Further observations on variation of lettuce mosaic virus in relation to lettuce (Lactuca sativa), and a discussion of resistance terminology

Two potyvirus isolates from endive, originating from southern France (Ls252) and from the Netherlands (Ls265), that were highly and poorly pathogenic on lettuce, respectively, were compared with a common isolate (Ls1) of lettuce mosaic virus (LMV) and with two highly deviant Greek isolates fromHelminthia (Picris) echioides (Gr4) and endive (Gr5), earlier recognized as LMV. The isolates could not be distinguished by particle morphology and serology, and were all identified as LMV. Leaf curling, plant stunting and necrosis were more characteristic of the virus than mosaic. The isolates studied varied considerably on differential host species and a range of lettce cultivars including pathotype differentials of Pink et al. [1992b]. Ls1 and Ls265 reacte largely as pathotype II, including the common strain of the virus, but Ls265 was least pathogenic on lettuce. Ls252 fitted pathotype IV and was very similar to LMV-E (the Spanish strain). The Greek isolates were very similar to each other in causing very severe symptoms on some non-lettuce hosts and a number of lettuce cultuvars. In lettuce variectal reaction Gr4 resembled pathotype I, but Gr5 severely affected Salinas 88, resistant to pathotypes I, II and III, and it appears to be a novel pathotype. Genetic interaction between lettuce and LMV is not following a simple yes-or-no pattern, and it is not a mere matter of resistance versus susceptibility. Adoption of a more realistic resistance terminology is proposed. None of the lettuce cultivars tested was resistant to the most pathogenic isolate Ls252, but resistance to it was found in 2 out of 12 wildLactuca species tested (Lactuca perennis andL. tatarica) while the symptomless plants ofL. perennis clearly reacted in ELISA.     

Tylose formation in elms after inoculation with Ceratocystis ulmi,a possible resistance mechanism

After inoculation of elms withCeratocystis ulmi tylose formation in vessels of the susceptible clone Belgica appeared to be delayed in comparison with tylose formation in the resistant clone 390. It is suggested that tylose formation may be a resistance mechanism in elms to Dutch elm disease.Samenvatting Na inoculatie van iepen metCeratocystis ulmi bleek de thyllenvorming te zijn vertraagd in de vaten van de vatbare kloon Belgica in vergelijking met de thyllenvorming in de resistente kloon 390 (Fig. 1). Thyllenvorming zou een resistentie mechanisme kunnen zijn tegen de iepziekte.     

Resistance to carbendazim in Pseudocercosporella herpotrichoides from Dutch wheat fields

Culm base samples were collected in August 1984, from winter wheat fields in Groningen, Flevoland and near Wageningen in Gelderland. In contrast to fields in the latter two areas, fields in Groningen were characterized by intensive wheat cultivation and yearly applications of benzimidazole fungicides for eyespot control. Seventy-seven isolates ofPseudocercosporella herpotrichoides were recovered. Forty-three percent of recovered isolates were carbendazim-resistant, all of which originated from fields in Groningen. Resistant isolates were detected among both rye- and wheat-types of the pathogen. Rye-type isolates were identified asP. herpotrichoides var.acuformis and those of the wheat-type asP. herpotrichoides var.herpotrichoides.Samenvatting Monsters van de halmbasis van tarwestro werden in augustus 1984 verzameld van percelen met wintertarwe in Groningen, Flevoland en bij Wageningen in Gelderland. In tegenstelling tot de percelen in de twee laatstgenoemde gebieden, waren de percelen in Groningen gekenmerkt door intensieve tarweteelt en door jaarlijkse bespuitingen met benzimidazool fungiciden tegen de oogvlekkenziekte. In totaal werden 77 isolaten vanPseudocercosporella herpotrichoides verkregen. Drieënveertig procent van de isolaten was resistant tegen carbendazim en alle resistente isolaten waren afkomstig van percelen in Groningen. Resistente isolaten werden zowel bij het rogge- als bij het tarwetype van het pathogeen aangetroffen. Isolaten van het roggetype werden geidentificeerd alsP. herpotrichoides var.acuformis en die van het tarwetype alsP. herpotrichoides var.herpotrichoides.     

The multiplication of three pathotypes of the potato root eelworm on different potato varieties

In two pot experiments the pathotypes, AB and ABCD ofHeterodera rostochiensis multiplied at lower rates on the potato variety Libertas than pathotype A. Pathotype ABCD multiplied at the same rate on a variety resistant against pathotype A as pathotype A on Libertas. Pathotype AB multiplied at a lower rate on Libertas and a variety resistant against A than A on Libertas. In a third experiment with the pathotypes A and ABCD and five potato varieties susceptible to all pathotypes, two varieties resistant to A and AB and three resistant to A significant differences between the rate of reproduction of the same pathotype on different susceptible varieties were found. The rate of reproduction of the pathotype ABCD on the varieties tested tended to be slightly lower than that of pathotype A on varieties susceptible to that pathotype.Samenvatting In twee potproeven vermenigvuldigden de pathotypen AB en ABCD vanHeterodera rostochiensis zich minder sterk op het aardappelras Libertas dan pathotype A. Pathotype ABCD vermenigvuldigde zich even sterk op een tegen pathotype A resistenteS. andigenum-hybride als pathotype A op Libertas. Pathotype AB vermenigvuldigde zich minder sterk op Libertas en een tegen A resistente variëteit dan A op Libertas.In een derde proef met de pathotypen A en ABCD en vijf voor alle pathotypen vatbare aardappelrassen, twee tegen A en AB resistente en drie tegen A resistente rassen, werden significante verschillen aangetoond tussen de vermenigvuldiging van dezelfde pathotypen op verschillende daarvoor gevoelige aardappelrassen (Tabel 1 en 2). Tabel 3 geeft de indruk dat pathotype ABCD zich op de getoetste rassen minder sterk vermeerdert dan A op de daarvoor vatbare rassen.     

Specific Behaviour of French Aphanomyces Euteiches Drechs. Populations For Virulence and Aggressiveness on Pea, Related to Isolates from Europe, America and New Zealand

The pathogenic variability of Aphanomyces euteiches on pea was investigated using a collection of 88 pea-infecting isolates from France and 21 isolates from Denmark, Sweden, Norway, USA, Canada and New Zealand. Aggressiveness and virulence were assessed by scoring the root symptoms on a differential set of six pea genotypes. Eleven virulence types were characterised. The virulence type I, previously described as virulent on the whole set, was predominant and included the most aggressive isolates of all geographical origins. The other types were much less prevalent, existing as one to five isolates. Three virulence types (III, IV and V) contained no French isolates. The type III, avirulent on MN313, was composed of American isolates only, and resembled the major group recently described in the USA. A wide range of aggressiveness was found within the virulence type I, and the French isolates appeared globally more aggressive than the foreign isolates. These findings indicate that isolates from the virulence type I should be used as references in breeding programs, and that pea lines PI180693 and 552 may be the most interesting resistance sources to date, despite their only partial resistance.     

Variation of chrysanthemum virus B

In an attempt to unravel the complexity of chrysanthemum virus B eight isolates were grafted onto the chrysanthemum indicator cultivars Fanfare, Good News, Mistletoe, and Pink Marble. Six symptomatologically different entities could be distinguished. However, withantisera prepared against two of the isolates, no serological differences were detected. It is concluded that chrysanthemum virus B consists of many strains and that the number of strains that can be distinguished depends on the number of indicator cultivars used. The Good News, Yellow Doty, Nightingale, Improved Bronze Daisy and September Morn, mosaic viruses described by Brierley and Smith (1958) as well as the vein mottle, dwarf mottle and necrotic mottle viruses named by Hollings and Stone (1967) should be considered as strains of chrysanthemum virus B.Samenvatting In een poging de complexiteit van het chrysantevirus B te ontrafelen werden acht isolaten geënt op de chrysante-indicatorrassen Fanfare, Good News, Mistletoe en Pink Marble. Zes symptomatologisch verschillende eenheden konden worden onderscheiden (Tabel 1; Fig. 1,2 en 3), die echter serologisch niet verschilden. De conclusie wordt getrokken dat van het virus vele stammen voorkomen en dat het anntal te onderscheiden stammen afhangt van het aantal indicatorrassen dat men gebruikt. Ook de vijf mozaïekvirussen van Brierley en Smith (1958) en de leden van het leaf mottling complex van Hollings en Stone (1967) dienen als stammen van het chrysantevirus B te worden beschouwd.Stationed at the Research Station for Floriculture, Aalsmeer.     

Structure of Melampsora larici-populina populations on wild and cultivated poplar

Populations of Melampsora larici-populina, causal agent of poplar leaf rust, have been collected during 1992–1994 in three types of sites: natural stands of Populus nigra in the south-east of France; cultivated poplar stands in the northern half of France, including mainly interspecific and exotic hybrids of poplar; and two overseas populations collected from Northern Ireland and Western USA. The pathotype of each rust isolate was determined by inoculating a differential set of poplar clones. The number of virulences per isolate and the frequency of each virulence in each population were compared, according to the sites and to the clones from which the isolates were collected.Several diversity indices were applied to the populations. The and Simpson indices were retained since they described best the richness and the evenness, respectively, of the populations, and they were the least sensitive to sample size. The complexity of races was lower in the natural stands of P. nigra than in the cultivated poplar stands. Populations of M. larici-populina collected on Populus × euramericana Robusta and P. nigra italica (Lombardy poplar) presented a high richness and evenness, confirming the value of these clones for describing race populations. Populations from Western USA presented a very low diversity, which is in accordance with the recent introduction of the pathogen in North America. These results suggest that the race populations of M. larici-populina are mainly influenced by the structure of the host populations.     

Heterogeneous Nature of a ‘new’ Pathotype of Melampsora Rust on Salix Revealed by AFLP

An outbreak of rust on Salix × mollissima (S. triandra × S. viminalis) Q83, an important biomass willow, was first observed at several locations in the UK in 1992. Rust collections obtained from Q83 in 1992 at Long Ashton (south west England), Markington (Northern England) and Loughgall (Northern Ireland), were tested for pathogenicity and examined using amplified fragment length polymorphism (AFLP). All collections showed the same pathogenicity patterns on the eight willow differentials and were assigned to f.sp. larici-epitea typica of Melampsora epitea. A total of 304 AFLP markers was scored for 54 rust isolates, 20 from Long Ashton, 20 from Markington and 14 from Loughgall. Cluster analysis placed the isolates into three distinct groups according to the collection sites. Within each site, Markington isolates were least variable, Nei & Li's similarity coefficients averaging 0.996. Average similarities within isolates from Long Ashton and Loughgall were 0.899 and 0.883, respectively. Average per-locus diversity within site (H_j), calculated using Shannon information index, was 0.014 in Markington, 0.24 in Long Ashton and 0.23 in Loughgall population. Most diversity (69.1%) was partitioned between populations. An analysis of molecular variance (AMOVA) attributed 85.8% of variance to between populations and 14.2% to the individuals within populations. The results suggest that, in 1992, this previously unknown pathotype was not spread from a common source but from separate sources. The AFLP analysis and early records on the host range of M. epitea indicate that the rust virulent to S. × mollissima may have existed in nature before 1992.     

Particle length of various isolates of potato virus S

The normal lengths of virus particles of five isolates of potato virus S, which in previous experiments had been detected to differ extremely in translocation speed in potato plants, were found to vary only from 641 to 655 m. However, these differences are not significant and the isolates are therefore considered to have a particle length of 650 m in agreement with Brandes' recent conclusion (1967).Samenvatting Er werd vastgesteld, dat de normale lengte van de virusdeeltjes van vijf isolaten van het aardappel-S-virus, waarvan in eerder uitgevoerde proeven was aangetoond dat ze met verschillende snelheden in aardappelplanten werden getransporteerd, varieerden van 641 tot 655 m. Aangezien de verschillen in de berekende lengten onbetrouwbaar zijn moet worden geconcludeerd dat de isolaten een gelijke normale lengte bezitten. Deze kan vanwege zijn overeenstemming met de gegevens van Brandes (1967) gesteld worden op 650 m.     

Some new races ofPuccinia striiformis

In the Netherlands in 1961 three new field races were found ofPuccinia striiformis, the causal fungus of yellow rust on wheat. Two of these, viz. the Falco race and the Opal race, together with the already known Etoile de Choisy race, constitute the so-called Nord-group of races, which is now clearly distinguished from the so-called Rubis-group. Most dangerous and interesting is the third of the new races. This so-called Cleo race proved to be the first to break both resistance to the Nord-group and resistance to the Rubis-group. On wheat seedlings in the greenhouse all three new races behaved like green-house race W(ageningen) 8=B(raunschweig) 55.Samenvatting Er zijn van de schimmelPuccinia striiformis Westend., de veroorzaker van de gele roest van tarwe, in Nederland in 1961 drie nieuwe veldfysio's gevonden. Dit zijn het Falco-fysio, het Opal-fysio en het Cleo-fysio. De eerste twee bleken met het reeds bekende Etoile de Choisy-fysio een groep te vormen, de z.g. Nord-groep, die zich duidelijk onderscheidt van de reeds beter bekende Rubis-groep van fysio's. Eerstgenoemde groep kenmerkt zich o.a. door de aantasting van het tarweras Nord, de laatstgenoemde door de aantasting van Rubis (zie tabel 1). Het belangrijkst en het gevaarlijkst is het Cleo-fysio. Dit bleek voor zover bekend het eerste en enige fysio te zijn dat zowel de resistentie tegen de Rubis-groep als die tegen de Nord-groep doorbreekt. Het heeft zodoende een zeer breed aantastingsspectrum. Van de tarwerassen die voorkomen in de 39ste Nederlandse Rassenlijst voor Landbouwgewassen (1964) zijn de volgende negen rassen vatbaar voor het Cleo-fysio met een aantastingsgraad van 6 tot 9 (Internationale Schaal): Cleo, Cappelle Desprez, Heine's VII, Hector, Stella, Sambo, Mado, Wodan en Jufy I. Bovendien bleek het Cleo-fysio in vergelijking met de andere veldfysio's goed te overwinteren en in 1964 reeds op vele plaatsen in Nederland aanwezig te zijn. Het Cleo-fysio maakt het des te noodzakelijker bij de selectie op resistentie tegen gele roest te werken op een brede genetische basis. Bij toetsing op tarwekiemplanten in de kas gedroegen de drie nieuwe fysio's zich alle als het kasfysio W(ageningen) 8 =B(raunschweig) 55 (zie tabel 2).     

Virulence inCeratocystis ulmi

Single-ascospore cultures ofCeratocystis ulmi from the progeny of a cross between a culture of compatibility type A from Belford, Massachusetts, U.S.A., and one of type B from Rotterdam, Netherlands, were morphologically heterogeneous and varied widely in virulence towards young elms inoculated in the greenhouse. Some cultures regularly caused typical wilt in callus cuttings ofUlmus hollandica Belgica (susceptible) but none in those ofU. carpinifolia Christine Buisman (resistant). Some cultures cuased wilt in trees of both clones; others in neither. All isolates produced abundant yeast-like cells in liquid culture. All isolated induced typical vascular discoloration except in the terminal inch of twigs. Similar progeny from a cross between cultures of types A and B, both from Naarden, Netherlands, however, were morphologically homogeneous and caused a fairly uniform level of symptoms in young trees ofU. hollandica Belgica (susceptible) and Commelin (moderately resistant), and in seedlings ofU. americana after inoculation in the nursery.Samenvatting Eén-ascosporecultures vanCeratocystis ulmi, verkregen uit de nakomelingen van een kruising tussen een cultuur van het compatibiliteitstype A uit Bedford, Massachusetts, V.S., en een type B uit Rotterdam, Nederland, waren morfologisch heterogeen en vertoonden grote verschillen in virulentie ten opzichte van jonge iepen, die in de kas waren geïnoculeerd. Enkele cultures veroorzaakten bij herhaling een typische verwelking bij callusstekken van de kloonUlmus hollandica Belgica (vatbaar), maar zij brachten geen verwelking teweeg bijUlmus carpinifolia Christine Buisman (resistent). Andere cultures veroorzaakten verwelking bij bomen van beide klonen, weer andere bij geen van beide. In alle gevallen ontstond na inoculatie de typische houtverkleuring, die echter uitbleef in de toppen van de twijgen (ca. 3cm). Alle cultures produceerden in een voedingsvloeistof een overvloed van gistachtige cellen.Eén-ascosporecultures, verkregen uit een kruising tussen de twee compatibiliteitstypen, beide afkomstig uit Naarden, Nederland, waren daarentegen morfologisch homogeen. Zij vertoonden bovendien vrijwel geen verschillen in virulentie na inoculatie in de kwekerij in jonge bomen vanU. hollandica Belgica en Commelin (respectivelijk vatbaar en matig resistent) en in zaailingen vanU. americana (vatbaar).A contribution of the Massachusetts Agricultural Experiment Station.     

Isolation and identification of pathogenic strains of tomato mosaic virus by host passage

Three isolates of tomato mosaic virus, A.8, SJ-64 and SL^a, assumed to contain the pathogenic strains 1 and 2, were each subjected to selection pressure by passage through different hosts in concurrent series. The sub-isolated obtained were tested at intervals on differential tomato lines heterozygous for each of the resistance genes Tm-1 and Tm-2 and for the combination of Tm-1 and Tm-2.Three preliminary passages throughSolanum pennellii followed by 22 passages through the tomato line CStMW-18 (Tm-1/Tm-1) resulted for A.8, SJ-64 and SL^a in sub-iolates, of strain 1. Three preliminary passages throughL. peruvianum P. I. 128655 followed by 22 passages through the tomato line Pérou-2 (Tm-2/Tm-2) resulted for A.8 and SL^a in sub-isolates of strain 2 and for SJ-64 in a sub-isolate of strain 1.2. Twenty passages through Pérou-2, followed by two additional passages through the tomato line Craigella Tm-1/+Tm-2/+ resulted for SJ-64 and SL^a in sub-isolates of strain 1.2 and for A.8 in a sub-isolate of strain 2.Samenvatting Op drie isolaten van het tomatemozaïekvirus, A.8, SJ-64 en SL^a, die naar alle waarschijnlijkheid de pathogene stammen 1 en 2 bevatten, werd selectiedruk uitgeoefend door een herhaalde waardplantpassage. Voor de afscheiding van stam 1 onderging elk van de isolaten eerst driemaal een passage doorSolanum pennellii en vervolgens 22 maal door de tomaatselectie CStMW-18 (Tm-1/Tm-1). Voor stam 2 passeerde elk isolaat eerst driemaalLycopersicon peruvianum P.I. 128655 en vervolgens 22 maal de tomaatselectie Pérou-2 (Tm-2/Tm-2). Op gezette tijden werden de sub-isolaten, verkregen uit de gelijktijdig verlopende passageproeven, getoetst op een differentiële reeks tomaatselecties. Deze waren heterozygoot en homozygoot voor de resistentiegenen Tm-1 en Tm-2 en voor de combinatie van Tm-1 en Tm-2.Voor elk van de isolaten A.8, SJ-64 en SL^a werden na passage door CStMW-18 sub-isolaten verkregen van stam 1. Terwijl voor A.8 en SL^a na passage door Pérou-2 sub-isolaten van stam 2 werden verkregen, leverde dezelfde passage voor SJ-64 een sub-isolaat, op van stam 1.2. Voor SJ-64 en SL^a gaven, twee passages door de tomaatselectie Craigella Tm-1/2 Tm-2/+ in aansluiting op 20 passages door Pérou-2 aanleiding tot de vorming van stam 1.2.In de discussie wordt op de mogelijke ontstaanswijze van stam 1.2 ingegaan.     

Injury to cut flowers of gladiolus by fluoridated water

The fluorine compound in fluoridated drinking water caused injury to the leaf tips as well as to the leaves and bracts of flower stalks of gladiolus, when cut leaves or cut flowers were placed with their bases in the water.In all cases the fluorine content of 4 cm long leaf tips was much higher when fluoridated water was used in place of non-fluoridated water.Samenvatting Aanzienlijk meer bladtopbeschadiging werd geconstateerd bij het voor HF zeer gevoelige gladioleras Sneeuwprinses wanneer bladstukken met de basis in gefluorideerd leidingwater zijn geplaatst dan wanneer deze in niet gefluorideerd leidingwater staan. Ook werden bij de als snijbloem gebruikte gladiolerassen Sneeuwprinses, Oscar en mary Housley de bladtoppen en de toppen van de schutblaadjes van de bloemen, door opname van gefluorideerd water meer beschadigd dan wanneer niet-gefluorideerd water werd toegepast.Zowel de bladstukken als de bladeren van de bloemstengels bevatten in de 4 cm lange bladtoppen een aanzienlijk hoger fluorgehalte wanneer deze geflurodeerd water hebben opgenomen dan wanneer niet gefluorideerd water opgenomen werd.     

Fitness of isolates of Sphaerotheca fuliginea resistant or sensitive to fungicides which inhibit ergosterol biosynthesis

Isolates ofSphaerotheca fuliginea resistant to fungicides which inhibit ergosterol biosynthesis (EBIs: bitertanol, fenarimol, imazalil) had been collected from glasshouses in the Netherlands. Fitness of these isolates was compared to that of isolates with a wild-type sensitivity to EBIs. Fitness parameters studied were germination of conidia, growth of germ tubes and mycelium, penetration, sporulation and competitive ability.In an experiment in which 10 EBI-resistant isolates were compared to 7 wild-type isolates, one or more values of fitness parameters for EBI-resistant isolates were slightly lower than those for the wild-type isolates. However, within the group of resistant isolates no relation existed between the degree of resistance to EBIs and the degree of fitness. In an experiment with fewer isolates but with more replicates, differences in fitness between EBI-resistant and wild-type isolates were not detected over a three-month period.In competition experiments in which no crowding was present, resistant isolates competed well with the wild-type isolate.It is concluded that the hypothesis that resistance to EBIs is unlikely to develop under practical conditions because of decreased fitness of EBI-resistant strains, does not seem to hold forS. fuliginea.Samenvatting Uit kassen in Nederland waren isolaten vanSphaerotheca fuliginea verzameld, die resistent waren tegen fungiciden die de ergosterol-biosynthese remmen (EBR's: bitertanol, fenarimol, imazalil). De fitness van deze isolaten werd vergeleken met die van isolaten met een wild-type gevoeligheid voor EBR's. De volgende fitness-parameters werden bestudeerd: sporekieming, groei van kiembuizen en mycelium, penetratie, sporulatie en competitievermogen.In een proef, waarin 10 EBR-resistente isolaten werden vergeleken met 7 wild-type isolaten, waren één of meer fitness-parameters iets lager dan die van de wild-type isolaten. Binnen de groep van de resistente isolaten bestond geen relatie tussen de mate van resistentie tegen EBR's en de waarden van de fitness-parameters. In een proef met iets minder isolaten maar met meer herhalingen in de tijd, werden geen verschillen in fitness waargenomen tussen de EBR-resistente en wild-type isolaten. In competitieproeven waarin een epidemie zich kon ontwikkelen, concurreerden de resistente isolaten goed met het wild-type isolaat.Er wordt geconcludeerd dat de hypothese dat resistentie tegen EBR's in de praktijk waarschijnlijk niet zou optreden vanwege een verminderde fitness van de EBR-resistente isolaten, niet van toepassing is opS. fuliginea.     

Pathogenic variation in poplar rust <Emphasis Type="Italic">Melampsora larici-populina</Emphasis> from England

Using a leaf disc method, 19 isolates of the poplar rust, Melampsora larici-populina , and one isolate of M.populnea from England were inoculated on to 25 poplar clones belonging to Populus nigra and P.trichocarpa, and hybrids between P. deltoides and P. nigra, P. deltoidesand P. trichocarpa, P.tacamahaca and P.trichocarpa, and P. alba and P. tremula. Disease was scored based on the pustule area and inoculum density. In terms of whether sporulating uredinia formed, the 19 isolates showed seven different patterns to the tested poplar clones. The majority of the rust isolates infected P. nigra P3090 and Vereecken, P.nigra×P. deltoides Casale and Tasman, P. tacamahaca×trichocarpa 36 and Balsam Spire, and P.trichocarpa Blom. Populus trichocarpa×P. deltoides 69039/4 was infected by only three isolates collected from southern England. No visible symptoms appeared on P. alba ×P. tremulaTower and P.trichcarpa×P. deltoides×P. deltoides76028/5 in inoculations with M. larici-populina isolates. Populus alba×P.tremula Tower was infected only by M. populnea. When M. larici-populina isolates were tested using AFLP, no differences were found either between isolates from different geographical regions or between those having narrow spectrum of virulence and those showing wide spectrum of virulence on the tested clones. The results suggest that the UK rust populations possess virulences which were found in races E1, E2, E3 and E4 in continental Europe and that rust having virulence patterns similar to race E4 has occurred in UK poplar plantations since 1996.     

Distribution of the FoToml gene encoding tomatinase in formae speciales of Fusarium oxysporum and identification of a novel tomatinase from F. oxysporum f. sp. radicis-lycopersici, the causal agent of Fusarium crown and root rot of tomato

The antifungal glycoalkaloid -tomatine accumulates in tomato plants and may protect plants from fungal infection. Fusarium oxysporum f. sp. lycopersici, the causal agent of vascular wilt of tomato, produces a tomatinase (FoToml) that degrades -tomatine to the nontoxic compounds tetrasaccharide lycotetraose and tomatidine. Induction of tomatinases and the distribution of FoToml homologs were examined among 30 strains belonging to 16 formae speciales of F. oxysporum. Tomatinase activity was found in 27 strains belonging to 15 formae speciales, but FoToml homologs (>98% sequence identity) were detected in only six strains belonging to four formae speciales. To identify tomatinases other than FoToml, -tomatine-inducible proteins of another tomato pathogen F. oxysporum f. sp. radicis-lycopersici were analyzed by two-dimensional gel electrophoresis. A protein with a molecular mass of 64kDa accumulated in the -tomatine-induced culture filtrates, and the protein had tomatinase activity, degrading -tomatine to lycotetraose and tomatidine.     

A revision of Billing's potential doublings table for fire blight prediction

In her fire blight prediction systems, E. Billing (1978, in: P.R. Scott & A. Bainbridge (Eds), Plant disease epidemiology, p. 159–166) has used the parameter potential doublings per day (PD) of the fire blight causing bacterium,Erwinia amylovora. Reconsideration of her calculations of PD revealed, however, that the PD values in Billing's table were underestimated. This leads to overestimation of the duration of incubation periods. A corrected PD table is presented. Sensitivity analyses indicated that day-of-the-year and latitude have little effect on the values in the PD table.Samenvatting Billing maakte in haar systemen waarin de ontwikkeling van bacterievuur wordt voor-speld, gebruik van de parameter potentiële verdubbelingen per dag (=PD) van de bacterievuur veroorzakende bacterieErwinia amylovora. Uit herberekeningen blijkt echter, dat de PD-waarden in Billings tabel onderschat worden. PD-onderschattingen leiden in haar systemen tot overschattingen van de duur van incubatieperioden. Een gecorrigeerde PD-tabel werd samengesteld. Uit gevoeligheidsanalyses bleek dat de dag van het jaar en de breedtegraad een gering effect hebben op de waarden in de PD-tabel.     

A greenhouse test for screening sugar-beet (Beta vulgaris) for resistance to beet necrotic yellow vein virus (BNYVV)

Small differences in activity between batches of purified beet necrotic yellow vein virus (BNYVV) were observed in ELISA. A four-parameter modelled dose-response curve of purified BNYVV was used for the conversion of ELISA values to virus concentrations. Seedlings of the susceptible cultivar Regina and the partially resistant cultivars Nymphe and Rima were tested for resistance to BNYVV in a mixture of sand and infested soil. Plants were grown in a green-house with low nutrient supply and at temperatures below the optimum of both the vectorPolymyxa betae and BNYVV. Root systems were small and consisted mainly of lateral roots. Significant differences in average virus concentrations were found between cultivars, both when using the complete root systems and when using either the top or the bottom part of the root systems. Average virus concentrations in Regina were always significantly higher than in Rima and higher than in Nymphe on all occasions except one (P<0.05). Differences between Nymphe and Rima were less evident. Variation between plants was greatest within Rima. The test described in this paper can be used for the discrimination of different cultivars and for the identification of individual plants with resistance to BNYVV.     

Virulence variation and DNA polymorphism in Sphaerotheca fuliginea, causal agent of powdery mildew of cucurbits

Strains of Sphaerotheca fuliginea, one of the causal agents of powdery mildew of cucurbits, were examined for differences in virulence, mating type and DNA polymorphism. The 28 strains were chosen to be diverse according to host and geographic origin. Characterization of virulence phenotypes was based on the expression of symptoms on 4 species of cucurbits and 6 cultivars of melon. Two pathotypes, capable of attacking either cucumber cv. Marketer and melon cv. IranH and squash cv. Diamant or cucumber cv. Marketer and melon cv. IranH were observed. Tests on melon cultivars revealed 3 races. In tests of sexual compatibility with reference strains, heterothallism was observed for all isolates. Frequency of the two mating types differed significantly in the population. DNA polymorphism was determined both by restriction fragment length polymorphism (RFLP) of the ribosomal internal transcribed spacers (ITS) and 5.8S DNA amplified by the polymerase chain reaction and by random amplified polymorphic DNA (RAPD). For any one of the 11 restriction enzymes tested all strains presented an identical pattern of ITS RFLP. RAPD analysis, using 22 primers which provided reproducible patterns, revealed a relatively low degree of polymorphism. Furthermore, cluster analysis based on RAPD data (152 markers) did not separate groups within the species S. fuliginea. No association could be found between virulence, mating type, geographical and host origin and RAPD patterns. The lack of association between phenotypic and molecular markers and the close fit to linkage equilibrium for the characters examined suggest that recombination may play a role in populations of S. fuliginea.     

Characterisation of the saponin hydrolysing enzyme avenacoside-α-l-rhamnosidase from the fungal pathogen of cereals, Stagonospora avenae

The fungal pathogen Stagonospora avenae f. sp. avenaria infects oat leaves, which contain the saponins avenacoside A and B. The avenacosides are glycosylated steroidal saponins that occur within oat leaves in a non-fungitoxic form and are converted upon damage or pathogen invasion to their antifungal form by a plant enzyme. It has previously been shown that oat-attacking isolates of S. avenae are able to hydrolyse the sugar chain at the C3 position of the avenacosides. This carbohydrate moiety is a branched chain that consists of one -{scL}-rhamnose and two or three -{scD}-glucose residues in avenacosides A and B, respectively. Removal of the -{scL}-rhamnose residue is sufficient to detoxify the avenacosides. This work describes the purification of the avenacoside-degrading -{scL}-rhamnosidase–and determination of peptide sequence from the protein which represents the first -{scL}-rhamnosidase thus characterised in a fungal plant pathogen.