On the basis of general situation and new characteristics of tobacco mosaic virus (TMV) in Wuxi tobacco area in 2011, the paper expounded the objec-tive reasons of TMV via systemic investigation, field experiment an...On the basis of general situation and new characteristics of tobacco mosaic virus (TMV) in Wuxi tobacco area in 2011, the paper expounded the objec-tive reasons of TMV via systemic investigation, field experiment and date sorting. Meanwhile, combined with mcteorological conditions and results of systemic inves-tigation, the study especially analyzed meteorological conditions, outbreak and prevalence regularity of TMV and control efficacy of chemical reagents against TMV. The results showed that climatic conditions were the main conditions affecting TMV. There were three occurrence peaks of TMV in 2011, as a result of meteorologi-cal conditions of the months from April to June. The peaks were concerned with a wide range of rainfall about half a month before, low temperature, high humidity and scant sunshine and temperature jump after rain. The results of control effects showed that the chemical reagents could obviously prevent TMV, but once tobacco plants were infected, spraying chemical reagents would not have effective control effect against TMV.展开更多
Using geostatistical method, the semi-variation function models of tobacco mosaic virus (TMV) in east-west and north-south directions were established, and the distribution pattern of TMV in large scale space was st...Using geostatistical method, the semi-variation function models of tobacco mosaic virus (TMV) in east-west and north-south directions were established, and the distribution pattern of TMV in large scale space was studied. The results showed that the distribution pattern of the disease in east-west and north-south directions belonged to linear model with abutment, and the spatial distribution pattern within the investigated areas was aggregated model. The spatial correlation distances in east-west and north-south directions were 29.953 4 and 47.813 8 km, and the spatial variabilities were 95.71% and 80.05%, respectively. This indicated that they had strong spatial correlation. Isoline map accessed by Kringing interpolation method could clearly reflect the spatial aggregated model.展开更多
[Objectives] This study was conducted to screen out suitable agents for controlling tobacco mosaic virus disease and the best control period in Zhangzhou tobacco area, providing a theoretical basis for the control of ...[Objectives] This study was conducted to screen out suitable agents for controlling tobacco mosaic virus disease and the best control period in Zhangzhou tobacco area, providing a theoretical basis for the control of virus diseases, thereby improving the quality of flue-cured tobacco and the income of tobacco farmers. [Methods] The effects on tobacco mosaic virus disease under the interaction between different agents and different application periods were investigated. The incidence of tobacco mosaic virus disease was investigated, and its control effect was analyzed. [Results] Different agents and different application periods had different control effects on tobacco mosaic virus disease. The incidence of tobacco mosaic virus disease: At 30 and 45 d after transplanting, the incidences of A2B1 treatment were the lowest, at 0.85%, 1.71%, respectively;and at 60 d after transplanting, the incidence of A3B1 treatment was the lowest, only 10.68%. The control effect: At 30 and 45 d after transplanting, A2B1 treatment had better control effects, reaching 79.39% and 73.06%, respectively. [Conclusions] 3% hypersensitive protein sprayed at 1 d before transplanting and 7 and 15 d after transplanting achieved the best effect, followed by 10% ningnanmycin sprayed at 1 d before transplanting and 7 and 15 d after transplanting. In tobacco production, it is recommended to apply 1 000 times dilution of 3% supersensitive protein microgranules for three times(at 1 d before transplanting and 7 and 15 d after transplanting), which can effectively prevent tobacco mosaic virus disease.展开更多
The effect of chitosan on the development of infection caused by Tobacco mosaic virus(TMV) in leaves of Nicotiana tabacum L. cv. Samsun has been studied. It was shown that the infectivity and viral coat protein conten...The effect of chitosan on the development of infection caused by Tobacco mosaic virus(TMV) in leaves of Nicotiana tabacum L. cv. Samsun has been studied. It was shown that the infectivity and viral coat protein content in leaves inoculated with a mixture of TMV(2 μg/mL) and chitosan(1 mg/mL) were lower in the early period of infection(3 days after inoculation), by 63% and 66% respectively, than in leaves inoculated with TMV only. Treatment of leaves with chitosan 24 h before inoculation with TMV also caused the antiviral effects, but these were less apparent than when the virus and polysaccharide were applied simultaneously. The inhibitory effects of the agent decreased as the infection progressed. Inoculation of leaves with TMV together with chitosan considerably enhanced the activity of hydrolases(proteases, RNases) in the leaves, in comparison with leaves inoculated with TMV alone. Electron microscope assays of phosphotungstic acid(PTA)-stained suspensions from infected tobacco leaves showed that, in addition to the normal TMV particles(18 nm in diameter, 300 nm long), these suspensions contained abnormal(swollen, "thin" and "short") virions. The highest number of abnormal virions was found in suspensions from leaves inoculated with a mixture of TMV and chitosan. Immuno-electron microscopy showed that "thin" virus particles, in contrast to the particles of normal diameter, lost the ability to bind to specific antiserum. It seems that the chitosan-induced activation of hydrolases stimulates the intracellular degradation of TMV particles and hence hydrolase activation may be considered to be one of the polysaccharide-mediated cellular defense mechanisms that limit virus accumulation in cells.展开更多
Tobacco mosaic virus(TMV) causes significant yield loss in susceptible crops irrigated with contaminated water. However, detection of TMV in water is difficult owing to extremely low concentrations of the virus. Here,...Tobacco mosaic virus(TMV) causes significant yield loss in susceptible crops irrigated with contaminated water. However, detection of TMV in water is difficult owing to extremely low concentrations of the virus. Here, we developed a simple method for the detection and quantification of TMV in irrigation water. TMV was reliably detected at concentrations as low as 10 viral copies/μL with real-time PCR. The sensitivity of detection was further improved using polyethylene glycol 6000(PEG6000, MW 6000) to concentrate TMV from water samples. Among the 28 samples from Shaanxi Province examined with our method, 17 were tested positive after virus concentration. Infectivity of TMV in the original water sample as well as after concentration was confirmed using PCR. The limiting concentration of TMV in water to re-infect plants was determined as 102 viral copies/mL. The method developed in this study offers a novel approach to detect TMV in irrigation water, and may provide an effective tool to control crop infection.展开更多
The AtTOM1 gene of Arabidopsis thaliana had been shown to be essential for the efficient multiplication of Tobacco mosaic virus(TMV) in A.thaliana.In this study,we cloned an AtTOM1-like gene from Nicotiana benthamiana...The AtTOM1 gene of Arabidopsis thaliana had been shown to be essential for the efficient multiplication of Tobacco mosaic virus(TMV) in A.thaliana.In this study,we cloned an AtTOM1-like gene from Nicotiana benthamiana named as NbTOM1.Sequence alignment showed that NbTOM1 is closely related to AtTOM1 homologues of N.tabacum and Lycopersicon esculentum with 97.2% and 92.6% nucleotide sequence identities,respectively.Silencing of NbTOM1 by a modified viral satellite DNA-based vector resulted in complete inhibition of the multiplication of TMV in N.benthamiana.The result suggests that inhibition of NbTOM1 via RNA silencing is a potentially useful method for generating TMV-resistant plants.展开更多
[Objectives] This study was conducted to explore the optimal period of TMV control and the physiological sites that interfere with TMV infection. [Methods] Proteome analysis was performed on the host tissues(tobacco p...[Objectives] This study was conducted to explore the optimal period of TMV control and the physiological sites that interfere with TMV infection. [Methods] Proteome analysis was performed on the host tissues(tobacco plants) at different time periods of viral infection, to verify the changes in the expression of differential protein genes and N and PR1-a in the photosynthetic pathway and porphyrin metabolism and chlorophyll metabolism pathways in proteome; and the tobacco plants were treated with different preparations. [Results] The expression levels of N and PR1-a in the tobacco leaves treated with preparation B reached the highest level, and the effects on the expression levels of the differential protein genes were also the most significant. The control effects of the preparations were tested by the half-leaf method, and the results showed that preparation B had a significant control effect against the early infection of the virus.[Conclusions] This study lays a foundation for the prevention and control of tobacco mosaic virus on crops.展开更多
Our previous reports showed that one opal mutation (UGA) and one ochre mutation (UAA) respectively located in the replicase and movement protein (MP) genes of the attenuated tomato mosaic virus K(ToMV-K) contribute to...Our previous reports showed that one opal mutation (UGA) and one ochre mutation (UAA) respectively located in the replicase and movement protein (MP) genes of the attenuated tomato mosaic virus K(ToMV-K) contribute to the viral attenuation. To explore a wider application of this attenuation pattern to other plant viruses, we have constructed three mutants which respectively contain one opal mutation of the replicase gene and/or one ochre mutation of the MP using PCR-mediated site-directed mutagenesis from a virulent tobacco mosaic virus isolated from China (TMV-Cv). Plant infection performed by in vitro transcripts revealed that the MP truncated mutant TMV-Cvmp and the replicase-MP truncated mutant TMV-Cvrase-mp were infectious on both local lesion (Nicotiana tabacum cv. Xanthi NC) and systemic (N. tabacum cv. K326) host plants, while the replicase truncated mutant TMV-Cvrase was non-infectious. The K326 plant infected by TMV- Cvrease-mp displayed only a little mild mosaic. By electronic microscopy (EM), plant re-inoculation, RNA Dot-blot, RT-PCR and sequencing we demonstrated that the progeny viruses of TMV-Cvmp and TMV-Cvrease-mp shared similar morphological character with TMV-Cv, owned the abilities to infect, replicate and propagate in the assayed plants, and maintained the mutated sites during infection. These data showed that both the opal and the ochre mutations are able to cooperatively induce the attenuated phenotypes of TMV-Cvrase-mp on plants, indicating that the mutation pattern of ToMV-K could be used to attenuate other virulent plant viruses.展开更多
In searching for more bioactive compounds, phytochemical investigations on the acetone extract of the leaves ofNicotiana tabacurn resulted in the isolation of two new sesquiterpenes, nicosesquiterpene A and B (1 and ...In searching for more bioactive compounds, phytochemical investigations on the acetone extract of the leaves ofNicotiana tabacurn resulted in the isolation of two new sesquiterpenes, nicosesquiterpene A and B (1 and 2), along with four known sesquiterpene derivatives (3-6). Structural elucidation of I and 2 was performed by spectral methods, such as HRMS, IR, UV, 1D and 2D NMR spectroscopy. Compounds 1 and 2 are the first naturally occurring pterosin-type sesquiterpene bearing an isopropyl moiety. Compounds 1-6 were also evaluated for their anti-tobacco mosaic virus (anti-TMV) activity. The results showed that compounds 1 and 2 exhibited high anti-TMV activity with inhibition rates of 36.7% and 45.6%, respectively, which is higher than that of positive control. The other compounds also showed potential activity with inhibition rates in the range of 22.7%-29.2%.展开更多
Tobacco mosaic virus(TMV)has been studied as a multi-functional agent for bone tissue engineering.An osteo-inductive effect of wild-type TMV has been reported,as it can significantly enhance the bone differentiation p...Tobacco mosaic virus(TMV)has been studied as a multi-functional agent for bone tissue engineering.An osteo-inductive effect of wild-type TMV has been reported,as it can significantly enhance the bone differentiation potential of bone marrow stromal cells both on a two-dimensional substrate and in a three-dimensional(3D)hydrogel system.A TMV mutant(TMV-RGD1)was created which featured the adhesion peptide arginyl-glycyl-aspartic acid(RGD),the most common peptide motif responsible for cell adhesion to the extracellular matrix,on the surface of the virus particle to enhance the bio-functionality of the scaffold material.We hypothesised that the incorporation of either wild-type TMV or TMV-RGD1 in the 3D hydrogel scaffold would induce bone healing in critical size defects of the cranial segmental bone.We have previously tested the virus-functionalised scaffolds,in vitro,with a hyaluronic acid-based system as an in-situ hydrogel platform for 3D cell encapsulation,culture,and differentiation.The results of these experiments suggested the potential of the virus-functionalised hydrogel to promote in vitro stem cell differentiation.The hydrogel-forming system we employed was shown to be safe and biocompatible in vivo.Here,we further explored the physiological responses regarding bone regeneration of a calvarial defect in both normal and osteoporotic ovariectomized rat models.Our results,based on histological analysis in both animal models,suggested that both wild-type TMV and TMV-RGD1 functionalised hydrogels could accelerate bone regeneration,without systemic toxicity,evaluated by blood counts.New bone formation was intensified by the incorporation of the RGD-mutant viral particles.This finding increased the potential for use of the rodshaped plant virus as a platform for the addition of powerful biofunctionality for tissue engineering applications.This study was approved by the Ethics Committee on Animal Use of the Zhenjiang Affiliated First People’s Hospital affiliated to Jiangsu University.展开更多
DNA methylation plays an important role in the epigenetic regulation of gene expression during plant growth,development,and polyploidization.However,there is still no distinct evidence in tobacco regarding the distrib...DNA methylation plays an important role in the epigenetic regulation of gene expression during plant growth,development,and polyploidization.However,there is still no distinct evidence in tobacco regarding the distribution of the methylation pattern and whether it contributes to qualitative characteristics.We studied the levels and patterns of methylation polymorphism at CCGG sites in 48 accessions of allotetraploid flue-cured tobacco,Nicotiana tabacum,using a methylation-sensitive amplified polymorphism(MSAP) technique.The results showed that methylation existed at a high level among tobacco accessions,among which 49.3% sites were methylated and 69.9% allelic sites were polymorphic.A cluster analysis revealed distinct patterns of geography-specific groups.In addition,three polymorphic sites significantly related to tobacco mosaic virus(TMV) resistance were explored.This suggests that tobacco breeders should pay more attention to epigenetic traits.展开更多
Based on reported TMV-U1 sequence, primers were designed and fragments covering the entire genome of TMV broad bean strain (TMV-B) were obtained with RT-PCR. These fragments were cloned and sequenced and the 5’ and 3...Based on reported TMV-U1 sequence, primers were designed and fragments covering the entire genome of TMV broad bean strain (TMV-B) were obtained with RT-PCR. These fragments were cloned and sequenced and the 5’ and 3’ end sequences of genome were confirmed with RACE. The complete sequence of TMV-B comprises 6 395 nucleotides (nt) and four open reading frames, which correspond to 126 ku (1 116 amino acids), 183 ku (1 616 amino acids), 30 ku (268 amino acids) and 17.5 ku proteins (159 amino acids). The complete nucleotide sequence of TMV-B is 99.4% identical to that of TMV-U1. The two virus isolates share the same sequence of 5’, 3’ non-coding region and 17.5 K ORF, and 6, 1 and 3 amino acid changes are found in 126 K protein, 54 K protein and 30 K protein, respectively. The possible mechanism on the infection of TMV-B in Vicia faba is discussed.展开更多
Tobacco mosaic virus (TMV) and Tomato mosaic virus (ToMV) are two closely related viruses in the genus Tobamovirus, but they induce obviously different sizes of necrotic lesions in tobacco plants containing the N gene...Tobacco mosaic virus (TMV) and Tomato mosaic virus (ToMV) are two closely related viruses in the genus Tobamovirus, but they induce obviously different sizes of necrotic lesions in tobacco plants containing the N gene. Comparison of the symptoms produced by TMV, ToMV and a chimaeric virus (T/OMP), in which the TMV movement protein (MP) gene was re- placed by the ToMV MP gene, showed T/OMP caused necrotic lesions that were similar in size to those of ToMV in tobacco plants containing the N gene. The coat protein and MP of the three viruses accumulated in planta with similar levels, and the replication level of TMV and T/OMP in protoplasts also had no difference. Comparison of the activities of defense-related enzymes (PAL, POD and PPO) induced by the three viruses also showed that the variability of enzyme activity induced by T/OMP was similar to that induced by TMV, but different from that induced by ToMV. The results indicate that the size difference of necrotic lesions induced by TMV and ToMV in to- bacco plants containing the N gene results from the functional difference of their MP genes.展开更多
Genetic linkage maps are essential for studies of genetics, genomic structure, and genomic evolution, and for mapping quantitative trait loci (QTL). Identification of molecular markers and construction of genetic link...Genetic linkage maps are essential for studies of genetics, genomic structure, and genomic evolution, and for mapping quantitative trait loci (QTL). Identification of molecular markers and construction of genetic linkage maps in tobacco (Nicotiana tabacum L.), a classical model plant and important economic crop, have remained limited. In the present study we identified a large number of single nucleotide polymorphism (SNP) markers and constructed a high-density SNP genetic map for tobacco using restriction site-associated DNA sequencing. In 1216.30 Gb of clean sequence obtained using the Illumina HiSeq 2000 sequencing platform, 99,647,735 SNPs were identified that differed between 203 sequenced plant genomes and the tobacco reference genome. Finally, 13,273 SNP markers were mapped on 24 high-density tobacco genetic linkage groups. The entire linkage map spanned 3421.80 cM, with a mean distance of 0.26 cM between adjacent markers. Compared with genetic linkage maps published previously, this version represents a considerable improvement in the number and density of markers. Seven QTL for resistance to cucumber mosaic virus (CMV) in tobacco were mapped to groups 5 and 8. This high-density genetic map is a promising tool for elucidation of the genetic bases of QTL and for molecular breeding in tobacco.展开更多
The recombinant clones of pTN and pNT have been constructed by exchanging the coding regions of the movement proteins (MP), coat proteins (CP) and 3’ noncoding regions between the cDNAs of the tobacco mosaic virus (C...The recombinant clones of pTN and pNT have been constructed by exchanging the coding regions of the movement proteins (MP), coat proteins (CP) and 3’ noncoding regions between the cDNAs of the tobacco mosaic virus (Chinese Isolate, TMV-Cv) and the attenuated tomato mosaic virus N14 genomes, and used as templates for in vitro runoff transcription. Their transcripts have been used for tobacco infection assays. The infection results show that the transcripts of pTN and pNT are infectious. Local lesions were observed in the leaves of Nicotiana tabacum cv. Samsun NN inoculated with pTN transcript, but were fewer than those in the same kind of plant induced by pTMV-Cv transcript. Systemic symptoms were also observed in N. tabacum cv. Huangmiaoyu induced by pTN transcript, but were slighter than those on the same kind of tobacco induced by pTMV-Cv transcript. Local lesions were shown in N. tabacum cv. Samsun NN inoculated with pNT transcript, but were more than those in the same kind of plant induced by pN14展开更多
Cucumber mosaic virus(CMV) is one of the most severe viral diseases transmitted by aphids infecting Solanum crops in China, causing great losses of crop yields and income in rural communities.The tobacco cultivars NC8...Cucumber mosaic virus(CMV) is one of the most severe viral diseases transmitted by aphids infecting Solanum crops in China, causing great losses of crop yields and income in rural communities.The tobacco cultivars NC82 and Taiyan 8 are closely related but differ in resistance to CMV.NC82 is susceptible to infection and Taiyan 8 is resistant, but the mechanisms underlying this difference in resistance are not clear.In this study, we conducted RNA sequencing to analyze changes in gene expression induced in the leaves of Taiyan 8 and NC82 upon systemic infection with CMV, compared with gene expression in the leaves of mock-inoculated plants.Leaves were sampled at one, three, eight, and 15 days after infection.In total, 3443 and 747 differentially expressed genes were identified in Taiyan 8 and NC82, respectively.Gene ontology and pathway enrichment analyses revealed that the different responses to CMV infection between cultivars were based on microtubulebased processes, pentose and glucuronate interconversions, plant–pathogen interaction,and hormone signal transduction pathways.Genes encoding pathogenesis-related proteins, disease-resistance proteins, lipoxygenase, cellulose synthase, an auxin response factor, and an ethylene receptor showed different expression patterns.The differences in gene expression following CMV infection likely contributed to the different resistance levels of these two tobacco cultivars.The comprehensive transcriptome dataset described here,which includes candidate response genes, will serve as a resource for further studies of the molecular mechanisms associated with tobacco defense responses against CMV.展开更多
Ten hybridomas were selected in 1984, which secreted monoclonal antibodies (McAb) specific to tobacco mosaic virus (TMV), and 14 TMV isolates were analysed by the 10 McAbs. Eight determinants were found in the coat pr...Ten hybridomas were selected in 1984, which secreted monoclonal antibodies (McAb) specific to tobacco mosaic virus (TMV), and 14 TMV isolates were analysed by the 10 McAbs. Eight determinants were found in the coat protein of the isolates, they divided the 14 TMV isolates into 6 antigenic types. At the展开更多
An intermediate expressing vector carrying the tobacco mosaic virus (TMV, Chinese common strain) coat protein (CP) gene was constructed by recombinant DNA techniques. The TMV-CP gene was transferred into the tobacco g...An intermediate expressing vector carrying the tobacco mosaic virus (TMV, Chinese common strain) coat protein (CP) gene was constructed by recombinant DNA techniques. The TMV-CP gene was transferred into the tobacco genome via Ti plasmid and a large number of regenerated plants, including both systemic and local lesion hosts for TMV, were obtained. Southern blot analysis revealed that 1-5 copies of the CP gene were integrated into the tobacco genome. RNA and protein analysis demonstrated that the TMV-CP gene was correctly expressed in the transgenic plants. The abundance of TMV-CP mRNA in total leaf RNA accounted for 0.005-0.01%, while the amount of coat proteins reached 0.05-0.2% of the total leaf soluble proteins. Virus challenge experiments showed that the symptom development of virus infection was markedly delayed and the replication as well as the spread of the virus was significantly inhibited in the transgenic plants expressing the TMV-CP gene. Three of these plants were completely protected展开更多
This paper reports a new transgenic tobacco line NC89 resistant to both tobaccomosaic virus(TMV)and cucumber mosaic virus(CMV).It deals with the details of field experi-ments of the whole process of segregation and se...This paper reports a new transgenic tobacco line NC89 resistant to both tobaccomosaic virus(TMV)and cucumber mosaic virus(CMV).It deals with the details of field experi-ments of the whole process of segregation and selection of the progenies of transgenic plant popu-lation.By using the gene transfer technics,some transgenic plants resistant to mosaic disease froma flue-cured tobacco cultivar NC89 were obtained in green house in 1989.Thereafter,throughfield inoculation and selection from the 2nd and 3rd generations,a homozygous line resistant toboth viruses and with good qualities was bred out.The inheretant stability of the resistance of thenew line was confirmed through the 4th to 6th generations.In 1992,the transgenic resistant linewas planted about 7000 hectares in Henan province.The relative effect of controlling mosaic dis-eases of tobacco was 50—70%.The out-put value increased 30—50% per unit area.This is a newexample of growing a transgenic virus-resistant Plant in large scale in China.展开更多
A facile approach to assembled virus film with tunable structure is presented.Rod-like tobacco mosaic virus (TMV) was selected as the prototype in this study for its anisotropic structural feature.TMV can either "...A facile approach to assembled virus film with tunable structure is presented.Rod-like tobacco mosaic virus (TMV) was selected as the prototype in this study for its anisotropic structural feature.TMV can either "lie down" or "stand up" on gold substrate by tuning the solution pH.A quartz crystal microbalance with dissipation monitoring was used to monitor the pH-dependent self-assembly behavior of TMV nanoparticles,and atomic force microscopy and single molecule force spectroscopy further confirmed the different assembly structures.展开更多
基金Supported by Project of Chongqing Tobacco Monopoly Bureau "Investigation and Research of Harmful Organisms in Tobacco of Chongqing City"
文摘On the basis of general situation and new characteristics of tobacco mosaic virus (TMV) in Wuxi tobacco area in 2011, the paper expounded the objec-tive reasons of TMV via systemic investigation, field experiment and date sorting. Meanwhile, combined with mcteorological conditions and results of systemic inves-tigation, the study especially analyzed meteorological conditions, outbreak and prevalence regularity of TMV and control efficacy of chemical reagents against TMV. The results showed that climatic conditions were the main conditions affecting TMV. There were three occurrence peaks of TMV in 2011, as a result of meteorologi-cal conditions of the months from April to June. The peaks were concerned with a wide range of rainfall about half a month before, low temperature, high humidity and scant sunshine and temperature jump after rain. The results of control effects showed that the chemical reagents could obviously prevent TMV, but once tobacco plants were infected, spraying chemical reagents would not have effective control effect against TMV.
基金Supported by Modern Tobacco Agriculture-Project of Dingzhai Base Unit
文摘Using geostatistical method, the semi-variation function models of tobacco mosaic virus (TMV) in east-west and north-south directions were established, and the distribution pattern of TMV in large scale space was studied. The results showed that the distribution pattern of the disease in east-west and north-south directions belonged to linear model with abutment, and the spatial distribution pattern within the investigated areas was aggregated model. The spatial correlation distances in east-west and north-south directions were 29.953 4 and 47.813 8 km, and the spatial variabilities were 95.71% and 80.05%, respectively. This indicated that they had strong spatial correlation. Isoline map accessed by Kringing interpolation method could clearly reflect the spatial aggregated model.
基金Supported by Science and Technology Project of Ganzhou Tobacco Company of Jiangxi Province(GSYJ[2016]9)
文摘[Objectives] This study was conducted to screen out suitable agents for controlling tobacco mosaic virus disease and the best control period in Zhangzhou tobacco area, providing a theoretical basis for the control of virus diseases, thereby improving the quality of flue-cured tobacco and the income of tobacco farmers. [Methods] The effects on tobacco mosaic virus disease under the interaction between different agents and different application periods were investigated. The incidence of tobacco mosaic virus disease was investigated, and its control effect was analyzed. [Results] Different agents and different application periods had different control effects on tobacco mosaic virus disease. The incidence of tobacco mosaic virus disease: At 30 and 45 d after transplanting, the incidences of A2B1 treatment were the lowest, at 0.85%, 1.71%, respectively;and at 60 d after transplanting, the incidence of A3B1 treatment was the lowest, only 10.68%. The control effect: At 30 and 45 d after transplanting, A2B1 treatment had better control effects, reaching 79.39% and 73.06%, respectively. [Conclusions] 3% hypersensitive protein sprayed at 1 d before transplanting and 7 and 15 d after transplanting achieved the best effect, followed by 10% ningnanmycin sprayed at 1 d before transplanting and 7 and 15 d after transplanting. In tobacco production, it is recommended to apply 1 000 times dilution of 3% supersensitive protein microgranules for three times(at 1 d before transplanting and 7 and 15 d after transplanting), which can effectively prevent tobacco mosaic virus disease.
基金supported by the Programs for Fundamental Research of the Presidium of the Russian Academy of Sciences"Molecular-cell biology"
文摘The effect of chitosan on the development of infection caused by Tobacco mosaic virus(TMV) in leaves of Nicotiana tabacum L. cv. Samsun has been studied. It was shown that the infectivity and viral coat protein content in leaves inoculated with a mixture of TMV(2 μg/mL) and chitosan(1 mg/mL) were lower in the early period of infection(3 days after inoculation), by 63% and 66% respectively, than in leaves inoculated with TMV only. Treatment of leaves with chitosan 24 h before inoculation with TMV also caused the antiviral effects, but these were less apparent than when the virus and polysaccharide were applied simultaneously. The inhibitory effects of the agent decreased as the infection progressed. Inoculation of leaves with TMV together with chitosan considerably enhanced the activity of hydrolases(proteases, RNases) in the leaves, in comparison with leaves inoculated with TMV alone. Electron microscope assays of phosphotungstic acid(PTA)-stained suspensions from infected tobacco leaves showed that, in addition to the normal TMV particles(18 nm in diameter, 300 nm long), these suspensions contained abnormal(swollen, "thin" and "short") virions. The highest number of abnormal virions was found in suspensions from leaves inoculated with a mixture of TMV and chitosan. Immuno-electron microscopy showed that "thin" virus particles, in contrast to the particles of normal diameter, lost the ability to bind to specific antiserum. It seems that the chitosan-induced activation of hydrolases stimulates the intracellular degradation of TMV particles and hence hydrolase activation may be considered to be one of the polysaccharide-mediated cellular defense mechanisms that limit virus accumulation in cells.
基金supported by the Key Technology Program of China National Tobacco Corporation (110200902046)111 Project from the Education Ministry of China, (No.B07049)the National High-tech R&D Program of China (no. 2012AA101504)
文摘Tobacco mosaic virus(TMV) causes significant yield loss in susceptible crops irrigated with contaminated water. However, detection of TMV in water is difficult owing to extremely low concentrations of the virus. Here, we developed a simple method for the detection and quantification of TMV in irrigation water. TMV was reliably detected at concentrations as low as 10 viral copies/μL with real-time PCR. The sensitivity of detection was further improved using polyethylene glycol 6000(PEG6000, MW 6000) to concentrate TMV from water samples. Among the 28 samples from Shaanxi Province examined with our method, 17 were tested positive after virus concentration. Infectivity of TMV in the original water sample as well as after concentration was confirmed using PCR. The limiting concentration of TMV in water to re-infect plants was determined as 102 viral copies/mL. The method developed in this study offers a novel approach to detect TMV in irrigation water, and may provide an effective tool to control crop infection.
基金Project supported by the Cultivation Fund of the Key Scientific and Technical Innovation Project, Ministry of Education of China (No. 705025) the National Natural Science Foundation of China (No. 30530520)
文摘The AtTOM1 gene of Arabidopsis thaliana had been shown to be essential for the efficient multiplication of Tobacco mosaic virus(TMV) in A.thaliana.In this study,we cloned an AtTOM1-like gene from Nicotiana benthamiana named as NbTOM1.Sequence alignment showed that NbTOM1 is closely related to AtTOM1 homologues of N.tabacum and Lycopersicon esculentum with 97.2% and 92.6% nucleotide sequence identities,respectively.Silencing of NbTOM1 by a modified viral satellite DNA-based vector resulted in complete inhibition of the multiplication of TMV in N.benthamiana.The result suggests that inhibition of NbTOM1 via RNA silencing is a potentially useful method for generating TMV-resistant plants.
文摘[Objectives] This study was conducted to explore the optimal period of TMV control and the physiological sites that interfere with TMV infection. [Methods] Proteome analysis was performed on the host tissues(tobacco plants) at different time periods of viral infection, to verify the changes in the expression of differential protein genes and N and PR1-a in the photosynthetic pathway and porphyrin metabolism and chlorophyll metabolism pathways in proteome; and the tobacco plants were treated with different preparations. [Results] The expression levels of N and PR1-a in the tobacco leaves treated with preparation B reached the highest level, and the effects on the expression levels of the differential protein genes were also the most significant. The control effects of the preparations were tested by the half-leaf method, and the results showed that preparation B had a significant control effect against the early infection of the virus.[Conclusions] This study lays a foundation for the prevention and control of tobacco mosaic virus on crops.
基金the National Natural Science Foundation of China.
文摘Our previous reports showed that one opal mutation (UGA) and one ochre mutation (UAA) respectively located in the replicase and movement protein (MP) genes of the attenuated tomato mosaic virus K(ToMV-K) contribute to the viral attenuation. To explore a wider application of this attenuation pattern to other plant viruses, we have constructed three mutants which respectively contain one opal mutation of the replicase gene and/or one ochre mutation of the MP using PCR-mediated site-directed mutagenesis from a virulent tobacco mosaic virus isolated from China (TMV-Cv). Plant infection performed by in vitro transcripts revealed that the MP truncated mutant TMV-Cvmp and the replicase-MP truncated mutant TMV-Cvrase-mp were infectious on both local lesion (Nicotiana tabacum cv. Xanthi NC) and systemic (N. tabacum cv. K326) host plants, while the replicase truncated mutant TMV-Cvrase was non-infectious. The K326 plant infected by TMV- Cvrease-mp displayed only a little mild mosaic. By electronic microscopy (EM), plant re-inoculation, RNA Dot-blot, RT-PCR and sequencing we demonstrated that the progeny viruses of TMV-Cvmp and TMV-Cvrease-mp shared similar morphological character with TMV-Cv, owned the abilities to infect, replicate and propagate in the assayed plants, and maintained the mutated sites during infection. These data showed that both the opal and the ochre mutations are able to cooperatively induce the attenuated phenotypes of TMV-Cvrase-mp on plants, indicating that the mutation pattern of ToMV-K could be used to attenuate other virulent plant viruses.
基金supported by the National Natural Science Foundation of China (Nos. 31360081 and 31400303)the Basic Research Foundation of Yunnan Province (Nos. 2014FB163 and 2014FD078)the Product Research Foundation of China Tobacco Yunnan Industrial Co., Ltd. (No. 2014CP01)
文摘In searching for more bioactive compounds, phytochemical investigations on the acetone extract of the leaves ofNicotiana tabacurn resulted in the isolation of two new sesquiterpenes, nicosesquiterpene A and B (1 and 2), along with four known sesquiterpene derivatives (3-6). Structural elucidation of I and 2 was performed by spectral methods, such as HRMS, IR, UV, 1D and 2D NMR spectroscopy. Compounds 1 and 2 are the first naturally occurring pterosin-type sesquiterpene bearing an isopropyl moiety. Compounds 1-6 were also evaluated for their anti-tobacco mosaic virus (anti-TMV) activity. The results showed that compounds 1 and 2 exhibited high anti-TMV activity with inhibition rates of 36.7% and 45.6%, respectively, which is higher than that of positive control. The other compounds also showed potential activity with inhibition rates in the range of 22.7%-29.2%.
基金This research was supported by the Thailand Research Fund and Office of the Higher Education Commission(No.MRG6180264)Chulalongkorn University,and the National Natural Science Foundation of China(No.21750110445).
文摘Tobacco mosaic virus(TMV)has been studied as a multi-functional agent for bone tissue engineering.An osteo-inductive effect of wild-type TMV has been reported,as it can significantly enhance the bone differentiation potential of bone marrow stromal cells both on a two-dimensional substrate and in a three-dimensional(3D)hydrogel system.A TMV mutant(TMV-RGD1)was created which featured the adhesion peptide arginyl-glycyl-aspartic acid(RGD),the most common peptide motif responsible for cell adhesion to the extracellular matrix,on the surface of the virus particle to enhance the bio-functionality of the scaffold material.We hypothesised that the incorporation of either wild-type TMV or TMV-RGD1 in the 3D hydrogel scaffold would induce bone healing in critical size defects of the cranial segmental bone.We have previously tested the virus-functionalised scaffolds,in vitro,with a hyaluronic acid-based system as an in-situ hydrogel platform for 3D cell encapsulation,culture,and differentiation.The results of these experiments suggested the potential of the virus-functionalised hydrogel to promote in vitro stem cell differentiation.The hydrogel-forming system we employed was shown to be safe and biocompatible in vivo.Here,we further explored the physiological responses regarding bone regeneration of a calvarial defect in both normal and osteoporotic ovariectomized rat models.Our results,based on histological analysis in both animal models,suggested that both wild-type TMV and TMV-RGD1 functionalised hydrogels could accelerate bone regeneration,without systemic toxicity,evaluated by blood counts.New bone formation was intensified by the incorporation of the RGD-mutant viral particles.This finding increased the potential for use of the rodshaped plant virus as a platform for the addition of powerful biofunctionality for tissue engineering applications.This study was approved by the Ethics Committee on Animal Use of the Zhenjiang Affiliated First People’s Hospital affiliated to Jiangsu University.
基金supported by the Program for High-Quality Tobacco Development of China (No. [2010]221)the Foundation of Science and Technology of Guizhou Province (No. J[2010]2251)the Program for Guizhou Tobacco Science of China (No. 200910)
文摘DNA methylation plays an important role in the epigenetic regulation of gene expression during plant growth,development,and polyploidization.However,there is still no distinct evidence in tobacco regarding the distribution of the methylation pattern and whether it contributes to qualitative characteristics.We studied the levels and patterns of methylation polymorphism at CCGG sites in 48 accessions of allotetraploid flue-cured tobacco,Nicotiana tabacum,using a methylation-sensitive amplified polymorphism(MSAP) technique.The results showed that methylation existed at a high level among tobacco accessions,among which 49.3% sites were methylated and 69.9% allelic sites were polymorphic.A cluster analysis revealed distinct patterns of geography-specific groups.In addition,three polymorphic sites significantly related to tobacco mosaic virus(TMV) resistance were explored.This suggests that tobacco breeders should pay more attention to epigenetic traits.
文摘Based on reported TMV-U1 sequence, primers were designed and fragments covering the entire genome of TMV broad bean strain (TMV-B) were obtained with RT-PCR. These fragments were cloned and sequenced and the 5’ and 3’ end sequences of genome were confirmed with RACE. The complete sequence of TMV-B comprises 6 395 nucleotides (nt) and four open reading frames, which correspond to 126 ku (1 116 amino acids), 183 ku (1 616 amino acids), 30 ku (268 amino acids) and 17.5 ku proteins (159 amino acids). The complete nucleotide sequence of TMV-B is 99.4% identical to that of TMV-U1. The two virus isolates share the same sequence of 5’, 3’ non-coding region and 17.5 K ORF, and 6, 1 and 3 amino acid changes are found in 126 K protein, 54 K protein and 30 K protein, respectively. The possible mechanism on the infection of TMV-B in Vicia faba is discussed.
基金This work was supported by China National Key Basic ResearchDevelopment Program(Grant No.G2000016204)the National Outstanding Youth Foundation from National Natural Science Foundation of China(Grant No.30125032).
文摘Tobacco mosaic virus (TMV) and Tomato mosaic virus (ToMV) are two closely related viruses in the genus Tobamovirus, but they induce obviously different sizes of necrotic lesions in tobacco plants containing the N gene. Comparison of the symptoms produced by TMV, ToMV and a chimaeric virus (T/OMP), in which the TMV movement protein (MP) gene was re- placed by the ToMV MP gene, showed T/OMP caused necrotic lesions that were similar in size to those of ToMV in tobacco plants containing the N gene. The coat protein and MP of the three viruses accumulated in planta with similar levels, and the replication level of TMV and T/OMP in protoplasts also had no difference. Comparison of the activities of defense-related enzymes (PAL, POD and PPO) induced by the three viruses also showed that the variability of enzyme activity induced by T/OMP was similar to that induced by TMV, but different from that induced by ToMV. The results indicate that the size difference of necrotic lesions induced by TMV and ToMV in to- bacco plants containing the N gene results from the functional difference of their MP genes.
基金supported by the Agricultural Science and Technology Innovation Program (ASTIP-TRIC01)
文摘Genetic linkage maps are essential for studies of genetics, genomic structure, and genomic evolution, and for mapping quantitative trait loci (QTL). Identification of molecular markers and construction of genetic linkage maps in tobacco (Nicotiana tabacum L.), a classical model plant and important economic crop, have remained limited. In the present study we identified a large number of single nucleotide polymorphism (SNP) markers and constructed a high-density SNP genetic map for tobacco using restriction site-associated DNA sequencing. In 1216.30 Gb of clean sequence obtained using the Illumina HiSeq 2000 sequencing platform, 99,647,735 SNPs were identified that differed between 203 sequenced plant genomes and the tobacco reference genome. Finally, 13,273 SNP markers were mapped on 24 high-density tobacco genetic linkage groups. The entire linkage map spanned 3421.80 cM, with a mean distance of 0.26 cM between adjacent markers. Compared with genetic linkage maps published previously, this version represents a considerable improvement in the number and density of markers. Seven QTL for resistance to cucumber mosaic virus (CMV) in tobacco were mapped to groups 5 and 8. This high-density genetic map is a promising tool for elucidation of the genetic bases of QTL and for molecular breeding in tobacco.
文摘The recombinant clones of pTN and pNT have been constructed by exchanging the coding regions of the movement proteins (MP), coat proteins (CP) and 3’ noncoding regions between the cDNAs of the tobacco mosaic virus (Chinese Isolate, TMV-Cv) and the attenuated tomato mosaic virus N14 genomes, and used as templates for in vitro runoff transcription. Their transcripts have been used for tobacco infection assays. The infection results show that the transcripts of pTN and pNT are infectious. Local lesions were observed in the leaves of Nicotiana tabacum cv. Samsun NN inoculated with pTN transcript, but were fewer than those in the same kind of plant induced by pTMV-Cv transcript. Systemic symptoms were also observed in N. tabacum cv. Huangmiaoyu induced by pTN transcript, but were slighter than those on the same kind of tobacco induced by pTMV-Cv transcript. Local lesions were shown in N. tabacum cv. Samsun NN inoculated with pNT transcript, but were more than those in the same kind of plant induced by pN14
基金funded by the Agricultural Science and Technology Innovation Program (ASTIP-TRIC01)the Science Foundation for Young Scientists of the Tobacco Research Institute of the Chinese Academy of Agricultural Sciences(2016A04)+2 种基金the National Natural Science Foundation of China (31301678)Fundamental Research Funds for Central Nonprofit Scientific InstitutionTobacco Genome Project of China National Tobacco Corporation (110201601028, 110201402006, 110201301009)
文摘Cucumber mosaic virus(CMV) is one of the most severe viral diseases transmitted by aphids infecting Solanum crops in China, causing great losses of crop yields and income in rural communities.The tobacco cultivars NC82 and Taiyan 8 are closely related but differ in resistance to CMV.NC82 is susceptible to infection and Taiyan 8 is resistant, but the mechanisms underlying this difference in resistance are not clear.In this study, we conducted RNA sequencing to analyze changes in gene expression induced in the leaves of Taiyan 8 and NC82 upon systemic infection with CMV, compared with gene expression in the leaves of mock-inoculated plants.Leaves were sampled at one, three, eight, and 15 days after infection.In total, 3443 and 747 differentially expressed genes were identified in Taiyan 8 and NC82, respectively.Gene ontology and pathway enrichment analyses revealed that the different responses to CMV infection between cultivars were based on microtubulebased processes, pentose and glucuronate interconversions, plant–pathogen interaction,and hormone signal transduction pathways.Genes encoding pathogenesis-related proteins, disease-resistance proteins, lipoxygenase, cellulose synthase, an auxin response factor, and an ethylene receptor showed different expression patterns.The differences in gene expression following CMV infection likely contributed to the different resistance levels of these two tobacco cultivars.The comprehensive transcriptome dataset described here,which includes candidate response genes, will serve as a resource for further studies of the molecular mechanisms associated with tobacco defense responses against CMV.
文摘Ten hybridomas were selected in 1984, which secreted monoclonal antibodies (McAb) specific to tobacco mosaic virus (TMV), and 14 TMV isolates were analysed by the 10 McAbs. Eight determinants were found in the coat protein of the isolates, they divided the 14 TMV isolates into 6 antigenic types. At the
基金Project supported by the Chinese National Science Committee and the World Laboratory of International Center for Science and Culture, Geneva, Switzerland.
文摘An intermediate expressing vector carrying the tobacco mosaic virus (TMV, Chinese common strain) coat protein (CP) gene was constructed by recombinant DNA techniques. The TMV-CP gene was transferred into the tobacco genome via Ti plasmid and a large number of regenerated plants, including both systemic and local lesion hosts for TMV, were obtained. Southern blot analysis revealed that 1-5 copies of the CP gene were integrated into the tobacco genome. RNA and protein analysis demonstrated that the TMV-CP gene was correctly expressed in the transgenic plants. The abundance of TMV-CP mRNA in total leaf RNA accounted for 0.005-0.01%, while the amount of coat proteins reached 0.05-0.2% of the total leaf soluble proteins. Virus challenge experiments showed that the symptom development of virus infection was markedly delayed and the replication as well as the spread of the virus was significantly inhibited in the transgenic plants expressing the TMV-CP gene. Three of these plants were completely protected
文摘This paper reports a new transgenic tobacco line NC89 resistant to both tobaccomosaic virus(TMV)and cucumber mosaic virus(CMV).It deals with the details of field experi-ments of the whole process of segregation and selection of the progenies of transgenic plant popu-lation.By using the gene transfer technics,some transgenic plants resistant to mosaic disease froma flue-cured tobacco cultivar NC89 were obtained in green house in 1989.Thereafter,throughfield inoculation and selection from the 2nd and 3rd generations,a homozygous line resistant toboth viruses and with good qualities was bred out.The inheretant stability of the resistance of thenew line was confirmed through the 4th to 6th generations.In 1992,the transgenic resistant linewas planted about 7000 hectares in Henan province.The relative effect of controlling mosaic dis-eases of tobacco was 50—70%.The out-put value increased 30—50% per unit area.This is a newexample of growing a transgenic virus-resistant Plant in large scale in China.
基金supported by the National Natural Science Foundation of China (20423003 & 20774097)Z.S. thanks the NSFC Fund for Creative Research Groups (50921062) for support+1 种基金the financial support from US NSF (DMR-0706431, CHE-0748690)US DoD, US DoE-BES, and the W. M. Keck Foundation
文摘A facile approach to assembled virus film with tunable structure is presented.Rod-like tobacco mosaic virus (TMV) was selected as the prototype in this study for its anisotropic structural feature.TMV can either "lie down" or "stand up" on gold substrate by tuning the solution pH.A quartz crystal microbalance with dissipation monitoring was used to monitor the pH-dependent self-assembly behavior of TMV nanoparticles,and atomic force microscopy and single molecule force spectroscopy further confirmed the different assembly structures.