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.     

Resistance of rose rootstocks to crown gall (Agrobacterium tumefaciens)

Various types of rose rootstocks were tested for their resistance to crown gall. The rootstock Iowa State University (ISU) 60–5 was the most resistant, followed by Brooks 48, Clarke 1957 and Welch. Rosa multiflora, R. noisettiana (Manetti) and Basye No. 3 were very susceptible. The inoculations were made with four isolates ofAgrobacterium tumefaciens (Smith et Townsend) Conn, respectively from aDahlia sp.,Rosa spp. andPrunus persica. It was found that the isolate fromDahlia was a different race to the isolates fromRosa andPrunus spp.Samenvatting Bij onderstammen van rozen, kunstmatig geïnoculeerd metAgrobacterium tumefaciens (Smith et Townsend) Conn., werden verschillen in resistentie tegen wortelknobbel gevonden. Het meest resistent was Iowa State University (ISU) 60–5, gevolgd door Brooks 48, Clarke 1957 en Welch, Zeer vatbaar warenRosa multiflora, R. noisettiana (Manetti) en Basye No. 3. De vier isolaten vanA. tumefaciens, gebruikt voor de inoculaties, waren respectievelijk afkomstig van eenDahlia sp.,Rosa spp. enPrunus persica. Het isolaat vanDahlia en de isolaten vanRosa enPrunus spp. behoorden tot twee verschillende fysiologische rassen. De vorming van tumoren was in sommige gevallen afhankelijk van de methode van inoculatie; inoculaties bij de stambasis waren meer succesvol dan in het midden van de stam.     

The Role of Trichoderma harzianum Protease in the Biocontrol of Botrytis cinerea

The role of protease of Trichoderma harzianum in the biocontrol of Botrytis cinerea was examined. Two isolates of T. harzianum were compared for their ability to produce protease in liquid culture medium and on the surface of bean leaves. The biocontrol agent T. harziaum T39 produced 58mU/ml of protease and T. harzianum NCIM1185 produced 54mU/ml on the 5th day of growth in liquid culture medium. On bean leaves, combinations of B. cinerea and T. harzianum isolates were examined for the synthesis of protease. The protease activities were 0.9 and 0.6mU/ml for T. harzianum T39 and NCIM1185, respectively, and 0.5mU/ml for B. cinerea alone after 48h of incubation. In the presence of T. harzianum T39 culture liquid containing protease, a 55% reduction in B. cinerea germination and a 80% reduction in the germ tube length were observed after 17h of incubation in vitro. When T. harzianum isolates were added to B. cinerea on bean leaves, increased synthesis of protease was observed (1.0 and 1.2mU/ml for T39 and NCIM1185, respectively). In the presence of T. harzianum NCIM1185 protease, although the rate of germination was reduced, B. cinerea attained 98% germination after 17h of incubation. The hydrolytic enzymes produced by B. cinerea, endo-polygalacturonase (PG) and exoPG were partially deactivated by protease from the T. harzianum isolates. Carboxymethyl cellulase was deactivated only by protease of NCIM1185. On the surface of bean leaves, the protease (obtained from liquid culture medium of T. harzianum isolates) resulted in 56–100% reduction of disease severity. The culture liquid containing protease synthesized on the surface of bean leaves treated with B. cinerea and with T. harzianum was collected and added to fresh leaves infected by B. cinerea. There was 56–100% and 30–75% reduction of disease severity with liquid droplet collected from the leaves treated with T. harzianum T39 and NCIM1185, respectively. Increased control of disease was obtained by combining the conidia of T. harzianum isolates with protease obtained from culture media. Protease inhibitors, trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane (E64), antipain hydrochloride, and a mixture of inhibitors, but not pepstatin A, fully or partially nullified the biocontrol effect of T39. T39 was found to be a poor producer of chitinase and -1,3-glucanase in vitro. These enzymes were not detected on leaves treated with T39. Involvement of protease in biocontrol of B. cinerea is suggested.     

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).     

Infection of Tomicus piniperda (Col., Scolytidae) with Canningia tomici sp. n. (Microsporidia, Unikaryonidae)

Canningia tomici sp. n. (Microsporidia, Unikaryonidae) infects the midgut epithelium, the gut muscules, Malpighian tubules, connective tissues, adipose tissues and the gonads of the pine shoot beetle, Tomicus piniperda (L.) (Coleoptera, Scolytidae). The infection is present in populations of Tomicus piniperda in Europe and in the United States. Uninucleate oval single spores occur in two sizes: 2.8±0.4× 1.4±0.4m and 3.8±0.3×2.0±0.2m. The polar filament of this microsporidium is fixed subapically in a flat anchoring disc. The thick posterior lamellae of the binary polaroplast are asymmetric due to the lateral fixation of the polar filament.     

Occurrence and etiology of death of young olive trees in southern Spain

New plantations of olive tree in southern Spain are being severely affected by wilt or dieback and death, which has been locally called Drying Syndrome. To determine the etiology of this problem, a study was carried out in samples of affected young trees collected during a seven year period (1989–1995), and in two field surveys in 1994–95 and 1996. Besides some insect damage and agronomic problems, the Drying Syndrome was associated with Verticillium wilt, winter frost and root rot fungi. Although Drying Syndrome can be distinguished from Verticillium wilt, the latter was included in this study, since, frequently, Verticillium wilt symptoms were unspecific and Verticillium dahliae could not be always isolated in the diagnostic work that preceded this study. Early winter frost caused a vascular necrosis and wilt of the young olive trees. This unusual and severe damage was related with the lack of frost hardiness due to warm temperatures during the previous autumn. Root rot fungi were very frequent in the samples of diseased olive trees of field or nursery origin, and they were the main cause of Drying Syndrome in the second field survey, when a heavy rainfall level occurred during winter. Pathogenicity tests showed that five fungal species (Cylindrocarpon destructans, Phytophthora megasperma, P. palmivora, Pythium irregulare and Sclerotium rolfsii) were pathogenic to olive trees and reproduced symptoms of Drying syndrome in rooted cuttings of cultivar Picual. Other fungal species associated with root rot of olive trees in the field or in the nurseries, including Fusarium acuminatum, F. eumartii, F. oxysporum, F. solani, Macrophomina phaseolina and Rhizoctonia solani, were weakly or not pathogenic. Pathogenicity of P. megasperma, P. palmivora and P. irregulare depended on soil water content, since isolates tested caused extensive root rot and sudden plant death only when the soil was continuously waterlogged. The high frequency of P. megasperma in waterlogged field soils and its pathogenicity dependence on soil water content suggest that this pathogen may play an important role in the well known sensitivity of young olive trees to root asphyxiation.     

Bud necrosis,a storage disease of tulips. I. Symptoms and the influence of storage conditions

The symptoms of bud necrosis in tulip bulbs are described. This disorder is neither contagious nor hereditary. It is found more often and more severely in certain cultivars (viz Red Champion and White Sail) than in others (viz Rose Copland) in which it appears less frequently or not at all. The disease originates in the stamens of the flower-producing main bud during dry storage between lifting and planting of the bulbs. The occurrence of the disease in sensitive cultivars is promoted by storage of the bulbs under conditions of poor ventilation (viz packed in cardboard boxes) and at higher temperatures later in the storage period (after September 1).The experimental results suggest that there are several pathogenic factors and that the combined action of these factors can lead to bud necrosis; the primary factor is probably of physiological nature.Samenvatting Kernrot is een afwijking in tulpebollen die een bloeibare maat hebben. De ziekte ontstaat tijdens de droge bewaring van de bollen en begint met afwijkingen aan de meeldraden. Bij de beschrijving is onderscheid gemaakt tussen symptomen welke zich tijdens de droge bewaring voordoen aan de knoppen waaruit zich de bloeiende spruit zal ontwikkelen en symptomen welke zich voordoen aan de bollen tijdens de bloei te velde. Naar de mate waarin de knoppen en planten afwijken, wordt gesproken van meeldraadnecrose (Fig. 1A en 2), bloemnecrose (Fig. 1B en 4A), spruitnecrose (Fig. 1C en 4B), kernnecrose (Fig. 1D en 4C) en bolnecrose (Fig. 4D).Het is gebleken, dat de ziekte bij bepaalde cultivars (Red Champion en White Sail) vaak voorkomt, bij andere minder vaak (Rose Copland) of in het geheel niet. Er kon worden vastgesteld, dat de ziekte niet besmettelijk of erfelijk is.In de proeven werd het ontstaan van de afwijking bij Red Champion en White Sail bevorderd door de bollen te verpakken in kartonnen dozen (ventilatiebeperking) en door de bollen later in het seizoen (na 1 september) te bewaren bij hogere temperaturen dan daarvoor. Op grond van de proefresultaten wordt aangenomen dat de primaire oorzaak van de afwijking van fysiologische aard is. De aanwezigheid van schimmels, bacteriën en mijten moet als tweede oorzaak worden beschouwd.     

Isolation of Phytophthora cactorum from soil in orchards and strawberry fields and differences in pathogenicity to apple

Phytophthora cactorum was isolated from soil of strawberry fields and from an apple and a pear orchard, using hard apples and pears as traps. Pears gave better isolation of the fungus. Isolates from diseased strawberry rhizomes and strawberry-field soil were much less pathogenic to stems and stem and branch pieces of apple variety Cox's Orange Pippin than isolates from apple stem, pear fruit, and orchard soils. Each group of soil isolates showed differences with respect to length and number of the resulting cankers in apple stem and branch pieces.Samenvatting De isolatie vanP. cactorum uit boomgaardgrond en uit grond an aardbeivelden slaagde het beste door gebruik te maken van harde perevruchten. De schimmel groeide uit de grond in de vruchten en kon door overbrengen van een stukje weefsel van de rand van de vruchtaantasting op aardappel-glucose-agar voedingsbodem geisoleerd worden (Tabel 1). Isolaties uit grond van aardbeivelden waren veel minder pathogeen dan isolaties uit appel- of pereboomgaardgrond, indien ze geïnoculeerd werden in ongeveer 35 cm lange stam- en takstukken van het appelras Cox's Orange Pippin, welke in vochtige turfmolm bij 20°C geplaatst waren (Tabel 2). Evenzo vertoonden isolaties uit aangetaste aardbeirhizomen een geringe pathogeniteit voor stammen en stam- en takstukken van Cox's Orange Pippin in vergelijking met isolaties uit appelstam en perevrucht (Tabel 2 en 3 en Fig. 1). De meeste sterk infektieuse isolaties veroorzaakten grote kankers, doch enkele waren maar weinig virulent.     

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.     

Yield of glasshouse tomatoes as affected by strains of tobacco mosaic virus

In the latter part of the summers of 1963, 1964 and 1965 the effect of an early inoculation with different strains of tobacco mosaic virus (TMV) upon yield was investigated. All experiments were carried out in the same glasshouse-compartment and with the tomato variety Moneymaker, the plants being grown in plastic pails containing ordinary potting soil. In 1963 a green strain, a distorting strain and a yellow (or aucuba) strain were used, all capable of causing local lesions onNicotiana tabacum White Burley, which reaction is characteristic for the tomato-type of TMV. Since unfavorable conditions affected control plants more than infected plants, the green strain caused no significant loss of yield. Plants infected by this strain yielded only 3% less weight of fruit than the control plants and even had a 12% greater number of fruits. Plants infected by either the distorting or the yellow strain yielded 43 and 65% respectively less weight of fruit and 32 and 48% respectively fewer fruits. In 1964 the same green strain of the tomato-type TMV was compared with a green strain of the tobacco-type TMV, which is characterized by a systemic reaction on White Burley. Plants infected by the tomato-type and tobacco-type green strains ylelded 16 and 21% respectively less weight of fruit than the control plants and 11 and 18% respectively fewer fruits. In 1965 the same green strain of the tomato-type TMV was used and compared with another green strain of the same virus, which in addition caused local lesions onN. glauca, and with a yellow ringspot strain, which showed the tomato-type reaction on White-Burley. Plants infected with these three strains yielded 14, 18 and 17% respectively less weight of fruit than the control plants, but only 2, 6 and 6% respectively fewer fruits. The relatively small reduction in the number of fruits and an appreciable delay of the harvest probably resulted from the fact that the plants in this experiment were inoculated in the seedling stage.Samenvatting In 1963,1964 en 1965 werd in de nazomer het effect van een vroege inoculatie met verschillende stammen van het tabaksmozalekvirus (TMV) op de opbrengst van tomaat nagegaan. Alle proeven werden uitgevoerd in dezelfde kas en met het tomateras Moneymaker; de planten werden opgekweekt in plastic emmers die met gewone potgrond gevuld waren.In 1963 werden een groene stam, een misvormende stam en een gele (of aucuba-)stam gebruikt, welke alle in staat zijn de voor het tomaattype TMV karakteristieke lokale lesies te veroorzaken opNicotiana tabacum White Burley. Aangezien ongunstige omstandigheden de controleplanten nadeliger beinvloedden dan de ge'infecteerde planten, veroorzaakte de groene stam geen duidelijk opbrengstverlies. Planten, die met deze stam geinfecteerd waren, brachten slechts 3% minder aan vruchtgewicht op dan de controleplanten en zelfs 12% meer vruchten. Planten, geïnfecteerd met de misvormende of met de gele stam, brachten respectievelijk 43 en 65% minder aan vruchtgewicht op en respectievelijk 32 en 48% minder vruchten.In 1964 werd dezelfde groene stam van het tomaattype vergeleken met een groene stam van het tabakstype TMV, welke gekarakteriseerd wordt door een systemische reactie op White Burley. Planten, geïnfecteerd met de groene stam van het tomaattype of tabakstype TMV, brachten respectievelijk 16 en 21% minder aan vruchtgewicht op dan de controleplanten en respectievelijk 11 en 18% minder vruchten.In 1965 werd opnieuw dezelfde groene stam van het tomaattype TMV gebruikt en vergeleken met een andere groene stam van hetzelfde virus, welke bovendien lokale necrotische lesies veroorzaakt opN. glauca, en met een gelekringvlekkenstam, welke eveneens de tomaattype-reactie vertoont op White Burley.Planten, welke met een van deze stammen waren geïnfecteerd, brachten respectievelijk 14, 18 en 17% minder aan vruchtgewicht op dan de controleplanten, maar slechts 2, 6 en 6% minder vruchten. De betrekkelijk geringe reductie in het aantal vruchten en een aanzienlijke oogstverlating waren waarschijnlijk het gevolg van het feit dat de planten in deze proef in het kiemplantstadium waren geïnoculeerd.Stationed at the Glasshouse Crops Research and Experiment Station, Naaldwijk.     

Evidence for Three Different Specific Saponin-detoxifying Activities in Botrytis cinerea and Cloning and Functional Analysis of a Gene Coding for a Putative Avenacinase

Saponins are typical phytoanticipins, i.e. preformed fungitoxic compounds suspected to play an important role in plant defense. Several phytopathogenic fungi are able to detoxify these compounds by deglycosylation, and this ability could be a factor of general importance for the successful colonization of saponin-containing plants. Botrytis cinerea, causing grey mould of many (especially greenhouse) crops and vegetables, among them several plants containing saponins, has been reported to deglycosylate and detoxify -tomatine. Here we show that it can also degrade avenacin, avenacosides and digitonin, and that it possesses more than one saponin-specific enzyme. Using the tomatinase gene (tom1) from Septoria lycopersici as probe, we have cloned and characterized a gene (sap1) coding for a 83kDa polypeptide which shows significant homology to tom1 and to the avenacinase gene of Gaeumannomyces graminis. Disruption of sap1 leads to loss of the ability to deglycosylate avenacin, whereas sap1 deletion mutants still can detoxify tomatine, digitonin and avenacosides, i.e. sap1 encodes an avenacinase. The putative product of this gene has been purified and characterized: it has a MW of 90,000, an isoelectric point (IP) of 5.2, and it has no activity against avenacosides. Since a tomatinase-deficient field isolate of B. cinerea, M3, also lacks the ability to deglycosylate digitonin, but can degrade avenacin and avenacosides, these data confirm the existence of at least three distinct activities of saponin-specific glycosidases in B. cinerea: a xylosidase (-tomatinase, digitoninase), and two different glucosidases (avenacinase and avenacosidase).     

Interactive Effects of Host, Pathogen and Mineral Nutrition on Grey Leaf Spot Epidemics in Uganda

Grey leaf spot incited by Cercospora zeae-maydis is a new devastating foliar disease of maize in East Africa. For effective control, elucidation of the most critical elements of the grey leaf spot disease pyramid is important. This study investigated the role of mineral nutrition, pathogen variability and host resistance in the epidemic. Trials were conducted under field and controlled environments. The 28 isolates used in the controlled environment varied significantly (P 0.05) in parasitic fitness measured indirectly as disease efficiency, but no infection pattern could be attributed to known C. zeae-maydis pathotypes. Data from field trials showed that host resistance and mineral nutrition significantly (P 0.05) affected disease efficiency, with highest disease development occurring in nitrogen-augmented plots. Exclusive phosphorus application had no clear effect on grey leaf spot epidemics but combined application with nitrogen significantly (P 0.05) reduced the predisposition effects of nitrogen to the disease. Overall, treated plots had less disease than unfertilised plots. Fertiliser application had no effect on sporulation capacity, while cultivars significantly affected it. Geographic differences in amount of disease were observed, suggesting environment influences on grey leaf spot incidence. The results suggest that the current grey leaf spot epidemics in East Africa are due to favourable cultivars, poor mineral nutrition and environmental interactions.     

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.     

Inoculation of cucumber roots with zoospores of mycoparasitic and plant pathogenic Pythium species: Differential zoospore accumulation, colonization ability and plant growth response

Hydroponically grown cucumber (Cucumis sativus) seedlings were inoculated with zoospores of 1 mycoparasitic (Pythium oligandrum) and 2 pathogenic (Pythium aphanidermatum and Pythium group F) Pythium spp. During the first 2 days after inoculation, all the Pythium spp. caused reduction in the root length. However, roots treated with Pythium oligandrum quickly reached the length of the control and on the 8th day, and for the rest of the experimental period, stimulation of root elongation was noted. Pythium oligandrum was not pathogenic on cucumber and no differences in the fresh weights of control and Pythium oligandrum inoculated plants were observed in the course of the experiment. Pythium group F and Pythium aphanidermatum were pathogenic on cucumber seedlings, but their pathogenicities differed. Thus, while Pythium group F had a constant, negative influence on root length and plant growth, measured as fresh weight, Pythium aphanidermatum caused generalized necroses of the root system, inhibiting consistently root elongation and plant growth and finally causing plant death. Moreover, the zoospores of 2 mycoparasitic species, Pythium oligandrum and Pythium periplocum, were not attracted to roots of cucumber and accumulated on the roots in very low numbers compared to those of the pathogenic species, Pythium aphanidermatum, which were strongly attracted and accumulated in large numbers. Finally, it was also found that Pythium oligandrum colonized the roots very poorly, while Pythium group F and Pythium aphanidermatum were significantly better root colonizers. The significance of these findings is discussed in relation to the ecology of Pythium species and biocontrol.     

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.     

Pelargonium flower-break and pelargonium line pattern viruses in the Netherlands; purification,antiserum preparation,serological identification,and detection in pelargonium by ELISA

Two viruses, detected frequently in the Netherlands in pelargonium, were identified by serology and test plant reactions. Antisera were prepared and an ELISA procedure was developed to detect the viruses in pelargonium.One of the viruses, PFBV-N, proved to be pelargonium flower-break virus. With the antiserum to PFBV-N, it could be detected reliably throughout the year inPelargonium zonale Springtime Irene.The other virus, PLPV-N, was serologically closely related to pelargonium line pattern virus (PLPV) and to pelargonium ring pattern virus (PRPV), as were an old virus isolate from Saturnus, collected in the Netherlands in 1971 (L128), and PLPV isolates from Yugoslavia (PLPV-Y) and Denmark (PLPV-D). There were only minor differences in host-plant reactions between the virus isolates. Based on these tests, PLPV and PRPV are considered as isolates of the same virus, for which, for practical reasons, the name pelargonium line pattern virus is proposed.PLPV could be reliably detected by ELISA inP. zonale Springtime Irene and Amanda throughout the year with only a few exceptions. InPelargonium peltatum Tavira, however, reslts were erratic due to uneven distribution of virus in the plant. Best results were obtained when petioles of fully expanded leaves were tested.     

Foliar Application of Acibenzolar-S-Methyl and Protection of Postharvest Rock Melons and Hami Melons from Disease

A pre-flowering foliar spray of the plant activator acibenzolar-S-methyl at 50mg/La.i. combined with a fruit dip in guazatine at 500mg/La.i. at harvest substantially decreased disease in stored melons. Major diseases occurring on rock melons and Hami melons were caused by Fusarium spp., Alternaria spp., Rhizopus spp. and Trichothecium sp. The treatment with acibenzolar-S-methyl alone was significantly effective in reduction of the disease severity in many but not all situations. The fungicide guazatine alone significantly decreased infection by Fusarium spp. but had a lesser effect on that caused by Alternaria spp. and Rhizopus spp.     

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.     

Enzymatic Maceration of Witloof Chicory by the Soft Rot Bacteria Erwinia carotovora Subsp. carotovora: The Effect of Nitrogen and Calcium Treatments of the Plant on Pectic Enzyme Production and Disease Development

Disease incidence of bacterial soft rot caused by Erwinia carotovora subsp. carotovora and activities of bacterial pectolytic enzymes were studied in witloof chicory. Disease incidence after forcing of the chicory heads was enhanced by the nitrogen treatment and reduced by the calcium treatment of the chicory plants prior to forcing. Significant differences in susceptibility to bacterial soft rot were found for the tested chicory cultivars. Disease severity after 96h ranged from 6% in Clause R2 to 100% in Bea and Tabor. Chicory cultivars Rumba and Salsa showed a final average severity of 65–70%. Activity of the pectolytic enzymes, polygalacturonase and pectate lyase increased in artificially inoculated chicory heads of cultivar Rumba and was not affected by calcium and nitrogen treatments of the plants. Polygalacturonase showed highest activity 48h after inoculation while pectate lyase activity increased throughout 72h of incubation. Maceration of the chicory tissue and bacterial growth increased continuously until 96h after inoculation, when more than 60% of the chicory heads was macerated by pectolytic enzymes of the bacteria. Enzyme activity of Erwinia carotovora subsp. carotovora grown on chicory cell wall extracts was influenced by nitrogen and calcium treatments of the chicory plants. The activity of polygalacturonase reached its highest level at 48h after incubation and pectate lyase activity peaked at 24h followed by a continuous decrease until 72h after inoculation.     

Fungal and plant gene expression during synchronized infection of tomato leaves by Botrytis cinerea

An inoculation procedure was developed to obtain efficient and synchronous infection on detached tomato leaves by Botrytis cinerea. In spray-inoculated leaves incubated at 20 °C, the infection process consisted of three phases: the formation of primary necrotic lesions (until 20 hpi), a quiescent phase (20-72 hpi), and the expansion of a proportion of the primary lesions (from 72 hpi onwards), resulting in full tissue maceration. At 4 °C, the infection progressed slowly but steadily without inducing necrotic responses in the host. The actin and -tubulin genes of B. cinerea were cloned, characterized and used as probes on blots containing RNAs from leaves at various stages of the infection. The genes displayed a similar expression pattern throughout the infection and the hybridization signal reflected the amount of fungal biomass. The actin mRNA accumulated to higher levels than the -tubulin mRNA. Tomato PR protein mRNAs (chitinase, -1,3-glucanase and PR-1) were induced during the infection, albeit with different kinetics and to different levels. At 20 °C, -1,3-glucanase and PR-1 mRNAs were induced more rapidly than chitinase mRNAs. At 4 °C, mRNAs encoding extracellular -1,3-glucanase and intracellular, as well as extracellular chitinase were hardly induced.