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 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.     

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.     

Effects of fusaric acid on cells from tomato cultivars resistant or susceptible toFusarium oxysporum f. sp.Lycopersici

Cell suspension cultures were set up from two tomato cultivars, one resistant, (Rio grande) and one susceptible (63.5) toFusarium oxysporum f. sp.lycopersici. Growth rates of the two cell cultures were comparable. Toxicity of fusaric acid, expressed as the fresh weight loss, was analyzed: It was significant in both cases after 10 h, but toxicity was twice as high for 63.5 suspension cells. In the same way, electrolyte leakage caused by fusaric acid was three times more important for 63.5 suspension cells. Moreover, fusaric acid treatment resulted in an acidification of the extracellular medium for 63.5 suspension cells (0.4 pH unit), whereas an alkalization was observed for Rio grande suspension cells (0.2 pH unit). Preliminary experiments suggest that fusaric acid was partially metabolized by Rio grande suspension cells, however, no detoxified forms of fusaric acid were detected either in cells or in culture filtrates. For these two tomato cultivars, the differences in sensitivity to fusaric acid of cultivated cells correspond to the differences in plant susceptibility toFusarium oxysporum f. sp.lycopersici.Abbreviations BAP 6-benzylaminopurine - conductivity - 2,4-D 2,4-dichlorophenoxyacetic acid - EtOAc ethyl acetate - FA fusaric acid - resistivity     

Diapause in the nematodeGlobodera pallida

Over a period of 12 months new and old cysts ofGlobodera pallida were hatched in potato root diffusate according to a novel nematode-response hatching protocol. In this protocol, cysts were set to hatch at the beginning of autumn and then left to indicate when their hatching ability was exhausted (when less than 100 juveniles/replicate/week emerged) before another batch of cysts was set to hatch. At any time of the year for the 12 months this experiment was conducted there were cysts hatching. After 12 months of hatching, eight hatching curves were obtained. Based on the hatching curves of the new and old cysts, diapause was shown to be present in new cysts in autumn, winter and early spring. However, diapause was absent in late spring and summer.Infectivity assays to distinguish between juveniles obtained in the periods when cysts were in diapause and when cysts had overcome their diapause failed to show any significant difference in their infectivity. There was no significant difference in the number of eggs in new and old cysts. Based on this observation, it was suggested that high emergence in old cysts may not be a result of few eggs in the cyst but rather due to absence of diapause. Also the presence of large numbers of eggs in old cysts even after being stored for 12 months outdoors in the soil does not support the theories of spontaneous hatching, micro-organism induced hatching or persistence of hatching factors in the soil.     

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.     

Phytophthora root rot of sweet pepper

Phytophthora capsici proved to be the causal agent of a root and crown rot of sweet pepper in the Netherlands.P. capsici was pathogenic on sweet pepper, tomato and sometimes on eggplant but not on tobacco Xanthi. Of these test plants only tomato was infected byP. nicotianae.No different symptoms in plants infected with eitherP. capsici orP. nicotianae were found. Dipping the roots of tomato and sweet pepper plants in a suspension ofP. capsici resulted in a more severe attack than pouring the suspension on the stem base.Resistance in tomato toP. nicotianae did not include resistance toP. capsici. A method to distinguishP. capsici fromP. nicotianae after isolation from soil is described. Both species were able to infect green fruits of tomato and sweet pepper.p. capsici survived in moist soil in the absence of a host for at least 15 months.Samenvatting Phytophthora capsici bleek de oorzaak te zijn van een voet-en wortelrot in paprika op twee bedrijven in 1977 in Nederland.P. capsici was pathogeen op paprika, tomaat en soms op aubergine maar niet op tabak Xanthi.P. nicotianae tastte van deze toetsplanten alleen tomaat aan. Verschillen in symptomen tussenP. nicotianae enP. capsici werden bij tomaat niet waargenomen.Het dompelen van de wortels in eenP. capsici suspensie gaf een ernstiger aantasting dan het begieten van de wortelhals met deze suspensie.Resistentie in tomaat tegenP. nicotianae bleek geen resistentie tegenP. capsici in te houden. P. capsici kan in grond worden aangetoond door groene paprikavruchten als vangsubstraat te gebruiken.P. capsici enP. nicotianae kunnen beide zowel vruchten van tomaat als paprika aantasten. P. capsici overleefde een periode van 15 maan den in vochtige grond waarop geen waardplant werd geteeld.     

Histology of roots of resistant and susceptible carnation cultivars from soil infested with fusarium oxysporum f.sp. dianthi

Fusarium wilt-resistant Novada and wilt-susceptible Early Sam carnations were planted in soil infested withFusarium oxysporum f.sp.dianthi, and their roots studied after five and ten weeks. Both in Novada and Early Sam, the extravascular tissue of undamaged young root parts were scarsely colonized. In roots of Novada, infected xylem vessels were usually occluded with gums and surrounded by phellem tissue. In mature parts of roots, the phellem surrounding occluded vessels often merged with the external phellem surrounding the vascular cylinder, after which the occluded vessels were shed from the roots. The phellem at the root surface appeared to be a strong barrier to fungal invasion. In roots of Early Sam carnations, as well as in a few roots of Novada carnations, the defence responses did not result in compartmentation of the fungus, and colonization and degradation of the vascular tissues followed. Diseased roots finally healthy. Shoots of Early Sam carnations, and eventually a few shoots of Novada carnations, were colonized and developed wilt symptoms.Samenvatting Resistente Novada en vatbare Early Sam anjers werden geplant in grond besmet metFusarium oxysporum f.sp.dianthi. Na vijf en tien weken werden de wortels van de planten bestudeerd. De extravasculaire delen van onbeschadigde jonge wortelgedeelten waren in beide cultivars nauwelijks gekoloniseerd. In de wortels van Novada waren geïnfecteerde houtvaten meestal verstopt door gommen en omgeven door kurkweefsel. In oudere wortelgedeelten sloot het kurkweefsel rond verstopte vaten vaak aan op het kurkweefsel aan de buitenkant van het vaatweefsel, waarna de aangetaste vaten werden uitgestoten uit de wortel. Het kurkweefsel aan het worteloppervlak leek een belangrijke barrière te vormen tegen het binnendringen van de schimmel. In de wortels van Early Sam en in enkele wortels van Novada mislukte het afweerproces, en werd het vaatweefsel door de schimmel gekoloniseerd en vervolgens afgebroken. Aangetaste wortels werden na afloop van tijd hol. De bovengrondse delen van de meeste Novada anjers werden niet gekoloniseerd en bleven gezond. De bovengrondse delen van de Early Sam anjers, en op den duur die van enkele Novada anjers, werden gekoloniseerd en verwelkten.     

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).     

Induced Resistance to Cyst and Root-knot Nematodes in Cereals by DL-β-amino-n-butyric Acid

Foliar sprays and soil drenches with DL--amino-n-butyric acid (BABA) reduced the number of Heterodera avenae and H. latipons cysts on wheat and barley. Foliar sprays of wheat with 8000mgl^–1 BABA reduced the number of H. avenae cysts by 90%, whereas 2000mgl^–1 BABA was enough to reduce the number of H. latipons cysts by 79%. Multiple spray treatments with 2000mgl^–1 BABA at 10-day intervals reduced the number of H. avenae cysts on wheat and barley. A soil drench of wheat with 125mgl^–1 BABA reduced the number of H. latipons cysts by 93% and H. avenae cysts by 43%. Second-stage juveniles of these nematodes penetrated and formed syncytia in wheat roots soil-drenched with BABA. More adult males of H. avenae were produced in BABA (<250mg1^–1)-treated wheat roots (~76%) than in untreated roots (27%). Soil drenches with higher concentrations of BABA inhibited development of adult males and females. Several chemical elicitors of induced resistance were tested for their ability to reduce the number of H. avenae cysts on wheat. Only BABA was found to be an effective resistance inducer. The number of egg masses of an unidentified Meloidogyne sp. root-knot nematode, which infects only monocots, was also reduced by 95% by a soil drench of wheat with 500mgl^–1 BABA. Development of this nematode inside the BABA-treated roots was also inhibited.     

Differences in pathogenesis observed among susceptible interactions of carnation with four races of Fusarium oxysporum f.sp. dianthi

Susceptible interactions of Early Sam carnations with races 1,2,4, and 8 ofFusarium oxysporum f. sp.dianthi differed in pathogenesis, both after stem and after root inoculation. Race 1 induced pallescence and withering of leaves. Affected vascular tissue had a uniform pallid to pale brown colour; though heavily colonized, it was not or virtually not degraded. Defence reactions developed only slowly. Race 2 induced yellowing, of the midribs in particular, and withering of leaves. Affected vascular tissue was white with dark brown margins. Colonized tissue was degraded to leave vascular cavities. At lower heights of colonization, many defence reactions developed, which sometimes resulted in localization of the pathogen. Race 4 induced a similar pathogenesis as race 2, except for less intensive defence reactions. Race 8 induced midrib lesions on, and pallescence, withering and necrosis of leaves. Affected vascular tissue had a uniform light brown colour. Degradation of colonized vascular tissues was rare; instead, many defence reactions were observed, even at high heights in the plants.Races 1, 2 and 4 ofF. oxysporum f. sp.dianthi did not induce disease symptoms in Novada carnations, known to be highly resistant to race 2. Stem-inoculated plants localized the infection close to the inoculation site; stems of root-inoculated plants remained unaffected. The localization response also occurred in Early Sam and Novada carnations stem-inoculated withF. oxysporum f. sp.lycopersici.Samenvatting Tussen interacties van Early Sam-anjers met fysio's 1, 2, 4 en 8 vanF. oxysporum f. sp.dianthi werden verschillen in ziekteontwikkeling gevonden na wortel-zowel als stengelinoculatie. Fysio 1 gaf verbleking en verdroging van de bladeren. Aangetast vaatweefsel was gelijkmatig vaal of lichtbruin van kleur, en werd hevig gekoloniseerd, maar vrijwel niet afgebroken. Afweerreacties kwamen slechts traag op gang. Fysio 2 gaf vergeling, in het bijzonder van de hoofdnerven, en verdroging van de bladeren. Aangetast vaatweefsel was wit met donkerbruine randen. Gekoloniseerd weefsel werd afgebroken, hetgeen leidde tot de vorming van holten in het vaatweefsel. In de lagere gekoloniseerde delen traden veel afweerreacties op, hetgeen soms lokalisatie van het pathogeen tot gevolg had. Fysio 4 gaf eenzelfde ziekteontwikkeling als fysio 2, maar minder afweerreacties. Fysio 8 gaf lesies bij de hoofdnerven, en verbleking, verdroging en necrose van bladeren. Aangetast vaatweefsel was gelijkmatig lichtbruin van kleur. Afbraak van gekoloniseerd vaatweefsel werd zelden waargenomen; veel afweerreacties vergezelden de kolonisatie tot hoog in de stengel.Inoculatie van Novada anjers met fysio's 1,2 en 4 vanF. oxysporum f. sp.dianthi had geen ziektesymptomen tot gevolg. Via de stengel geïnoculeerde planten lokaliseerden de infectie ter hoogte van het inoculatiepunt; de stengels van via de wortels geïnoculeerde planten waren onaangetast. De lokalisatiereactie trad ook op in Early Sam en Novada anjers na inoculatie via de stengel metf. oxysporum f. sp.lycopersici.     

The β-tubulin Gene is a Useful Target for PCR-based Detection of an Albino Ophiostoma piliferum Used in Biological Control of Sapstain

The potential of Cartapip, an albino Ophiostoma piliferum, as a biocontrol agent against sapstain in logs has been tested in Germany. To detect the albino strain in field-tested wood, the usefulness of the -tubulin gene as a target region for developing PCR-based assays was evaluated with 102 strains of O. piliferum and 31 strains of other wood-inhabiting species. A partial -tubulin gene sequence of O. piliferum strains from different geographic origins was amplified by PCR and analyzed by restriction enzyme digestions and DNA sequencing. Variation in size and nucleotide sequences was found in intron regions indicating that intraspecific variation is present in the -tubulin gene. Consequently, -tubulin gene-derived PCR methods using PCR–RFLP patterns generated by HinfI and SpeI and sequence-specific primers Cat1 and Cat2, were developed and their specificity for Cartapip was accessed with field-tested logs and lumber. The -tubulin gene-based PCR methods were found to be valuable tools for rapid and reliable identification of Cartapip in field-tested logs and lumber in Germany. Specificity tests against other wood-inhabiting species and wild type O. piliferum strains from diverse nations showed that the Cat1 and Cat2 primers have potential to be used in other European countries, New Zealand, Alberta and British Columbia.     

Changes in susceptibility of bean leaves (Phaseolus vulgaris) to Sclerotinia sclerotiorum and Botrytis cinerea by pre-inoculative ozone exposures

The effects of ozone on the susceptibility of leaves ofPhaseolus vulgaris toSclerotinia sclerotiorum andBotrytis cinerea have been investigated. Seedlings of one ozone-sensitive (Pros) and five relatively ozone-insensitive cultivars (Gamin, Precores, Groffy, Narda, Berna) were exposed to different ozone concentrations (0, 120, 180 and 270 g m^–3) for 8 h. One day after the exposures, primary leaves were detached and immediately inoculated with spores of either pathogen suspended in water or in a 62.5 mM KH₂PO₄ (Pi) solution. Visible ozone injury differed between the cultivars and increased with increasing ozone concentration. On the leaves of non-exposed plants, spores of the pathogens suspended in water caused very few lesions, whereas fungal pathogenicity was stimulated by addition of Pi to the inoculum. Ozone-injured leaves of all cultivars exhibited lesions after inoculation of the leaves with the pathogens suspended in water, and the number of lesions was positively correlated with the level of ozone injury for either pathogen and cultivar. The increase in susceptibility of bean leaves in response to increasing ozone concentrations was greater forB. cinerea than forS. sclerotiorum when spores were suspended in water, but was similar when the spores were suspended in Pi.In general, the number of lesions following inoculation with spores in Pi increased with increasing ozone concentration. However, the number of lesions in the ozone-insensitive Groffy was reduced by an exposure to 120 g m^–3 but increased with higher concentrations. This pattern of susceptibility response to the pathogens was not found in the other ozone-insensitive cultivars and, thus, did not appear to be related to the inherent ozone-insensitivity in bean.     

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.     

Influence of Inoculum Density of Races 0 and 5 of Fusarium oxysporum f. sp. ciceris on Development of Fusarium Wilt in Chickpea Cultivars

Artificial inoculation experiments were carried out at 25°C to determine the effects of inoculum density of Fusarium oxysporum f.sp. ciceris races 0 (Foc-0) and 5 (Foc-5) and susceptibility of chickpea cultivars P-2245 and PV-61 on development of Fusarium wilt. Foc-5 proved much more virulent than Foc-0. Increasing the inoculum density of F. oxysporum f.sp. ciceris caused an exponential reduction in disease incubation period and a monomolecular increase of disease incidence and the area under the disease intensity progress curve. The extent of these effects was highest in the most conducive P-2245/Foc-5 combination and decreased in the less susceptible PV-61 and for the less virulent Foc-0, in that order. For P-2245/Foc-5, the highest disease intensity was attained with 6 chlamydospores g^–1 of soil, the lowest inoculum density in the study. One thousand chlamydospores g^–1 of soil of the same race were needed to attain a comparable disease intensity in PV-61. Twenty thousand chlamydospores g^–1 of soil of Foc-0 were required for maximum disease intensity in P-2245.The disease intensity curves were adequately described by the Gompertz model. Using this model, a response surface for disease intensity was developed, in which the model parameters are expressed as a function of both time from inoculation and inoculum density. This response surface confirmed that the final amount of disease intensity increases in a monomolecular relationship with increasing inoculum density and showed that the relative rate of disease progress increases exponentially with increasing inoculum density of the pathogen.     

Application of enzymes during bulb tissue extraction for detection of lily symptomless virus by ELISA in Lilium spp

Lily symptomless virus (LSV) was readily detected with the enzyme-linked immunosorbent assay (ELISA) in leaves ofLilium Mid-Century hybrids cvs Enchantment and Destiny andLilium speciosum cv. Brabander. In bulbs of cv. Enchantment LSV could be detected without any additional treatment of the extracts, but for reliable results in cv. Destiny it was necessary to pre-incubate the bulb extracts with cellulase or hemicellulase.Samenvatting In de bladeren van secundair geïnfecteerde planten van de leliecultivars Enchantment, Destiny en Brabander kon LSV met ELISA gemakkelijk worden aangetoond. In bladextracten van Enchantment kon nog ca. 100 ng LSV/ml extract worden teruggevonden. Bij toepassing van de standaard-methode van extraheren (2 g weefsel malen in 4 ml extractiebuffer) kon ook in de bolschubben van Enchantment LSV worden aangetoond; bij Destiny gelukte dat aanzienlijk minder goed. Wijzigingen in de extractie-procedure of een speciale behandeling van het standaard-extract (dialyseren, verhitten tot 50°C etc.) leidde niet tot een betere aantoonbaarheid van het virus met ELISA. Enzymen als hemicellulase, cellulase en pectinase, toegevoegd aan het standaard-extract van bolschubben van Destiny vóór de toetsing (incubatie gedurende 18 h bij kamertemperatuur), verbeteren de aantoonbaarheid van het LSV aanzienlijk.     

Chemical and Biological Treatments for Control of Gummy Stem Blight of Greenhouse Cucumbers

Experiments were conducted to determine the effects of chemical and biological treatments on gummy stem blight of cucumber caused by Didymella bryoniae in vitro and under greenhouse conditions. Eleven strains of Bacillus subtilis, strain AGB10 of B. cereus, and strain B8Fr of Enterobacter agglomerans produced antagonistic zone against D. bryoniae in vitro. Of four experiments conducted, the chemical treatments Nova,¹ kresoxim-methyl, and azoxystrobin controlled the disease in three experiments and the biological treatments E. agglomerans (B8Fr), B. subtilis (AGS-4), and lysozyme in one experiment when applied as sprays on lesions caused by D. bryoniae on cucumber plants under greenhouse conditions. Fruit rot of cucumber was significantly reduced when the fruit was treated with Nova or kresoxim-methyl. These results suggest the potential of azoxystrobin, kresoxim-methyl, E. agglomerans (B8Fr), and B. subtilis (AGS-4) applied post-inoculation to control gummy stem blight on greenhouse cucumbers.     

Phytophthora syringae,geïsoleerd uit bast van appel en peer

Summary In 1960 and 1961Phytophthora syringae Klebahn was isolated repeatedly from diseased bark of apple (Cox's Orange Pippin, James Grieve) and pear (Beurré Hardy).As far as the author is aware, natural infection of apple- and pear bark byPhytophthora syringae has not hitherto been reported, either in The Netherlands or elsewhere.gedetacheerd bij het Proefstation voor de Fruitteelt in de volle grond, Wilhelminadorp     

Metabolism of phaseollin by different races of Colletotrichum lindemuthianum

The ability of four races of the bean pathogenColletotrichum lindemuthianum to metabolize the phytoalexin phaseollin in shake cultures was compared. Apart from some differences in the rate of conversion, all races metabolized the phytoalexin in the same way. Phaseollin was first converted to 6a-hydroxyphaseollin, and this product was further metabolized to 6a, 7-dihydroxyphaseollin. No metabolites of the latter compound could be detected.6a, 7-Dihydroxyphaseollin was as inhibitory as phaseollin to race 11, but was only slightly inhibitory to races 1, 2 and 1.Samenvatting Een vergelijkend onderzoek werd verricht naar het vermogen van vier fysiologische rassen van het bonepathogeenColletotrichum lindemuthianum om het fytoalexine phaseolline om te zetten.In schudculturen waaraan 10 g phaseolline/ml was toegevoegd, werd dit door alle fysio's op gelijke wijze omgezet, hoewel met verschillende snelheid (Fig. 1). Steeds werd phaseolline eerst omgezet tot 6a-hydroxyphaseolline, en dit produkt vervolgens tot een verbinding die geïdentificeerd kon worden als 6a, 7-dihydroxyphaseolline. Hierna konden geen verdere produkten worden aangetoond.6a, 7-Dihydroxyphaseolline was even fungitoxisch als phaseolline voor fysio 11, maar was slechts weinig fungitoxisch voor de fysio's 1, 2 en 1 (Tabel 1).De verschillen in omzettingssnelheid van phaseolline en in gevoeligheid voor phaseolline ee zijn omzettingsprodukten die tussen de fysio's gevonden zijn, zijn onvoldoende om de fysiospecifieke interacties tussen de boon en de verschillende fysio's vanC. lindemuthianum te verklaren.     

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.