The diversity of Botryosphaeriaceae species associated
with “Botryosphaeria dieback” of grapevine was investigated
in 18 vineyards in Sardinia, Italy. Lasiodiplodia isolates
obtained from different woody hosts including holm oak,
sweet orange and broom bush in Italy, Algeria and Tunisia
were also characterized. Morphological and cultural characteristics
as well as ITS and EF1-α sequence data were used to
identify the fungal isolates. Forty-eight botryosphaeriaceous
isolates were obtained from 113 symptomatic grapevine samples,
from which ten species were identified. Diplodia seriata
was the dominant species (25 % of isolates), followed by
Neofusicoccum parvum (21.7 %). Two species, Diplodia
olivarum and D. africana are reported for the first time on
grapevine. In addition, two new species namely Lasiodiplodia
mediterranea sp. nov. from grapevine, holm oak and sweet
orange and Lasiodiplodia exigua sp. nov. from broom bush
are described. In artificial inoculation experiments conducted
on excised green grapevine shoots and lignified canes as well
as holm oak seedlings, L. mediterranea was shown to be an
aggressive pathogen
Two isolates of Neofusicoccum australe belonging to ITS haplotypes H4 and H1 and associated with grapevine cordon dieback and branch dieback of Phoenicean juniper, respectively, have been shown to produce in vitro structurally different secondary metabolites. From the strain BOT48 of N. australe (haplotype H4) a new cyclohexenone oxide, namely, cyclobotryoxide, was isolated together with 3-methylcatechol and tyrosol. Cyclobotryoxide was characterized as (1S,5R,6S)-5-hydroxy-3-methoxy-4-methyl-7-oxabicyclo[4.1.0]hept-3-en-2-one by spectroscopic, optical, and chemical methods. The strain BL24 (haplotype H1) produced tyrosol along with botryosphaerone D and (3S,4S)-3,4,8-trihydroxy-6-methoxy-3,4-dihydro-1(2H)-naphthalenone. The metabolites obtained from both strains were tested at four concentrations on leaves of grapevine cv. Cannonau, holm oak, and cork oak by the leaf puncture assay. Cyclobotryoxide proved to be the most phytotoxic compound. Tyrosol and cyclobotryoxide were also tested on detached grapevine leaves at concentrations of 0.25 and 0.5 mg/mL. Only cyclobotryoxide was found to be active in this bioassay.
Sphaeropsis sapinea was repeatedly isolated in Sardinia from symptomatic samples of the upper part of declining pine (Pinus radiata) plants. Observed symptoms mainly consisted of foliage chlorosis, drying of needles and cankers on branches. The S. sapinea strains were shown to produce phytotoxic metabolites in culture filtrates. Three metabolites were isolated for the first time from this fungus and identified by their spectroscopic and optical properties as R-())-mellein, (3R,4R)-4-hydroxymellein and (3R,4S)-4-hydroxymellein. When assayed for phytotoxic and antifungal activities on host and non-host plants and on some phytopathogenic fungi, the R-())-mellein showed significant activity, while the other two 3,4-dihydroisocoumarins showed only a synergic activity in both tests.
A new antifungal 6-substituted 2H-pyran-2-one, named viridepyronone, has been isolated from a cultural filtrate of a strain of Trichoderma viride showing antagonistic activity in vitro toward Sclerotium rolfsii, which is the causal agent of crown and stem rot of artichoke. Viridepyronone was characterized as 6-(4-oxopentyl)-2H-pyran-2-one 2 with spectroscopic methods. Bioassays showed that viridepyronone had a good antifungal activity against S. rolfsii, and its minimum inhibitory concentration (over 90% inhibition) was found to be 196 microg/mL. This is the first report of viridepyronone produced by any species of fungi.
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