Verticillium wilt of sainfoin

A wilt disease of sainfoin caused by Verticillium Dahliae Kleb. is described, and it is shown that the fungus can penetrate sainfoin seedlings through unwounded roots as well as through ruptures caused by the emergence of lateral rootlets. V. Dahliae was isolated from naturally infected soil only in June, July and August, although another species, V. nigrescens , was obtained throughout the year.
Comparative studies of the longevity of cultures of V. Dahliae, V: albo-atrum and V. nigrescens proved that all are viable for at least 3 years on agar media. On sterilized wheat grains V. Dahliae dies within 8 weeks after inoculation, V. albo-atrum and V. nigrescens within 12 weeks, while the hyaline variants of the first two remain viable for 6 months.
Evidence was obtained that in artificially inoculated soil V. Dahliae persists mainly as microsclerotia. The fungus may also exist in the soil as hyaline mycelium or conidia, but only for a relatively short time.
The incidence of this disease in sainfoin is reduced by an increase in soil-water content, but is unaffected by the application of lime to the soil.     

Identification des acides caféoylshikimiques des racines du palmier dattier, principaux composés fongitoxiques vis-à-vis de Fusarium oxysporum f. sp. albedinis

5-caffeoylshikimic acid and its isomeres of position (dactylifric acid) found in the roots of date-palms ( Phoenix daetylijera L.) inhibit the germination of conidia and the growth of the mycelium of Fusarium oxysporum f. sp. albedinis . The acid content accumulated in resistant cultivars are toxic to fungi, whereas that accumulated in sensitive cultivars does not seem to affect growth and development of the parasite. This well demonstrates the implication of caffeoylshikimic acid in the resistance of the date-palm to this fusarium disease. Moreover, these results will be of great use, especially for screening resistant genotypes obtained from directed crossings or from in vitro cultures.     

Effects of Leaf Wetness Duration and Inoculum Level on Resistance of Wheat Genotypes to Pyrenophora tritici-repentis

Percent leaf necrosis and lesion length on wheat genotypes increased markedly with increasing duration of leaf wetness (up to 24h or 48 h) following inoculation with Pyrenophora tritici-repentis. A long wetting duration favoured less disease development on resistant (Fink's'), and moderately resistant (Bon/YR/3/F3570/KAL/BB) genotypes than on susceptible Glenlea. No significant difference in percent necrosis was detected among the upper three leaf positions within a genotype. A long wetnessduration had a varying effect on the resistance of wheat genotypes, depending upon the inoculum level. Increasing the inoculum level along with the leaf wetness period increased the per cent leaf necrosis on all three wheat genotypes tested. However, the, ranking of the genotype for resistance did not alter even after prolonged duration of leaf wetness (up to 96 h) and/or high inoculum level (12000 conidia/ml water). Various post-inoculation wet-periods in combination with high conidia concentrations in inoculum should be used in identifying highly resistant germplasm in breeding populations at the seedling stage of the wheats.     

Effects of Leaf Wetness Duration and Inoculum Level on Resistance of Wheat Genotypes to Pyrenophora tritici-repentis

Percent leaf necrosis and lesion length on wheat genotypes increased markedly with increasing duration of leaf wetness (up to 24h or 48 h) following inoculation with Pyrenophora tritici-repentis. A long wetting duration favoured less disease development on resistant (Fink's'), and moderately resistant (Bon/YR/3/F3570//KAL/BB) genotypes than on susceptible Glenlea. No significant difference in per cent necrosis was detected among the upper three leaf positions within a genotype. A long wetness duration had a varying effect on the resistance of wheat genotypes, depending upon the inoculum level. Increasing the inoculum level along with the leaf wetness period increased the per cent leaf necrosis on all three wheat genotypes tested. However, the ranking of the genotype for resistance did not alter even after prolonged duration of leaf wetness (up to 96 h) and/or high inoculum level (12000 conidia/ml water). Various post-inoculation wet-periods in combination with high conidia concentrations in inoculum should be used in identifying highly resistant germplasm in breeding populations at the seedling stage of the wheats.     

Differential Activation of Expression of a Suberization-Associated Anionic Peroxidase Gene in Near-Isogenic Resistant and Susceptible Tomato Lines by Elicitors of Verticillium albo-atratrum

We tested whether the expression of the suberization-associated anionic peroxidase gene is involved in the timely appearance of the vascular suberized coating involved in the resistance of a tomato line to Verticillium albo-atrum. The mRNA for this peroxidase appeared at a higher level one day earlier in wound-healing fruits of the resistant tomato line than in a near-isogenic susceptible line. Cell cultures from the resistant line, when treated with low levels (nanograms per milliliter) of fungal elicitor, generated the peroxidase mRNA and this apparent activation of the peroxidase gene expression could be detected in minutes, whereas the cells from the susceptible line hardly responded.     

An analysis of responses of resistant and of susceptible tomato plants to Verticillium infection

Comparative studies were made of the responses of resistant and of susceptible Gem tomato plants to infection by Verticillium albo-atrum. When inoculated through roots, there were striking differences in their responses. In susceptible plants, the foliar symptoms and amount of mycelium in the stem increased rapidly for some time. Then the mycelium started to disappear from the stem; this was accompanied by a check to the normal progress of symptoms, and by the formation of tyloses. In resistant plants, a limited invasion of the root and lower stem was accompanied by rapid and extensive tylosis. The mycelium soon disappeared from the stem and the plant then recovered from the initially mild symptoms. There was an inverse relationship between the amount of mycelium and the extent of tylosis in infected plants. The growth of susceptible plants was markedly reduced by infection. Total leaf area was much less because the newly produced leaves did not expand normally. The root system in infected plants was smaller because there were many fewer tertiary roots. In resistant plants infection stimulated growth. Tomato cuttings inoculated with conidia reacted similarly to root-inoculated plants. Hyphae grew well in the vascular system of susceptible cuttings whereas in resistant cuttings the pathogen started to grow but soon disappeared. Detached leaves of susceptible plants, inoculated through cut ends, wilted more than did leaves from resistant plants. It is suggested that resistance is mainly of the active type that develops after infection.     

WILT OF LUCERNE CAUSED BY SPECIES OF VERTICILLIUM

A wilt disease of lucerne caused by species of Verticillium is described: at twenty-eight disease areas in England and Wales the pathogen was V. albo-atrum , while at one site it was V. dahliae . Both pathogens form superficial conidia on the basal regions of infected stems. It is shown that V. albo-atrum is introduced into a new area in contaminated plant material harvested with the seed from an infected crop. Rapid secondary spread of the disease follows the dissemination of spores from infected stems, and by contact of these and transported fragments of diseased tissues with the wounded surfaces of recently cut lucerne plants. It is recommended that seed should be collected only from healthy crops and that machinery, footwear, etc. should be disinfected before leaving a site of infection.
Manurial trials showed that the incidence of wilt induced by V. albo-atrum was very severe under all soil conditions tested, whereas V. dahliae is a virulent pathogen only to plants in soil rich in superphosphate. Generally the more vigorous the growth of the lucerne–in soils rich in potash and hoof and horn–the more rapid is both the onset of wilt and the resulting secondary spread of the disease throughout the crop.     

Reduced Anthracnose Stalk Rot in Resistant Maize is Associated with Restricted Development of Colletotrichum graminicola in Pith Tissues

Resistance to anthracnose stalk rot (ASR) in maize was investigated for its effects on the development of Colletotrichum graminicola. ASR and fungal presence in pith tissues of resistant and susceptible genotypes, inoculated at time intervals after wounding in the first internodes, were assessed by rating tissue discoloration and by quantifying ergosterol production using high performance liquid chromatography (HPLC) and fungal recovery from tissues, respectively. Slices (30 μm thick) of pith cores (2 mm diam) of first internodes at late‐whorl and kernel blister stages were also inoculated with a suspension of fungal conidia immediately, 2 or 6 h after slicing. Fungal development was observed in tissues by light microscopy. ASR was markedly reduced in resistant genotypes when compared to susceptible genotypes and when inoculation was delayed after stalk wounding. Ergosterol content in tissues was associated with extent of discoloration due to ASR and fungal recovery. Conidial germination, germ tube elongation, appressorium formation and penetration of cortical cells were all markedly delayed in resistant genotypes, in both resistant and susceptible maize at vegetative stages, and by wound healing. C. graminicola macerated more rapidly and to a greater extent pith tissues of susceptible than resistant genotypes. Resistance mediated by maize genotype and ontogeny, and wound healing is expressed at early stages and subsequent events of host–pathogen interaction. Fungal structural development in detached pith tissues and the rapidity and extent of pith maceration in susceptible when compared to resistant genotypes was indicative of genotypic reaction to ASR in maize in the field. Laboratory inoculation and observation of detached pith tissues could be a useful and accurate tool for rapid screening of maize germplasm to identify ASR resistant genotypes that will function well in the field even where pathogen ingress occurs via wounds.     

Kinetics and Histopathology of the Cacao-Ceratocystis cacaofunesta Interaction

Theobroma cacao L., the chocolate tree, is an important tropical tree-crop that provides sustainable economic and environmental benefits to some of the poorest and most ecologically sensitive areas of the world. Ceratocystis wilt (CW), caused by the fungus Ceratocystis cacaofunesta, is a highly dangerous disease capable of killing the cacao plant. Histopathological studies of the host–pathogen interaction are an effective resource to study pathogenesis which will ultimately lead to a better elucidation of the mechanisms of host resistance. We analyzed the development of C. cacaofunesta reproductive structures, the pattern of fungal colonization and the anatomical responses of cacao resistant (TSH 1188) and susceptible (CCN 51) genotypes to CW. Results showed that conidia germination started within 4 h after inoculation (hai) in agar, by 12 hai conidia germination was 80 %. The resistant genotype showed little or no fungal colonization compared to the susceptible genotype. Host reactions were characterized by greater quantities of gels, gums and tyloses in the susceptible genotypes compared to the resistant. The susceptible genotype had a large number of cells surrounding the xylem vessels that were infused with dark brown phenolic compounds. The host defense reaction observed near the xylem in the resistant genotype, suggest that a mechanism involved with resistance to C. cacaofunesta.     

Enzyme activities in cell suspension cultures of two hop cultivars after elicitation by a fungal culture filtrate

The activities of some key enzymes of the secondary metabolism in cell suspension cultures of two Humulus lupulus cultivars, namely a Verticillium albo-atrum susceptible (Hallertauer Mittelfrüh) and a resistant cultivar (Wye Target), were measured under normal growth conditions and after the addition of a culture filtrate of the fungus V. albo-atrum as elicitor. A marked change was observed upon elicitation in the phenylalanine ammonia lyase activity found for Wye Target, being almost 2-3 times higher than the control during the first hours after elicitation.     

Effect of elicitation on the peroxidase activity in some cell suspension cultures of hop,t Humulus lupulus

The effect of elicitation on peroxidase activity in cell suspension cultures of one variety and four cultivars of the hop plant, Humulus lupulus L., was studied. The peroxidase activity was measured using guaiacol (for all the cultures), ascorbate (for cultures of 'Olympic') and hop α-acids (for cultures of 'Olympic' and 'Wye Target') as substrates. Elicitation of the cell suspension cultures by preparations of the fungus t Verticillium albo-atrum (filtrate and homogenate) resulted in some cases in an increase of the peroxidase activity. The spectrum of activity towards the three substrates was changed by the elicitation. Since different extraction buffers resulted in different profiles of activity for the substrates, some enzyme specificity is supposed to be present. This revised version was published online in June 2006 with corrections to the Cover Date.     

Adhesion of the Entomopathogenic Fungus Beauveria (Cordyceps) bassiana to Substrata

The entomopathogenic fungus Beauveria bassiana produces at least three distinct single-cell propagules, aerial conidia, vegetative cells termed blastospores, and submerged conidia, which can be isolated from agar plates, from rich broth liquid cultures, and under nutrient limitation conditions in submerged cultures, respectively. Fluorescently labeled fungal cells were used to quantify the kinetics of adhesion of these cell types to surfaces having various hydrophobic or hydrophilic properties. Aerial conidia adhered poorly to weakly polar surfaces and rapidly to both hydrophobic and hydrophilic surfaces but could be readily washed off the latter surfaces. In contrast, blastospores bound poorly to hydrophobic surfaces, forming small aggregates, bound rapidly to hydrophilic surfaces, and required a longer incubation time to bind to weakly polar surfaces than to hydrophilic surfaces. Submerged conidia displayed the broadest binding specificity, adhering to hydrophobic, weakly polar, and hydrophilic surfaces. The adhesion of the B. bassiana cell types also differed in sensitivity to glycosidase and protease treatments, pH, and addition of various carbohydrate competitors and detergents. The outer cell wall layer of aerial conidia contained sodium dodecyl sulfate-insoluble, trifluoroacetic acid-soluble proteins (presumably hydrophobins) that were not present on either blastospores or submerged conidia. The variations in the cell surface properties leading to the different adhesion qualities of B. bassiana aerial conidia, blastospores, and submerged conidia could lead to rational design decisions for improving the efficacy and possibly the specificity of entomopathogenic fungi for host targets.     

In vitro selection for Fusarium wilt resistance in Gladiolus

Cormels pieces of four Fusarium susceptible Gladiolus cultivara (Friendship, Peter Pears, Victor Borge and Novalux) formed friable calli when cultured in vitro on Murashige and Skoog basal medium containing various concentrations of auxin and cytokinin. The friable calli established cell suspensions. Plantlet regeneration was obtained from the control callus, control cell suspension derived callus and in vitro selected Fusarium oxysporum Schlecht. resistant cell-lines of Friendship. The in vitro cormlets showed 85-95% germination after breaking dormancy of 8weeks at 4℃. Cell suspensions of all four Gladiolus cultivara were found to be highly sensitive to fusaric acid. Gradual Increase in fusaric acid concentrations to the cell-suspension cultures decreased cell growth considerably. One albino plant was found from the second generation of the In vitro selected cell line of Friendship. The albino plant was found to be highly susceptible to F. oxysporum. The cormlets of all in vitro selected call lines of Friendship were inoculated with a conidial suspension of the F. oxysporum before planting and were also sprayed with the same spore suspension for further characterization when the height of plants was about 6cm. The four selected cell lines showed the same response whether or not they were Inoculated with conidia of the F. oxysporum. Plantlets of all of the selected call lines exhibited significant growth as compared with the control after application of conidia of the F.oxysporum.     

Common Bean Reaction to Fusarium oxysporum f. sp. Phaseoli, the Cause of Severe Vascular Wilt in Central Africa

During the September‐December season of 1990, severe symptoms of Fusarium wilt were for the first time observed on a popular climbing bean (Phaseolus vulgaris L.) cultivar. G 2333. introduced within the previous 5 years. Seventy‐three bean genotypes were screened for resistance lo the disease, using artificial inoculation. The effect of inoculation density on the reaction of four selected genotypes was also investigated. Of the 29 climbing bean genotypes evaluated, 19 were resistant, including 11 of the 15 pre‐release or released cultivars. Of the 44 bush bean cultivars evaluated, 28 were resistant, five were intermediate and 11 were susceptible. All susceptible cultivars showed vascular discoloration. In both susceptible and resistant genotypes, the fungus spread almost equally from the entry points in inoculated roots to the base of the plants, but colonization and vertical spread within the vascular system were markedly less in resistant than in susceptible cultivars. At 20 and 30 cm above soil level, the fungus was only recovered from susceptible cultivars. Increasing inoculum density from 10² to 10⁷ conidia/ml did not affect the resistance of cultivars RWR 950 and G 685 but. in the susceptible cultivars G 2333 and MLB‐48‐89 A. it resulted in early appearance, high incidence and severity of the disease.     

Adhesion of the entomopathogenic fungus Beauveria (Cordyceps) bassiana to substrata

The entomopathogenic fungus Beauveria bassiana produces at least three distinct single-cell propagules, aerial conidia, vegetative cells termed blastospores, and submerged conidia, which can be isolated from agar plates, from rich broth liquid cultures, and under nutrient limitation conditions in submerged cultures, respectively. Fluorescently labeled fungal cells were used to quantify the kinetics of adhesion of these cell types to surfaces having various hydrophobic or hydrophilic properties. Aerial conidia adhered poorly to weakly polar surfaces and rapidly to both hydrophobic and hydrophilic surfaces but could be readily washed off the latter surfaces. In contrast, blastospores bound poorly to hydrophobic surfaces, forming small aggregates, bound rapidly to hydrophilic surfaces, and required a longer incubation time to bind to weakly polar surfaces than to hydrophilic surfaces. Submerged conidia displayed the broadest binding specificity, adhering to hydrophobic, weakly polar, and hydrophilic surfaces. The adhesion of the B. bassiana cell types also differed in sensitivity to glycosidase and protease treatments, pH, and addition of various carbohydrate competitors and detergents. The outer cell wall layer of aerial conidia contained sodium dodecyl sulfate-insoluble, trifluoroacetic acid-soluble proteins (presumably hydrophobins) that were not present on either blastospores or submerged conidia. The variations in the cell surface properties leading to the different adhesion qualities of B. bassiana aerial conidia, blastospores, and submerged conidia could lead to rational design decisions for improving the efficacy and possibly the specificity of entomopathogenic fungi for host targets.     

Developmental gene regulation in Aspergillus nidulans

The Ascomycete fungus Aspergillus nidulans reproduces asexually by differentiating conidiophores and conidia. Gene regulation during asexual reproduction was investigated by comparing poly(A) RNA populations derived from somatic hyphae, conidiating cultures and purified conidia. Single-copy and complementary DNA hybridization experiments showed that vegetative cells contained 5600–6000 diverse, average-sized poly(A) RNA sequences distributed into three prevalence classes. cDNA hybridization experiments indicated that a significant proportion of the poly(A) RNA derived from either conidiating cultures or spores consisted of sequences absent from somatic hyphae. To assess accurately the degree to which the poly(A) RNA populations differed, cDNA preparations were isolated which were complementary to sequences present only in conidia or in conidiating cultures. Hybridization of these cDNAs with poly(A) RNA from conidiating cultures showed that approximately 18.5% of the poly(A) RNA mass comprised 1300 diverse sequences not present in somatic cells. Of these, about 300 were present only in conidia. The remainder were accumulated specifically during sporulation, but were absent from spores. Analogous experiments showed that the great majority of the poly(A) RNA sequences accumulated by vegetative hyphae were also present in conidiating cultures. Thus, cell differentiation during A. nidulans asexual reproduction involves the accumulation of many new poly(A) RNA sequences, but not the loss of preexisting ones.     

The protective role of pigments against UV rays in fungi isolated from the mesosphere

Five among six species of microorganisms isolated from the mesosphere contained pigments which made them more resistant to the action of UV as compared to pigmentless microorganisms in the atmosphere of Earth. UV irradiation in the atmosphere is supposed to select resistant pigmented forms, so that they predominate in the mesosphere. To confirm this assumption, mutants of Aspergillus niger, Penicillium notatum and Circinella muscae were sported by irradiating them four times and then subjecting to stepwise selection. These mutants either synthesized pigments at a very low rate or did not produce them at all. No significant differences were found by studying the biomass, mycelium and sporeforming organs of the parent cultures and their mutants. However, their resistance to UV was not the same. Addition of the pigment apsergillin, isolated from the conidia of Aspergillus niger, to a suspension of the pigmentless (mutant) conidia of Penicillium notatum, the spores of Circinella muscae, and the vegetative cells of Micrococcus albus, before their irradiation with UV, considerably increased their resistance to this factor.     

Microcycle conidiation and the conidial properties in the entomopathogenic fungus Metarhizium acridum on agar medium

Conidiation of the entomopathogenic fungus Metarhizium acridum on agar media was investigated. M. acridum CQMa102 exhibits two different conidiation patterns on agar media: normal conidiation in which conidia are formed on extended hyphae and microcycle conidiation in which conidiation occurs directly after conidia germination. Microcycle conidiation resulted in a mass of conidia produced via budding by accelerated development at the inoculation site. The mean total conidial yield (conidiation at day 10) was 4–5-fold greater after microcycle conidiation than during normal conidiation. Insect pathology assays indicated that microcycle conidia produced on SYA agar were as effective as normal aerial conidia against the locust. Ultraviolet (UV)-resistance tests showed no significant differences between the two types of cell propagules. However, microcycle conidia were more heat resistant than normal aerial conidia, and accumulated higher levels of trehalose in response to heat induction compared to normal aerial conidia.     

Glycosylphosphatidylinositol-anchored Ecm33p influences conidial cell wall biosynthesis in Aspergillus fumigatus

ECM33 encodes a glycosylphosphatidylinositol-anchored protein whose orthologs in yeast are essential for sporulation. Aspergillus fumigatus Ecm33p is unique and has an apparent mass of 55 kDa. Disruption of A. fumigatus ECM33 results in a mutant with several morphogenetic aberrations, including the following: (i) a defect in conidial separation, (ii) an increase in the diameter of the conidia of the mutant associated with an increase in the concentration of the cell wall chitin, (iii) conidia that were sensitive to the absence of aeration during long-term storage, and (iv) conidia that were more resistant to killing by phagocytes, whereas the mycelium was more easily killed by neutrophils.     

The Occurrence of Cell Wall Appositions in Flag Leaves of Spring Wheats, Susceptible and Partially Resistant to Wheat Leaf Rust

In two experiments, the presence of cell wall appositions in flag leaves of spring wheat genotypes susceptible and partially resistant to wheat leaf rust was studied. More cell wall appositions were observed near aborted infection structures than in estabhshed colonies. There was not a marked difference in the number of cell wall appositions per colony between susceptible and partially resistant genotypes. More cell wall appositions per unit area colony were present in partially resistant genotypes. It was concluded that the low number of cell wall appositions could not be responsible for the observed difference in colony size between susceptible and partially resistant genotypes. Partial resistance in wheat to wheat leaf rust can be divided into two phases. The first phase is pre-haustorial and results in a reduction of the number of colonizing infection structures. In the second phase a post-haustorial retardation of fungal growth rate occurs. The latter appears to be the more important phase.