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Dwarf Virus (dwarf + virus)
Kinds of Dwarf Virus Selected AbstractsUltrastructural and Immunocytochemical Studies on Effects of Barley Yellow Dwarf Virus , Infection on Fusarium Head Blight, Caused by Fusarium graminearum, in Wheat PlantsJOURNAL OF PHYTOPATHOLOGY, Issue 1 2006Y. Liu Abstract The interactions between barley yellow dwarf virus (BYDV) and Fusarium head blight (FHB), caused by Fusarium graminearum, were studied in the two winter wheat cultivars (cvs.), Agent (susceptible to FHB) and Petrus (moderately resistant to FHB), using ultrastructural and immunocytochemical methods. Infections of wheat plants of both cvs. by BYDV increased susceptibility to FHB. BYDV infection caused numerous cytological changes in lemma tissue of both cvs. such as formation of vesicles in the cytoplasm, degradation of fine structures of chloroplasts of both cvs. and accumulation of large starch grains in the chloroplasts. Electron microscopical studies showed that the development of F. graminearum on spike surfaces was not affected in BYDV-infected plants. After penetration and intercellular growth in lemma tissue, defence responses to Fusarium infections were markedly reduced in BYDV-diseased plants compared to the tissue of virus-free plants. At sites of contact of fungal cells with host tissue, depositions of cell wall material were distinctly less pronounced than in tissues of virus-free plants of cv. Petrus. Detection of , -1,3-glucanases and chitinases in lemma tissue of cv. Agent revealed no appreciably increased accumulation of both defence enzymes in F. graminearum -infected virus-free and BYDV-infected tissues compared to the non-infected control tissue. On the other hand, in cv. Petrus, infection with F. graminearum induced a markedly enhanced activity of both enzymes 3 days after inoculation. The increase of both enzyme activities was less pronounced in BYDV-infected plants than in tissue exclusively infected with F. graminearum. Cytological studies suggest that in contrast to the susceptible cv. Agent postinfectional defence responses may play still an important role in the resistance of the moderately resistant cv. Petrus to FHB. [source] Bean Yellow Dwarf Virus replicons for high-level transgene expression in transgenic plants and cell culturesBIOTECHNOLOGY & BIOENGINEERING, Issue 2 2006Xiuren Zhang Abstract A novel stable transgenic plant expression system was developed using elements of the replication machinery of Bean Yellow Dwarf Virus (BeYDV). The system contains two transgenes: 1) The BeYDV replicon vector with an expression cassette flanked by cis -acting DNA elements of BeYDV, and 2) The viral replication initiator protein (Rep) controlled by an alcohol-inducible promoter. When Rep expression was triggered by treatment with ethanol, it induced release of the BeYDV replicon from stably integrated T-DNA and episomal replication to high copy number. Replicon amplification resulted in substantially increased transgene mRNA levels (up to 80-fold) and translation products (up to 10-fold) after induction of Rep expression by ethanol treatment in tobacco NT1 cells and leaves of whole potato plants. Thus, the BeYDV stable transformant replicon system is a powerful tool for plant-based production of recombinant proteins. © 2005 Wiley Periodicals, Inc. [source] Aphid vectors of Barley yellow dwarf virusEPPO BULLETIN, Issue 3 2005Article first published online: 19 DEC 200 First page of article [source] The effect of drought stress and temperature on spread of barley yellow dwarf virus (BYDV)AGRICULTURAL AND FOREST ENTOMOLOGY, Issue 3 2000I. N. Smyrnioudis Summary 1 The effect of drought stress and temperature on the dispersal of wingless aphids Rhopalosiphum padi (L.) and the pattern of spread of BYDV (barley yellow dwarf virus) within wheat plants in controlled environment chambers was quantified. Combinations of three different drought stress levels, unstressed, moderate and high stress level, and three different temperatures, 5 ± 1 °C, 10 ± 1 °C, and 15 ± 1 °C, were investigated. 2 With increased temperature there was an increase in the mean distance of visited plants from the point of release and in the number of plants visited and infected with BYDV. Drought stress had no effect on mean distance moved by aphids at any temperature or on plants infected with virus at 10 °C and 5 °C. When plants were drought stressed, the numbers of plants visited and infected were greater at 15 °C than at 10 °C and 5 °C. 3 A greater proportion of plants visited by aphids was infected with BYDV when plants were stressed than when not stressed. At 15 °C a greater proportion of these plants was infected than at lower temperatures. There was no difference between treatments in the numbers of aphids present at the end of the experiment. 4 It is concluded that drought stress and temperature are of considerable importance in virus spread. [source] Virus Resistance in Cereals: Sources of Resistance, Genetics and BreedingJOURNAL OF PHYTOPATHOLOGY, Issue 9 2009Frank Ordon Abstract In cereals, soil-borne viruses transmitted by the plasmodiophorid Polymyxa graminis (e.g., Barley mild mosaic virus, Barley yellow mosaic virus or Soil-borne cereal mosaic virus), have increased in importance due to the increase of the acreage infested and because yield losses cannot be prevented by chemical measures. Due to global warming, it is also expected that insect transmitted viruses vectored by aphids (e.g., Barley yellow dwarf virus, Cereal yellow dwarf virus), leafhoppers (Wheat dwarf virus) or mites (e.g., Wheat streak mosaic virus), will become much more important even in cooler regions. The environmentally most sound and also most cost effective approach to prevent high yield losses caused by these viruses is breeding for resistance. Therefore, in contrast to other reviews on cereal viruses, this study briefly reviews present knowledge on cereal-infecting viruses and emphasizes especially the sources of resistance or tolerance to these viruses and their use in molecular breeding schemes. [source] Screening for Barley yellow dwarf virus -Resistant Barley Genotypes by Assessment of Virus Content in Inoculated SeedlingsJOURNAL OF PHYTOPATHOLOGY, Issue 6 2006Abstract The content of Barley yellow dwarf virus (BYDV) in roots and leaves of barley seedling plants differing in their level of resistance was assessed by quantitative ELISA 1,42 days after inoculation with the strain of BYDV (PAV). High virus accumulation in roots and low concentration in leaves was characteristic of the period 9,15 days after inoculation. In leaves, the differences in virus content between resistant and susceptible genotypes became significant after 15 days and resistance to virus accumulation was better expressed 30,39 days after inoculation. Roots of resistant materials exhibited evident retardation of virus accumulation and the greatest difference in virus content between resistant and susceptible plants was detected 9 days after inoculation. By these criteria, the selected winter and spring barley cultivars and lines (in total 44 materials) fell in to five groups according to field reactions and the presence or absence of the Yd2 resistance gene. There were highly significant and positive relations between ELISA values and 5-year field data on symptomatic reactions and grain-yield reductions due to infection. Using the described method, resistant and moderately resistant genotypes (both Yd2 and non- Yd2) were significantly differentiated from susceptible genotypes. The possible use of this method in screening for BYDV resistance is discussed. [source] Ultrastructural and Immunocytochemical Studies on Effects of Barley Yellow Dwarf Virus , Infection on Fusarium Head Blight, Caused by Fusarium graminearum, in Wheat PlantsJOURNAL OF PHYTOPATHOLOGY, Issue 1 2006Y. Liu Abstract The interactions between barley yellow dwarf virus (BYDV) and Fusarium head blight (FHB), caused by Fusarium graminearum, were studied in the two winter wheat cultivars (cvs.), Agent (susceptible to FHB) and Petrus (moderately resistant to FHB), using ultrastructural and immunocytochemical methods. Infections of wheat plants of both cvs. by BYDV increased susceptibility to FHB. BYDV infection caused numerous cytological changes in lemma tissue of both cvs. such as formation of vesicles in the cytoplasm, degradation of fine structures of chloroplasts of both cvs. and accumulation of large starch grains in the chloroplasts. Electron microscopical studies showed that the development of F. graminearum on spike surfaces was not affected in BYDV-infected plants. After penetration and intercellular growth in lemma tissue, defence responses to Fusarium infections were markedly reduced in BYDV-diseased plants compared to the tissue of virus-free plants. At sites of contact of fungal cells with host tissue, depositions of cell wall material were distinctly less pronounced than in tissues of virus-free plants of cv. Petrus. Detection of , -1,3-glucanases and chitinases in lemma tissue of cv. Agent revealed no appreciably increased accumulation of both defence enzymes in F. graminearum -infected virus-free and BYDV-infected tissues compared to the non-infected control tissue. On the other hand, in cv. Petrus, infection with F. graminearum induced a markedly enhanced activity of both enzymes 3 days after inoculation. The increase of both enzyme activities was less pronounced in BYDV-infected plants than in tissue exclusively infected with F. graminearum. Cytological studies suggest that in contrast to the susceptible cv. Agent postinfectional defence responses may play still an important role in the resistance of the moderately resistant cv. Petrus to FHB. [source] Combined thermotherapy and cryotherapy for efficient virus eradication: relation of virus distribution, subcellular changes, cell survival and viral RNA degradation in shoot tipsMOLECULAR PLANT PATHOLOGY, Issue 2 2008QIAOCHUN WANG SUMMARY Accumulation of viruses in vegetatively propagated plants causes heavy yield losses. Therefore, supply of virus-free planting materials is pivotal to sustainable crop production. In previous studies, Raspberry bushy dwarf virus (RBDV) was difficult to eradicate from raspberry (Rubus idaeus) using the conventional means of meristem tip culture. As shown in the present study, it was probably because this pollen-transmitted virus efficiently invades leaf primordia and all meristematic tissues except the least differentiated cells of the apical dome. Subjecting plants to thermotherapy prior to meristem tip culture heavily reduced viral RNA2, RNA3 and the coat protein in the shoot tips, but no virus-free plants were obtained. Therefore, a novel method including thermotherapy followed by cryotherapy was developed for efficient virus eradication. Heat treatment caused subcellular alterations such as enlargement of vacuoles in the more developed, virus-infected cells, which were largely eliminated following subsequent cryotherapy. Using this protocol, 20,36% of the treated shoot tips survived, 30,40% regenerated and up to 35% of the regenerated plants were virus-free, as tested by ELISA and reverse transcription loop-mediated isothermal amplification. Novel cellular and molecular insights into RBDV,host interactions and the factors influencing virus eradication were obtained, including invasion of shoot tips and meristematic tissues by RBDV, enhanced viral RNA degradation and increased sensitivity to freezing caused by thermotherapy, and subcellular changes and subsequent death of cells caused by cryotherapy. This novel procedure should be helpful with many virus,host combinations in which virus eradication by conventional means has proven difficult. [source] Field trial of serially passaged isolates of BYDV-PAV overcoming resistance derived from Thinopyrum intermedium in wheatPLANT BREEDING, Issue 3 2006F. Chain Abstract Barley yellow dwarf disease (BYDD) is one of the main viral diseases of small grain cereals. This disease, reported on numerous plant species of the Poaceae family, is caused by a complex of viral species including the species Barley yellow dwarf virus -PAV (BYDV-PAV, family Luteoviridae, genus Luteovirus), frequently found in western Europe. Resistance sources towards BYDD are scarce. Indeed, breeding-resistant genotypes is a long and expensive process. Thus, estimating the durability of the resistance genes before the achievement of selection would be an asset for breeders. One isolate of BYDV-PAV has been serially passaged on two hosts, ,Zhong ZH' and ,TC14', carrying a gene for partial resistance. The resulting viral population showed an increase of the speed of development of the infection in controlled conditions. In this study, these viral populations were evaluated in a 3-year field trial, including a susceptible host, ,Rendezvous', and a host carrying the resistance gene of ,TC14' in a ,Rendezvous' background, to assess the effect of serial passages in field conditions. Results indicate that isolates issued from serial passages on hosts carrying a gene for partial resistance induced increased damage in field conditions when compared with the initial isolate. Yield losses are mainly due to a decrease of the number of kernels per square metre. The interest on using partial resistance gene to control BYDD is discussed. [source] Identification of wheat,Thinopyrum intermedium 2Ai-2 ditelosomic addition and substitution lines with resistance to barley yellow dwarf virusPLANT BREEDING, Issue 2 2006Z. S. Lin Abstract Among the regenerated plants derived from immature hybrid embryos of wheat,Thinopyrum intermedium disomic addition line Z6 × common wheat variety ,Zhong8601', a plant with a telocentric chromosome and barley yellow dwarf virus (BYDV) resistance was obtained. The telocentric chromosome paired with an entire Thinopyrum chromosome to form a heteromorphic bivalent at meiotic metaphase I. Genomic in situ hybridization showed that the telosome originated from Th. intermedium. Two ditelosomic additions and one disomic substitution were identified among the offspring of the plant. Two random amplified polymorphic DNA molecular markers were identified among 150 random primers used to detect the different arms of the alien chromosome. These might be useful for developing translocation lines with BYDV resistance. [source] A generic RT-PCR assay for the detection of LuteoviridaePLANT PATHOLOGY, Issue 3 2010A. Chomi This study, using RT-PCR, is the first comprehensive assessment since 1991 of a generic detection method for the Luteoviridae. Thirteen Luteoviridae species were detected using three separate sets of low-degeneracy generic primers with RT-PCR to amplify 68-, 75- and 129/156-bp regions of the Luteoviridae coat-protein gene. Species detected include all members of the genus Luteovirus [Barley yellow dwarf virus (BYDV) -PAV, BYDV-PAS, BYDV-MAV (129 and/or 156 bp amplicons), Soybean dwarf virus, Bean leafroll virus (68 bp amplicon)] and eight of nine species from the genus Polerovirus [Beet western yellows virus, Beet chlorosis virus, Beet mild yellowing virus, Turnip yellows virus, Potato leafroll virus, Cucurbit aphid-borne yellows virus, Cereal yellow dwarf virus-RPV (68-bp amplicon) and Sugarcane yellow leaf virus (75-bp amplicon)]. These primers were not able to detect Carrot red leaf virus, Sweet potato leaf speckling virus (both belong to unassigned Luteoviridae) and Pea enation mosaic virus-1 (genus Enamovirus). A synthetic positive control containing all primer sequence priming sites was designed to facilitate this method as a generic tool for use with a variety of host plants. The Luteoviridae primers described in this study present a simple infection-detection tool of benefit to biosecurity authorities in nursery-stock surveillance, disease management or outbreak prevention, and may also be useful in detection of as-yet undiscovered species within the Luteovirus and Polerovirus genera. [source] Characterization of Potato rough dwarf virus and Potato virus P: distinct strains of the same viral species in the genus CarlavirusPLANT PATHOLOGY, Issue 6 2006C. Nisbet In an attempt to resolve whether two putative carlaviruses, potato rough dwarf virus (PRDV) and potato virus P (PVP), reported infecting potato in Argentina and Brazil, respectively, are the same virus in the genus Carlavirus, they were characterized using nucleotide sequence analysis, serology and bioassay. For PRDV and PVP, sequences of 2016 and 1492 bp, respectively, were obtained. Similarity searches of coat-protein amino acid sequences and the presence of the nucleic-acid-binding protein exhibiting the characteristic motif C-X2 -C-X11 -C-X4 -C, highly conserved in carlaviruses, showed PRDV and PVP to be members of the genus Carlavirus. Further analysis showed that they were the same virus species since the full coat-protein sequence, translated from the nucleotide sequence, exceeded the 84% minimum identity required for members of the same species within the genus Carlavirus, at 90·8% identity in an ungapped alignment of 304 amino acids. Moreover, the viruses could not be distinguished using polyclonal antibodies raised to PRDV and PVP in ELISA. They could, however, be distinguished using an anti-PVP monoclonal antibody, which did not react to PRDV, and by differences in the susceptibility of plant species and potato cultivars to infection and/or symptom expression following mechanical inoculation. Host plants Datura metel, Nicandra physalodes and Nicotiana edwardsonii were reliably infected systemically with PVP, but not with PRDV. Although all potato cultivars tested were infected, there were marked differences between cultivars in the symptoms produced. The cultivars Ditta and King Edward showed symptoms (including stunting and mottle) when infected with PRDV, but not when infected with PVP. Based on the results, it is proposed that PRDV is a strain of PVP within the genus Carlavirus. For detection of the viruses in postentry quarantine, it is recommended that PRDV or PVP polyclonal antibodies are used, together with inoculation to Nicotiana occidentalis N. megalosiphon -P1. For differentiation of PRDV and PVP, the PVP monoclonal antibody may be used together with inoculation to D. metel and N. physalodes (systemic infection with PVP but not PRDV, determined using ELISA). [source] Occurrence and distribution of Raspberry bushy dwarf virus in commercial Rubus plantations in England and WalesPLANT PATHOLOGY, Issue 6 2001D. J. Barbara Serological surveys for Raspberry bushy dwarf virus (RBDV) made between 1995 and 1997 and covering ,,10% of the commercial farms growing Rubus (red raspberry and hybrid berries) in England and Wales showed that this virus was present on approximately one-quarter of all farms and in approximately one-sixth of all plots tested. RBDV was found in all of the four main raspberry cultivars being grown at that time (Autumn Bliss, Glen Moy, Glen Prosen and Leo), in Loganberry and in Tayberry. Fifteen RBDV genotypes (including two that appeared to be mixed) were identified using RT-PCR/RFLPs, but the majority of genotypes were found only rarely. Of the RBDV isolates tested, two genotypes each comprised 12·5% and another 46·4%. None of the three most common genotypes was associated solely with single Rubus cultivars and vice versa. It is suggested that two separate outbreaks of RBDV are occurring in England and Wales. One outbreak comprises the most frequent genotype combined with one of the moderately frequent genotypes; this outbreak is largely confined to the main growing areas and is either spreading between farms or coming from multiple local sources. Circumstantial evidence suggests that these isolates (and hence this first outbreak) are of the RB pathotype. The second outbreak consists of the other moderately frequent genotype and those genotypes which are less common. These genotypes appear to be more scattered across England and Wales and seem more likely to be coming from local sources and not to be spreading naturally between commercial farms. [source] First record of Beet western yellows virus, Chickpea chlorotic dwarf virus and Faba bean necrotic yellows virus affecting faba bean (Vicia faba) crops in IraqPLANT PATHOLOGY, Issue 6 2001NEW DISEASE REPORT No abstract is available for this article. [source] Vectors and alternative hosts of Tobacco yellow dwarf virus in southeastern AustraliaANNALS OF APPLIED BIOLOGY, Issue 1 2010P. Tr, bicki Factors that determine the epidemiology of Tobacco yellow dwarf virus (TbYDV), including alternative host plants and insect vector(s), were assessed over three consecutive growing seasons at four field sites in Northeastern Victoria in commercial tobacco growing properties. In addition, these factors were assessed for one growing season at three bean growing properties. Overall, 23 leafhopper species were identified at the 7 sites, with Orosius orientalis as the predominant leafhopper. Of the leafhoppers collected, only O. orientalis and Anzygina zealandica tested positive for TbYDV by polymerase chain reaction (PCR). The population dynamics of O. orientalis was assessed using sweep net sampling over three growing seasons and a trimodal distribution was observed. Despite large numbers of O. orientalis occurring early in the growing season (September,October), TbYDV was only detected in these leafhoppers between late November and end of January. The peaks in the detection of TbYDV in O. orientalis correlated with the observation of disease symptoms in tobacco and bean and were associated with warmer temperatures and lower rainfall. Spatial and temporal distribution of vegetation at selected sites was determined using quadrat sampling. Of the 40 plant species identified, TbYDV was detected only in four dicotyledonous species, Amaranthus retroflexus, Phaseolus vulgaris, Nicotiana tabacum and Raphanus raphanistrum. The proportion of host and non-host availability for leafhoppers was associated with climatic conditions. [source] Incidence of cereal and pasture viruses in New Zealand's native grassesANNALS OF APPLIED BIOLOGY, Issue 1 2010C. Delmiglio This study provides evidence for frequent and multiple invasions of New Zealand's native grasses by exotic cereal and pasture viruses. Fifteen native and three exotic grasses from 29 North Island and six South Island sites were surveyed for the presence of viruses using enzyme-linked immunosorbent assay (ELISA). Barley yellow dwarf virus and Cereal yellow dwarf virus (BYDV, CYDV: Luteoviridae) and Cocksfoot mottle virus (CoMV, Sobemovirus) are widespread throughout New Zealand. CoMV, previously considered to have a natural host range restricted to Dactylis and Triticum, was detected in Poa anceps, P. cita, Festuca novae-zelandiae, and Chionochloa rubra. New virus host reports include BYDV-PAV in Microlaena stipoides and Dichelachne crinita; BYDV-MAV in P. cita, F. novae-zelandiae and Hierochloe redolens; and CYDV-RPV in P. cita and M. stipoides. Nominal logistic regression analyses showed a correlation between the presence of exotic grass species and virus incidence. Host range experiments for BYDV-PAV and CoMV were performed with selected native and exotic grasses, and the results are discussed in context of the field-survey findings. [source] Cryotherapy of shoot tips: a technique for pathogen eradication to produce healthy planting materials and prepare healthy plant genetic resources for cryopreservationANNALS OF APPLIED BIOLOGY, Issue 3 2009Q.C. Wang Abstract Cryotherapy of shoot tips is a new method for pathogen eradication based on cryopreservation techniques. Cryopreservation refers to the storage of biological samples at ultra-low temperature, usually that of liquid nitrogen (,196°C), and is considered as an ideal means for long-term storage of plant germplasm. In cryotherapy, plant pathogens such as viruses, phytoplasmas and bacteria are eradicated from shoot tips by exposing them briefly to liquid nitrogen. Uneven distribution of viruses and obligate vasculature-limited microbes in shoot tips allows elimination of the infected cells by injuring them with the cryo-treatment and regeneration of healthy shoots from the surviving pathogen-free meristematic cells. Thermotherapy followed by cryotherapy of shoot tips can be used to enhance virus eradication. Cryotherapy of shoot tips is easy to implement. It allows treatment of large numbers of samples and results in a high frequency of pathogen-free regenerants. Difficulties related to excision and regeneration of small meristems are largely circumvented. To date, severe pathogens in banana (Musa spp.), Citrus spp., grapevine (Vitis vinifera), Prunus spp., raspberry (Rubus idaeus), potato (Solanum tuberosum) and sweet potato (Ipomoea batatas) have been eradicated using cryotherapy. These pathogens include nine viruses (banana streak virus, cucumber mosaic virus, grapevine virus A, plum pox virus, potato leaf roll virus, potato virus Y, raspberry bushy dwarf virus, sweet potato feathery mottle virus and sweet potato chlorotic stunt virus), sweet potato little leaf phytoplasma and Huanglongbing bacterium causing ,citrus greening'. Cryopreservation protocols have been developed for a wide variety of plant species, including agricultural and horticultural crops and ornamental plants, and can be used as such or adjusted for the purpose of cryotherapy. [source] Effect of sowing date and straw mulch on virus incidence and aphid infestation in organically grown faba beans (Vicia faba)ANNALS OF APPLIED BIOLOGY, Issue 2 2009H. Saucke Abstract The effect of sowing date on aphid infestation and the incidence of aphid-transmitted viruses were investigated in organically managed, small-scale field experiments with two faba bean cultivars over 3 years (2002,04). As an additional factor, straw mulch was applied in 2 of the 3 years shortly before the start of vector activity in May. Virus incidence was determined using enzyme-linked immunosorbent assay and immunoelectron microscopy. Aphid flight activity was monitored using standard yellow water traps. Bean colonising aphids were assessed throughout the vegetation period by counting the number of plants infested with Acyrthosiphon pisum, Megoura viciae and Aphis fabae. Pea enation mosaic virus and bean yellow mosaic virus were the most abundant aphid-transmitted viruses, being detected in 22,54% and 9,69%, respectively, of the total number of virus-infected plants analysed per year. Further aphid-transmitted viruses found in faba bean were bean leaf roll virus, beet western yellows virus, clover yellow vein virus (in 2002) and soybean dwarf virus (in 2004). A. pisum was the predominant aphid species colonising faba bean plants. Early sowing compared with late sowing led to a significant reduction of the total virus incidence in faba bean in all 3 years. However, significantly decreased levels of A. pisum colonisation as a result of early sowing were observed only in 1 year and one cultivar. Irrespective of sowing date, straw mulching had no significant effects on virus incidence and aphid colonisation. Compared with late sowing, early sowing significantly increased bean yield in all 3 years and kernel weight in 2 years, whereas straw mulching had no effect on yield. [source] Fluctuations in concentration of two potyviruses in garlic during the growing period and sampling conditions for reliable detection by ELISAANNALS OF APPLIED BIOLOGY, Issue 1 2002C I DOVAS Summary To optimise sampling conditions for the detection by ELISA of Onion yellow dwarf virus (OYDV) and Leek yellow stripe virus (LYSV), the most important viral pathogens of garlic worldwide, relative virus concentrations were determined during the growing period and in different leaf parts by DAS-ELISA. Both viruses were found to have uneven distributions in garlic plants, with the tips of the two latest fully developed leaves showing the highest concentrations and the oldest leaves the lowest concentrations. The tips of the youngest leaves were found to have higher virus concentrations than their middle and basal sections. In the older leaves, viruses were distributed more uniformly in the three leaf sections. In the oldest leaves virus levels in the leaf tips were significantly decreased. The concentrations of OYDV and LYSV increased until March, whereas later on they decreased. During storage of leaf samples at 6°C for 15 days, a loss was found of both virus antigens of more than 80%, and during 109 days of storage at ,30°C a loss of more than 90% was found. [source] A DNA replicon system for rapid high-level production of virus-like particles in plantsBIOTECHNOLOGY & BIOENGINEERING, Issue 4 2009Zhong Huang Abstract Recombinant virus-like particles (VLPs) represent a safe and effective vaccine strategy. We previously described a stable transgenic plant system for inexpensive production and oral delivery of VLP vaccines. However, the relatively low-level antigen accumulation and long-time frame to produce transgenic plants are the two major roadblocks in the practical development of plant-based VLP production. In this article, we describe the optimization of geminivirus-derived DNA replicon vectors for rapid, high-yield plant-based production of VLPs. Co-delivery of bean yellow dwarf virus (BeYDV)-derived vector and Rep/RepA-supplying vector by agroinfiltration of Nicotiana benthamiana leaves resulted in efficient replicon amplification and robust protein production within 5 days. Co-expression of the P19 protein of tomato bush stunt virus, a gene silencing inhibitor, further enhanced VLP accumulation by stabilizing the mRNA. With this system, hepatitis B core antigen (HBc) and Norwalk virus capsid protein (NVCP) were produced at 0.80 and 0.34 mg/g leaf fresh weight, respectively. Sedimentation analysis and electron microscopy of transiently expressed antigens verified the efficient assembly of VLPs. Furthermore, a single replicon vector containing a built-in Rep/RepA cassette without P19 drove protein expression at similar levels as the three-component system. These results demonstrate the advantages of fast and high-level production of VLP-based vaccines using the BeYDV-derived DNA replicon system for transient expression in plants. Biotechnol. Bioeng. 2009;103: 706,714. © 2009 Wiley Periodicals, Inc. [source] | ||||||||||