Tuscia Germplasm Bank

Pteridophytes spore banks are a promising ex situ conservation tool used to increase the chances of survival of ferns, in fact that large quantities of germplasm with high genetic variation can be conserved in a small space with low economic and technical costs. However, methods to maintain the viability of chlorophyllous spores during storage are less understood.

The aim of this study was to investigate the influence of long term storage on the viability of Royal Fern spores, which were stored under different conditions derived from various combinations of temperature and degrees of hydration. Survival and germination tests were performed after 1 and 28 months of storage. Our results showed the highest survival percentages for spores stored under Normal humidity at subzero temperatures (T = ⩽ −20 °C). These spores received no pre-treatment, dehydration, or cryoprotectants, which resulted in fast germination and gametophyte development which seemed to be stimulated by low temperatures.

Batt. is an threatened orchid species of the Italian flora, with a centralwestern stenomediterranean distribution, that is sporadic in the western part of the distribution area of the more common L. abortivum (L.) Sw., an eurimediterranean species. It occurs in Italy only with a few populations in Tuscany, Latium, Umbria, Sicily and Sardinia [1], often with L. abortivum [2], [3], [4] from which it is easily recognizable only during anthesis for the denser inflorescence spike, the ribbon-like lip without differentiation in epychile and hypochile, and for the spur that is very short or absent [5].

A modern method, geometric morphometrics, was used to clarify
the taxonomic position of the European Ophioglossum species: O. azoricum, O. lusitanicum, and O. vulgatum. The identification of these taxa by traditional methods is rather difficult, due to different taxonomic interpretations. Sterile leaf shapes were investigated using a landmark-based method and the Fourier analysis of outlines. Both methods highlight the shape and the base of the leaf as an important diagnostic character.

Nel Viterbese l'unica popolazione, costituita da numerosi esemplari, si trova nella zona umida del Bagnaccio (piana di Viterbo, 300 m s.l.m.) dove è presente una sorgente idrotermale calda che alimenta un'area acquitrinosa inondata per alcuni mesi all'anno con un prato incolto di elevato interesse floristico (6). Attualmente le acque sorgive sono convogliate all'interno di vasche per la balneazione, pertanto la popolazione di O. palustris rischia di scomparire.

Taxonomy and distribution of Lupinus L. sect. Albus are discussed and updated with new data from Italy. The study of natural populations of Lupinus in the central Tyrrhenian Italy was based on the observations (in situ and ex situ) of the combination of the diagnostic characters (colour of corolla and seeds) mainly used by other authors, as well as the dehiscence of the pods, the dimension, shape and permeability of the seeds, according to the agronomic and genetic studies widely reported in the literature, to discriminate between the wild form (Lupinus albus subsp. graecus) and those escaped from cultivation (L. albus subsp. albus). Our results (1) led us to confirm the rank of subspecies to the wild form, L. albus L. subsp. graecus (Boiss. & Spruner) Franco & P. Silva, (2) confirm the presence of hybrids forms in Italy and (3) update the distribution area of subsp. graecus in Italy, including Apulia (Gargano).

L. albus subsp. graecus can be considered as a flag species of the interesting form of the edaphophilous “steppe”, rare in central Italy, confirming its value as an indicator of habitat “naturality”.

Ex situ approaches represent important conservation tools to increase the chances of many ferns survival, as they are very sensitive to environmental disturbances. This work presents the first results of in vitro reproduction of Dryopteris tyrrhena, an endangered western Mediterranean endemism with a highly fragmented distribution area, using spores taken from exsiccata. Our results highlight the value of herbarium specimens in pteridophyte conservation, providing a useful method to reproduce species that are threatened or extinct in the wild, at least at a local level, and to plan eventual reintroductions using the native germplasm.

The present study is a first approach to in vitro asymbiotic reproduction of Orchis palustris, an endangered species due to the fragmentation of its distribution area and the reduction and alteration of its suitable habitat. Within a project of the Tuscia Germplasm Bank (Viterbo, Italy) for the ex situ conservation of wild orchids, seeds were collected from a population near the thermal spring at Bagnaccio (Viterbo, Italy). We investigated the effects of the maturity level of the seeds on the percentages of germination and on the development of protocorms, culturing mature seeds and immature ones collected from green pods. The percentages of seed germination and the developmental stages of protocorms were evaluated up to 10 months after sowing.In the cultures of immature seeds was possible to observe all the developmental stages of orchid seed germination, with a maximum percentage of germination of 33%. On the contrary has not been obtained any germination in the cultures of mature seeds.

"Dryopteris tyrrhena is a Western Mediterranean endemism, a silicicolous species presents in a few populations in Spain,
France, and Italy, where it is in regression. Its current distribution leads to hypothesize an Ibero-Provencal-Tyrrhenian origin, while the fragmentation of its distribution area, and the distances between the stations, suggest a very ancient origin, suggesting that it is a relict species of the Tertiary flora of the Mediterranean mountains. The present study on in vitro reproduction of D. tyrrhena has been undertaken to learn more about the characteristics of the reproductive cycle of this species for ex situ conservation and to obtain more information on the biological factors that may have contributed to the fragmentation of its distribution area. The spores were collected from 2 Italian populations from Liguria (Riomaggiore, La Spezia) and Tuscany (Capraia Island, Livorno), and in vitro reproduction started in the summer of 2008. The results show a significant difference between the 2 populations: in the time and percentage of germination, in the gametophytes growth and, especially, in the sporophytes development."

The increasing emphasis on terrestrial orchid conservation has led to conservation actions for a wide range of threatened Mediterraneanspecies. Many terrestrial orchids are currently at great risk for extinction as a result of a multiplicity of threatening processes. We focus onorchid seed germination capabilities in vitro, specifically germination capability of a threatened species, Anacamptis palustris, comparedto three other more common species (A. laxiflora, A. morio, and A. papilionacea), and also discuss its potential impact on orchid distributionand conservation. Asymbiotic germination tests were performed with mature seeds using BM-1 medium. In vitro seed germinationand protocorm developmental stages were evaluated up to 20 weeks after sowing. Significant differences in seedling development weredetected among the species, and a correlation was evident between the rarity of the species and their germinability. Thus, the presence ofintrinsic, biological factors that affect and limit the distribution of A. palustris may exist.

"This paper aims to illustrate a project for the ex situ conservation of Epipactis placentina Bongiorni & Grünanger in Latium (central Italy), where it is very rare and highly localized in a few stations in the beech forests of the north-central area of the region. The project was carried out through collaboration between the G.I.R.O.S. association (Italian Group for Wild Orchids Research) and the Tuscia Germplasm Bank (University of Tuscia, Italy).
In the first phase, the following actions were undertaken: 1) population census and monitoring, 2) seed collection, and 3) long term germplasm conservation. In a second phase of the project, other actions will be carried out: 4) asymbiotic germination tests, 5) reproductive biology studies, and 6) in vitro reproduction."