Tobacco rattle virus serotypes and associated nematode vector species of Trichodoridae in the bulb-growing areas in the Netherlands

Soil samples from the coastal bulb-growing areas in the provinces of North- and South-Holland and the North-East Polder in the Netherlands were examined for trichodorid nematodes and tobacco rattle virus (TRV) serotypes. At least one of a total of eight species of Trichodoridae, of whichParatrichodorus pachydermus was most prevalent, was found in 93% of the samples from the provinces of North- and South-Holland and TRV, including four serotypes, was obtained from 49% of these samples. In the North-East Polder one of three species of trichodorids, of whichP. teres occurred most frequently, was present in 72% of the samples, and TRV of one serotype was obtained from 28% of these samples. The TRV isolates recovered from these samples reacted serologically with one of four antisera to strains of TRV. Virus transmitted byP. pachydermus reacted to the PRN-, byTrichodorus viruliferus to the RQ-, byP. teres to the N5- and byT. similis, to the TS-antiserum, respectively.     

Characterization and vector relation of a serologically distinct isolate of tobacco rattle tobravirus (TRV) transmitted byTrichodorus similis in northern Greece

A virus isolated from diseased tobacco plants growing in Macedonia, northern Greece, had host range and physico-chemical properties typical of a tobravirus. Although it was serologically unrelated to any of the ten tobravirus isolates tested, it reacted in spot hybridization tests with a probe derived from RNA-1 of tobacco rattle virus (TRV) strain SYM. Therefore, the isolate probably represents a previously undescribed serotype of TRV. Male, female and juvenileTrichodorus similis nematodes recovered from the rhizosphere of the diseased tobacco plants transmitted TRV in each of three laboratory experiments. In two of these experiments 50% and 54%, respectively, of the nematodes transmitted virus toPetunia hybrida bait plants, whereas only 18% transmitted virus toNicotiana tabacum plants in a third test. Ultrathin sections of the feeding apparatus of individual nematodes, which had transmitted virus, were examined by electron microscopy. Virus particles were observed, retained as a monolayer in the apices of the oesophageal lumen and as a group of particles within a matrix in the open part of the lumen.     

The transmission by nematodes of tobraviruses is not determined exclusively by the virus coat protein

The coat protein gene of the nematode non-transmissible, SP5 isolate of pea early-browning tobravius was replaced with that of the highly nematode transmissible, PPK20 isolate of tobacco rattle tobravirus. Plants were infected with the recombinant virus when mechanically inoculated and the virus invaded the plants systemically. However, although the PPK20 isolate of TRV was transmitted by nematodes from these plants, the recombinant virus was not transmitted. Therefore, the virus coat protein is not the exclusive determinant of nematode transmission.     

侵染牡丹的烟草脆裂病毒的检测鉴定及序列分析

为明确江苏省牡丹上烟草脆裂病毒(Tobacco rattle virus,TRV)的发生情况,利用ELISA和RT-PCR方法对采集自扬州市的40份牡丹叶片样品进行了检测鉴定,测定了其中一个分离物Peony-11中TRV RNA1分子的部分蛋白编码区序列,并结合Gen Bank中已报道的相关序列对其进行了多样性和系统发生分析。结果显示,江苏省扬州市牡丹上TRV的检出率高达62.5%;序列分析表明本研究获得的TRV牡丹分离物Peony-11与Gen Bank中其它63个分离物的核苷酸序列一致率为90.5%~99.7%;系统发生和遗传距离分析表明TRV可以分成2个组10个亚组,组间、亚组间具有较为清晰的地理和寄主特异性,其中Peony-11分离物位于亚组I-1。     

The association between serotypes of tobraviruses and Trichodorus and Paratrichodorus species1

New procedures developed for studying virus transmission by Trichodorus and Paratrichodorus species permit accurate characterization of the virus and identification of the nematode species. Results obtained using this virus transmission system show that serologically distinct isolates (serotypes) of tobraviruses are transmitted by different vector species. Furthermore, one species may be associated with the same virus serotype in several countries. These results suggest that a high degree of specificity occurs between tobraviruses and their vector nematodes. These new methods also permit the application of stringent criteria which are considered necessary to establish that a Trichodorus or Paratrichodorus species is a vector of a tobravirus. Only 11 of 36 published associations between Trichodorus or Paratrichodorus and tobraviruses fulfil these criteria.     

Notched leaf inGladiolus spp., caused by viruses of the tobacco rattle virus group

Symptoms of notched leaf in gladiolus are described in this paper. The disease can occur either in patches in the field (only on sandy or light sandy loam soils) or in plants scattered throughout a field planted with a certain stock (independent of soil type). Experiments showed that this disease is caused by tobacco rattle virus (TRV), which is transmitted byTrichodorus pachydermus Seinhorst andT. similis Seinhorst. Gladiolus corms planted in infested soil show a more or less serious reduction of growth and sometimes typical symptoms of notched leaf. In experiments these notched leaf symptoms could be reproduced in the first season only whenTrichodorus from infested soil was able to infect the young sprouts on top of the corms. If infection took place through the roots, the growth of the plants was not or only slightly retarded and the typical notched leaf symptoms appeared in the offspring in the following season. At least two serologically different strains of TRV could be isolated from affected gladiolus. These serotypes were not found to be typical for a certain location as in some cases both were isolated from the same infested field. In fields where potatoes showed stem-mottle and/or spraing, gladiolus was affected by notched leaf.Samenvatting Kartelblad in gladiolen kenmerkt zich door slecht uitgegroeide, misvormde planten, waarbij langs de randen en de nerven van de bladeren necrotische strepen en karakteristieke kartel- en zaagranden optreden (fig. 1). De planten komen meestal niet tot bloei. Alle overgangen tussen normale planten en die met de beschreven symptomen komen voor.In de knollen en kralen van de aangetaste planten zijn geen kenmerkende symptomen waar te nemen. In het gewas te velde kunnen deze aangetaste planten, in variërende percentages, verspreid tussen de gezonde planten van een partij voorkomen, terwijl aangrenzende partijen geheel gezond kunnen zijn. In deze gevallen, die op alle grondsoorten worden aangetroffen, is de partij één of meer seizoenen eerder geïnfecteerd. In andere gevallen worden planten als zojuist beschreven pleksgewijs in het gewas aangetroffen (fig. 2). Dergelijke aantastingen komen uitsluitend voor op de lichtere gronden en hierbij wordt eveneens een abnormale ontwikkeling van het wortelstelsel waargenomen (fig. 3).In proeven kon worden aangetoond dat de ziekte veroorzaakt wordt door virussen van de ratelvirusgroep (zie tabel 1 tot en met 6). In de grond aanwezige aaltjes van het geslachtTrichodorus (T. pachydermus Seinhorst,T. similis Seinhorst en mogelijk nog andere soorten) brachten het virus op de waardplant over (tabellen 2, 5 en 6).In het seizoen waarin infectie plaatsvond, ontstonden uitsluitend kenmerkende kartelblad-symptomen indien aaltjes, afkomstig van besmette grond, de mogelijkheid werd gegeven jonge, nauwelijks uitgegroeide spruiten aan te tasten (fig. 5, tabel 6). Infectie in de wortels veroorzaakt geen of slechts een geringe groeistoornis, maar in de nakomelingschap van deze planten komen in het volgende seizoen de typische symptomen van kartelblad voor (fig. 4, tabel 3, 5 en 6).Op een aantal gronden waar aardappelen waren geïnfecteerd door stengelbont of kringerigheid, werden gladiolen door kartelblad aangetast. In proeven waar gladiolen en aardappelen door TRV op dezelfde percelen waren geïnfecteerd, bleek een opmerkelijke overeenkomst in het verloop van kartelbladaantasting bij gladiolen en stengelbontaantasting bij aardappelen te bestaan (tabel 4).Bij het onderzoek van verschillende TRV-isolaties uit gladiolen met kartelblad-symptomen en uit andere waardplanten die afkomstig waren van met TRV enTrichodorus sp. besmette gronden, konden tenminste twee verschillende serotypen van het TRV worden aangetoond. Isolaties van het ene serotype bleken nauw verwant te zijn met een TRV-isolatie uit tabak White Burley, waartegen een antiserum is vervaardigd doorMaat (1963). Vertegenwoordigers van het andere serotype reageerden alle serologisch positief met een antiserum bereid tegen een TRV-isolatie uit gladiool. Beide serotypen bleken niet specifiek te zijn voor bepaalde plaatsen, daar beide verspreid door het land blijken voor te komen en in sommige gevallen uit eenzelfde perceel konden worden geïsoleerd. In een volgende publikatie zal hierop nader worden ingegaan.A preliminary report on this subject has been published in Nematologica 10 (1964):69–70.     

Sensitivity of<Emphasis Type="Italic">Botrytis cinerea</Emphasis> from greenhouse vegetables to DMIs and fenhexamid

Two hundred isolates ofBotrytis cinerea were collected from greenhouse vegetables between 2003 and 2006 to determine their baseline sensitivity to triadimefone, penconazole, tebuconazole and fenhexamid. Mean values of 50% effective concentrations (EC₅₀) of inhibiting growth were 4.853±5.102, 0.41±0.215, 0.19±0.099 and 0.36±0.891 mgl ⁻¹, respectively (mean±SD). Individuals ofB. cinerea in the population differed by a factor (EC₅₀ of the least sensitive isolate/EC₅₀ of the most sensitive isolate) of 6625, 20, 603 and 1800, respectively. Naturally fenhexamid-resistant isolates were detected with an unexpected high frequency of 10% although the pathogen population had never been exposed to this fungicide. The resistance level (mean EC₅₀ of resistant isolates / mean EC₅₀ of sensitive isolates) was 19.5. These naturally resistant isolates also were resistantin vivo, and there was no significant difference in growth rate, conidial production or pathogenicity ability between naturally resistant and wild sensitive isolates. These results indicated that there was a potential risk of practical resistance if fenhexamid was applied alone. Negative cross-resistance was observed between fenhexamid and tebuconazole in 90% of the naturally resistant isolates. Moreover, an obvious synergism of the antifungal activity of fenhexamid by tebuconazole was demonstrated in some of the naturally fenhexamid-resistant isolates. http://www.phytoparasitica.org posting May 9, 2007.     

Gut bacteria associated with the Pacific Coast wireworm,<Emphasis Type="Italic">Limonius canus</Emphasis>, inferred from 16s rDNA sequences and their implications for control

A multitude of bacteria have been isolated from the guts of several insect species. Some of these have been modified to interfere with the development of the host insect or with the development and transmission of plant and animal pathogens transmitted by the host insect. We surveyed the gut flora of the Pacific Coast wireworm,Limonius canus LeConte, a serious pest of potato, at two sites in Oregon and Washington. Isolates were obtained from surface-sterilized triturated larvae by dilution plating on standard media. A rich diversity of species was found in 86 isolates, including spore-formers, non-spore-formers and aerobic and facultatively anaerobic species collected on four sampling dates at each location. Twenty-one of the isolates were identified to species based on rDNA sequence (nine distinct species). An additional 34 isolates were identified to genus from the sequence data while six isolates could be assigned only to family based on sequence comparisons. Twenty-seven additional isolates were identified to species (9), genus (17) or family (1) based on side-by-side morphological comparisons with isolates identified from rDNA sequence. The most frequently isolated bacterium wasBacillus megaterium, followed byRahnella aquatilis. A naturally occurring bacterium found in the gut and/or environment of a targeted insect that is modified to express toxins or other detrimental substances could provide certain advantages (such as persistence and recycling) over inundatively applied microbial control agents, particularly within soil habitats. The hypothesis that these species or others from the survey represent candidates for genetic modification to provide control options forL. canus is discussed.     

Viruses transmitted by nematodes1

The transmission of plant viruses by nematodes is remarkable in involving only two distinct groups of viruses, nepo viruses and tobraviruses, and being limited to longidorid, Paratrichodorus and Trichodorus nematodes respectively. Tobraviruses and their associated vector nematodes are not discussed here. Only 11 of the 36 described nepoviruses are transmitted by nematodes, and 6 of these 11 viruses are present in Europe naturally associated with 8 virus-vector longidorids. Specific relationships exist between the serologically distinct viruses and their vector nematode species. Specificity is largely determined by the virus coat protein and by an inherited ability of the nematode to retain virus particles at specific sites within its oesophagus. This specific relationship can be quite subtle, extending to populations of vector nematodes and also to virus isolates which apparently are serologically indistinguishable. Several serological and/or symptomatological variants of nepoviruses may be present at a field site in association with one or more vector nematode species. The exposure of different crops and new cultivars to these virus and vector combinations will probably result in the occurrence of further nematode-transmitted virus diseases. New methods for suppressing damage to crops caused by these diseases are required including the likely use of transgenic resistant cultivars.     

Identification and properties of a deviant isolate of the broad bean yellow band serotype of pea early-browning virus from faba bean (Vicia faba) in Algeria

A virus, isolated from faba bean (Vicia faba) obtained from Algeria, was readily recognized as a tobravirus by its particle sizes and morphology. Pea (Pisum sativum) and French bean (Phaseolus vulgaris) characteristically reacted to the isolate like pea early-browning virus (PEBV), but faba bean,Antirrhinum majus, Nicotiana rustica, andN. tabacum reacted with line-pattern symptoms which were unusually brilliant on theNicotiana species. In electronmicroscope decoration tests, the isolate did not react with an antiserum to the Dutch type strain of PEBV, but with one to the broad bean yellow band (BBYB) serotype from Italy. It resembles this serotype in reaction on faba bean, but seems to differ appreciably onN. rustica, N. tabacum, andPetunia hybrida. It is described as a deviant isolate of the BBYB serotype of PEBV.All thirteen faba-bean genotypes tested were found to be susceptible to the Algerian isolate and two Dutch type strain isolates of the virus, and to react with erratic line-pattern symptoms to the Algerian isolate only. All ten genotypes of chickpea (Cicer arietinum) tested reacted hypersensitively, and four out of ten genotypes of lentil (Lens culinaris) were susceptible to the virus but reacted differentially to the three isolates. Seed transmission of PEBV, including the new isolate, in faba bean is confirmed (9% for the Algerian isolate, and over 45% for one of the Dutch type strain isolates), and seed transmission of the virus in a non-legume (N. rustica, 4%) is herewith first reported. This is the first report on the occurrence of the BBYB serotype of PEBV outside Italy, and of PEBV outside Morocco in North Africa.     

Transmission efficiency of five tobravirus strains byParatrichodorus teres

Nematodes from a population of virus-freeParatrichodorus teres were allowed to feed on roots ofPetunia hybrida andNicotiana tabacum cv. White Burley plants infected with five strains of tobacco rattle or pea early-browning tobraviruses and were subsequently tested for virus transmission to roots of virus-freeP. hybrida andN. tabacum cv. White Burley plants. Virus transmission was not correlated with serotypes. Two strains of TRV with different serotypes were efficiently transmitted, whereas two other strains of TRV and one strain of PEBV with serotypes similar to transmitted ones were not transmitted at all. The two efficiently transmitted strains had originally been obtained fromP. hybrida roots which had been exposed to viruliferousP. teres in laboratory bait tests, whereas the three non-transmitted strains had been obtained from infected plant material collected in the field.     

Virus transmission by host-specific strains ofOlpidium bornovanus andOlpidium brassicae

Zoospores of 12 isolatesO. bornovanus from geographically diverse sites and representing the three host specific cucurbit strains were tested as vectors for seven viruses using watermelon bait plants and the in vitro acquisition method. All isolates of the cucumber, melon, and squash strains transmitted melon necrotic spot carmovirus (MNSV) and cucumber necrosis tombusvirus (CNV) but none transmitted petunia asteroid mosaic tombusvirus (PAMV) or tobacco necrosis necrovirus (TNV). The isolates varied as vectors of three other carmoviruses: cucumber leaf spot virus (CLSV); cucumber soil borne virus (CSBV); and squash necrosis virus (SqNV). All cucumber isolates transmitted CLSV and SqNV but not CSBV. Some of the melon isolates transmitted CLSV and SqNV but none transmitted CSBV. Two squash isolates transmitted CSBV and SqNV but not CLSV. Two isolates ofO. brassicae transmitted only TNV and a third did not transmit any of the viruses. The species of bait plant sometimes affected transmission. The most efficient vector strains ofO. bornovanus, as determined by reducing zoospores and virus in the inoculum, were the cucumber strain for CLSV; the cucumber strain for CNV if cucumber was the bait plant or melon strain if watermelon was the bait plant; and the squash strain for SqNV. The plurivorous strain ofO. brassicae was the most efficient vector of TNV.Olpidium bornovanus is the first vector reported for CSBV and is confirmed as a vector of SqNV. It is proposed that all carmoviruses may have fungal vectors.Ligniera sp. did not transmit any of the viruses in one attempt.Abbreviations CLSV cucumber leaf spot virus - CNV cucumber necrosis virus - CSBV cucumber soil borne virus - MNSV melon necrotic spot virus - PAMV petunia asteroid mosaic virus - SqNV squash necrosis virus - TNV tobacco necrosis virus - TBSV tomato bushy stunt virus     

VCG and AFLP analyses identify the same groups in the causal agents of mango malformation in Brazil

The causal agents of mango malformation disease in Brazil are a new Fusarium lineage in the Gibberella fujikuroi species complex and Fusarium sterilihyphosum; however information on the genetic and geographical diversity of these pathogens in Brazil is missing. Vegetative compatibility group (VCG) and amplified fragment length polymorphism (AFLP) analyses were used to measure the genetic diversity within these populations. Both techniques identified the same genetic groups. Six VCG and AFLP groups were identified amongst isolates of the new lineage from Brazil. FB-VCG 1/AFLP I was the most widespread group, found in seven of the 13 sites sampled. The second most frequent group was recovered from three sites. The remaining four groups were recovered from single-sites. We think that this lineage represents a genetically and geographically diverse indigenous population that reproduces clonally. In F. sterilihyphosum, group FS-VCG 1/AFLP VII was found at three sites in the southeast region of Brazil. FS-VCG 2/AFLP VIII contained isolates from South Africa but not from Brazil. Fusarium mangiferae isolates from India and South Africa formed one group, while isolates from Egypt and the USA formed a second group. F. sterilihyphosum at present is represented by a small population that might have been introduced only once into a restricted area. The clonal nature of the observed populations suggests that these fungi either occur naturally on indigenous hosts and have jumped to the introduced mango host (introduced in Brazil) or that they originated with mango and went through a severe population bottleneck when they were introduced to Brazil from India or Southeast Asia.     

Nematicidal potential of materials from <Emphasis Type="Italic">Medicago</Emphasis> spp.

The nematicidal effect of soil amendments with dry top and root material from Medicago sativa and/or Medicago arborea was evaluated on the root-knot nematode Meloidogyne incognita and on the cyst nematode Globodera rostochiensis in potting mixes. All amendments suppressed root and soil population densities of both nematode species compared to non-treated and chemical controls. The suppressiveness of M. sativa differed between top and root material and among the amendment rates. In field conditions soil amendments with 20 or 40 t ha⁻¹ of a pelleted M. sativa meal increased tomato crop yield and reduced soil population densities and root galling by M. incognita. It is suggested that saponins were at least partly responsible for the nematicidal activity.     

The effect of DD and dazomet treatments on nematode populations at various depths in the soil and on the transmission of soil-borne tobacco rattle virus to gladiolus

The relation between log dosage of DD injected at 15 cm depth or of dazomet applied to the soil surface (all in November 1971) and probit mortality ofRotylenchus and trichodorids in the top 20 cm of a field on sandy soil was found to be linear. Dosage increase efficiencies of both chemicals against both nematode species were medium to high. Superficial application of dazomet was very effective against the nematodes that would have survived if only a low dosage of DD had been injected at 15 cm depth. Injection of 40 ml or 80 ml DD per m² at 15 cm depth killed all nematodes between 20 cm and 60 cm deep. Gladiolus planted in the spring of 1972 grew better, flowered earlier and produced more weight of corms on treated than on untreated plots. The poor growth on the untreated plots cannot be ascribed to direct damage by nematodes or to the effect of TRV transmitted to the plants by the viruliferous trichodorids occurring in these plots in high densities. Symptoms of TRV infection in plants grown in 1973 from the corms harvested in the 1972 experimental field showed that only DD treatments had reduced the rate of TRV transmission considerably. However, even the highest dosages of DD had only reduced it from 26% (on untreated plots) to about 8%. Most probably, this residual TRV infection was due to transmission by trichodorids that had survived in soil layers below 60 cm depth. Therefore, soil treatment with nematicides, cannot prevent TRV transmission to gladiolus sufficiently where viruliferous trichodorids occur at great depths, as is the case in many sandy soils having a low water table.     

Interactions Between the Potato Cyst Nematode Globodera rostochiensis and Diseases Caused by Rhizoctonia solani AG3 in Potatoes Under Field Conditions

Findings from 2 years of field experiments investigating the relationship between Globodera rostochiensis and Rhizoctonia solani on unique field sites are reported. In 2000, a field experiment was positioned on land that had previously been used for experimental work investigating integrated potato cyst nematode (PCN) management methods. This study had produced an ‘untypical’ mosaic of PCN population densities ranging from 5 to 221 eggs g⁻¹ soil. In 2001, the field experiment was conducted on a different field site and overlaid on a focus of G. rostochiensis population densities ranging from 11 to 108 eggs g⁻¹ soil. In each experiment, potatoes (cv. Désirée) were grown in plots with similar population densities of G. rostochiensis that were either uninoculated or inoculated with R. solani. A series of potato plant harvests were undertaken to investigate the effects of nematode infestation on the incidence and severity of R. solani diseases and the associated development of plants. In both experiments, a clear relationship was found between the density of G. rostochiensis juveniles present in potato roots and the incidence of stolons infected by R. solani, 6 weeks after planting. For the first time this interaction has been determined under field conditions. The results of the study suggest that the interaction between nematode and fungus is indirect and possible mechanisms are discussed.     

Molecular and Pathotype Analysis of the Rice Blast Fungus in North Vietnam

Rice blast caused by Pyricularia oryzae is a devastating disease worldwide. In Vietnam, rice blast is especially severe in the Red River Delta in the North. The genetic diversity of 114 P. oryzae isolates collected from rice in 2001 in the Red River Delta and nine additional Vietnamese P. oryzae isolates was analysed using Amplified Fragment Length Polymorphism (AFLP). DNA similarity and cluster analysis based on 160 polymorphic AFLP markers showed twelve different AFLP genetic groups among the 123 field isolates. Isolates collected from japonica hosts clustered separately from indica host isolates with at least 60% dissimilarity with little evidence for gene flow between the two populations. In the 2001 population originating from indica hosts, three genetic groups were predominant and represented 99% of the isolates sampled. One predominant clonal lineage represented 59% of the 2001 indica host population and was found in eleven provinces in the Red River Delta of North Vietnam. Significant genotype flow could be demonstrated between the indica population south of Red river and the indica population north of Red river. There was significant linkage disequilibrium between the AFLP loci within the indica population, indicating that this is not a random mating population. Pathogenicity tests of 25 isolates selected from the 12 AFLP groups on a set of 29 differential rice lines revealed two avirulent isolates and 23 pathotypes. Different combinations of known resistance genes were found to have potential for blast resistance breeding for North Vietnam. First two authors contributed equally     

Pine Wilt Disease Caused by the Pine Wood Nematode: The Induced Resistance of Pine Trees by the Avirulent Isolates of Nematode

Since the beginning of the 20th century, pine trees in Japan have been seriously damaged by the pine wilt disease. This disease is caused by the pine wood nematode, Bursaphelenchus xylophilus, which is transmitted by the Japanese pine sawyer, Monochamus alternatus. The control of disease depends to a large extent on chemicals, but the public is now demanding environmentally friendly control methods. The virulence of B. xylophilus varies very widely. Pre-inoculation of young pine trees in a nursery with avirulent B. xylophilus has induced systemic resistance of trees against a subsequent inoculation with virulent B. xylophilus. This induced resistance was considered a hopeful means for developing a biological control for the disease. The induced resistance by the avirulent nematodes was also expressed in mature pine trees in a forest where the disease was naturally epidemic. However, the effects of induced resistance were not satisfactory for practical biological control. Since the inoculation with higher concentrations of the avirulent B. xylophilus induced the resistance more effectively, the pre-inoculation method will need to be improved to develop the biological control. The induced resistance of pine trees by avirulent B. xylophilus should be one of the candidate biological control methods against pine wilt disease. This induced resistance also provides an experimental system to clarify physiological interactions between the nematodes and pine trees.     

Line pattern in Limonium latifolium caused by tobacco rattle virus

Tobacco rattle virus (TRV) was isolated from plants ofLimonium latifolium showing bright yellow or red line patterns and ringspots on the leaves. It was proved that this virus, designated TRV-Lim, was the causal agent of the disease. In its reactions onNicotiana clevelandii it resembled a yellow strain of TRV from Oregon (USA), but the symptoms inN. glutinosa, N. megalosiphon, N. tabacum andPetunia hybrida were more comparable to those caused by socalled unstable variants of TRV. Dilution end-point was 10^–6–10^–7, thermal inactivation at 76–80°C, and ageing in vitro 55–60 days. The purified virus suspension contained particles of three normal lengths, 70, 102, and 194 nm. The virus sedimented as three components with average sedimentation coefficients of 129, 161 and 206 S, respectively. In purified suspensions TRV-Lim had two different buoyant densities. A serological relationship was found with TRV isolated from Europe and Brazil.Samenvatting Tabaksratelvirus (TRV) werd geïsoleerd uitLimonium latifolium planten die heldergeel of rood figuurbont op de bladeren vertoonden. Er werd aangetoond dat dit virus, aangeduid als TRV-Lim, de ziekteverwekker was. De reacties van dit virus opNicotiana clevelandii deden denken aan die van een gele stam van TRV afkomstig uit Oregon (VS), maar de symptomen opN. glutinosa, N. megalosiphon, N. tabacum enPetunia hybrida vertoonden meer gelijkenis met die welke veroorzaakt worden door de zogenaamde onstabiele varianten van TRV. De verdunningsgrens was 10^–6–10^–7, de inactiveringstemperatuur 75–80°C en de houdbaarheid in vitro 55–60 dagen. De gezuiverde virussuspensie bevatte deeltjes met drie normale lengtes, nl. 70, 102 en 194 nm. Het virus sedimenteerde, als drie componenten met gemiddelde sedimentatiecoëfficiënten van respectievelijk 129, 161 en 206 S. In gezuiverde suspensie vertoonde TRV-Lim twee verschillende zweefdichtheden. Het virus was serologisch verwant aan TRV-isolaten uit Europa en Brazilië.