The diversity of fungi associated with the gut of Pantala flavescens larvae was investigated using a culture-dependent method and molecular identification based on an analysis of the internally transcribed spacer sequence. In total, 48 fungal isolates were obtained from P. flavescens larvae. Based on phylogenetic analyses, the fungal isolates were grouped in 5 classes and 12 different genera. Fourteen bacterial 16S rDNA sequences derived from total genomic DNA extractions of fungal mycelia were obtained. The majority of the sequences were associated with Proteobacteria (13/14), and one Bacillaceae (1/14) was included. Leclercia sp., Oceanobacillus oncorhynchi and Methylobacterium extorquens, were reported for the first time as bacterial endosymbionts in fungi. High-performance liquid chromatography (HPLC) analysis indicated that bacterial symbionts produced specific metabolites and also exerted an inhibitory effect on fungal metabolites. The biological activity of the fungal culture extracts against the pathogenic bacteria Staphylococcus aureus (ATCC 6538), Bacillus subtilis (ATCC 6633) and Escherichia coli (ATCC 8739) was investigated, and 20 extracts (42%) exhibited antibacterial activity against at least one of the tested bacterial strains. This study is the first report on the diversity and antibacterial activity of symbiotic fungi residing in the gut of P. flavescens larvae, and the results show that these fungi are highly diverse and could be exploited as a potential source of bioactive compounds.
Aroma is one of the most important attributes of orange wine quality. The volatile compounds in orange wine mainly derive from oranges, yeast fermentation and compounds released from odourless glycosidic precursors present in the orange. In this study, free volatile compounds in orange juice and wine made from Citrus sinensis (L). Osbeck cv. Washington Sanguine were analysed by SPME-GC-MS. Bound fractions were isolated and extracted with methanol and Amberlite XAD-2 resin and then hydrolysed by almond β β β β-glucosidase. Totals of 31 and 19 free volatiles were identified in orange juice and wines, respectively. Terpenes were the most abundant compounds in orange juice, while esters were quantitatively the dominant group in orange wine and most of them were compounds newly formed during fermentation, such as isoamyl acetate, ethyl hexanoate, ethyl benzoate, diethyl succinate, ethyl decanoate and ethyl laurate. In total, 11 and three released bound volatiles were found in orange juice and wine, respectively, and most of them were not found in free form. Only ethyl 3-hydroxyhexanoate and cis-carveol were found present in both the free and bound forms of orange juice.
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