Infection dynamics of Fusarium mangiferae, causal agent of mango malformation disease.

Conditions affecting germination and growth of Fusarium mangiferae, causal agent of mango malformation disease, were studied in vitro. Both conidial germination and colony growth required temperatures >5 degrees C and reached a peak at 28 and 25 degrees C, respectively. A minimum 2-h wetness period was required for conidial germination, reaching a peak after 8 h of wetness. High incidence of fungal colonization in buds, predominantly the apical buds, was detected compared with inoculated leaves. The pathogen was detected in the roots of inoculated soil 19 weeks postinoculation but not in aboveground parts of the plants, and symptoms of the disease were not observed, either. Dry, malformed inflorescence debris serving as a source of inoculum caused significantly higher colonization (52 and 20%) of inoculated buds, compared with that (0%) of the untreated controls. Incidence of sampled leaf disks bearing propagules of F. mangiferae from an infected orchard peaked in June and July and decreased during the following months, whereas airborne infections on 1-month-old branches was the highest in May and June, corresponding with inoculum availability released from infected inflorescences. Colonization pattern, determined in naturally infected vegetative and woody branches, was significantly higher in node sections than in the internode sections. This study sheds light on infection dynamics, colonization patters, and the disease cycle of F. mangiferae in mango.

Interaction of the mite Aceria mangiferae with Fusarium mangiferae, the causal agent of mango malformation disease.

The role of the mango bud mite, Aceria mangiferae, in carrying conidia of Fusarium mangiferae, vectoring them into potential infection sites, and assisting fungal infection and dissemination was studied. Following the mite's exposure to a green fluorescent protein-marked isolate, conidia were observed clinging to the mite's body. Agar plugs bearing either bud mites or the pathogen were placed on leaves near the apical buds of potted mango plants. Conidia were found in bud bracts only when both mites and conidia were co-inoculated on the plant, demonstrating that the mite vectored the conidia into the apical bud. Potted mango plants were inoculated with conidia in the presence or absence of mites. Frequency and severity of infected buds were significantly higher in the presence of mites, revealing their significant role in the fungal infection process. Conidia and mite presence were monitored with traps in a diseased orchard over a 2-year period. No windborne bud mites bearing conidia were found; however, high numbers of windborne conidia were detected in the traps. These results suggest that A. mangiferae can carry and vector conidia between buds and assist in fungal penetration but does not play a role in the aerial dissemination of conidia between trees.

Inoculum availability and conidial dispersal patterns of Fusarium mangiferae, the causal agent of mango malformation disease.

Inoculum availability and conidial dispersal patterns of Fusarium mangiferae, causal agent of mango malformation disease, were studied during 2006 and 2007 in an experimental orchard. The spatial pattern of primary infections in a heavily infected commercial mango orchard corresponded with a typical dispersal pattern caused by airborne propagules. Malformed inflorescences were first observed in mid-March, gradually increased, reaching a peak in May, and declined to negligible levels in August. The sporulation capacity of the malformed inflorescences was evaluated during three consecutive months. Significantly higher numbers of conidia per gram of malformed inflorescence were detected in May and June than in April. Annual conidial dissemination patterns were evaluated by active and passive trapping of conidia. A peak in trapped airborne conidia was detected in May and June for both years. The daily pattern of conidial dispersal was not associated with a specifically discernable time of day, and an exponential correlation was determined between mean relative humidity (RH) and mean number of trapped conidia. Higher numbers of conidia were trapped when RH values were low (<55%). This is the first detailed report on airborne dispersal of F. mangiferae, serving as the primary means of inoculum spread.

A tale of three acaropathogenic fungi in Israel: Hirsutella, Meira and Acaromyces.

We review published and unpublished studies conducted in Israel with six acaropathogenic fungi, assayed in order to control the citrus rust mite, Phyllocoptruta oleivora (Ashmead) (CRM). Hirsutella thompsonii Fisher was introduced twice, killed 80-90% of the exposed mites, but due to its requirements for near-saturation humidities was deemed unsuitable for local outdoors conditions. Hirsutella kirchneri (Rostrup) Minter et al. and Hirsutella necatrix Minter et al. were also introduced and assayed against CRM and spider mites, but their efficacy was unsatisfactory. Three indigenous fungi found to be associated with mites, Meira geulakonigii, Meira argovae and Acaromyces ingoldii--all three recently described by Boekhout, Gerson, Scorzetti & Sztejnberg--were assayed against several mites. Meira geulakonigii killed 80-90% of several spider mites and of the CRM, and caused some mortality of Iphiseius degenerans (Berlese), one out of three phytoseiid predators assayed. Mortality was not due to parasitization; extracts from the media in which the fungi had developed caused considerable mite death, suggesting that it was a result of fungal toxins. Data from a field study indicated that spraying blastoconidia of M. geulakonigii on grapefruits infested by CRM significantly reduced pest-incurred damage from 23 to 13%. Applying qRT-PCR methodology indicated that M. geulakonigii was endophytic within sealed grapefruit flowers and in the flavedo of the fruits' peel. Neither in the laboratory nor in the field was any evidence ever obtained that this fungus damaged the plants, leading us to hypothesize that M. geulakonigii serves as a "body guard" of grapefruits (and perhaps other plants as well). All three fungi suffered very little mortality after being exposed to various insecticides and acaricides that are in current local use (with the exception of sulfur). The ability of M. geulakonigii to reduce mite numbers without affecting the host plant, the minimal fungal effect on some predatory mites, its endophytic nature along with the apparent tolerance of M. geulakonigii to many insecticides and acaricides, suggest that this fungus could be suitable for integrated pest management (IPM) program.

Antagonistic effects of the endophytic fungus Meira geulakonigii on the citrus rust mite Phyllocoptruta oleivora.

AIMS: The fungus Meira geulakonigii has been shown to reduce populations of citrus rust mite (CRM; Phyllocoptruta oleivora) on citrus leaves and fruits, in both the field and laboratory. However, attempts to isolate the fungus from leaves and fruits have been unsuccessful. The aims of this study were therefore to determine whether M. geulakonigii is a citrus endophyte, and to assess possible mechanisms involved in its mite-antagonist activity. METHODS AND RESULTS: A quantitative real-time PCR and regular PCR approaches were developed to detect M. geulakonigii in both the field and laboratory. The fungus was detected throughout. Different methods revealed that M. geulakonigii is an endophyte, which colonizes both the peel of grapefruits. Applications of conidia protected the grapefruits against CRM, and fungal secretions extracted from growth media caused 100% CRM mortality. CONCLUSIONS: Meira geulakonigii is a beneficial endophyte of grapefruits that colonizes the fruit's peel, and protects it from CRM. SIGNIFICANCE AND IMPACT OF THE STUDY: Findings from this study demonstrate the endophytic nature of M. geulakonigii in its interaction with grapefruits. In addition, a molecular approach was developed to specifically detect the fungus inside the grapefruit peel. This approach can be used to assess the natural occurrence of M. geulakonigii in grapefruit.

Isolation and Characterization of a Cold-Tolerant Strain of Fusarium proliferatum, a Biocontrol Agent of Grape Downy Mildew.

ABSTRACT A cold-tolerant strain of the mycoparasite Fusarium proliferatum was isolated following UV mutagenesis of the G6 strain, which is a biocontrol agent of grape downy mildew. The isolated strain (designated 1505) exhibited radial growth two to threefold that of the parent strain when grown at 13 degrees C, which is generally suboptimal for growth of Fusarium spp., but desirable for its host, Plasmopara viticola. This rapid growth was correlated with improved biological control of P. viticola, determined by a detached-leaf assay. Even though radial growth of strain 1505 at higher temperatures was slower than that of G6 and the strain failed to conidiate, there was no reduction in biocontrol efficacy. Significantly higher levels of extracellular beta-glucosidase and endo-1,4-beta-glucanase activity were measured in the culture filtrate of strain 1505 relative to that of strain G6. A DNA-mediated transformation procedure that included the introduction of antibiotic resistance and a GUS reporter gene system was adapted for F. proliferatum. Using the GUS-engineered strains, we demonstrated that both G6 and 1505 exhibit the characteristic coiling and penetration of host structures.

The Mycoparasite Ampelomyces quisqualis Expresses exgA Encoding an exo-beta-1,3-Glucanase in Culture and During Mycoparasitism.

ABSTRACT Ampelomyces quisqualis, a mycoparasite of fungi causing powdery mildews, exhibited high levels of extracellular exo-beta-1,3-glucanase activity in culture compared with Neurospora crassa and Gliocladium roseum. A. quisqualis culture filtrates affected powdery mildew caused by Sphaerotheca fusca in a manner indicative of cell wall degradation, as determined by microscopic examination. A gene encoding an exo-beta-1,3-glucanase in A. quisqualis, designated exgA, was isolated and sequenced. The predicted polypeptide deduced from exgA had 46, 42, and 30% identity with amino acid sequences of Trichoderma harzianum exo-beta-1,3-glucanase and Cochliobolus carbonum EXG1 (both encoding exo-beta-1,3-glucanase) and T. harzianum bng13.1 (encoding an endo-beta-1,3-glucanase), respectively. The exgA gene had a predicted molecular mass of 84 kDa and a pI of 4.79. The gene was expressed during the late stages of growth in culture, and transcription was induced by fungal cell wall components. Transcript levels for exgA were present during late stages of hyperpar-asitism and were abundant along A. quisqualis mycelium and were slightly less abundant in A. quisqualis pycnidia.

The chsA gene, encoding a class-I chitin synthase from Ampelomyces quisqualis.

Degenerate oligodeoxyribonucleotide primers, designed on the basis of conserved regions of the chitin synthase gene family, were used to amplify a fragment of the Ampelomyces quisqualis (Aq) chsA gene. Subsequently, the PCR product was used as a probe in order to identify and isolate genomic clones harboring the entire chsA gene. Aq chsA is 2786-nt long, has one intron and encodes a 910-amino-acid polypeptide belonging to the class-I chitin synthases. Low-stringency Southern hybridizations to Aq genomic DNA provided evidence for the presence of additional DNA fragments resembling chsA in the fungal genome, suggesting the presence of a multigene family of chitin synthases in Aq.