Traits associated with virulence to the aphid Macrosiphoniella sanborni in eighteen isolates of Verticillium lecanii

Eighteen isolates of the entomopathogenic fungus Verticillium lecanii from various world-wide locations, from insect hosts and soil were bioassayed against the aphid Macrosiphoniella sanborni in the laboratory. Virulence ranged from an isolate which achieved 100% mortality and LT 50 value (adults) of 3 days ± 0· 2 at 24 ° C, compared with the least virulent isolates causing less than 10% mortality over 14 days (when treated with an inoculum of 1 × 10⁶ conidiospores/ml). All isolates produced extracellular protease and lipase, irrespective of their virulence. A number of traits were frequently associated with the expression of virulence including fast germination, high sporulation rate, an absence of extracellular amylase activity and high extracellular chitinase activities. Large spore size was not strongly associated with virulence. There were exceptions in each variate studied, suggesting that overall expression of virulence is a result of the total complex of these and other traits still to be determined.     

Protein Changes in the Vascular Wilt Disease of Lucerne caused by Verticillium albo-atrum R. et B

Field trials confirmed that, whereas the Du Puits variety oflucerne was highly susceptible to Verticillium albo-atrum, ‘Gaetula’hybrids showed slight resistance and certain ‘Swedish’material was moderately resistant. Symptom-free ‘Gaetula’and Swedish plants were used to produce uninfected clonal materialby root propagation. Comparisons of the proteins of such plants with seed-grown DuPuits produced no evidence of a correlation between proteinpattern before inoculation and differences in susceptibilityand resistance. Disc electrophoretic comparisons of infected host material showedthat the pathogen gradually induced a change in the shoot proteinsof Du Puits whereas in the resistant host, after an initialmarked change, there was a development towards normality. Bythe terminal stages of wilting there was a noticeable fall inthe numbers of proteins in both roots and shoots of Du Puits,a finding confirmed both by double diffusion and immunoelectrophoresis.At the same time there was good evidence for a slight increasein protein bands in the resistant material after inoculation. After feeding tritium-labelled leucine to infected Du Puitsplants, a small protein component of fungal origin was detectedin the host. Labelled components from both healthy and infectedDu Puits plants were separated on Sephadex G.25 and compared.     

Factors affecting the resistance of cold-stored carrots to Botrytis cinerea

The secondary phloem parenchyma of cold-stored turgid roots of carrot (Daucus carota) is capable of localizing mycelial infection by Botrytis cinerea, producing a dark resistant lesion. The percentage of roots exhibiting this reaction declined with increasing time in cold-store: when freshly harvested and wound-inoculated in October 1974, 99%of roots resisted invasion, whereas only 5% of those stored until March 1975 did so. The surface dimensions of resistant lesions did not increase between 33 and 55 days after inoculation. However, the surface dimensions and mean weights of lesions (arising from inoculations performed at different times over the course of the storage season) were both larger with increasing time in storage of roots prior to inoculation. The ability of the root tissue to localize infection was reduced if the roots lost 5–10% or more of their fresh weight before inoculation, resulting progressively in susceptibility. Compared with roots wound-inoculated using mycelial disks, there was an overall reduction in infection when carrots were wound-inoculated using conidia or when conidia or mycelial disks were inoculated onto the apparently undamaged surface of roots.     

Germination and effect of reduced humidity on expression of pathogenicity in Verticillium lecanii against the glasshouse whitefly Trialeurodes vaporariorum

Fifteen isolates of the entomopathogenic fungus Verticillium lecanii, from various hosts and locations, were bioassayed in the laboratory at 19 ± 1°C and > 95% r.h. against fourth instar scales of Trialeurodes vaporariorum. Highly pathogenic isolates generally originated from whitefly; however, there was no correlation between conidiospore size or germination rate on agar, and pathogenicity. LT₅₀ values ranged from 5.7 to > 9 days and LC₅₀ (7 days) from 1.3 × 10⁵ to 4.2 × 10⁷ spores/ml. The effect of periods in low humidity (70% r.h.) following 16 or 96 h at > 95% r.h. after immersion of whitefly scales on tomato leaf discs in a suspension of 1 × 10⁶ spores/ml, was compared for five isolates. Progressively longer periods at 70% r.h. following 16 h at > 95% r.h., reduced significantly (P < 0.001) the pathogenicity of all isolates. In these conditions isolate A was least affected by low-humidity transfer. The high pathogenicity of isolate A was associated with its more rapid development on the host cuticle during the first 16 h in high humidity. When inoculated whitefly scales were transferred to 70% r.h. after a preliminary 96 h incubation in high humidity, all five isolates achieved a higher level of pathogenicity compared with their transfer to low humidity after 16 h high humidity. Thus the screening of V. lecanii isolates in limiting humidity conditions provided a more critical assessment of their pathogenicity and of their potential success in whitefly control relevant to the glasshouse environment.     

Incipient infections caused by Botrytis cinerea in carrots entering storage

A new marking method has been developed to follow the movements and dispersal of cabbage root flies during their most active period, 2–7 days after emergence, using one of thirty-four combinations of four head markings with fluorescent powders and six body markings with oil-soluble dyes. The method permits simultaneous replication of dispersal experiments and rapid detection of marked flies. It requires no expensive equipment and provides a permanent record of the results.     

Endogenous Elicitor Activity from Damaged Carrot Root Tissue and Induced Resistance to Botrytis cinerea

A peptide-containing fraction (mol. wt. c 5000 Da) extractedfrom carrot root tissues damaged by slicing or freeze-thawing,induced active defence mechanisms in carrot slices against Botrytiscinerea. Endogenous elicitor activity was present in homogenatesof fresh and freeze-thawed tissues and those treated with germinationfluid, but was absent in autoclaved tissue. The detection ofthe elicitor in homogenates within 2h of treatment, suggeststhat it is released or activated during the early stages ofcell damage. Botrytis cinerea, carrot, induced resistance, endogenous elicitor     

Competitive interaction between strains of Verticillium lecanii on two insect hosts

Differences were observed in the pathogenicities of two strains of Verticillium lecanii (strain numbers 1.72 and 19.79) to the whitefly Trialeurodes uaporariorum and the aphid Macrosiphoniella sanborni. The mean pathogenicity of a mixture of the two strains was intermediate to that of the individual strains, attributable to a competitive interaction during infection. Differences between strains were observed in the production of conidia in vitro and on host insect cadavers. When hosts were infected with the dual-strain suspension, conidia of strain 19.79 only were recovered from T. vaporariorum whereas conidia from both strains were recovered from M. sanborni.     

A rapid method to estimate the resistance of cold stored carrot roots to Botrytis cinerea

Inoculating whole carrot roots at 4°C with mycelial/agar discs of the grey mould fungus Botrytis cinerea gives a measure of their resistance and hence storage potential to this pathogen, but results are not obtained for at least 33 days. In the present investigation a more rapid method was used which involved inoculating root slices with spore suspensions containing 5 × 10^3–5 × 10⁶ spores/cm³ at 24°C. Resistance was assessed visually and by a chitin estimation 48 h after inoculation. Both methods were used to measure the resistance of cold stored carrot roots during two storage seasons, 1976/77 and 1977/78. In each season there was a particular inoculum level which most clearly recorded the increasing susceptibility of roots with time in store. In 1976/77 this was 1 × 10⁵ spores/cm³ whereas in 1977/78 it was the lower inoculum concentration of 5 × 10⁴ spores/cm³, suggesting the roots were generally more susceptible in 1977/78 than the previous season. This was in accord with the results from the whole root method of assessment. A chitin estimation proved to be the more sensitive method of assessment for inoculum potential experiments.     

The Induced Resistance Response of Carrot Root Slices to Heat-killed Conidia and Cell-free Germination Fluid of Botrytis cinerea Pers. ex Pers.: 2. Nuclear Migration, Nucleolar Volume and Uptake of Tritiated Uracil

Sixty-eight per cent of nuclei in the cells of the upper fourlayers of carrot slices treated with heat-killed conidia ofBotrytis cinerea for 6 h followed by inoculation with live sporesfor 18 h, migrated to the cell face nearest to the treated surface,compared with 46 per cent in cells of control slices showinga wound-healing response only. Nucleolar volumes in the surfacecell layers of control slices increased from a mean of 1.0 µm³to 3.8 µm³ over 24 h, and in ‘induced’ slicesto 7.28 µm³. Using a 40 min pulse of [5–³H]uracil,there was an increase within 15 h of slicing in the number oflabelled nuclei in cells from control slices undergoing healing.Within 8 h after treatment of slice surfaces with heat-killedconidia, there was an accelerated incorporation of label into‘nuclear’ RNA. Slices from roots cold-stored for12 months failed to show an induction response and nucleolarvolumes did not increase more than in control slices. Theseresults are discussed in relation to active defence mechanismsin plant tissue. Botrytis cinerea, carrot, induced resistance, nuclear migration, nucleolar volume, RNA incorporation