Ink disease is a devastating disease of chestnut (Castanea sativa) worldwide, caused by Phytophthora species. The only management measures of this disease are chemical and agronomic interventions. This work focuses on the evaluation of the in vitro antagonistic capacity of 20 isolates of Trichoderma spp. selected in a diseased chestnut orchard in Tuscan Apennines (San Godenzo, Italy) for the biocontrol of Phytophthora xcambivora. Each Trichoderma isolate was tested to investigate pathogen inhibition capability by antagonism in dual cultures and antibiosis by secondary metabolites production (diffusible and Volatile Organic Compounds). The six most performing isolates of Trichoderma spp. were further assessed for their aptitude to synthesize chitinase, glucanase and cellulase, and to act as mycoparasite. All six selected isolates displayed the capability to control the pathogen in vitro by synergistically coupling antibiosis and mycoparasitism at different levels regardless of the species they belong to, but rather, in relation to specific features of the single genotypes. In particular, T. hamatum SG18 and T. koningiopsis SG6 displayed the most promising results in pathogen inhibition, thus further investigations are needed to confirm their in vivo efficacy.

Abies nebrodensis (Lojac.) Mattei is an endemic species of the north-west of Sicily located in an 84 ha area in the Madonie Regional park. The current population is limited to 30 relic adult trees and a fluctuating number of juveniles of natural regeneration. The species is defined as “Critically Endangered” in the Italian list of threatened plants and is classified as CR-D in the 2000 IUCN Red List of Threatened Species. This article reports the key action undertaken by the LIFE4FIR project aimed at preserving A. nebrodensis, and the results obtained so far in three years of activity. OpenArrays SNPs genotyping revealed a high rate of inbreeding in the natural population and that the adult trees are genetically related. Controlled cross-pollination was consequently performed to increase the genetic variability of the progeny. Outbred offspring are currently being grown in the nursery. Reforestation has been planned by using 4000 selected outbred seedlings in 10 areas within Madonie Park to create re-diffusion cores. Support and protection of the relic population have been implemented through regular phytosanitary surveys, as well as new fencing and video surveillance systems against grazing and wild herbivores. A seedbank and cryobank for the long-term germplasm conservation have been established.

Ocimum basilicum (sweet basil) is an economically important aromatic herb; in Italy, approximately 1000 ha of “Genovese-type” basil are grown annually in greenhouses and open fields and are subjected to Downy Mildew (DM) disease, caused by Peronospora belbahrii, leading to huge crop losses. Mutation of the Susceptibility (S) gene DMR6 (Downy Mildew Resistant 6) has been proven to confer a broad-spectrum resistance to DM. In this work, an effective Genome Editing (GE) approach mediated by CRISPR/Cas9 in O. basilicum ‘Italiko’, the élite cultivar used to produce “Pesto Genovese D.O.P”, was developed. A highly efficient genetic transformation method mediated by A. tumefaciens has been optimized from cotyledonary nodes, obtaining 82.2% of regenerated shoots, 84.6% of which resulted in Cas9+ plants. Eleven T0 lines presented different type of mutations in ObDMR6; 60% were indel frameshift mutations with knock-out of ObDMR6 of ‘FT Italiko’. Analysis of six T1 transgene-free seedlings revealed that the mutations of T0 plants were inherited and segregated. Based on infection trials conducted on T0 plants, clone 22B showed a very low percentage of disease incidence after 14 days post infection. The aromatic profile of all in vitro edited plants was also reported; all of them showed oxygenated monoterpenes as the major fraction.

Forest decline is a widespread phenomenon on coasts throughout the Mediterranean basin. Pathogenic fungi are amongst the main causes of forest tree diseases. Diplodia species, having a cosmopolitan distribution, are well-known as pathogens of woody plant hosts including Pinus spp. In recent years, symptoms of shoot dieback, necrosis, twig blight, canopy transparency and trunk cankers have been observed on Pinus pinea trees in Tunisian forests. Nevertheless, this has been less well-studied in North Africa and especially in Tunisia. The purpose of this study is to report the occurrence of Diplodia species causing dieback on P. pinea in two forests in northeastern and the northern Tunisia. A collection of fifty-eight isolates were obtained from symptomatic branches of P. pinea trees. Three Diplodia species were identified: D. africana, D. pseudoseriata and D. scrobiculata by means of morphological characteristics, and phylogenetic analysis. Their incidence on P. pinea trees appeared to be significantly correlated to the dendrometric parameters and ecological factors. A pathogenicity test was conducted on 3-years-old P. pinea seedlings, confirmed the virulence of three Diplodia species. Based on the length of vascular necrosis in the wood of the P. pinea inoculated stem, D. scrobiculata was the most virulent species. Data from the present study provide the first report of the species of Diplodia associated with P. pinea dieback in Tunisia.