The hidden world within plants: metatranscriptomics unveils the complexity of wood microbiomes.

The importance of plants as complex entities influenced by genomes of the associated microorganisms is now seen as a new source of variability for a more sustainable agriculture, also in the light of ongoing climate change. For this reason, we investigated through metatranscriptomics whether the taxa profile and behaviour of microbial communities associated with the wood of 20-year-old grapevine plants are influenced by the health status of the host. We report for the first time a metatranscriptome from a complex tissue in a real environment, highlighting that this approach is able to define the microbial community better than referenced transcriptomic approaches. In parallel, the use of total RNA enabled the identification of bacterial taxa in healthy samples that, once isolated from the original wood tissue, displayed potential biocontrol activities against a wood-degrading fungal taxon. Furthermore, we revealed an unprecedented high number of new viral entities (~120 new viral species among 180 identified) associated with a single and limited environment and with potential impact on the whole holobiont. Taken together, our results suggest a complex multitrophic interaction in which the viral community also plays a crucial role in raising new ecological questions for the exploitation of microbial-assisted sustainable agriculture.

Isolation, molecular characterization and virome analysis of culturable wood fungal endophytes in esca symptomatic and asymptomatic grapevine plants.

Europe is the world largest grape producer, but in recent years, the report of diseases due to infection by grapevine trunk pathogens (GTPs) is becoming one of the main constraints for viticulture. Among grapevine trunk diseases (GTDs), esca syndrome is one of the most complex, characterized by simultaneous infection of several fungi, which leads to important reduction in yield and quality. Previous characterization of fungal isolates associated with esca syndrome leads to the recognition of at least three important players: Phaeocremonium minimum, Phaemoniella chlamydospora and Fomitiporia mediterranea. Here we isolated and characterized molecularly fungal endophytes directly from field wood tissues of plants showing or not esca symptoms. In addition, to better characterize such collection, a deep RNA sequencing (100 M reads in paired-end) to screen for mycovirus presence was performed. Thirty-nine viral genomes were detected, 38 of which were putative new viral species; some of these viruses infected GTPs, including P. minimum and F. mediterranea. In this work, we reported for the first time a curated collection of grapevine fungal endophytes identifying the associated mycoviruses some of which could be employed in future biotechnological exploitation as biological control agents for sustainable plant protection.

The presence of melatonin in grapevine (Vitis vinifera L.) berry tissues.

Melatonin has been reported in a variety of food plants and, consequently, in a number of plant-derived foodstuffs. In grapevine (Vitis vinifera L.) products, it was found in berry exocarp (skin) of different cultivars and monovarietal wines. Herein, we assessed, by means of mass spectrometry, the occurrence of melatonin in all berry tissues (skin, flesh, and seed) at two different phenological stages, pre-véraison and véraison. We detected the highest melatonin content in skin, at pre-véraison, whereas, at véraison, the highest levels were reported in the seed. Furthermore, during ripening, melatonin decreased in skin, while increasing in both seed and flesh. The relative concentrations of melatonin in diverse berry tissues were somewhat different from those of total polyphenols (TP), the latter measured by the Folin-Ciocalteau assay, and more abundant in seed at pre-véraison and in exocarp at véraison. The highest antiradical activity, determined by both DPPH (2,2-diphenyl-1-pycryl hydrazyl) and ABTS [(2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)] radical-scavenging assay, was reported at pre-veráison in seed. To the best of our knowledge, we reported, for the first time, the occurrence of melatonin in grape seeds.

From vineyard to glass: agrochemicals enhance the melatonin and total polyphenol contents and antiradical activity of red wines.

Resistance inducers are a class of agrochemicals, including benzothiadiazole and chitosan, which activate the plant own defence mechanisms. In this work, open-field treatments with plant activators were performed on two red grape (Vitis vinifera L.) varieties cultivated in different localities, Groppello (Brescia, Lombardia, Italy) and Merlot (Treviso, Veneto, Italy). Treatments were carried out every 10 days until the véraison and, after harvesting, experimental wines (microvinificates) were prepared. In general, both melatonin and total polyphenol content, determined by mass spectrometry and Folin-Ciocalteu assay, respectively, were higher in wines produced from grapes treated with resistance inducers than in those obtained from untreated control and conventional fungicide-treated grapes. Accordingly, antiradical power of wines derived from plant activator-treated grapes, measured by both DPPH (2,2-diphenyl-1-picrylhydrazyl) and the ABTS [(2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)] radical-scavenging assay, was higher than in their counterparts. To the best of our knowledge, this is the first report on the effects of agrochemicals on the melatonin content of red wine.