In the summer of 2021, the new fencing system to protect the 30 trees of the population and the established natural renewal was completed. 1800 chestnut poles (2.50 meters high) were used, and over 3.5 kilometers were enclosed with netting and metal wires. The protected area was expanded to cover a total surface of 14000 m2 around the Abies nebrodensis plants.
Phytosanitary investigations on trees in the natural population have been completed. These investigations included inspections of the foliage, sampling of branches affected by disorders (blight, needle reddening and loss, chlorosis) and isolation of fungal microorganisms have been carried out. Outcomes showed that the disturbances observed are related to the severe environmental conditions which Abies nebrodensis is experiencing, and, together with the fungal community observed on the symptomatic and healthy needles (consisting mainly of weak pathogens and saprophytes,) indicate that the species is able to cope with the restrictions of its habitat.
The planting out of the selected seedlings in the 10 areas chosen for the reforestation has started in spring 2023. The figures below show the repopulation nuclei set up in Favarotti (Isnello) and Mandarini (Petralia Sottana). Two-meter-high fences provide protection for the 300 Abies nebrodensis plants in the installations.
In the photo below, a newly planted Abies nebrodensis seedling is circled in red, while the arrows indicate the two broom plants (Spartium junceum) placed inside the hole to protect the fir seedling.
The inauguration of the Seed Bank and Cryobank developed within the framework of the LIFE4FIR project took place on July 21, 2023, in the presence of the Mayor of Polizzi Generosa, Gandolfo Librizzi, and the Councilor for Environmental Policies, Gaetano Bellavia, as well as researchers from IBE-CNR and the University of Palermo who developed the protocols and procedures. The biobank is managed by an employee of the Madonie Park Authority who has previously participated in a specific training session for this purpose.
The activities of the LIFE4FIR project also include the implementation of a video surveillance system which, on the one hand, represents a deterrent for any illegal activities and, on the other, allows the monitoring of wild herbivores.
The plant consists of several video cameras (powered by solar panels and batteries) which have been positioned in ertain strategic points of the Abies nebrodensis trail.
The system was completed and activated in June 2022. Below are some photos taken during the installation of the cameras and the start-up of the system.
During March-April 2022 the trees of the natural population were propagated by grafting, in order to set up the clonal orchard.
Thanks to the protocol developed within the LIFE4FIR project, a much higher success and survival rate of the seedlings was obtained compared to previous experiences.
The room dedicated to the seedbank and the cryo-bank was set up at the MAN (Museum of Abies nebrodensis) in Polizzi Generosa.
Instrumentation and safety devices were installed and the seed bank went into operation at the beginning of 2022. Seeds batches are progressively added to the collection stored at -18 degrees Celsius. The cryobank is expected to be fully operational in Spring 2023.
Germination of seedlings in the nursery
The seedlings obtained from the controlled crosses carried out in spring of 2020 have recently germinated and are growing at the ‘Piano Noce’ forest nursery. The development of the seedlings is constantly monitored by UNIPA researchers and DRSRT technicians. These plants will be used to create new plantations in the reforestation and re-diffusion plots identified for the species.
The system of fences to protect the Abies Nebrodensis is under construction
The technicians and workers of the DRSRT are engaged in setting up the new fencing system, to replace the old, deteriorated fences. The new fences are higher than the previous ones and surround a wider area around the trees of the natural population, allowing to protect the recently surveyed natural regeneration from wild herbivores and anthropogenic pressure.
The LIFE4FIR project is currently working on the project action A 1.4 ‘Seed conservation at low temperature (-18 ° C), application of cryopreservation protocols’.
The activities relate the developing of techniques for the conservation at low temperatures of seeds, pollen, isolated embryos and lines of embryogenic callus of Abies nebrodensis. These activities are functional to the implementation of the sperm bank and cryobank.
X-ray screening of seeds.
The vain seed problem is the main obstacle in the germination of Abies nebrodiensis seeds. X-ray analysis is a technique developed by IBE-CNR for the rapid selection of seeds with embryo and empty seeds of Abies nebrodensis (Figure 1). This procedure represents a mass screening of the seeds to be used for germination tests or to be stored at -18 ° C.
Figure 1 and Figure 2. Mature seeds of Abies nebrodiensis observed under X-rays for the selection of seeds with embryo (top frames) and empty seeds (bottom frames)
Figure 3. Mature seeds of Abies nebrodiensis with embryo, seen using X-rays
Prove di germinazione e coltura
In vitro germination tests of seeds stored at -18 ° C and embryos extracted from mature seeds and kept in a refrigerator (4 ° C) were carried out at the CNR-IBE and Unipa-Saaf laboratories, placing the isolated embryos on filter paper and then on different culture substrates (Figures 4,5 and 6).
Figure 4. Isolation of mature embryos from Abies nebrodensis seeds
Figure 5. In vitro germination of an Abies nebrodensis embryo
Figure 6. Germination and in vitro development of Abies nebrodensis embryos
In vitro culture tests of immature embryos isolated from cones collected in July are underway at the two laboratories, in order to obtain direct or indirect embryogenesis. The embryogenic callus, obtained from the excision of the embryos extracted from the seeds of the immature cones from plants 6, 8, 10, 15, 16 and 27 shows a rather slow growth. Various culture media containing different basic salts and plant growth hormones are being tested. A better growth of calluses seems to be favored by the contribution of cytokinins.
Germination tests are also underway on Abies nebrodensis pollen samples stored at low temperatures (Fig. 7).
Figure 7: Abies nebrodensis pollen grain in the germination phase
The remote sensing via drone of the LIFE4FIR project
At the beginning of October 2020 the survey of the Abies nebrodensis natural are was carried out using a drone equipped with multispectral cameras. Remote sensing allows the monitoring of the health of trees and habitat through the measure of the radiation reflected by the vegetation.
During the drone flight, panoramic shots of the habitat and of the individual trees of the Abies nebrodensis population were acquired.
Tree n. 8
Trees n. 10 ed 11
Tree n. 12
Trees n. 16 e 17
Tree n. 19
Phytosanitary monitoring and laboratory activities of LIFE4FIR project
Phytosanitary surveys were carried out in November 2019 and in October 2020. Trees were subjected to a careful visual examination to evaluate their vegetative condition based on observations of the crown shape and transparency, turning foliage, presence of declining, desiccated or damaged parts, occurrence of lesions.
Laboratory analysis for LIFE4FIR monitoring activities
Laboratory analysis of collected samples were conducted through: observations under stereomicroscope, isolation and culture of fungal colonies and their genetic characterization through PCR amplification and sequencing of target loci for their identification. This allowed identification of the fungal microflora associated with the observed alterations and to detect the possible presence of harmful pathogens.