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.     

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.     

Winter-type california tobacco rattle virus in its relation to the infected cell

Particles of California tobacco rattle virus (CTRV) were observed in thin sections of vascular elements of leaves showing oak leaf symptoms. At no time during this study were particles observed in mesophyl cells of these leaves. A peculiar chloroplast breakdown is described.     

Detection of tobacco rattle virus in nematodes by reverse transcription and polymerase chain reaction

An assay, based on amplification of cDNA synthesized from genomic viral RNA, has been developed to detect tobacco rattle virus in infected plant material and viruliferous nematodes. A range of different TRV strains could be detected using the procedure developed. The presence of one to three viruliferous nematodes in a nematode suspension was sufficient for the detection of TRV. The minimum amount of purified virus detectable in the assay was 15 fg, indicating an increased sensitivity of the PCR-based assay as compared to serological detection methods, like ELISA. A dot-blot hybridization procedure was developed for the detection of the PCR products, making agarose gel electrophoresis dispensable.     

Separation of long and short particles of tobacco rattle virus with polyethylene glycol

Two methods to separate long and short particles of tobacco rattle virus with polyethylene glycol 6,000 (PEG) are described. The first is based on specific precipitation, the second on specific solubilization of particles with different lengths at different PEG concentrations. The results of the separations are comparable to, or better than those obtained by sucrose-gradient centrifuging. The advantage of the methods using PEG is that no expensive equipment is required.Samenvatting Twee methoden om lange en korte deeltjes van het tabaksratelvirus met behulp van polyethyleenglycol 6000 (PEG) te scheiden worden beschreven. De ene is gebaseerd op specifieke precipitatie, de andere op specifieke oplosbaarheid van deeltjes met verschillende lengten bij verschillende PEG-concentraties. De resultaten van de scheidingen zijn even goed als of beter dan die van scheidingen verkregen door centrifugeringen op een suikergradiënt. Het voordeel van de PEG-methoden is gelegen in het feit dat er geen kostbare apparatuur voor nodig is.     

Tobamoviruses of pepper,eggplant and tobacco: Comparative host reactions and serological relationships

Using test plants and serology six tobamoviruses of pepper (FO, Ob, P8, P11, P14 and SL) and one of eggplant (A1) were compared with common tobacco mosaic virus (TMV-WU1). WU1, A1 and FO were closely similar in their reactions inCapsicum spp. as were P14 and SL. Ob, P11 and P8 were also similar in this respect except inC. frutescens Tabasco in which P8 differed from Ob and P11.Using micro-precipitation tests the virus strains could be roughly divided into three serological groups: Group I consisted of WU1, group II of A1, FO, P8, P14 and SL, and group III of P11 and Ob. With ELISA group II was further divisible into two subgroups, including A1 and FO, and P8, P14 and SL.It was concluded that similarities of strains in their reactions inCapsicum spp., were not necessarily confirmed by their serological relationships.Samenvatting Zes tobamovirussen uit peper (FO, Ob, P8, P11, P14 en SL) en één uit aubergine (A1) konden met behulp van toetsplanten alle van elkaar worden onderscheiden. In hun reacties inCapsicum-soorten, kwamen A1 en FO sterk overeen met elkaar en met het gewone tabaksmozaïekvirus (WU1). Ob, P11 en P8, die in dit opzicht onderling veel overeenkomst vertoonden, verschilden duidelijk van alle andere. Hetzelfde gold voor P14 en SL.Ook met behulp van de micro-precipitatietoets konden de virusstammen in groepen worden ingedeeld. Groep I werd gevormd door WU1, groep II door A1, FO, P8, P14 en SL en groep III door P11 en Ob. Met behulp van ELISA kon groep II worden onderverdeeld in twee ondergroepen, namelijk A1 en FO enerzijds en P8, P14 en SL anderzijds.De nauwe serologische verwantschap van A1 met FO is conform de grote overeenkomst in waardplantreacties. Hetzelfde geldt voor P11 en Ob, wanneer we alleen hun reacties inCapsicum-soorten beschouwen. P8 echter, die wat het laatste betreft meer op Ob en P11 lijkt, vertoonde serologisch meer overeenkomst met P14 en SL. WU1 verschilde serologisch zeer sterk van alle andere onderzochte virusstammen.Geoconcludeerd kan worden dat de waargenomen overeenkomst tussen de onderzochte virusstammen in hun reacties inCapsicum-soorten niet altijd gesteund wordt door hun serologische verwantschappen.Guestworker from September 1981 till January 1982 as a fellow of the International Agricultural Centre, Wageningen, the Netherlands, from the Vegetable Crops Research Institute, Budapest, Hungary.Seconded to the Glasshouse Crops Research and Experiment Station, Naaldwijk, the Netherlands.     

Acquisition and subsequent transmission of tobacco rattle virus isolates by Paratrichodorus and Trichodorus nematode species

Isolates of the PRN serotype of tobacco rattle virus (TRV) were transmitted with different efficiencies by the nematode vectorParatrichodorus pachydermus. Virus isolates which belonged to other serotypes were not acquired and/or transmitted by this vector, nor were PRN serotype isolates which had been obtained from naturally infected potato plants and maintained by mechanical transmission in the glasshouse for several years. PRN serotype TRV isolates from the Netherlands or from Scotland were equally well transmitted by initially virus-freeP. pachydermus populations from either country. Allowing a naturally viruliferous nematode population access for 3 weeks to uninfected or TRV-infected roots resulted in an increased proportion of the trichodorid population transmitting TRV.     

Detection of tobacco rattle virus in different parts of tulip by ELISA and cDNA hybridisation assays

Different parts of tulips cv. Apeldoorn were assayed for the presence of tobacco rattle virus (TRV) by means of ELISA, cDNA hybridisation and immuno-electron microscopy. Assays were periodically performed during the growing season and upon storage of the bulbs, During the growing season in the field the relative TRV concentrations detected by ELISA and cDNA were highest mainly in the basal stem-parts and basal leaf-parts, respectively. When, during storage, infected bulbs were divided into a number of sections, TRV could be detected only in some of the sections, irrespective of the test used. However, nearly all sprouts of infected bulbs, stored at 5°C for 7 months, appeared to contain detectable amounts of TRV upon testing with ELISA and cDNA. Thus, testing of sprouts may offer a possibility to develop a routine test for TRV in tulip bulbs in due course.Samenvatting Verschillende delen van tulp cv. Apeldoorn werden getoetst op de aanwezigheid van tabaksratelvirus (TRV) met behulp van ELISA, cDNA-hybridisatie en immuno-elektronemicroscopie. Tijdens het groeiseizoen en de bewaring van de bollen werden regelmatig toetsingen uitgevoerd. Gedurende het groeiseizoen op het veld werden de relatief hoogste TRV concentraties voornamelijk gevonden in het basale deel van de stengel en het okselgedeelte van het blad met respectievelijk ELISA en cDNA-hybridisatie. TRV bleek gelokaliseerd aanwezig te zijn in een of meer stukjes van een gedeelde bol, onafhankelijk van de gebruikte toetsmethode. Bijna alle spruiten van geïnfecteerde bollen die gedurende 7 maanden bij 5°C bewaard waren, bleken bij het toetsen met behulp van ELISA en cDNA aantoonbare hoeveelheden van TRV te bevatten. Het toetsen van spruiten biedt de mogelijkheid te zijner tijd een routinetoets voor TRV in tulpebollen te ontwikkelen.     

Detection of tobacco rattle virus by ELISA and test plants in main sprouts of tulip bulbs during storage at different temperatures

The detectability of tobacco rattle virus (TRV) in the main sprouts of primarily and secondarily infected tulip bulbs of cv. Apeldoorn stored at different temperatures from the lifting in July up to February is described. Detection by ELISA was not affected by the size of the main sprout, nor by the size of the bulbs. The rates of TRV-infected bulbs found by ELISA were highest during storage at 13, 9 or 5°C continuously, and when temperatures were lowered from 20 or 17°C to 5°C in October. The percentages detected via test plants, but undetectable by ELISA were also lowest at these temperatures. The unfavourable effect of continuous storage at 20, 17, or 2°C as expressed in low ELISA absorbances not significantly different from the mean value of healthy bulbs, was largely overcome during long storage by the change of temperature down to 5°C from 20 and 17°C or upwards from 2°C. The reverse from 5°C upwards to 17 and 20°C affected the detectability by ELISA unfavourably. The rate of detection via test plants in the main sprout and in the small sprouts from different positions in bulbs was only possible at low percentages.The effect of some factors, like different temperatures during storage, detectability of different TRV serotypes, interference of irregular occurrence of TRV in the removed scale and basal-plate tissue with the main sprout, and variable recurrence of TRV in progeny bulbs, is discussed in view of its impact on routine testing of bulbs during storage.     

Grapevine red blotch virus is sporadically present in a germplasm collection in Northern Italy

Journal of Plant Diseases and Protection - Limited information is available on the distribution of grapevine red blotch virus (GRBV) in grape-growing regions outside North America. In this study,...     

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.     

Molecular diagnostics of some trichodorid nematodes and associated Tobacco rattle virus

Several Paratrichodorus and Trichodorus (trichodorid) nematode species are natural vectors of Tobacco rattle virus (TRV) and cause economically important diseases, especially in potato and ornamental bulbous crops. Identification of trichodorid species based on morphological characters is laborious, time-consuming, and requires the services of highly trained personnel. Molecular diagnostics for trichodorid nematodes, using the ribosomal DNA repeat unit, were successfully developed to distinguish two Paratrichodorus and two Trichodorus species. The complete sequences of the 18S genes and the ITS-1 regions for these species were obtained and species-specific primers successfully designed for them. An RT-PCR assay was developed utilizing isolate-specific primers that amplify serologically distinguishable strains of TRV in individual trichodorid nematodes. The primers were based on the highly conserved RNA-1 segment of the bipartite genome and also on different parts of the RNA-2 segment of the virus genome.     

Viruses and virus diseases of grapevine in Palestine

Surveys were carried out in vineyards in the main grapevine-growing areas of Palestine (Hebron, Bethlehem, Gaza, Jerusalem, Ramallah, Jenin, Jericho and Nablus) to assess the presence and incidence of virus and virus-like diseases. Leafroll symptoms were observed in Bethlehem, Ramallah and Jerusalem in native and imported cultivars, with higher rates in the red-fruited Shami, Beitoni and Smari. Rugose-wood symptoms were also observed in local and foreign cultivars, especially on grafted vines with a high incidence in Bethlehem. Fanleaf symptoms were rarely observed, while phytoplasma-induced symptoms were observed in Jenin, Jericho and Bethlehem on cvs Biadi, Superior Seedless and Beitoni. ELISA tests showed that 463 out of 566 (82%) tested vines were infected by at least one virus. GVA was the prevailing virus (66.1%), followed by GLRaV-1 (45.6%), GLRaV-3 (21.7%), GFkV (15.7%) and GLRaV-2 (8.3%). GVB and GFLV were also detected to a lesser extent, their incidence ranging between 3.7 and 1.2%, whereas GLRaV-7 was detected in a single vine of cv. Sultanina of foreign origin. Vineyards in the Bethlehem area were particularly badly damaged (97.5%), and some local cultivars were totally (Jandali, Marrawi and Shoyoukhi) or heavily infected (Zaini, Biadi and Shami). ELISA testing of 69 young rootstock mother plants showed a relatively high incidence of virus infection (20.3%). Vein necrosis and vein mosaic diseases were also ascertained on graft-inoculated 110R and Vitis riparia indicator plants, whereas no viruses other than GFLV were mechanically transmitted from about 200 vines onto inoculated herbaceous hosts.     

Viruses and virus diseases of grapevine in Tunisia

Mature canes were collected from vines in the main grapevine-growing areas in Tunisia (Cape Bon, Bizerte, Ben Arous), from commercial vineyards and mother-plant plots, to assess the presence of virus and virus-like diseases. Biological (mechanical transmission onto herbaceous hosts and grafting onto indicator woody plants) and serological detection (ELISA) methods were applied. ELISA showed that 96.4% of 669 vines tested were infected, most of them (88.1%) by at least two viruses. Grapevine leafroll-associated 3 closterovirus (GLRaV-3) was the most widespread virus (87.9%), followed by grapevine A vitiviras (GVA, 69.4%), grapevine fleck virus (GFkV, 51.9%), grapevine leafroll-associated 1 closterovirus (GLRaV-1, 36.8%), grapevine leafroll-associated 2 closterovirus (GLRaV-2, 19.1%), grapevine fan leaf nepovirus (GFLV, 18.2%) and grapevine B vitiviras (GVB, 14.8%). ELISA tests yielded negative results for grapevine leafroll-associated 7 closterovirus (GLRaV-7) and potato X potexvirus (PVX). The highest infections were found in Bizerte and Cape Bon regions (100 and 99.2%), and in vineyards aged over 20 years (98.5%) as compared with the younger ones (81.1%). Rootstocks in mother-plant plots were practically free from all the viruses tested (1 plant infected out of 81), whereas severe infections were found in Vitis vinifera mother plants (67.4% of 341 samples), in particular table grapes (92.6%) compared with wine grapes (47.9%). In these mother-plant plots, the prevailing viruses were GLRaV-3 (41.3%), followed by GFkV (36.7%), GVA (27.9%), GLRaV-1 (17%) and GLRaV-2 (15.2%). GFLV and GVB were far more limited (1.5 and 0.6%, respectively). The presence of vein necrosis and vein mosaic was ascertained by transmission onto 110R and Vitis riparia indicators, whereas only GFLV was mechanically transmitted onto herbaceous hosts (from about 20% of the samples).     

Viruses and virus diseases of grapevine in Egypt

Surveys for virus and virus-like diseases were carried out in commercial vineyards of the main grapevine-growing areas of Egypt along the river Nile and in recently reclaimed desert lands. The only symptoms observed and identified with reasonable confidence in the field were those of leafroll disease in red-berried cultivars. No virus was transmitted to herbaceous hosts by mechanical inoculation from glasshouse-forced cuttings of about 300 vines (40% of total samples). By contrast, ELISA tests showed that 78% of the assayed European vines (521 out of 664) were infected by one (29%) or more (49%) viruses. Grapevine virus A (GVA) was the most widespread virus (67.9% infection), followed by Grapevine leafroll-associated virus 3 (GLRaV-3) (55.9% infection). All the other viruses tested for were scarcely represented, i.e. Grapevine leafroll-associated virus 1 (GLRaV-1) 1.8% infection, Grapevine leafroll-associated virus 2 (GLRaV-2) 1.4% infection, Grapevine virus B (GVB) (0.6% infection) and Grapevine fleck virus (GFkV) (0.2% infection), or, like Grapevine fanleaf virus (GFLV), were totally absent. The infection rate of native cultivars (86%) was particularly heavy. 'Banaty Abiad' and 'Romy Ahmer', the two major Egyptian cultivars, had infection levels of 78% and 89%, respectively, and 'Fayoumy', the most important cultivar in the Fayoum area, had 96% infection. Totally infected were the tested samples of several minor native cultivars such as 'Farg El-Tair', 'Siwi Abiad', 'Ta'afi', 'Romy Abiad', 'Eswid El-Wady', 'Edkawy' and 'Bez El-Anza'. Slightly better was the sanitary situation of imported European grapevine cultivars (60% infection) and of American rootstocks (11.5% infection). In rootstocks, infection rate by GVA and GLRaV-3 was 5.5%, whereas GVB and GLRaV-1 were only sporadically detected.