Molecular analysis of the fungal microbiome associated with the olive fruit fly Bactrocera oleae
Abstract A molecular approach was used to investigate the fungal microbiome associated with Bactrocera oleae a major key pest of Olea europea , using the ITS2 region of the ribosomal DNA (rDNA) as barcode gene. Amplicons were cloned and a representative number of sequenced fragments were used as barcode genes for the identification of fungi. The analysis of the detected sequence types (STs) enabled the identification of a total of 34 phylotypes which were associated with 10 fungal species, 3 species complexes and 8 genera. Three phylotypes remained unresolved within the order Saccharomycetales and the phylum Ascomycota because of the lack of closely related sequences in GenBank. Cladosporiu…
Diversity and distribution of Phytophthora species in protected natural areas in Sicily
: The aim of this study was to investigate the occurrence, diversity, and distribution of Phytophthora species in Protected Natural Areas (PNAs), including forest stands, rivers, and riparian ecosystems, in Sicily (Italy), and assessing correlations with natural vegetation and host plants. Fifteen forest stands and 14 rivers in 10 Sicilian PNAs were studied. Phytophthora isolations from soil and stream water were performed using leaf baitings. Isolates were identified using both morphological characters and sequence analysis of the internal transcribed spacer (ITS) region. A rich community of 20 Phytophthora species from eight phylogenetic clades, including three new Phytophthora taxa, was …
Metabarcoding Analysis of Fungal Diversity in the Phyllosphere and Carposphere of Olive (Olea europaea)
The fungal diversity associated with leaves, flowers and fruits of olive (Olea europaea) was investigated in different phenological stages (May, June, October and December) using an implemented metabarcoding approach. It consisted of the 454 pyrosequencing of the fungal ITS2 region and the subsequent phylogenetic analysis of relevant genera along with validated reference sequences. Most sequences were identified up to the species level or were associated with a restricted number of related taxa enabling supported speculations regarding their biological role. Analyses revealed a rich fungal community with 195 different OTUs. Ascomycota was the dominating phyla representing 93.6% of the total…
A Metabarcoding Survey on the Fungal Microbiota Associated to the Olive Fruit Fly.
The occurrence of interaction between insects and fungi is interesting from an ecological point of view, particularly when these interactions involve insect pests and plant pathogens within an agroecosystem. In this study, we aimed to perform an accurate analysis on the fungal microbiota associated to Bactrocera oleae (Rossi) through a metabarcoding approach based on 454 pyrosequencing. From this analysis, we retrieved 43,549 reads that clustered into 128 operational taxonomic units (OTUs), of which 29 resulted in the “core” associate fungi of B. oleae. This fungal community was mainly represented by sooty mould fungi, such as Cladosporium spp., Alternaria spp. and Aureobasidium spp., by pl…
A molecular method to assess Phytophthora diversity in environmental samples
Current molecular detection methods for the genus Phytophthora are specific to a few key species rather than the whole genus and this is a recognized weakness of protocols for ecological studies and international plant health legislation. In the present study a molecular approach was developed to detect Phytophthora species in soil and water samples using novel sets of genus-specific primers designed against the internal transcribed spacer (ITS) regions. Two different rDNA primer sets were tested: one assay amplified a long product including the ITS1, 5.8S and ITS2 regions (LP) and the other a shorter product including the ITS1 only (SP). Both assays specifically amplified products from Phy…
Molecular analysis ofPhytophthoradiversity in nursery-grown ornamental and fruit plants
The genetic diversity of Phytophthora spp. was investigated in potted ornamental and fruit tree species. A metabarcoding approach was used, based on a semi-nested PCR with Phytophthora genus-specific primers targeting the ITS1 region of the rDNA. More than 50 ITS1 sequence types representing at least 15 distinct Phytophthora taxa were detected. Nine had ITS sequences that grouped them in defined taxonomic groups (P. nicotianae, P. citrophthora, P. meadii, P. taxon Pgchlamydo, P. cinnamomi, P. parvispora, P. cambivora, P. niederhauserii and P. lateralis) whereas three phylotypes were associated to two or more taxa (P. citricola taxon E or III; P. pseudosyringae, P. ilicis or P. nemorosa; and…
First report of neofusicoccum batangarum as causal agent of scabby cankers of cactus pear (Opuntia ficus-indica) in minor islands of sicily
Cactus pear (Opuntia ficus-indica, Cactaceae), native to Mexico, is a multipurpose crop. About 90% of Italian production of cactus pear fruit is from Sicily. In 2013, a disease of cactus pear was noticed in minor islands of Sicily, Lampedusa and Linosa (Pelagie archipelago), Favignana (Aegadian archipelago), and Ustica, where cactus pear is grown as living fences. Symptoms were on flattened stems functioning as leaves (cladodes) and included radially expanding cankers, up to 20 cm in diameter, concentric, crusty, silvery areas, with minute, black dots (pycnidia erumpent from epidermis) and a leathery, brown halo. A milky to buff colored exudate, caking on contact with air, oozed from active…