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1.
Isolation and Partial Characterization of a Tenuivirus from Wheat in Iran   总被引:1,自引:0,他引:1  
A tenuivirus, tentatively designated Iranian wheat stripe virus (IWSV), was transmitted by the delphacid Unkanodes tanasijevici and induced dwarfing, striping and yellowing symptoms in wheat. It could also infect barley, oat, rice, rye, sorghum and a number of other gramineous species. The virus was purified from wheat by chloroform clarification and differential and density-gradient centrifugation. It formed several layers in density-gradient columns. Purified virus preparations had a UV absorption spectrum typical of nucleoproteins and contained flexuous supercoils of 8.7 nm and fine filaments of 4.3 nm width. The antiserum produced against the virus reacted with infected plant sap in agar-gel diffusion and enzyme-linked immunosorbent assay (ELISA). The virus could be readily detected in individual viruliferous planthoppers by ELISA. IWSV was serologically related to rice hoja blanca virus but not to maize stripe virus. The relationship of IWSV with other tenuiviruses is discussed.  相似文献   

2.
The effect of co-infection by maize streak virus (MSV) and maize stripe virus (MStV) on plant growth and grain yield was investigated in a susceptible variety of maize (Zea mays), ZS 5206, in Mauritius. Under natural conditions MSV, transmitted by the leafhopper Cicadulina mbila, was normally established before MStV, which is vectored by the planthopper Peregrinus maidis; as a result, MStV symptoms were often partially or completely masked by those of MSV, making MStV detection by symptomatology very unreliable. MSV and MStV were diagnosed by ELISA and MStV by a novel method of detecting the MStV-coded non-capsid protein. The maize hybrid ZS 5206 was inoculated with either MSV, MStV or both, at two stages in the growth cycle (3–5 or 7–10 leaf stage). A greater reduction in plant growth was observed in plants inoculated singly with MStV (80% and 29% for first and second stage, respectively) than with MSV (50% and 23%, respectively). No cobs were produced by plants singly infected with MStV at the first stage, or co-infected with MSV and MStV at both stages; however, marginal grain production was recorded in plants singly infected with MSV at the first stage (91% reduction), or infected either with MSV or MStV, at the second stage (65% and 80% reduction, respectively). In maize hybrid ZS 5206, MStV is more virulent than MSV; co-infection by both viruses causes greater reductions in plant growth and grain yield than single infection by either virus at a given stage of plant development. In the event of co-infection by MSV and MStV, yield losses can be erroneously attributed to MSV only if the symptoms of MStV are masked by those of the former and if adequate methods for MStV detection are not used.  相似文献   

3.
As previously reported, narcissus latent virus (NLV) has flexuous filamentous particles measuring c. 650 nm × 13 nm, is manually transmissible to Nicotiana clevelandii and Tetragonia expansa, and is transmitted by the aphid Myzus persicae following brief acquisition access periods. In contrast to previous reports the virus particle protein has an apparent mol. wt of c. 45 kD. Moreover, infected cells in N. clevelandii leaves contain cytoplasmic inclusion bodies resembling those of potyviruses. In vitro translation of NLV RNA produced only one major product (mol. wt c. 25 kD) which was not precipitated by antisera to virus particle protein or to cytoplasmic inclusion protein. Antisera to 12 potyviruses and nine carlaviruses failed to react with sap containing NLV particles. Similarly antiserum to NLV particles did not react with particles of seven potyviruses or four carlaviruses. A weak reaction was detected between NLV particles and antiserum to particles of maclura mosaic virus (MMV), a virus which resembles NLV in particle morphology and particle-protein size, and in inducing pinwheel inclusions. The cytoplasmic inclusion proteins (CIPs) of NLV, MMV and from narcissus plants with yellow stripe symptoms were serologically inter-related. These proteins were also serologically related to, and had mol. wt similar to, the CIP of members of the potyvirus group. Particles with the size and antigenic specificity of those of NLV were found consistently in narcissus plants with yellow stripe disease. Narcissus latent and narcissus yellow stripe viruses therefore seem to be synonymous and, together with MMV, have properties distinct from those of any previously described virus group.  相似文献   

4.
A mechanically transmissible virus with isometric particles c. 32 nm in diameter, was isolated from infected watermelons and sweet melons in the People's Democratic Republic of Yemen. Purified virus preparations contained two major sedimenting components with sedimentation coefficients of 61S and 117S. In isopycn ic centrifugation in CsCl the particles formed a single band of buoyant density 1.39 g cm-3. Preparations of virus particles comprised of a single polypeptide of mol. wt c. 22 000 and ssRNA of mol. wt 2.1 × 106. The virus was serologically related to three of six subgroups of tymoviruses tested. The name melon rugose mosaic virus is proposed for this newly described virus.  相似文献   

5.
Groundnut plants with chlorotic rosette disease contain a manually transmissible virus, groundnut rosette (GRV), which is also transmitted in the persistent (circulative) manner by aphids (Aphis craccivora), but only from plants that are co-infected with a manually non-transmissible luteovirus, groundnut rosette assistor virus (GRAV). Strains of GRV from plants with chlorotic or green forms of rosette are called GRV(C) and GRV(G) respectively. An isolate of GRV(C) from Nigeria remained infective in Nicotiana clevelandii leaf extracts for 1 day at room temperature and for 15 days at 4d?C, but lost infectivity after 1 day at -20d?C or after dilution to 10--4. Its infectivity and longevity in vitro were not altered by addition of 1 mg/litre bentonite to the extraction buffer. Infectivity in leaf extracts was abolished by treatment with 50% (v/v) ether, 10% (v/v) chloroform or 8% (v/v) n-butanol, but not by treatment for 30 min with RNase A at up to 100 ng/ml. In attempts to purify GRV(C), nearly all the infectivity from N. clevelandii extracts was found in the pellets from centrifugation at 65 000 g for 1. 5 h; infectivity also occurred in a cell membrane fraction that collected at the top of a 30% sucrose ‘cushion’ containing 4% polyethylene glycol and 0.2 M NaCI. However, no virus-like particles were found in either type of preparation by electron microscopy. Nucleic acid preparations made directly from GRV(C)-infected N. clevelandii leaves were very infective; this infectivity was totally inactivated by treatment for 30 min with RNase A at 10 ng/ml in buffers of both low and high ionic strength and was therefore attributed to ssRNA. When nucleic acid preparations were electrophoresed in gels no virus-specific bands were visible but the position of the infectivity indicated that the infective ssRNA has an apparent mol. wt of c. 1.55 × 106. A similar mol. wt was indicated by the rate of sedimentation of the infective ssRNA in sucrose gradients. Preparations of dsRNA made from GRV(C)-infected N. clevelandii leaves contained a species of mol. wt c. 3.0 × 106; in addition some dsRNA preparations contained an abundant component of mol. wt c. 0.6 × 106 together with several other components of intermediate mol. wt. Similar patterns of bands were observed in dsRNA preparations made from Nigerian-grown groundnut material infected with GRV(C) alone, or with GRV(C) + GRAV, or with GRV(G) + GRAV. The properties of GRV closely resemble those of two other viruses that depend on luteoviruses for transmission by aphids, carrot mottle virus and lettuce speckles mottle virus.  相似文献   

6.
提纯的水稻条纹病毒(云南宜良分离物)在电镜下的形态为多型性,但主要是宽8-10 nm,长80-250的分枝丝状体,有些为直径3 nm或8 nm的开环环状体,有些为13 nm宽,130-190 nm长的丝状体,但其基本结构应是直径3 nm、长度不等的丝状体.经聚丙烯酰胺凝胶电泳分析,vRNA4编码的病害特异蛋白(SP)分子量为19.9 kDa,而vcRNA3编码的外壳蛋白(CP)约为33.6 kDa.在非变性条件下,RSV的4条ssRNAs大小分别为3.0×106(ssRNA1)、1.2×106(ssRNA2)、0.9×106(ssRNA3)和0.8×106 Da(ssRNA4),有时出现一条大小为0.58×106 Da的单链RNA(ssRNA5);而4条dsRNAs的分子量分别为4.9×106(dsRNA1)、2.8×106(dsRNA2)、2.0×106(dsRNA3)和1.7×106 Da(dsRNA4).利用制备电泳分离提纯的外壳蛋白免疫家兔,得到了高特异性的抗血清.A蛋白夹心ELISA检测结果表明,RSV-CP与水稻草状矮化病毒(RGSV)CP抗血清有微弱的反应,但与RSV、RGSV的SP抗血清没有反应,而RSV-CP抗血清与RSV-SP及RGSV的SP、CP都无血清学关系,这个结果表明RGSV与RSV之间在进化上具有一定的亲缘关系.  相似文献   

7.
An expression library was created and 2304 clones sequenced from a monogyne colony of Solenopsis invicta. The primary intention of the project was to utilize homologous gene identification to facilitate discovery of viruses infecting this ant pest that could potentially be used in pest management. Additional genes were identified from the ant host and associated pathogens that serve as an important resource for studying these organisms. After assembly and removal of mitochondrial and poor quality sequences, 1054 unique sequences were yielded and deposited into the GenBank database under Accession Nos. EH412746 through EH413799. At least nine expressed sequence tags (ESTs) were identified as possessing microsatellite motifs and 15 ESTs exhibited significant homology with microsporidian genes. These sequences most likely originated from Thelohania solenopsae, a well-characterized microsporidian that infects S. invicta. Six ESTs exhibited significant homology with single-stranded RNA viruses (3B4, 3F6, 11F1, 12G12, 14D5, and 24C10). Subsequent analysis of these putative viral ESTs revealed that 3B4 was most likely a ribosomal gene of S. invicta, 11F1 was a single-stranded RNA (ssRNA) virus contaminant introduced into the colony from the cricket food source, 12G12 appeared to be a plant-infecting tenuivirus also introduced into the colony as a field contaminant, and 3F6, 14D5, and 24C10 were all from a unique ssRNA virus found to infect S. invicta. The sequencing project illustrates the utility of this method for discovery of viruses and pathogens that may otherwise go undiscovered.  相似文献   

8.
The properties of Elderberry latent virus (ELV) and Pelargonium ringspot virus (PelRSV) were compared. The viruses were largely indistinguishable in herbaceous host range and symptomatology, particle morphology, sedimentation coefficient and RNA profiles and size. They were also very closely related serologically with SDI differences in agarose gel double‐diffusion tests of 1 to 3. Purified virus particle preparations of each virus contained isometric particles c. 30 nm in diameter that sedimented as a major component with an sO20W of 112–115S. Purified virus particle preparations contained a major and a minor ssRNA species that in polyacrylamide gel electrophoresis (PAGE) had estimated sizes of c. 3.8 kb and c. 1.6 kb respectively. Plants of Chenopodium quinoa infected with ELV or PelRSV each contained three dsRNA species of c. 3.8, 2.6 and 1.8 kbp, although the smallest of these species was not evident in all preparations. Protein from purified virus particle preparations contained a major polypeptide that, in SDS‐PAGE, had an estimated Mr of 40 000 (40K). However, after storage of purified virus particles for 7–10 days, protein preparations from PelRSV particles also contained an additional major polypeptide of estimated Mr of 37 000 that is probably derived by degradation of the 40K protein; this additional component was not observed in freshly prepared preparations of ELV. Neither virus was found to be related serologically to 16 other viruses with isometric particles and similar properties. These data, together with the recent finding by other researchers that the smallest RNA species is a sub‐genomic RNA, suggests that both viruses are members of the genus Carmovirus, and that PelRSV is a minor variant of ELV. However, the taxonomic status of these two viruses is discussed in relation to recent brief reports comparing the nucleotide and amino acid sequences of these two viruses.  相似文献   

9.
A Scottish isolate of cocksfoot streak virus (CSV-S) was found to have flexuous filamentous particles which, in sap of infected cocksfoot plants, had a modal length of 712 nm. It was transmitted from infected to healthy cocksfoot plants in a non-persistent manner by Myzus persicae and by mechanical inoculation of infective sap extracts containing an anti-oxidant. Apart from cocksfoot, mechanical inoculation of infective sap succeeded in infecting only four of 22 plant species tested. The infectivity of sap extracts containing 0.2% thioglycerol was lost after heating for 10 min at 55oC but not 50oC, storage at room temperature for 48 but not 24 hours, and after diluting 10-2to 10-3. Highly purified preparations of CSV-S particles sedimented as a single component with a sedimentation coefficient of 139S and had a buoyant density in rubidium bromide of 1.31 g/cm3. Virus particles were composed of one protein and one ssRNA species with estimated Mr of 31 000 and 3.2 times 106respectively. In ELISA, an antiserum prepared to CSV-S detected the virus in all aerial parts of infected cocksfoot plants and, when present in the ratio of 1 infected leaf: 1000 healthy leaves. Both CSV-S-infected and -uninfected cocksfoot also contained a previously undescribed virus with isometric particles c. 30 nm in diameter. This virus, named cocksfoot cryptic virus (CCV), was seed-borne in two cvs of cocksfoot tested and its particles contained two dsRNA species of estimated Mr of 1.14 times 106and 1.27 times 106. Despite the fact that particles of CSV-S were largely free from CCV particles following exclusion chromatography on agarose beads prior to immunisation, immunoelectron microscopy (IEM) showed that the antiserum prepared to CSV-S also contained some antibodies to CCV. Evidence from IEM suggested a possible distant serological relationship of CCV to ryegrass and beet (BCV 1 or BCV 2, or both) cryptoviruses, all members of sub-group A of crypto viruses.  相似文献   

10.
Flexuous thread‐like virus particles c. 650–700 nm in length were isolated from brusca (Senna pallida) plants showing stunting, mosaic, vein yellowing and leaf malformation. The virus was mechanically transmitted to healthy Senna pallida, Cassia obovata and Cassia emarginata L. plant species. Virus particles sedimented in sucrose density gradients as one component, with a bouyant density of 1.2 g cm?3 in caesium chloride equilibrium gradients. Virions contained a molecule of ssRNA with an apparent size of 6.4 kb. The dsRNA pattern showed one main band of about 12 kb, and two subgenomic dsRNA of c. 10 and c. 5.4 kb. Analyses of purified virus preparations by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE) resolved two coat protein subunits, with mol. wt of c. 28 000 and 26 000 daltons. In Western blotting the virus coat proteins reacted with an homologous polyclonal antiserum and with an antiserum to Lettuce infectious yellow virus. Electron microscopic observations of cells from infected plants showed the accumulation of cytoplasmic vesiculate inclusion bodies and crystalline aggregates of virus particles within phloem tissue. Some of the physicochemical and ultrastructural properties of this virus resemble those of a Closterovirus; however, differences show it to be sufficiently distinct from any previously reported viruses. We proposed the name of Senna chlorotic stunt (SeCSV) for this virus.  相似文献   

11.
The germination stimulants for root parasitic plants Striga and Orobanche produced by sorghum (Sorghum bicolor (L.) Moench), maize (Zea mays L.), and pearl millet (Pennisetum typhoideum Rich.) were examined. Characterization of strigolactones in the root exudates from the plants grown hydroponically was conducted by comparing retention times of germination stimulants on reverse phase high performance liquid chromatography (HPLC) with those of synthetic standards, and by using HPLC linked with tandem mass spectrometry (LC/MS/MS). All the plants tested, except for a sorghum cultivar Swarna, were found to exude two major stimulants, 5-deoxy-strigol, which is known as a branching factor for arbuscular mycorrhizal (AM) fungi, and an isomer of strigol, tentatively named sorghumol. Swarna was found to exude 5-deoxy-strigol and strigol. These results imply that 5-deoxy-strigol is one of major germination stimulants of gramineous plants and that major stimulants may differ even among cultivars within the same species.  相似文献   

12.
抗水稻条纹叶枯病毒核酶的设计,克隆及体外活性测定   总被引:10,自引:0,他引:10  
为探索控制水稻条纹叶枯病毒(Ricestripevirus,RSV)设计合成了特异切割该病毒RNA保守区及编码病害特异性蛋白(DiseaseSpecificProtein,DSP)基因的核酶,核酶基因的长度均为40个碱基,用化学合成方法合成其正链及与其3'-末端互补的15个碱基引物,用TagDNA多聚酶合成其互补链。双链DNA直接插入克隆载体PGEM3zf(+)的Smal位点。序列测定表明,克隆得到的核酶序列与设计的核酶序列完全一致。经SP6RNA多聚酶体外转录得到核酶RNA。当核酶RNA与以同样方法转录得到的靶基因RNA混合反应,可得到预期结果相同的切割片段,表明两种核酶在体外均具有特异性切割活性。  相似文献   

13.
From earlier studies with the Ustilago maydis virus and other dsRNA viruses it is known that discrete dsRNA segments typical of each virus are obtained by extraction. A variation exists with respect to the encapsidation of these segments among different viruses. The encapsidation of the genome in individual particles of the Ustilago virus was examined by electron microscopy after disruption of virus particles. The study included the P6 wild-type and 2 mutants containing only part of the genome. The results indicate that most virus particles of the wild-type and the mutants contain up to 12–14×106 daltons of dsRNA. Since the largest extracted molecule is 3.2×106D these findings suggest that an individual particle may contain more than one segment of dsRNA. Free linear molecules that exceed in size the extracted segments were also found following the disruption of each of the 3 virus types examined. Thus, the viral genome seen segmented after extraction is organized as a concatamer in the capsid and each virus particle can contain an entire viral genome consisting of each type of the segments seen after extraction, a repeat of a single segment or a random assortment. In each case the information may be organized as a concatamer.  相似文献   

14.
Plant RNA virus-based guide RNA (gRNA) delivery has substantial advantages compared to that of the conventional constitutive promoter-driven expression due to the rapid and robust amplification of gRNAs during virus replication and movement. To date, virus-induced genome editing tools have not been developed for wheat and maize. In this study, we engineered a barley stripe mosaic virus (BSMV)-based gRNA delivery system for clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9-mediated targeted mutagenesis in wheat and maize. BSMV-based delivery of single gRNAs for targeted mutagenesis was first validated in Nicotiana benthamiana. To extend this work, we transformed wheat and maize with the Cas9 nuclease gene and selected the wheat TaGASR7 and maize ZmTMS5 genes as targets to assess the feasibility and efficiency of BSMV-mediated mutagenesis. Positive targeted mutagenesis of the TaGASR7 and ZmTMS5 genes was achieved for wheat and maize with efficiencies of up to 78% and 48%. Our results provide a useful tool for fast and efficient delivery of gRNAs into economically important crops.  相似文献   

15.
Nitrate concentration and nitrate reductase activity (NRA) were studied in the leaves of soybean (Glycine max), groundnut (Arachis hypogaea and cowpea (Vigna unguiculata) and sorghum (Sorghum bicolor), pearl millet (Pennisetum americanum) and maize (Zea mays) at three nitrogen fertiliser levels in two field experiments. Higher nitrate concentrations were detected in the leaves of groundnut, cowpea and pearl millet than in sorghum and maize. Nitrate content in the leaves and leaf NRA were not related across crop species, nor was a generalised pattern of leaf NRA and leaf nitrate observed within legumes or within cereals. Nitrogen application resulted in higher nitrate availability in the leaves, with varied leaf NRA.  相似文献   

16.
A fijivirus causing minor enations, stunting, leaf notching, seed head deformity and excess tillering of Digitaria spp. was transmitted from naturally infected Digitaria ciliaris to D. ciliaris, D. decumbens and Urochloa panicoides by the planthopper Sogatella kolophon; 40–70% of insects transmitted after an incubation period of 15–21 days, and continued to transmit for up to 30 more days until death. Symptoms developed in test plants 30–50 days after inoculation. Sogatella longifurcifera failed to transmit the virus under similar conditions. Virus particles were present in roots, stems and leaves of infected plants, and particles were found in regular arrays and random aggregates in fat body cells of transmitting insects. Viroplasm and tubular structures were associated with these particles. Extracts from infective insects contained 10-segment dsRNA when analysed by polyacrylamide gel electrophoresis. Virus survives over winter in planthoppers and D. ciliaris seedlings in frost-free areas of coastal Queensland, but infected plants have debilitated root systems and compete poorly with healthy plants.  相似文献   

17.
During last decades, stripe rust has emerged as a major disease of wheat causing considerable yield loss in northern western plain and northern hill zones of India. Considering significant impact of the disease on wheat crop, field experiments were conducted during rabi seasons of 2013 and 2015 to evaluate the effect of different abiotic factors in different varieties (HD 2967, RSP 561, Agra Local and PBW 343) on the progress and spread of the disease as well as development of a predictive model to predict the disease initiation and spread in the field. Statistical analysis of data revealed that existing of low temperature (10–12 °C), high relative humidity (90%) along with intermittent rainfall was found conducive for disease onset. Thermic variables (atmospheric, canopy and soil temperature) along with age of crop in the selected varieties showed significant positive correlation with disease severity. Step-wise regression showed high R2 of 0.919, 0.885, 0.967 and 0.956 for the predicative model of stripe rust in RSP 561, HD 2967, Agra Local and PBW 343, respectively.  相似文献   

18.
Partially purified virus preparations from sporophores of Agaricus bisporus affected with LaFrance disease had up to a 15-fold-higher RNA-dependent RNA polymerase activity than did comparable preparations from healthy sporophores. Enzyme activity was dependent upon the presence of Mg2+ and the four nucleoside triphosphates and was insensitive to actinomycin D, α-amanitin, and rifampin. The 3H-labeled enzyme reaction products were double-stranded RNA (dsRNA) as indicated by CF-11 cellulose column chromatography and by their ionic-strength-dependent sensitivity to hydrolysis by RNase A. The principal dsRNA products had estimated molecular weights of 4.3 × 106 and 1.4 × 106; they corresponded in size and hybridized to the major dsRNAs detected in the virus preparation by ethidium bromide staining. Cs2SO4 equilibrium centrifugation of the virus preparation resolved a single peak of RNA polymerase activity that banded with a 35-nm spherical virus particle containing dsRNAs with molecular weights of 4.3 × 106 and 1.4 × 106. The data suggest that the RNA-dependent RNA polymerase associated with the 35-nm spherical virus is a replicase which catalyzes the synthesis of the genomic dsRNAs.  相似文献   

19.
Maize stripe virus (MStV) is a potentially threatening virus disease of maize in the tropics. We mapped quantitative trait loci (QTLs) controlling resistance to MStV in a maize population of 157 F(2:3) families derived from the cross between two maize lines, Rev81 (tropical resistant) and B73 (temperate susceptible). Resistance was evaluated under artificial inoculations in replicated screenhouse trials across different seasons in Réunion Island, France. Composite interval mapping was employed for QTL detection with a linkage map of 143 microsatellite markers. Heritability estimates across seasons were 0.96 and 0.90 for incidence and severity, respectively, demonstrating a high genotypic variability and a good control of the environment. Three regions on chromosomes 2L, 3 and 5, with major effects, and another region on chromosome 2S, with minor effects, provided resistance to MStV in Rev81. In individual seasons, the chr2L QTL explained 60-65% of the phenotypic variation for disease incidence and 21-42% for severity. The chr3 QTL, mainly associated with incidence and located near centromere, explained 42-57% of the phenotypic variation, whereas the chr5 QTL, mainly associated with severity, explained 26-53%. Overall, these QTLs explained 68-73% of the phenotypic variance for incidence and 50-59% for severity. The major QTLs on chr2 and 3 showed additive gene action and were found to be stable over time and across seasons. They also were found to be included in genomic regions with important clusters of resistance genes to diseases and pests. The major QTL on chr5 appeared to be partially dominant in favour of resistance. It was stable over time but showed highly significant QTL x season interactions. Possible implications of these QTLs in different mechanisms of resistance against the virus or the insect vector are discussed. The prospects for transferring these QTLs in susceptible maize cultivars and combining them with other resistances to virus diseases by conventional or marker-assisted breeding are promising.  相似文献   

20.
A mechanically transmissible soil-borne virus causing peanut clump disease in Upper Volta is described. It infected mainly species of Chenopodia-ceae and was propagated in Chenopodium amaranticolor. Infectivity was lost from sap of C. amaranticolor after 10 min at 64 °C, and after dilution to 10-5 but not io-4. A purification procedure is described. The particles are rod-shaped and of two predominant lengths, 190 and 245 nm. The virus is not serologically related to tobacco rattle, pea early-browning, or soil-borne wheat mosaic viruses, or to a virus associated with a rhizomania-like disease of beet.  相似文献   

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