the transgenic plants t0 exceeds that of the sr1 reference line by 1.5 to 4 times. under conditions of salt stress (200, 300 mm nacl) t1-generation transgenic plants in ...201527055296
overexpression of an apocynum venetum dead-box helicase gene (avdh1) in cotton confers salinity tolerance and increases yield in a saline field.soil salinity is a major environmental stress limiting plant growth and productivity. we have reported previously the isolation of an apocynum venetum dead-box helicase 1 (avdh1) that is expressed in response to salt exposure. here, we report that the overexpression of avdh1 driven by a constitutive cauliflower mosaic virus-35s promoter in cotton plants confers salinity tolerance. southern and northern blotting analyses showed that the avdh1 gene was integrated into the cotton genome and express ...201526779246
isolation and characterization of "gmscream" promoters that regulate highly expressing soybean (glycine max merr.) genes.to increase our understanding of the regulatory components that control gene expression, it is important to identify, isolate and characterize new promoters. in this study, a group of highly expressed soybean (glycine max merr.) genes, which we have named "gmscream", were first identified from rna-seq data. the promoter regions were then identified, cloned and fused with the coding region of the green fluorescent protein (gfp) gene, for introduction and analysis in different tissues using 3 tool ...201526706070
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 ...201526674677
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 ...201526546319
functional analysis of cotton leaf curl kokhran virus/cotton leaf curl multan betasatellite rna silencing suppressors.in south asia, cotton leaf curl disease (clcud) is caused by a complex of phylogenetically-related begomovirus species and a specific betasatellite, cotton leaf curl multan betasatellite (clcumub). the post-transcriptional gene silencing (ptgs) suppression activities of the transcriptional activator protein (trap), c4, v2 and βc1 proteins encoded by cotton leaf curl kokhran virus (clcukov)/clcumub were assessed in nicotiana benthamiana. a variable degree of local silencing suppression was observ ...201526512705
complementary dna cloning of the pear 1-aminocyclopropane-1-carboxylic acid oxidase gene and agrobacterium-mediated anti-sense genetic transformation.the aim of the present study was to genetically modify plantlets of the chinese yali pear to reduce their expression of ripening-associated 1-aminocyclopropane-1-carboxylic acid oxidase (aco) and therefore increase the shelf-life of the fruit. primers were designed with selectivity for the conserved regions of published aco gene sequences, and yali complementary dna (cdna) cloning was performed by reverse transcription quantitative polymerase chain reaction (pcr). the obtained cdna fragment cont ...201526460204
comparative study of non-enveloped icosahedral viruses size.now, as before, transmission electron microscopy (tem) is a widely used technique for the determination of virions size. in some studies, dynamic light scattering (dls) has also been applied for this purpose. data obtained by different authors and using different methods could vary significantly. the process of tem sample preparation involves drying on the substrate, which can cause virions to undergo morphology changes. therefore, other techniques should be used for measurements of virions size ...201526545232
inducible and constitutive expression of an elicitor gene hrip1 from alternaria tenuissima enhances stress tolerance in arabidopsis.hrip1 is a novel hypersensitive response-inducing protein secreted by alternaria tenuissima that activates defense responses and systemic acquired resistance in tobacco. this study investigates the role that hrip1 plays in responses to abiotic and biotic stress using transgenic arabidopsis thaliana expressing the hrip1 gene under the control of the stress-inducible rd29a promoter or constitutive cauliflower mosaic virus 35s promoter. bioassays showed that inducible hrip1 expression in rd29a∷hrip ...201525120219
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 ...201525281563
multiplex, construct-specific, and real-time pcr-based analytical methods for bt rice with cry1ac gene.qualitative and quantitative analytical methods based on pcr for bacillus thuringiensis (bt) rice hybrid, namely, mrp 5401 bt expressing a modified version of the bt cry1ac gene, are reported here. multiplex pcr assays were developed to target the cry1ac transgene, cauliflower mosaic virus (camv) 35s promoter, agrobacterium tumefaciens nopaline synthase (nos) terminator, the neomycin phosphotransferase ii (nptll) marker gene, and an endogenous a-tubulin (tuba) gene in bt rice. the 3.178 kb regio ...201522468358
a vamp-associated protein, pva31 is involved in leaf senescence in arabidopsis.vamp-associated proteins (vaps) are highly conserved among eukaryotes. here, we report a functional analysis of one of the vaps, pva31, and demonstrate its novel function on leaf senescence in arabidopsis. the expression of pva31 is highly induced in senescence leaves, and localizes to the plasma membrane as well as the ara7-positive endosomes. yeast two-hybrid analysis demonstrates that pva31 is interacted with the plasma membrane localized-vamp proteins, vamp721/722/724 but not with the endoso ...201525897470
[construction of transgenic tobacco expressing tomato ggps2 gene and analysis of its low light tolerance].to explore the influence of low light on the synthesis of carotenoids, chlorophyll and the adaptability of transgenic plants with tomato solanum lycopersicon l. ggps2 gene, we constructed a vector containing a ggps2 gene with green fluorescent protein (gfp) as report gene under the control of a cauliflower mosaic virus 35s promoter and introduced it into tobacco nicotiana tabacum l. cv. wisconsin 38 by agrobacterium tumefaciens-mediated transformation. pcr analysis of the dna from kanamycin resi ...201526571690
a novel reference plasmid for the qualitative detection of genetically modified rice in food and feed.rice is one of the most important food crops in the world. genetically modified (gm) technology has been used in rice to confer herbicide tolerance and pathogen or insect resistance. china invests heavily in research on gm rice. by the end of 2014, at least 250 transgenic rice lines had been developed in china. to monitor the presence of gm rice in food and feed, we collected information on foreign elements from 250 transgenic rice lines and found 5 elements, including the agrobacterium tumefaci ...201526495318
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 ...201526278008
new multiplex pcr methods for rapid screening of genetically modified organisms in foods.we present novel multiplex pcr methods for rapid and reliable screening of genetically modified organisms (gmos). new designed pcr primers targeting four frequently used gmo specific sequences permitted identification of new dna markers, in particular 141 bp fragment of cauliflower mosaic virus (camv) 35s promoter, 224 bp fragment of agrobacterium tumefaciens nopaline synthase (nos) terminator, 256 bp fragment of 5-enolppyruvylshikimate-phosphate synthase (epsps) gene and 258 bp fragment of cry1 ...201526257724
transgene expression in tick cells using agrobacterium tumefaciens.ticks transmit infectious agents to humans and other animals. genetic manipulation of vectors like ticks could enhance the development of alternative disease control strategies. transgene expression using the phytopathogen agrobacterium tumefaciens has been shown to promote the genetic modification of non-plant cells. in the present work we developed t-dna constructs for a. tumefaciens to mediate transgene expression in hela cells as well as rhipicephalus microplus tick cells. translational fusi ...201526188857
a simple and efficient genetic transformation method of ganoderma weberianum.in this study, the agrobacterium tumefaciens-mediated transformation method for ganderma weberianum has been established. driven by the cauliflower mosaic virus (camv) 35s promoter, the hygromycin phosphotransferase (hpt), β-glucuronidase (uida), and enhanced green fluorescent protein (egfp) genes have been efficiently expressed in transgenic mycelia and spores. the transformation system was composed of the growing mycelia, a. tumefaciens strain gv3101, and the expression vector pbi-h1, harborin ...201525655754
simple screening strategy with only water bath needed for the identification of insect-resistant genetically modified rice.an informative, with simple instrument needed, rapid and easily updated strategy for the identification of insect-resistant genetically modified (gm) rice has been described. such strategy is based on a parallel series of loop-mediated isothermal amplification (lamp) reactions targeting the rice endogenous gene sucrose phosphate synthase (sps), the top two most frequently used genetic elements (agrobacterium tumefaciens nopaline synthase terminator (nos) and cauliflower mosaic virus 35s promoter ...201525582220
cloning and expression of tnf related apoptosis inducing ligand in nicotiana tabacum.molecular farming has been considered as a secure and economical approach for production of biopharmaceuticals. human tnf related apoptosis inducing ligand (trail) as a promising biopharmaceutical candidate has been produced in different expression hosts. however, little attention has been paid to molecular farming of the trail in spite of numerous advantages of plant expression systems. therefore, in this study the cytoplasmic production of the trail was tackled in nicotiana tabacum using agrob ...201525561925
an efficient plant regeneration and agrobacterium-mediated genetic transformation of tagetes erecta.tagetes erecta, l. an asteraceous plant of industrial and medicinal value, contains important compounds like pyrethrins, thiophenes and lutein, possessing immense potential for insecticidal, nematicidal and nutraceutical activities. considering the importance and demand for these natural compounds, genetic manipulation of this crop for better productivity of secondary metabolites holds great significance. a rapid and reproducible direct regeneration and genetic transformation system is the prere ...201525504508
novel reference molecules for quantitation of genetically modified maize and soybean.new quantitation methods based on a real-time polymerase chain reaction (pcr) technique were developed for 5 lines of genetically modified (gm) maize, including mon810, event176, bt11, t25, and ga21, and a gm soy, roundup ready. oligonucleotide dna, including specific primers and fluorescent dye-labeled probes, were designed for pcrs. two plasmids were constructed as reference molecules (rms) for the detection of gm maize and gm soy. the molecules contain the dna sequences of a specific region f ...201512374407
expression and functional analyses of microrna417 in arabidopsis thaliana under stress conditions.micrornas (mirnas) are 20-25 nucleotides-long rna transcripts that are capable of suppressing target gene expression by either mrna cleavage or translational repression. although many recent reports propose that mirnas play diverse roles in growth, development, morphogenesis, and stress responses of plants, the biological roles of many mirnas remain to be verified. here, we investigated stress-responsive expression patterns and functional roles of mirna417 in seed germination and seedling growth ...201617845858
overexpression of endoplasmic reticulum omega-3 fatty acid desaturase gene improves chilling tolerance in tomato.an endoplasmic reticulum-localized tomato omega-3 fatty acid desaturase gene (lefad3) was isolated and characterized with regard to its sequence, response to various temperatures and function in transgenic tomato plants. northern blot analysis showed that lefad3 was expressed in all organs tested and was markedly abundant in roots. meanwhile, the expression of lefad3 was induced by chilling stress (4 degrees c), but inhibited by high temperature (40 degrees c). the transgenic plants were obtaine ...201619648018
[structure and function of the genome of the cauliflower mosaic virus and its use in genetic engineering]. 20162692300
recombination-based generation of the agroinfectious clones of peanut stunt virus.full-length cdna clones of peanut stunt virus strain p (psv-p) were constructed and introduced into nicotiana benthamiana plants via agrobacterium tumefaciens. the cdna fragments corresponding to three psv genomic rnas and satellite rna were cloned into pgreen binary vector between cauliflower mosaic virus (camv) 35s promoter and nopaline synthase (nos) terminator employing seamless recombinational cloning system. the plasmids were delivered into a. tumefaciens, followed by infiltration of hosts ...201627659243
development of cotton leaf curl virus resistant transgenic cotton using antisense ßc1 gene.cotton leaf curl virus (clcuv) is a serious pathogen causing leaf curl disease and affecting the cotton production in major growing areas. the transgenic cotton (gossypium hirsutum cv. coker 310) plants were developed by using βc1 gene in antisense orientation gene driven by cauliflower mosaic virus-35s promoter and nos (nopaline synthase) terminator and mediated by agrobacterium tumefaciens transformation and somatic embryogenesis system. molecular confirmation of the transformants was carried ...201627081361
spike-dip transformation of setaria viridis.traditional method of agrobacterium-mediated transformation through the generation of tissue culture had limited success for setaria viridis, an emerging c4 monocot model. here we present an efficient in planta method for agrobacterium-mediated genetic transformation of s. viridis using spike dip. pre-anthesis developing spikes were dipped into a solution of agrobacterium tumefaciens strain agl1 harboring the β-glucuronidase (gus) reporter gene driven by the cauliflower mosaic virus 35s (camv35s ...201626932666
a determinant of disease symptom severity is located in rna2 of broad bean wilt virus 2.broad bean wilt virus 2 (bbwv2), which belongs to the genus fabavirus, is a destructive pathogen of many economically important horticultural and ornamental crops. in this study, we constructed infectious full-length cdna clones of two distinct isolates of bbwv2 under control of the cauliflower mosaic virus 35s promoter. bbwv2-pap1 isolated from paprika (capsicum annuum var. gulosum) induces severe disease symptoms in various pepper varieties, whereas bbwv2-rp1 isolated from red pepper (capsicum ...201626428303
transgenic bt cotton driven by the green tissue-specific promoter shows strong toxicity to lepidopteran pests and lower bt toxin accumulation in seeds.a promoter of the pnzip (pharbitis nil leucine zipper) gene (1.459 kb) was cloned from pharbitis nil and fused to the gus (β-glucuronidase) and bacillus thuringiensis endotoxin (cry9c) genes. several transgenic pnzip::gus and pnzip::cry9c cotton lines were developed by agrobacterium-mediated transformation. strong gus staining was detected in the green tissues of the transgenic pnzip::gus cotton plants. in contrast, gus staining in the reproductive structures such as petals, anther, and immature ...201626728504
cloning and expressing trypsin modulating oostatic factor in chlorella desiccata to control mosquito larvae.the insect peptide hormone trypsin modulating oostatic factor (tmof), a decapeptide that is synthesized by the mosquito ovary and controls the translation of the gut's trypsin mrna was cloned and expressed in the marine alga chlorella desiccata. to express aedes aegypti tmof gene (tmfa) in c. desiccata cells, two plasmids (pyes2/tmof and pydb4-tmfa) were engineered with pkylx71 dna (5 kb) carrying the cauliflower mosaic virus (camv) promoter 35s(2) and the kanamycin resistant gene (neo), as well ...201626440910
development of a new vector using soybean yellow common mosaic virus for gene function study or heterologous protein expression in soybeans.a new vector using soybean yellow common mosaic virus (sycmv) was constructed for gene function study or heterologous protein expression in soybeans. the in vitro transcript with a 5' cap analog m7gpppg from an sycmv full-length infectious vector driven by a t7 promoter infected soybeans (psycmvt7-full). the symptoms observed in the soybeans infected with either the sap from sycmv-infected leaves or psycmvt7-full were indistinguishable, suggesting that the vector exhibits equivalent biological a ...201626569351
a model for intracellular movement of cauliflower mosaic virus: the concept of the mobile virion factory.the genomes of many plant viruses have a coding capacity limited to <10 proteins, yet it is becoming increasingly clear that individual plant virus proteins may interact with several targets in the host for establishment of infection. as new functions are uncovered for individual viral proteins, virologists have realized that the apparent simplicity of the virus genome is an illusion that belies the true impact that plant viruses have on host physiology. in this review, we discuss our evolving u ...201626687180
salicylic acid treatment and expression of an rna-dependent rna polymerase 1 transgene inhibit lethal symptoms and meristem invasion during tobacco mosaic virus infection in nicotiana benthamiana.host rna-dependent rna polymerases (rdrs) 1 and 6 contribute to antiviral rna silencing in plants. rdr6 is constitutively expressed and was previously shown to limit invasion of nicotiana benthamiana meristem tissue by potato virus x and thereby inhibit disease development. rdr1 is inducible by salicylic acid (sa) and several other phytohormones. but although it contributes to basal resistance to tobacco mosaic virus (tmv) it is dispensable for sa-induced resistance in inoculated leaves. the lab ...201626757721
myb transcription factor isolated from raphanus sativus enhances anthocyanin accumulation in chrysanthemum cultivars.a myb transcription factor gene, rsmyb1, from radish was introduced into the chrysanthemum cultivars 'peach nd', 'peach red', and 'vivid scarlet' under the control of the cauliflower mosaic virus 35s promoter. presence of rsmyb1 in transgenic lines was confirmed using polymerase chain reaction (pcr). results of reverse-transcription-pcr analysis revealed that the expression of rsmyb1 was stable in all transgenic lines and could enhance the expression levels of three key biosynthetic genes (f3h, ...201628330149
photoelectrochemical camv35s biosensor for discriminating transgenic from non-transgenic soybean based on sio2@cdte quantum dots core-shell nanoparticles as signal indicators.a methodology for detection of the cauliflower mosaic virus 35s(camv35s) promoter was developed to distinguish transgenic from non-transgenic soybean samples by using photoelectrochemical (pec) biosensor. in this pec biosensing system, the as-prepared gold nanoparticles-reduced graphene oxide acted as a nanocarrier to immobilize the thiol-functional probe (probe1), and the sio2@cdte quantum dots (qds) core-shell nanoparticles tagged with the amino-functional probe (probe2) acted as signal indica ...201627769398
a robotic platform for high-throughput protoplast isolation and transformation.over the last decade there has been a resurgence in the use of plant protoplasts that range from model species to crop species, for analysis of signal transduction pathways, transcriptional regulatory networks, gene expression, genome-editing, and gene-silencing. furthermore, significant progress has been made in the regeneration of plants from protoplasts, which has generated even more interest in the use of these systems for plant genomics. in this work, a protocol has been developed for autom ...201627768035
infectious clones of the crinivirus cucurbit chlorotic yellows virus are competent for plant systemic infection and vector transmission.cucurbit chlorotic yellows virus (ccyv), a recently identified bipartite crinivirus, causes economic losses in cucurbit plants. ccyv is naturally transmitted only by whitefly bemisia tabaci. here we constructed full-length cdna clones of ccyv (rna1 and rna2) fused to the t7 rna polymerase promoter and the cauliflower mosaic virus 35s promoter. ccyv replicated and accumulated efficiently in cucumis sativus protoplasts transfected with in vitro transcripts. without rna2, rna1 replicated efficientl ...201626982585
overexpression of nakr3 enhances salt tolerance in arabidopsis.salinity is a major abiotic stress in agriculture. here, we report that sodium potassium root defective3 (nakr3), which encodes a heavy metal-associated domain protein, is involved in salt tolerance in arabidopsis. the results of quantitative reverse transcription-polymerase chain reaction analysis revealed that nakr3 was induced by high salinity and osmotic stresses, but not by cu(2+) stress. transient expression of nakr3-gfp in arabidopsis protoplasts showed that the nakr3 protein was localize ...201626909945
noncoding rnas of plant viruses and viroids: sponges of host translation and rna interference machinery.noncoding sequences in plant viral genomes are well-known to control viral replication and gene expression in cis. however, plant viral and viroid noncoding (nc)rna sequences can also regulate gene expression acting in trans, often acting like 'sponges' that bind and sequester host cellular machinery to favor viral infection. noncoding sequences of small subgenomic (sg)rnas of barley yellow dwarf virus (bydv) and red clover necrotic mosaic virus (rcnmv) contain a cap-independent translation elem ...201626900786
tolerance to excess-boron conditions acquired by stabilization of a bor1 variant with weak polarity in arabidopsis.boron (b) is a metalloid that is essential for plant growth but is toxic when present in excess. arabidopsis bor1 is a borate exporter, facilitating b translocation from root to shoot under limited-b conditions. bor1 shows stele side polar localization in the plasma membrane of various root cells, presumably to support b translocation toward the stele. bor1 is degraded under high-b supply through vacuolar sorting via ubiquitination at the k590 residue to prevent the accumulation of b to a toxic ...201626870730
35s promoter methylation in kanamycin-resistant kalanchoe (kalanchoe pinnata l.) plants expressing the antimicrobial peptide cecropin p1 transgene.transgenic kalanchoe plants (kalanchoe pinnata l.) expressing the antimicrobial peptide cecropin p1 gene (cecp1) under the control of the 35s cauliflower mosaic virus 35s rna promoter and the selective neomycin phosphotransferase ii (nptii) gene under the control of the nopaline synthase gene promoter were studied. the 35s promoter methylation and the cecropin p1 biosynthesis levels were compared in plants growing on media with and without kanamycin. the low level of active 35s promoter methylat ...201627682168
a viral satellite dna vector-induced transcriptional gene silencing via dna methylation of gene promoter in nicotiana benthamiana.virus-induced gene silencing (vigs) has been widely used for plant functional genomics study at the post-transcriptional level using various dna or rna viral vectors. however, while virus-induced transcriptional gene silencing (vitgs) via dna methylation of gene promoter was achieved using several plant rna viral vectors, it has not yet been done using a satellite dna viral vector. in this study, a viral satellite dna associated with tomato yellow leaf curl china virus (tylccnv), which has been ...201627422476
expression of cadr enhances its specific activity for cd detoxification and accumulation in arabidopsis.cadmium (cd) is a transition metal that is highly toxic in biological systems. anthropogenic emissions of cd have increased biogeochemical cycling and the amount of cd in the biosphere. here we studied the utility of a bacterial cd-binding protein, cadr, for the remediation of cd contamination. cadr was successfully targeted to chloroplasts using a constitutive cauliflower mosaic virus (camv) 35s promoter or a shoot-specific chl a/b-binding protein 2 gene (cab2) promoter and an rbcs (small subun ...201627382127
the role of plasmodesma-located proteins in tubule-guided virus transport is limited to the plasmodesmata.intercellular spread of plant viruses involves passage of the viral genome or virion through a plasmodesma (pd). some viruses severely modify the pd structure, as they assemble a virion carrying tubule composed of the viral movement protein (mp) inside the pd channel. successful modulation of the host plant to allow infection requires an intimate interaction between viral proteins and both structural and regulatory host proteins. to date, however, very few host proteins are known to promote viru ...201627339685
gene expression and promoter analysis of a novel tomato aldo-keto reductase in response to environmental stresses.the functional role of an uncharacterized tomato (solanum lycopersicum) aldo-keto reductase 4b, denoted as slakr4b, was investigated. the gene expression of tomato slakr4b was detected at a high level in the senescent leaves and the ripening fruits of tomato. although d-galacturonic acid reductase activities tended to be higher in tomato slakr4b-overexpressing transgenic tobacco by-2 cell lines than those in control cell lines, slakr4b gene expression was not well correlated with l-ascorbic acid ...201627337067
detection of herbaceous-plant pararetrovirus in lichen herbarium samples.cauliflower mosaic virus (camv) - a plant pararetrovirus that naturally causes diseases in brassicaceae and solanaceae plant hosts worldwide - has been detected by pcr for the first time in herbarium samples of usnea sp. lichens. the virus's presence in these lichens did not result in any micro- or macromorphological changes, and the herbarium records were classified as representative for the distinct species. sequence analyses classified all the detected viruses into one lineage of camv isolate ...201627265470
ectopic expression of jatropha curcas apetala1 (jcap1) caused early flowering in arabidopsis, but not in jatropha.jatropha curcas is a promising feedstock for biofuel production because jatropha oil is highly suitable for the production of biodiesel and bio-jet fuels. however, jatropha exhibits a low seed yield as a result of unreliable and poor flowering. apetala1 (ap1) is a floral meristem and organ identity gene in higher plants. the flower meristem identity genes of jatropha have not yet been identified or characterized. to better understand the genetic control of flowering in jatropha, an ap1 homolog ( ...201627168978
viral protein suppresses oxidative burst and salicylic acid-dependent autophagy and facilitates bacterial growth on virus-infected plants.virus interactions with plant silencing and innate immunity pathways can potentially alter the susceptibility of virus-infected plants to secondary infections with nonviral pathogens. we found that arabidopsis plants infected with cauliflower mosaic virus (camv) or transgenic for camv silencing suppressor p6 exhibit increased susceptibility to pseudomonas syringae pv. tomato (pst) and allow robust growth of the pst mutant hrcc-, which cannot deploy effectors to suppress innate immunity. the impa ...201627120694
generation of novel floral traits using a combination of floral organ-specific promoters and a chimeric repressor in torenia fournieri lind.in this study, we attempted to develop a new biotechnological method for the efficient modification of floral traits. because transcription factors play an important role in determining floral traits, chimeric repressors, which are generated by attaching a short transcriptional repressor domain to transcription factors, have been widely used as effective tools for modifying floral traits in many plant species. however, the overexpression of these chimeric repressors by the cauliflower mosaic vir ...201627107289
a chimeric repressor of petunia ph4 r2r3-myb family transcription factor generates margined flowers in torenia.the development of new phenotypes is key to the commercial development of the main floricultural species and cultivars. important new phenotypes include features such as multiple-flowers, color variations, increased flower size, new petal shapes, variegation and distinctive petal margin colourations. although their commercial use is not yet common, the transgenic technologies provide a potentially rapid means of generating interesting new phenotypes. in this report, we construct 5 vectors which ...201627089475
identification and functional characterization of the nac gene promoter from populus euphratica.the penac1 promoter is a non-tissue-specific and stress-inducible promoter containing a ga-responsive element and a myb recognition sequence that are responsible for induced expression patterns. nac transcription factors play vital roles in complex signaling networks during plant stress responses. promoters as crucial molecular switches are involved in the transcriptional regulation of gene activities dynamic network controlling a variety of biological processes, such as developmental processes, ...201627084679
comment on "large bottleneck size in cauliflower mosaic virus populations during host plant colonization" by monsion et al. (2008). 201627078639
ft overexpression induces precocious flowering and normal reproductive development in eucalyptus.eucalyptus trees are among the most important species for industrial forestry worldwide. however, as with most forest trees, flowering does not begin for one to several years after planting which can limit the rate of conventional and molecular breeding. to speed flowering, we transformed a eucalyptus grandis × urophylla hybrid (sp7) with a variety of constructs that enable overexpression of flowering locus t (ft). we found that ft expression led to very early flowering, with events showing flor ...201626132805
heterologous expression of chloroplast-localized geranylgeranyl pyrophosphate synthase confers fast plant growth, early flowering and increased seed yield.geranylgeranyl pyrophosphate synthase (ggps) is a key enzyme for a structurally diverse class of isoprenoid biosynthetic metabolites including gibberellins, carotenoids, chlorophylls and rubber. we expressed a chloroplast-targeted ggps isolated from sunflower (helianthus annuus) under control of the cauliflower mosaic virus 35s promoter in tobacco (nicotiana tabacum). the resulting transgenic tobacco plants expressing heterologous ggps showed remarkably enhanced growth (an increase in shoot and ...201625644367
camv-35s promoter sequence-specific dna methylation in lettuce.we found 35s promoter sequence-specific dna methylation in lettuce. additionally, transgenic lettuce plants having a modified 35s promoter lost methylation, suggesting the modified sequence is subjected to the methylation machinery. we previously reported that cauliflower mosaic virus 35s promoter-specific dna methylation in transgenic gentian (gentiana triflora × g. scabra) plants occurs irrespective of the copy number and the genomic location of t-dna, and causes strong gene silencing. to conf ...201626373653
a temperature-tolerant multiplex elements and genes screening system for genetically modified organisms based on dual priming oligonucleotide primers and capillary electrophoresis.high throughput screening systems are the preferred solution to meet the urgent requirement of increasing number of genetically modified organisms (gmos). in this study, we have successfully developed a multiplex gmo element screening system with dual priming oligonucleotide (dpo) primers. this system can detect the cauliflower mosaic virus 35s (camv 35s), terminator of nopaline synthase gene (nos), figwort mosaic virus 35s (fmv 35s) promoter, neomycin phosphotransferaseii (nptii), bt cry 1ab, p ...201728372191
three cis-regulatory motifs, auxre, mycrs1 and mycrs2, are required for modulating the auxin- and mycorrhiza-responsive expression of a tomato gh3 gene.auxin is well known to be a key regulator that acts in almost all physiological processes during plant growth, and in interactions between plants and microbes. however, to date, the regulatory mechanisms underlying auxin-mediated plant-arbuscular mycorrhizal (am) fungi symbiosis have not been well deciphered. previously we identified a gh3 gene, slgh3.4, strongly responsive to both auxin induction and mycorrhizal symbiosis. here, we reported a refined dissection of the slgh3.4 promoter activity ...201728339724
genome sequence of cauliflower mosaic virus identified in earwigs (doru luteipes) through a metagenomic approach.here we report the first complete genome sequence of a cauliflower mosaic virus from brazil, obtained from the gut content of the predator earwig (doru luteipes). this virus has a genome of 8,030 nucleotides (nt) and shares 97% genome-wide identity with an isolate from argentina.201728302781
selective autophagy limits cauliflower mosaic virus infection by nbr1-mediated targeting of viral capsid protein and particles.autophagy plays a paramount role in mammalian antiviral immunity including direct targeting of viruses and their individual components, and many viruses have evolved measures to antagonize or even exploit autophagy mechanisms for the benefit of infection. in plants, however, the functions of autophagy in host immunity and viral pathogenesis are poorly understood. in this study, we have identified both anti- and proviral roles of autophagy in the compatible interaction of cauliflower mosaic virus ...201728223514
fluorescent "on-off-on" switching sensor based on cdte quantum dots coupled with multiwalled carbon nanotubes@graphene oxide nanoribbons for simultaneous monitoring of dual foreign dnas in transgenic soybean.with the increasing concern of potential health and environmental risk, it is essential to develop reliable methods for transgenic soybean detection. herein, a simple, sensitive and selective assay was constructed based on homogeneous fluorescence resonance energy transfer (fret) between cdte quantum dots (qds) and multiwalled carbon nanotubes@graphene oxide nanoribbons (mwcnts@gonrs) to form the fluorescent "on-off-on" switching for simultaneous monitoring dual target dnas of promoter cauliflow ...201728182975
comparison of the impact of viral and plant-derived promoters regulating selectable marker gene on maize transformation and transgene expression.the choice of promoter regulating the selectable marker gene impacts transformation efficiency, copy number and the expression of selectable marker and flanking genes in maize. viral or plant-derived constitutive promoters are often used to regulate selectable marker genes. we compared two viral promoters, cauliflower mosaic virus (camv 35t) and sugarcane bacilliform virus (scbv) with two plant promoters, rice actin1 (osact1) and maize ubiquitin 1 (zmubi1) to drive aryloxyalkanoate dioxygenase ( ...201728160062
a dna probe based on phosphorescent resonance energy transfer for detection of transgenic 35s promoter dna.a qds-dna nano-probe was made by combining mn-doped zns room-temperature phosphorescence (rtp) quantum dots (qds) and dna. then an rtp sensor for quantitative detection of genetically-modified mark sequence cauliflower mosaic virus 35s promoter (ca mv 35s) dna was built on basis of phosphorescent resonance energy transfer (pret). the underlying principles were that a qds-dna water-soluble nano-probe was built by connecting single-strand dna to the surfaces of qds via a ligand exchange method. th ...201728088748
ectopic expression of an apple cytochrome p450 gene mdcypm1 negatively regulates plant photomorphogenesis and stress response in arabidopsis.cytochrome p450s play an important role in plant growth and are involved in multiple stresses response. however, little is known about the functions of cytochrome p450s in apple. here, a malus × domestica cytochrome p450 monooxygenase 1 gene, mdcypm1, was identified and subsequently cloned from apple 'gala' (malus × domestica). to verify the functions of mdcypm1, we generated transgenic arabidopsis plants expressing the apple mdcypm1 gene under the control of the cauliflower mosaic virus 35s pro ...201728073698
synthetic introns help identify sequences in the 5' utr intron of the glycine max polyubiquitin (gmubi) promoter that give increased promoter activity.specific sequences within the leader intron of a soybean polyubiquitin gene stimulated gene expression when placed either within a synthetic intron or upstream of a core promoter. the intron in the 5' untranslated region of the soybean polyubiquitin promoter, gmubi, seems to contribute to the high activity of this promoter. to identify the stimulatory sequences within the intron, ten different sequential intronic sequences of 40 nt were isolated, cloned as tetrameric repeats and placed upstream ...201728070655
generation of gene-edited chrysanthemum morifolium using multi-copy transgenes as targets and markers.the most widely used gene editing technology-the crispr/cas9 system-employs a bacterial monomeric dna endonuclease known as clustered regularly interspaced short palindromic repeats (crispr)-associated protein 9 (cas9) and single-guide rna (sgrna) that directs cas9 to a complementary target dna. however, introducing mutations into higher polyploid plant species, especially for species without genome information, has been difficult. chrysanthemum morifolium (chrysanthemum) is one of the most impo ...201728049122
use of a novel metal indicator to judge loop-mediated isothermal amplification for detecting the 35s promoter.loop-mediated isothermal amplification (lamp) is a widely used isothermal nucleic acid amplification method. here we developed a new closed-tube colorimetric method for judging lamp with a novel metal indicator. first, the metal indicator, acid chrome blue k (acbk), was evaluated in the lamp reaction with various combinations of reaction reagents, such as reaction buffer, dntp mixtures, primer mixtures, or mg(2+). we found that the solution color of the lamp reaction with acbk changed from red t ...201727873004
electroactive crown ester-cu(2+) complex with in-situ modification at molecular beacon probe serving as a facile electrochemical dna biosensor for the detection of camv 35s.a novel electrochemical dna biosensor has been facilely constructed by in-situ assembly of electroactive 4'-aminobenzo-18-crown-6-copper(ii) complex (abc-cu(2+)) on the free terminal of the hairpin-structured molecule beacon. the 3'-sh modified molecule beacon probe was first immobilized on the gold electrode (aue) surface through self-assembly chemistry of au-s bond. then the crow ester of abc was covalently coupled with 5'-cooh on the molecule beacon, and served as a platform to attach the cu( ...201727829553
barley hvpaphy_a as transgene provides high and stable phytase activities in mature barley straw and in grains.the phytase purple acid phosphatase (hvpaphy_a) expressed during barley seed development was evaluated as transgene for overexpression in barley. the phytase was expressed constitutively driven by the cauliflower mosaic virus 35s-promoter, and the phytase activity was measured in the mature grains, the green leaves and in the dry mature vegetative plant parts left after harvest of the grains. the t2 -generation of hvpaphy_a transformed barley showed phytase activity increases up to 19-fold (29 0 ...201727633382
construction of a full-length infectious cdna clone of cowpea mild mottle virus.infectious cdna clones are an important tool to study the molecular and cellular process of rna virus infection. in vitro and in vivo transcription systems are the two main strategies used in the generation of infectious cdna clones for rna viruses. this study describes the first generation of a full-length infectious cdna clone of cowpea mild mottle virus (cpmmv), a carlavirus. the full-length genome was synthesized by overlap extension pcr of two overlapping fragments and cloned in a puc-based ...201727730428
overexpression of populus trichocarpa cyp85a3 promotes growth and biomass production in transgenic trees.brassinosteroids (brs) are essential hormones that play crucial roles in plant growth, reproduction and response to abiotic and biotic stress. in arabidopsis, atcyp85a2 works as a bifunctional cytochrome p450 monooxygenase to catalyze the conversion of castasterone (cs) to brassinolide (bl), a final rate-limiting step in the br biosynthetic pathway. here, we report the functional characterizations of ptcyp85a3, one of the three atcyp85a2 homologous genes from populus trichocarpa. ptcyp85a3 share ...201728258966
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