Symbiotic Fungi of an Ambrosia Beetle Alter the Volatile Bouquet of Cork Oak Seedlings.

In Portugal, fungal symbionts of the ambrosia beetle Platypus cylindrus affect tree vigor of cork oak (Quercus suber) and are linked with the cork oak decline process. Fungal symbionts play crucial roles in the life history of bark and ambrosia beetles and recent work indicates complex interactions on the fungal and plant metabolic level. Colonized trees may respond with an array of currently unknown volatile metabolites being indicative of such interactions, acting as infochemicals with their environment. In this study, we examined volatile organic compounds (VOCs) of cork oak seedlings wound inoculated with strains of three fungal associates of P. cylindrus (Raffaelea montetyi, R. quercina, and Ceratocystiopsis sp. nov.) over a 45-day period by means of thermodesorption gas chromatography-mass spectrometry techniques. Fungal strains induced largely quantitative but species-specific changes among the 58 VOCs characterized. Overall, monoterpenes-the major volatiles of cork oak foliage-were significantly reduced, possibly a result of fungal biotransformation. Acetophenone, sulcatone, and nonanal-volatiles known for mediating ambrosia beetle behavior-increased in response to fungal inoculation. Qualitative VOC profiles of excised tissue of wood lesions (21 VOCs) and pure fungal cultures (60 VOCs) showed little overlap with seedling VOCs, indicating their plant-derived but fungal-induced origin. This chemoecological study expands on the limited knowledge of VOCs as infochemicals emitted from oak trees threatened by oak decline in relation to beetle-vectored ophiostomatoid fungi. It opens new avenues of research to clarify mutualistic or pathogenic aspects of these complex symbiotic interactions and develop new control strategies for P. cylindrus, including its mycobiota.

Microbiome Associated with the Mycangia of Female and Male Adults of the Ambrosia Beetle Platypus cylindrus Fab. (Coleoptera: Curculionidae).

The ambrosia beetle Platypus cylindrus Fab. (Coleoptera: Curculionidae) is a major cork oak pest in Portugal. Female and male beetles have different roles in host tree colonization and are both equipped with prothoracic mycangia for fungal transportation. Despite a known beneficial role of bacteria in ambrosia beetles, information on bacterial composition associated with prothoracic mycangia structures is scarce. Bacterial community from mycangia of P. cylindrus male and female beetles collected from cork oak galleries was investigated by means of 16S metagenomics. Mycangia anatomical structure was also explored with histological techniques and X-ray computed microtomography to highlight evidence supporting biological sexual dimorphism. A bacterial community with highly diverse bacterial taxa with low abundances at the genus level was revealed. Lactobacillales, Leptotrichia, Neisseria, Rothia, and Sphingomonadaceae were significantly more abundant in males, while Acinetobacter, Chitinophagaceae, Enterobacteriaceae, Erwiniaceae, Microbacteriaceae, and Pseudoclavibacter were more abundant in females. Additionally, a core bacteriome of five genera was shared by both sexes. Histological examination revealed visible connections linking external and internal tissues in females, but none in males. Overall, these results provide the first insights into sexual differentiation for bacteria in a Platypodinae beetle species, identifying key patterns of bacteria distribution in the context of beetle ecology and functional behavior.

Diel rhythms in the volatile emission of apple and grape foliage.

This study investigated the diel emission of volatile organic compounds (VOCs) from intact apple (Malus x domestica Borkh., cv. Golden Delicious) and grape (Vitis vinifera L., cv. Pinot Noir) foliage. Volatiles were monitored continuously for 48 h by proton transfer reaction - time of flight - mass spectrometry (PTR-ToF-MS). In addition, volatiles were collected by closed-loop-stripping-analysis (CLSA) and characterized by gas chromatography-mass spectrometry (GC-MS) after 1 h and again 24 and 48 h later. Fourteen and ten volatiles were characterized by GC-MS in apple and grape, respectively. The majority of these were terpenes, followed by green leaf volatiles, and aromatic compounds. The PTR-ToF-MS identified 10 additional compounds and established their diel emission rhythms. The most abundant volatiles displaying a diel rhythm included methanol and dimethyl sulfide in both plants, acetone in grape, and mono-, homo- and sesquiterpenes in apple. The majority of volatiles were released from both plants during the photophase; whereas methanol, CO2, methyl-butenol and benzeneacetaldehyde were released at significantly higher levels during the scotophase. Acetaldehyde, ethanol, and some green leaf volatiles showed distinct emission bursts in both plants following the daily light switch-off. These new results obtained with a combined analytical approach broaden our understanding of the rhythms of constitutive volatile release from two important horticultural crops. In particular, diel emission of sulfur and nitrogen-containing volatiles are reported here for the first time in these two crops.