Some properties of the virus causing the mosaic of sweet peas in Czechoslovakia

Identification trials were carried out to determine what virus causes a mosaic disease of sweet peas in Czechoslovakia. The found properties of the identified sweet pea mosaic virus, the character of its transmission and its host range prove that sweet peas in Czechoslovakia are attacked by common pea mosaic virus (CPMV). Some insignificant differences in properties between our virus isolate and CPMV were observed in the course of the determination of properties of our virus isolate on various host plants. The possible existence of more strains of CPMV is discussed. The transmission of sweet pea mosaic to the plants ofPhaseolus vulgaris L. was negative. For this reason bean yellow mosaic virus (BYMV) was eliminated as a possible pathogen of our virus isolate. At the same time the indicator plantsChenopodium giganteum Don. andChenopodium guinoa Willd. with eight developed leaves were established to be most suitable for the determination of the properties of the isolate by the half-leaf test. The transmission of the virus isolate by seeds was not proved.     

Serological studies of Bratislava mosaic virus of grape vine

Bratislava mosaic virus was transmitted mechanically toChenopodium quinoa. The virus was isolated by means of gradient ultracentrifugation and the, proteins were distributed on an automatic recording photometer. Single fractions were taken with a fraction collector and used as antigen. At the same time, antigen from leaves of the variety Sylván zelený infected with the Bratislava mosaic virus was prepared by means of the gradient ultracentrifugation. The antisera were obtained from rabbits immunized with individual antigens. The antisera were tested with the saps ofC. quinoa infected with the Bratislava mosaic virus, by the method of double diffusion into agar according to Ouchterlony, and with adjusted saps from suckers infected with the Bratislava mosaic virus, by the method of double diffusion into agar according to Oudin. A specific reaction can be obtained in both cases only under the assumption that all conditions mentioned in the study are strictly kept.     

Interaction of cucumber mosaic virus and potato virus y with tobacco mosaic virus

A study was performed on the interaction of cucumber mosaic virus (CMV) of potato virus Y (PVY) with tobacco mosaic virus (TMV). Interference was evaluated using tobacco plantsNicotiana tabacum cv. Java responding to CMV and PVY with a systemic infection and to TMV with local necrotic lesions. The decrease in TMV — induced lesion number gave evidence of a decrease in susceptibility caused by the previous infection with CMV or PVY, the decrease of lesion enlargement demonstrated a decreased TMV reproduction in the plants previously infected with CMV or PVY. The interference concerned was incomplete, as evaluated from reproduction of the challenging TMV and from the decrease in susceptibility of the host to TMV brought about by the first infection with CMV or PVY.     

Purification and conservation of infective sugar beet mosaic virus

Sugar beet mosaic virus (SBMV) was precipitated by polyethyleneglycol (PEG) 6000 from the cell sap of infected sugar beet leaves. After centrifugation and addition of dextrane T 10 the virus was lyophilized. Its infectious activity was demonstrated by mechanical transmission toChenopodium quinoa Willd. and to sugar beet. Stability of infectious activity of the lyophilized virus was verified.     

Identification by immunoprecipitation of cauliflower mosaic virus in vitro major translation product with a specific serum against viroplasm protein

A highly specific antiserum was prepared against purified cauliflower mosaic virus viroplasm-protein (VmP). A virus specific in vitro major translation product (TPmaj), encoded by the 19S poly(A)⁺ RNA fraction from cauliflower mosaic virus infected turnip leaves, was recognized by this antiserum. The N-terminal sequence of TPmaj corresponds to the sequence following the first in-phase initiation codon in gene VI of the cauliflower mosaic virus genome. Both VmP and TPmaj have blocked termini and probably start from the same AUG codon.     

Purification and properties of arabis mosaic and tomato bushy stunt viruses isolated from lilac (Syringa vulgaris L.)

The paper gives more detailed characteristics of Arabis mosaic virus (AMV) and tomato bushy stunt virus (TBSV) isolated from lilac, the latter being identified in lilac (from plants suffering from yellow ring disease) for the first time. The isolate of TBSV from lilac, from which an antiserum with a titre of 1024 was prepared, is closely related to the artichoke strain. Information is given about two types of ringspot disease and about chlorotic ringspot of lilac. Whereas in the leaves of lilac suffering from ringspot disease (of ring mosaic type) the presence of AMV was demonstrated, the sap transmission from the leaves diseased with ringspot of linepattern (and wave-like mosaic) type failed; from the leaves affected by chlorotic ringspot a mixture of AMV and cherry leaf roll virus was identified. In addition, the polyetiological nature of “spring” mosaic and necrotic mosaic of lilac, in which bacteriumPseudomonas syringae van Hall, was found is dealt with. The TBSV was also identified in the isolate of necrotic mosaic.Additional index words: Lilac ringspot, chlorotic ringspot, yellow ring, “spring” mosaic, necrotic mosaic, cherry leaf roll virus,Pseudomonas syringae van Hall.     

Natural infections ofSisymbrium loeselii Jusl. with cabbage black ringspot and tobacco mosaic viruses

Investigating weeds for viruses in ruderal localities of Greater Prague two forms of mosaic diseases inSisymbrium loeselii Jusl. were demonstrated (green and yellowish-green mosaic). Transmission tests carried out on differential host plants showed that the green mosaic is caused by cabbage black ringspot virus (CBRV) and the yellowish green by mixed infection of CBRV and tobacco mosaic virus (TMV). TMV—isolate is characterized as an unusual necrotic strain; its capability to reproduce in cruciferous plant in nature is unique. It was ascertained that green mosaic was commonly spread overSisymbrium plants in ruderal ***DIRECT SUPPORT *** A01GP029 00004 associations on Prague territory; epidemiological significance of this discovery is discussed.     

Proof by synthesis of Tobacco mosaic virus

Background

Synthetic biology is a discipline that includes making life forms artificially from chemicals. Here, a DNA molecule was enzymatically synthesized in vitro from DNA templates made from oligonucleotides representing the text of the first Tobacco mosaic virus (TMV) sequence elucidated in 1982. No infectious DNA molecule of that seminal reference sequence exists, so the goal was to synthesize it and then build viral chimeras.

Results

RNA was transcribed from synthetic DNA and encapsidated with capsid protein in vitro to make synthetic virions. Plants inoculated with the virions did not develop symptoms. When two nucleotide mutations present in the original sequence, but not present in most other TMV sequences in GenBank, were altered to reflect the consensus, the derivative synthetic virions produced classic TMV symptoms. Chimeras were then made by exchanging TMV capsid protein DNA with Tomato mosaic virus (ToMV) and Barley stripe mosaic virus (BSMV) capsid protein DNA. Virus expressing ToMV capsid protein exhibited altered, ToMV-like symptoms in Nicotiana sylvestris. A hybrid ORF6 protein unknown to nature, created by substituting the capsid protein genes in the virus, was found to be a major symptom determinant in Nicotiana benthamiana. Virus expressing BSMV capsid protein did not have an extended host range to barley, but did produce novel symptoms in N. benthamiana.

Conclusions

This first report of the chemical synthesis and artificial assembly of a plant virus corrects a long-standing error in the TMV reference genome sequence and reveals that unnatural hybrid virus proteins can alter symptoms unexpectedly.     

A yellow mosaic disease of horse chestnut (Aesculus spp.) caused by apple mosaic virus

A severe foliar yellow mosaic disease was observed in horse chestnut trees (Aesculus carnea and A. hippocastanum). Reactions in woody indicator plants grafted with diseased horse chestnut suggested the presence of an ilarvirus. Virus isolates obtained by mechanical inoculation of herbaceous test plants reacted with antisera to apple mosaic virus but not with antisera to its serotype prunus necrotic ringspot virus, or to prune dwarf virus. Yellow mosaic was induced in horse chestnut seedlings grafted with tissues from herbaceous hosts infected with horse chestnut isolates or with the European plum line pattern isolate of apple mosaic virus. Virus was detected by enzyme-linked immunosorbent assay (ELISA) in embryo and endosperm of immature seed from infected trees but not in mature seed, or progeny seedlings. Strawberry latent ringspot virus was detected in one of six A. hippocastanum trees with a leaf vein yellows disease.     

Reproduction of sugar beet mosaic and tobacco mosaic viruses in anthocyanized beet plants

During our studies on the interaction of anthocyanins and plant virus diseases, reproduction of sugar beet mosaic (SBMV) and tobacco mosaic viruses (TMV) was investigated. Experiments were carried out in leaves of sugar beet,Beta vulgaris cv. Dobrovicka N and its spontaneous anthocyanized mutant. SBMV induces a systemic infection while TMV is responsible for primary local symptoms in sugar beet leaves only. Our quantitative analyses onAmaranthus caudatus L. andChenopodium quinoa Wilid. showed a significant decrease in concentration of SBMV in juice extracted from anthocyanized beet plants as compared with extracts from normal green infected plants. Significant differences were also obtained when SBMV — containing juice was tested in mixtures with healthy extracts from anthocyanized and normal green plants. Also the intensity of TMV symptoms in beet leaves was considerably decreased in leaves of antho-eyanized plants.     

Diagnosis of Symptomless Yellow mosaic begomovirus Infection in Pigeonpea by Using Cloned Mungbean yellow mosaic India virus as Probe

One isolate of Mungbean yellow mosaic India virus (MYMIV) of mungbean plants from Sri Ganganagar, Rajasthan, designated as MYMIV-Mg was isolated and DNA-A and DNA-B, the two full length bipartite genomic components of this virus, were cloned. The [α-³²P] labeled diagnostic probes specific to these cloned DNA-A and -B of MYMIV-Mg were used to detect the virus infection in infected plants by nucleic acid spot hybridization (NASH) test. The NASH tests detected the MYMIV infection and concentration of viral titre in susceptible, moderately susceptible, resistant and symptomless genotypes of pigeonpea (Cajanus cajan) plants. Fourteen genotypes of pigeonpea were tested against five naturally occurring MYMIV variants viz.,.MYMIV Bg, -MgD, -MoL, -Mg and -Pp1 through viruliferous whitefly (Bemisia tabaci) transmission in greenhouse condition. Disease incidence and severity of MYMIV in different pigeonpea genotypes varied with the variants of MYMIV. Many genotypes of pigeonpea did not produce visible yellow mosaic symptoms after inoculation with MYMIV variants MYMIV-Bg, -MbD and -MoL, although, majority of the symptomless genotypes were found to be infected by MYMIV, as viral DNA was detected by NASH test.     

Studies of some ways in which carrot mosaic virus can be transmitted

The transmission of carrot mosaic virus (CMV) by the crude sap to 11 varieties of plants from 4 families was demonstrated. From these plants the virus could be transferred back to a healthy carrot cultivated from the seed in isolation. The incubation time required for the appearance of the symptoms of CMV was 7–20 days. The plants on which mosaic or spot symptoms appeared on the leaves after transfer by the sap at temperatures below 15°C remains habitually healthy after the transfer of virus at higher temperatures. The results of the mechanical inoculation of CMV by the crude infectious sap to young carrots cultivated from seeds differentiated this virus fromApium virus 1, which after mechanical inoculation causes chlorosis of the youngest carrot leaves in contrast to CMV. A further differentiation of CMV fromApium virus 1 is shown by the fact that CMV can be transferred only to the familyDaucaceae. It differs in this fromApium virus 1 which is transferred exclusively to this family (Köhler, Klinkowski 1954). CMV is differentiated fromCucumis virus 1/Doolittle Smith by some different host plants.     

Expression of cauliflower mosaic virus gene I in Saccharomyces cerevisiae

Cauliflower mosaic virus (CaMV) gene I encodes a 40-kDa protein, P1, which is thought to be involved in the cell-to-cell movement of the virus. In order to investigate its functioning, P1 was expressed in Saccharomyces cerevisiae transformed by an expression vector containing CaMV gene I. When produced in yeast, P1 was 40 kDa in size and not N-glycosylated.     

Evolutionary relationship of alfalfa mosaic virus with cucumber mosaic virus and brome mosaic virus

The amino acid sequences of the non-structural protein (molecular weight 35,000; 3a protein) from three plant viruses — cucumber mosaic, brome mosaic and alfalfa mosaic have been systematically compared using the partial genomic sequences for these three viruses already available. The 3a protein of cucumber mosaic virus has an amino acid sequence homology of 33.7% with the corresponding protein of brome mosaic virus. A similar protein from alfalfa mosaic virus has a homology of 18.2% and 14.2% with the protein from brome mosaic virus and cucumber mosaic virus, respectively. These results suggest that the three plant viruses are evolutionarily related, although, the evolutionary distance between alfalfa mosaic virus and cucumber mosaic virus or brome mosaic virus is much larger than the corresponding distance between the latter two viruses.     

High Plains wheat mosaic virus: An enigmatic disease of wheat and corn causing the High Plains disease

Brief historyIn 1993, severe mosaic and necrosis symptoms were observed on corn (maize) and wheat from several Great Plains states of the USA. Based on the geographical location of infections, the disease was named High Plains disease and the causal agent was tentatively named High Plains virus. Subsequently, researchers renamed this virus as maize red stripe virus and wheat mosaic virus to represent the host and symptom phenotype of the virus. After sequencing the genome of the pathogen, the causal agent of High Plains disease was officially named as High Plains wheat mosaic virus. Hence, High Plains virus, maize red stripe virus, wheat mosaic virus, and High Plains wheat mosaic virus (HPWMoV) are synonyms for the causal agent of High Plains disease.TaxonomyHigh Plains wheat mosaic virus is one of the 21 definitive species in the genus Emaravirus in the family Fimoviridae.VirionThe genomic RNAs are encapsidated in thread‐like nucleocapsids in double‐membrane 80–200 nm spherical or ovoid virions.Genome characterizationThe HPWMoV genome consists of eight single‐stranded negative‐sense RNA segments encoding a single open reading frame (ORF) in each genomic RNA segment. RNA 1 is 6,981‐nucleotide (nt) long, coding for a 2,272 amino acid protein of RNA‐dependent RNA polymerase. RNA 2 is 2,211‐nt long and codes for a 667 amino acid glycoprotein precursor. RNA 3 has two variants of 1,439‐ and 1,441‐nt length that code for 286 and 289 amino acid nucleocapsid proteins, respectively. RNA 4 is 1,682‐nt long, coding for a 364 amino acid protein. RNA 5 and RNA 6 are 1,715‐ and 1,752‐nt long, respectively, and code for 478 and 492 amino acid proteins, respectively. RNA 7 and RNA 8 are 1,434‐ and 1,339‐nt long, code for 305 and 176 amino acid proteins, respectively.Biological propertiesHPWMoV can infect wheat, corn (maize), barley, rye brome, oat, rye, green foxtail, yellow foxtail, and foxtail barley. HPWMoV is transmitted by the wheat curl mite and through corn seed.Disease managementGenetic resistance against HPWMoV in wheat is not available, but most commercial corn hybrids are resistant while sweet corn varieties remain susceptible. Even though corn hybrids are resistant to virus, it still serves as a green bridge host that enables mites to carry the virus from corn to new crop wheat in the autumn. The main management strategy for High Plains disease in wheat relies on the management of green bridge hosts. Cultural practices such as avoiding early planting can be used to avoid mite buildup and virus infections.     

Resistance of transformed and non-transformed oilseed rape cv. HM-81 to the infection with cauliflower mosaic,turnip yellow mosaic and turnip mosaic viruses

Resistance of transformed and non-transformed spring oilseed rape cv. HM-81 to the infection with cauliflower mosaic virus (CaMV), turnip yellow mosaic virus (TYMV) and turnip mosaic virus (TuMV) was studied, to determine the influence of transformation on susceptibility of plants to viruses. For experiments the non-segregating R 1 generation of primary transformant HM-81-JZ and control plants of cv. HM-81 were used. The primary transformant was obtained by inoculation of stems withAgrobacterium rhizogenes 15834. All transformed plants of R 1 generation had typically „transformed“ phenotype. No significant differences were revealed in the resistance of both transformed and non-transformed plants to each virus, as proved by qualitative and quantitative ELISA and visual evaluation of symptoms. Transformed plants infected with turnip yellow mosaic virus showed significantly lower reduction of green mass yield than non-transformed. In the case of CaMV and TuMV infection reduction of yield of transformed and non-transformed plants was almost the same.     

Response of maize (Zea mays L.) lines carrying Wsm1, Wsm2, and Wsm3 to the potyviruses Johnsongrass mosaic virus and Sorghum mosaic virus

Maize dwarf mosaic disease is one of the most important viral diseases of maize (Zea mays L.) throughout the world. It is caused by several virus species in the family Potyviridae, genus Potyvirus, including Maize dwarf mosaic virus (MDMV), Sugarcane mosaic virus (SCMV), Johnsongrass mosaic virus (JGMV) and Sorghum mosaic virus (SrMV). Resistance to another member of the family Potyviridae, Wheat streak mosaic virus (WSMV), is conferred by three alleles (Wsm1, Wsm2, Wsm3) in the maize inbred line Pa405, and these or closely linked genes were previously shown to confer resistance to the potyviruses MDMV and SCMV. In this study, we assessed whether Wsm alleles are linked to resistance to JGMV and SrMV. Near isogenic lines (NILs) carrying one or two of the Wsm alleles introgressed into the susceptible line Oh28 and F1 progeny from NIL × Oh28 were tested for their response to JGMV and SrMV. Our results indicate that Wsm1 provides resistance to both JGMV and SrMV in a dose-dependent manner. Wsm2 and Wsm3 each provide limited resistance, and combining Wsm alleles enhances that resistance.     

Expression of bottom component RNA of cowpea mosaic virus in cowpea protoplasts

Upon inoculation of cowpea protoplasts with the bottom component of cowpea mosaic virus, at least six virus-induced proteins (with sizes of 170, 110, 87, 84, 60, and 32 kilodaltons) are synthesized, but not the capsid proteins (37 and 23 kilodaltons). These bottom-component-induced proteins were studied with respect to their genetic origin and mode of synthesis. The analyses were based on their electrophoretic peptide patterns resulting from partial digestion with Staphylococcus aureus protease V8. Comparison of the peptide patterns of the virus-induced proteins with those of the cowpea mosaic virus RNA-coded polypeptides produced in rabbit reticulocyte lysate showed that the 170- and 32-kilodalton polypeptides, which are the first viral products in cowpea mosaic virus-infected cells, were actually coded by the bottom component RNA of the virus. The 110-, 87-, and 84-kilodalton polypeptides, and possibly the 60-kilodalton polypeptide, appeared to have amino acid sequences in common with the 170-kilodalton polypeptide, demonstrating that they were virus coded as well. The results indicated that cowpea mosaic virus bottom component RNA was translated in vivo into a single 200-kilodalton polyprotein from which probably all bottom-component-specific proteins arose by three successive cleavages.     

Transmission of carrot mosaic virus by aphids

The transmission of the carrot mosaic virus (CMV) by the aphidsAcyrtJiosiphon pisum HARRÍS,Cavariela aegopodii SCOP, andMyzus persicae SULZ was proved experimentally. It was observed simultaneously that CMV has a non-persistent character. CMV can be transmitted already 2 min after acquisition feeding by the aphidsMyzus persicae SDLZ andCavariella aego-podii Scop. When the time of acquisition feeding is prolonged to 4 min, CMV is transmitted also by aphidAcyrthosiphon pisum HAREÍs. The host range of the investigated virus wasalso determined and its transmission to 8 plant species, belonging to 4 families, was achieved. On the basis of studies of the vector virus relationship and of the host range, further proof was given for the different character of the Australian Carrot motley dwarf virus, theApivm virus 1 Roland and CMV. The experiments showed that preliminary starving of the aphids for 1 h increases their ability to transmit the virus by 3–3%.