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.     

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.     

Biochemical properties of the pathogenesis-related proteins from tobacco

This review describes the discovery and identification of the pathogenesis-related proteins (PRs) from tobacco. In crude leaf extracts the PRs are distinguished from the proteins in uninfected plants by their solubility at pH 3, resistance to a range of proteases, and mobility in polyacrylamide gels upon electrophoresis (PAGE) in non-denaturing conditions. PAGE has been used as a qualitative and semi-quantitative assay for PRs, and their migration in gels made from different acrylamide concentrations has been used to identify charge and size isomers and electrophoretically identical PRs in different tobacco cultivars. The subunit composition and molecular weight (mol. wt) of the four PRs identified first in Xanthi-nc were determined by SDS-PAGE; staining the gels has shown that these same four proteins in Samsun NN did not contain carbohydrate, lipid or nucleic acid, nor were they isozymic forms of twenty five enzymes known to increase in activity following infection with TMV. Evidence suggests that most of the PRs in Xanthi-nc and Samsun NN are extracellular.The purification of several PRs from Xanthi-nc, Samsun NN and other tobaccos is described, as well as their mol. wt, subunit and amino acid composition. PRs 1a, b and c consist of a single polypeptide and have similar mol. wt and amino acid compositions. Antisera prepared against purified Xanthi-nc b₁ protein have been used to determine serological relationships between PRs and form the basis of a very sensitive quantitative assay using ELISA. The regulation of synthesis of some PRs has been shown to involve translational control.     

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).     

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.     

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.     

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.     

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.     

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     

The ash wilt disease: A preliminary investigation of wood anatomy

A macroscopical, microscopical and SEM examination of wood samples, taken from diseased ash trees, was undertaken to probe into the cause of an ash wilt disease considered to be identical with dieback. The symptoms, particularly the obstruction of the vessels by tyloses and the occurrence of fungal hyphae in one specimen, bear a strong resemblance to those found in Dutch elm disease.Samenvatting Houtmonsters van zieke essen werden macroscopisch, microscopisch en met de SEM onderzocht om de oorzaak van de esseverwelkingsziekte op te sporen. Deze ziekte, ook wel aangeduid met essensterven, vertoont grote overeenkomsten met de in de Verenigde Staten van Amerika en in Groot-Brittanië al langer bekende ash dieback. De gevonden verschijnselen, in het bijzonder de verstopping van vaten door thyllen en het vóórkomen van schimmeldraden in een van de monsters, vertonen veel overeenkomsten met karakteristieke symptomen van de iepeziekte en van de eikeverwelking.     

The relationship of leaf wetness duration and disease progress of glume blotch,caused byStagonospora nodorum,in winter wheat to standard weather data

Almost 50% of the variation in leaf wetness duration can be explained by maximum and minimum temperatures, rainfall and hours with relative humidity above 90% on a daily basis. All of these parameters can be estimated from a standard weather station. If variables related to wind are added the level of explanation increases to 69–76%. Leaf wetness duration explained up to 42% of the rate of disease increase (RDI) forS. nodorum. Leaf wetness duration was accumulated over a 5-day window period and correlated with rate of disease increase after a 7-day lag period. Standard weather variables could explain 20–34% of the disease increase. The relevance of these statistical models to disease prediction is discussed.     

Colonization and histopathology of susceptible and resistant carnation cultivars infected with Fusarium oxysporum f. sp. dianthi

Stems of the susceptible Early Sam and resistant Novada carnations were inoculated with a conidial suspension ofFusarium oxysporum f. sp.dianthi. Stem segments of either cultivar were sampled regularly and used for determination of fungal growth and for microscopical investigation.Early Sam showed typicalFusarium wilt symptoms and its stems were colonized intensively. The observed vascular browning appeared to be caused by discolouration of primary walls of infected vessels and surrounding cells. Vessels were rarely occluded with gel. Cell wall degradation led to the formation of stem cavities. Hyperplasia of xylem parenchyma was not seen.In Novada, fungal colonization remained low throughout the experiment. Macroscopic symptoms were absent except for longitudinal bursts in the stem, which appeared to be caused by hyperplasia of xylem parenchyma bordering infection. Vascular gelation occurred in the infected tissues, causing some vascular browning also. Xylem vessel regeneration was observed in the hyperplastic layer. Cavities were not formed, and wall discolouration was rare. Vascular gelation is considered part of theFusarium wilt resistance mechanism. It is followed by xylem vessel regeneration, which expresses a general plant response to vascular dysfunction rather than being part of the resistance mechanism.Although of different origin, vascular browning as such occurs in both susceptible and resistant interactions. In breeding for resistance, care should hence be taken with the current use of browning as an indication of disease.Samenvatting Anjers van de vatbare cultivar Early Sam en de resistente cultivar Novada werden geïnoculeerd met een conidiënsuspensie vanFusarium oxysporum f. sp.dianthi. Van beide cultivars werden regelmatig stengeldelen geoogst om deze microscopisch te onderzoeken en om de schimmelgroei te bepalen.Early Sam vertoonde de voor deze verwelkingsziekte kenmerkende symptomen en werd intensief gekoloniseerd. Aan het vaatweefsel waargenomen bruinkleuring bleek veroorzaakt te worden door verkleuring van de primaire wanden van geïnfecteerde vaten en de hen omringende cellen. Zelden trad er in de vaten gomvorming op. Celwandafbraak veroorzaakte de vorming van holten in de stengel. Hyperplasie van het houtparenchym werd niet waargenomen.In Novada bleef de schimmelgroei gedurende het hele experiment beperkt. Macroscopisch waren er enkel lengtescheuren in de stengel te zien, die veroorzaakt bleken te worden door hyperplasie van aan de infectie grenzend houtparenchym. In het geïnfecteerde vaatweefsel optredende gomvorming veroorzaakte ook enige bruinkleuring. In het hyperplastische weefsel werd regeneratie van houtvaten waargenomen. In de stengel werden geen holten gevormd, en verkleuring van de celwanden kwam weinig voor. De vorming van gommen in de houtvaten maakt waarschijnlijk deel uit van het resistentiemechanisme. De daarop volgende houtvatregeneratie is eerder een algemene reactie van de plant op vaatverstopping dan een deel van het resistentiemechanisme.Vaatverbruining, zij het van verschillende oorsprong, komt voor in zowel vatbare als resistente interacties. Om die reden moet men in de resistentieveredeling bij de anjer voorzichtig zijn met het gebruik van bruinkleuring als ziekteïndicatie.     

Race-specific aspects of polygenic resistance of barley to leaf rust,Puccinia hordei

Partial resistance of barley to leaf rust,Puccinia hordei, is characterized by a reduced rate of epidemic development in spite of a susceptible infection type. Barley cultivars vary greatly for partial resistance and its components. In a test for interaction between host cultivars and pathogen isolates most variation was of a horizontal nature. However, in the combination between Julia and the rust isolate 18, a differential interaction (vertical effect) occured; Julia had lost a small part of its partial resistance. The same interaction was found for latent period (LP), the most important component of partial resistance. Julia showed a shortened LP for isolate 18. Genetic analyses revealed, that Julia carries a polygene for longer LP not present in the other cultivars. The effect of this polygene appears to be broken by isolate 18 indicative for a gene-for-isolate relation, and even for a gene-forgene relation.Samenvatting Partiële resistantie van gerst voor dwergroest,Puccinia hordei, is gekenmerkt door een vertraagde epidemie-opbouw ondanks een vatbaar infectietype. Gerstrassen variëren sterk in partiële resistentie en haar componenten. In een toets ter bestudering van de interactie tussen waardplant en pathogeenisolaat bleek de meeste variatie horizontaal van aard te zijn. In de combinatie van Julia met isolaat 18 werd echter een differentiële interactie waargenomen; Julia had een klein deel van zijn partiële resistentie verloren. Dezelfde interactie werd waargenomen voor de belangrijkste component van partiële resistentie, de latentieperiode (LP); Julia had een iets verkorte LP voor isolaat 18. Genetische analyses toonden aan, dat Julia een polygen voor langere LP bevat; dit gen is niet aanwezig in de andere bestudeerde rassen. Er wordt verondersteld dat het effect van dit polygen door isolaat 18 is doorbroken. Dit wijst op een gen-om-isolaat en zelfs op een gen-om-gen-relatie.     

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.     

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.     

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.     

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.     

Comparison of the effects of salicylic acid and ethephon with virus-induced hypersensitivity and acquired resistance in tobacco

The induction of a hypersensitive reaction in Samsun NN tobacco by tobacco mosaic virus (TMV) at 20°C leads to the development of both localized and systemic acquired resistance, and is associated with the appearance of pathogenesis-related proteins (PR's) and large increases in peroxidase activity and ethylene production. Salicylic acid (SA) induced a similar resistance in treated plant parts and occasionally also in untreated upper leaves of plants of which three lower leaves had been injected. SA also induced the same four PR's, but these were confined to the treated leaves. Thus, the connection between the presence of PR's and the reduction of TMV multiplication and spread may not be direct.In contrast to TMV, SA did not stimulate ethylene production and hardly increased peroxidase activity. Induction of acquired resistance and PR's by SA developed equally well at 20°C and at 32°C. However, pricking leaves with needles moistened with the ethylene-releasing compound ethephon mimicked TMV infection in inducing acquired resistance and PR's in both treated and untreated leaves at 20°C, but not at 32°C. Ethephon increased peroxidase activity at both temperatures, but only at 20°C dit it induce changes in both the anodic and the cathodic isoenzymes that were similar to those induced as a result of TMV infection. SA induced PR's and reduced TMV multiplication in Samsun tobacco, and inhibited virus spread in Samsun NN at 32°C.These observation indicate that neither the induction of PR's, nor the development of acquired resistance is temperature-sensitive. On the other hand, the effects of ethephon are temperature-sensitive in the same way as the hypersensitive response to TMV. It can thus be hypothesized that ethylene, produced naturally during the hypersensitive reaction of tobacco to TMV, leads to the temperature-sensitive synthesis or release of a presumably benzoic acid-type compound that functions as the natural inducer of PR's and acquired resistance. Although vanillic acid has been shown to accumulate in hypersensitively reacting tobacco leaves, it produced none of the effects of SA, and thus cannot be the natural inducer.Samenvatting Inductie van een overgevoeligheidsreactie in Samsun NN-tabak door tabaksmozaïekvirus (TMV) bij 20°C leidt tot de ontwikkeling van een verworven resistentie die zowel lokaal als systemisch werkzaam is, en gaat samen met het verschijnen van pathogenesis-related proteins (PR's) en sterke toename in de activitieit van peroxidase en de produktie van ethyleen. Salicylzuur (SA) induceerde een vergelijkbare resistentie in behandelde plantedelen en af en toe ook in niet behandelde bovenbladeren van planten waarvan drie onderbladeren waren ingespoten. SA induceerde ook dezelfde vier PR's, maar deze waren beperkt tot de behandelde bladeren. Er bestaat dus geen directe samenhang tussen de aanwezigheid van PR's en de remming van de vermeerdering en uitbreiding van TMV in de plant.In tegenstelling tot TMV stimuleerde SA de ethyleenproduktie niet en verhoogde het de peroxidaseactiviteit nauwelijks. Inductie van verworven resistentie en PR's door SA trad even goed op bij 32°C als bij 20°C. Net als infectie met TMV leidde aanprikken van bladeren met naalden die gedoopt waren in een oplossing van ethefon — waaruit in het blad ethyleen vrijkomt — echter tot inductie van verworven resistentie en PR's in zowel behandelde als onbehandelde bladeren bij 20°C, maar niet bij 32°C. Ethefon verhoogde de peroxidaseactiviteit bij beide temperaturen, maar alleen bij 20°C induceerde het veranderingen in zowel de anodische als de kathodische isoënzymen die vergelijkbaar waren met die welke geïnduceerd werden als gevolg van infectie met TMV. SA induceerde PR's en verminderde de vermenigvuldiging van TMV in Samsun tabak, en remde de uitbreiding van het virus in Samsun NN bij 32°C.Deze waarnemingen tonen dat noch de inductie van PR's, noch de ontwikkeling van verworven resistentie een temperatuurgevoelig proces is. Daarentegen zijn de effecten van ethefon op dezelfde wijze temperatuurgevoelig als de overgevoeligheidsreactie op TMV. Men kan daarom veronderstellen dat ethyleen, dat op natuurlijke wijze geproduceerd wordt tijdens de overgevoeligheidsreactie van tabak op TMV, aanleiding geeft tot een temperatuurgevoelig proces, namelijk de synthese of het vrijkomen van een verbinding, vermoedelijk een benzoëzuurderivaat, dat fungeert als de natuurlijke inductor van PR's en verworven resistentie. Hoewel is aangetoond dat vanillinezuur zich ophoopt in overgevoelig reagerende tabaksbladeren, veroorzaakte deze verbinding geen enkel van de effecten van SA. Vanillinezuur kan dus niet de natuurlijke inductor zijn.     

Some coat protein constituents from strawberry latent ringspot virus expressed in transgenic tobacco protect plants against systematic invasion following root inoculation by nematode vectors

The coding sequences in RNA2 for the coat proteins (CP) of strawberry latent ringspot virus (SLRSV) were modified and amplified using polymerase chain amplification reactions (PCR) to facilitate their expression inAgrobacterium tumefaciens-transformedNicotiana tabacum Xanthi-nc. The coding sequences for the smaller capsid protein (S, 29kDa) and that for the theoretical precursor of L and S (P, 73kDa) had ATG initiation codon sequences added at the 5-proximal Ser/Gly (S/G) cleavage site in the unmodified sequence. The sequence coding for the larger of the two proteins of mature SLRSV capsids (L, 44kDa) had an ATG codon added at its 5 S/G site and a TAG stop codon sequence added at the 3-proximal S/G site. The P, L and S proteins were expressedin planta to a maximum concentration of 0.01 % of total extractable proteins but did not assemble into virus-like particles. When challenged by mechanical inoculation with virus particles or viral RNA, and compared with control plants, tobacco plants (primary transgenic clones or S1 and S2, kanamycin-resistant seedlings) expressing the virus capsid subunits separately, or their precursor, decreased the accumulation of SLRSV particles in inoculated leaves and fewer plants became invaded systemically. In experiments in which the roots of seedlings were exposed to SLRSV-carrying vector nematodes (Xiphinema diversicaudatum), SLRSV was detected in the roots of non-transformed control tobacco plants (6/20) and in transgenic tobacco expressing the L protein (7/40), but not in any of 25 tobacco plants expressing the S protein or in 35 expressing the P protein. This is the second example of CP-mediated resistance to virus inoculation by nematode vectors.     

Scytalidium Wilt of Citrus

In July 1998, a sudden wilt of Star Ruby grapefruit (Citrus paradisi Macf.) occurred in Israel in a region with a warm Mediterranean climate. The wilt of the middle and upper canopy of main limbs was accompanied by gum oozing from the affected branches. The bark of these branches attained a dark colour and the epidermis sloughed off easily revealing a mass of black powder, resulting from copious sporulation of dark conidia. Both the bark and the xylem were intensively colonized with mycelium. The fungus was identified as Scytalidium lignicola Pesante, based on the characteristic mixture of some colourless and dark conidia, produced in branched chains by conversion of the vegetative hyphae. Artificial inoculations induced typical disease symptoms and the fungus was reisolated, thus confirming Koch's postulates. Similar symptoms appeared in Star Ruby interim segments which had been cut and top grafted with various citrus cultivars. This is the first detailed report of pathogenicity of S. lignicola to citrus trees. It is postulated that the disease developed after predisposition of Star Ruby grapefruit trees by comprehensive pruning followed by extremely hot conditions.