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.     

Induced resistance in tomato plants against Fusarium wilt invoked by Fusarium oxysporum f.sp. dianthi

Tomato plants, susceptible toFusarium oxysporum f. sp.lycopersici, were inoculated by immersing the roots in a conidial suspension ofF. oxysporum f. sp.lycopersici race 1,F. oxysporum f. sp.dianthi race 2 or a mixture of both fungi. Plants inoculated withF. oxysporum f. sp.lycopersici showed disease symptoms after 2 weeks, whereas plants inoculated withF. oxysporum f. sp.dianthi or a mixture of both fungi remained symptomless for over 7 weeks, the duration of the experiment. In another experiment root systems of plants were split and each half was separately inoculated. One half was firstly inoculated withF. oxysporum f. sp.dianthi or treated with water, followed after a week by a second inoculation of the other half withF. oxysporum f. sp.lycopersici or by a water treatment. The disease symptoms in the half firstly inoculated withF. oxysporum f. sp.dianthi were significantly delayed, compared to plants of which that half had been treated with water. BecauseF. oxysporum f. sp.dianthi reduced disease symptoms caused byF. oxysporum f. sp.lycopersici without any direct interaction with this pathogen, it is concluded thatF. oxysporum f. sp.dianthi is able to induce resistance againstF. oxysporum f. sp.lycopersici in tomato plants.     

Further studies on the activity of cell wall degrading enzymes of aggressive and non-aggressive isolates of Ophiostoma ulmi

In vitro activity of glycosidases and exo-glycanases in 10 aggressive and 10 non-aggressive isolates of Ophiostoma ulmi were compared. Enzyme preparations of aggressive isolates released more arabinose and xylose from cell walls of elm wood than those of non-aggressive isolates. Significantly more rhamnose was released as well by enzyme preparations of aggressive isolates, although some of non-aggressive isolates released comparable amounts.     

Genetic Variation for Seed Yield in Perennial Ryegrass (Lolium perenne L.)

In perennial ryegrass seed yield is low and unreliable and little is known about the seed-yielding capacity of different cultivars. Therefore, genetic variation for seed yield was studied for therefore, genetic variation for seed yield was studied for three years at two locations. Twelve trials consisting of drilled plots of nine diploid, late-flowering cultivars in four replications were harvested for seed. Significant gentic differences for seed yield and seed weight weight were detected. The ranking of the cultivars for seed yield differed from that for seed weight. The levels of seed yield and seed weight were affected by such environmental factors as year, soil type and crop production year, but interactions of these factors with cultivars were generally not significant. Variation in seed yield was more related to variation in seed number than to variation in seed weight. There were no significant cultivar differences for yield stability. The highest yielding cultivar was superior over a wide range of environments and the seed yield of the poorest cultivar was on average only 64% of that of the best cultivar.     

Effects of white clover cultivar and companion grass on winter survival of seedlings in autumn-sown swards

The aim was to study the effects of white clover cultivar and combinations with perennial ryegrass cultivars on seedling establishment in autumn‐sown swards and on winter survival of seedlings. Large‐leaved white clover cv. Alice and small‐leaved white clover cv. Gwenda, and an erect and a prostrate perennial ryegrass cultivar were sown in autumn in pure stands and as four binary grass‐clover mixtures. Mixtures of white clover cv. Huia and Aberherald with perennial ryegrass were also sown. Companion grasses had no significant impact on the establishment of white clover. The number of seedlings of white clover cv. Alice in mixtures (335 m^?2) was higher than cv. Gwenda (183 m^?2) and pure swards had similar white clover population densities as mixed swards. White clover cv. Huia tended to have more seedlings than Aberherald (355 and 205 m^?2 respectively). No stolons were produced prior to a severe winter, because of the late sowing date. Winter survival of clover seedlings was 0·56 in mixtures and 0·69 in pure stands, irrespective of white clover or companion grass cultivar. Stolon development of white clover in autumn is often considered essential for overwintering survival and spring growth. In this study, there was considerable survival of the non‐stoloniferous tap‐rooted seedlings of all four clover cultivars despite a severe winter.     

Control of Dutch elm disease by the sterol biosynthesis inhibitors fenpropimorph and fenpropidin

Sterol biosynthesis inhibitors that inhibit the yeast-hyphae conversion inOphiostoma ulmi suppressed Dutch elm disease development in two elm clones. After curative treatment with fenpropimorph-sulphate of 27 Vegeta elms which had previously been inoculated withO. ulmi, 25 trees did not show disease symptoms by the end of the second season. All 41 control trees, inoculated withO. ulmi only, were clearly diseased. In an experiment with Commelin elms three fenpropimorph salts and thiabendazole were compared. Injection of the trees three weeks after inoculation withO. ulmi gave by the end of the second season no symptoms of Dutch elm disease in any of the trees injected with fenpropimorph-phosphate or thiabendazole, and in most trees injected with fenpropimorph-acetate or-sulphate. Similar treatments with the free base of fenpropimorph and fenpropidin-sulphate were less effective due to insufficient uptake of the fenpropimorph emulsion and phytotoxicity of fenpropidin-sulphate, respectively. Injection of fenpropimorph-sulphate or thiabendazole six weeks after inoculation withO. ulmi did not result in significant differences from the control group inoculated withO. ulmi only.Fenpropimorph-phosphate and-sulphate completely suppressed Dutch elm disease upon injection of only 7.5 or 10 g per tree (average tree diameter 28 cm). Residue analyses showed only a slow decrease in concentration of the fungicide over two growing seasons and an apparent transport into the new annual ring, other prerequisites for a possible future use for control of Dutch elm disease.Samenvatting De iepeziekte kan onderdrukt worden door sterolbiosyntheseremmers die de overgang vanOphiostoma ulmi van de gistvorm in de hyfevorm remmen. Aan het eind van het tweede seizoen na een curatieve behandeling van 27 Vegeta iepen met fenpropimorfsulfaat bleken 25 bomen geen symptomen van iepeziekte te vertonen. Alle controlebomen, die alleen metO. ulmi geïnoculeerd waren, waren duidelijk ziek. In een proef met Commelin iepen werden drie fenpropimorfzouten en thiabendazool vergeleken. De zouten werden drie weken na de inoculatie metO. ulmi geïnjecteerd. Aan het eind van het tweede seizoen vertoonden geen van de bomen die met fenpropimorffosfaat of thiabendazool geïnjecteerd waren en slechts enkele bomen die met fenpropimorfacetaat of-sulfaat geïnjecteerd waren iepeziektesymptomen. Behandelingen met fenpropimorf (vrije base) en fenpropidinsulfaat werkten minder goed door de slechte opname van de fenpropimorfemulsie en de fytotoxiciteit van fenpropidin. Injectie met fenpropimorfsulfaat of thiabendazool zes weken na inoculatie leidde niet tot significante verschillen met de controlegroep die alleen metO. ulmi geïnoculeerd was.Een dosis fenpropimorffosfaat of-sulfaat van 7.5 of 10 g per boom met een gemiddelde boomdiameter van 26 cm bleek de iepeziekte volledig te kunnen onderdrukken. Uit residue-onderzoek bleek dat de concentratie van het fungicide gedurende de twee groeiseizoenen slechts langzaam afnam en dat het middel naar de nieuwe jaarring werd getransporteerd, twee voorwaarden voor een toepassing op praktijkschaal van fenpropimorf voor de bestrijding van de iepeziekte.     

Induction of mutants of Fusarium oxysporum f. sp. lycopersici with altered virulence

Mutants ofFusarium oxysporum f. sp.lycopersici were obtained by UV irradiation. The mutants of race 1 and race 2 caused disease symptoms on plants with resistance genes against the corresponding wild type strains. Mutants of race 1 of the pathogen were stable, whereas mutants of race 2 lost the ability to cause disease symptoms in plants carrying the 1–2 resistance gene, after prolonged maintenenance on potato dextrose agar. Mutants of race 1 resembled race 2 in pathogenicity and they were vegetatively compatible with race 2, but no longer with race 1. These results suggest that the isolated strains with an altered virulence pattern have mutations in loci involved in avirulence.     

Pseudomonas for biological control of dutch elm disease. I. Labeling,detection and identification of pseudomonas isolates injected into elms; comparison of various methods

To understand the mechanisms involved in biological control of Dutch elm disease byPseudomonas, data were needed on the distribution of the introduced bacteria within elm and on the development of the bacterial population over a period of time.As traditional biochemical identification techniques are not suitable for distinguishment between individualPseudomonas isolates, three alternative approaches were compared.

Competition in perennial ryegrass–white clover mixtures under cutting. 1. Dry-matter yield, species composition and nitrogen fixation

Persistence of white clover (Trifolium repens) in mixtures was studied in a long-term experiment. Mixtures of two cultivars of perennial ryegrass (Lolium perenne) with contrasting growth habits and three white clover cultivars differing in leaf size were sown in 1991 and evaluated at two cutting frequencies. During 1995 and 1996 mixtures with large-leaved white clover cv. Alice had the highest dry-matter (DM) content, clover and N yield, and the highest white clover content, and mixtures with medium-leaved Retor the lowest, whereas mixtures with small-leaved Gwenda yielded most grass DM. In 1995 averaged over cutting treatments and mixtures, the mixtures yielded 11·8 t DM ha^?1 with a white clover content of 0·6; the apparent N fixation was 393 kg N ha^?1. In 1996 these values declined to 8·5 t DM ha^?1, 0·48 white clover and 236 kg N ha^?1. There was no significant effect of cutting frequency on DM yield or white clover content, whereas the effects of grass cultivar were not consistent. In spring there was a peak in the DM production of the mixtures, coinciding with a peak in production of the grass component. However, in summer and autumn the seasonal pattern of DM production of the mixtures was similar to that of the white clover component. Both cultivars of perennial ryegrass showed the same seasonal response, but the seasonal growth pattern of white clover differed slightly between clover cultivars and cutting treatments. In later years only one cutting frequency was imposed, and no yield measurements were taken. White clover was judged to have performed well during 1997; the clover content in September was very high (0·76), whereas in October 1998 it was 0·45. Mixtures with Alice contained most white clover. Despite fluctuations in white clover content during 1991–98, all clover cultivars had persisted 7 years after sowing, irrespective of companion grass cultivar, at both cutting treatments.     

Production in vitro of phytotoxic compounds by non-aggressive and aggressive isolates of Ophiostoma ulmi, the Dutch elm disease pathogen

Development in shake culture of two non-aggressive and two aggressive isolates of the Dutch elm disease fungus, Ophiostoma ulmi, was monitored in a time course study. The time sequences for the production of total biomass, spores, polysaccharides and phytotoxic compounds were followed, but none of them showed a clear correlation with aggressiveness. The same parameters were examined for nine non-aggressive and 11 aggressive isolates after 10 days growth. Of two phytotoxic compounds, a glycopeptide and the protein cerato-ulmin, only the latter was generally produced in larger quantities by aggressive isolates. However, variability in production and a number of exceptions to the trend made cerato-ulmin production an uncertain criterion for assessing the aggressiveness of an individual isolate.Immunofluorescence of sections of infected elms treated with an anti-glycopeptide serum showed intense fluorescence only in the cell walls of the fungus. Apparently the glycopeptide is abundantly present in or on fungal cell walls.The roles of the different compounds in relation to Dutch elm disease symptom development are discussed.