Marco Porceddu

Interchangeable effects of gibberellic acid and temperature on embryo growth, seed germination and epicotyl emergence in Ribes multiflorum ssp. sandalioticum (Grossulariaceae).

Morphophysiological dormancy was investigated in seeds of Ribes multiflorum Kit ex Roem et Schult. ssp. sandalioticum Arrigoni, a rare mountain species endemic to Sardinia (Italy). There were no differences in imbibition rates between intact and scarified seeds, suggesting a lack of physical dormancy, while methylene blue solution (0.5%) highlighted a preferential pathway for solution entrance through the raphe. Embryos were small at seed dispersal, with an initial embryo:seed ratio (E:S) of ca. 0.2 (embryo length, ca. 0.5 mm), whereas the critical E:S ratio for germination was three times longer (ca. 0.6). Gibberellic acid (GA(3), 250 mg · l(-1)) and warm stratification (25 °C for 3 months) followed by low temperature (<15 °C) enhanced embryo growth rate (maximum of ca. 0.04 mm · day(-1) at 10 °C) and subsequent seed germination (radicle emergence; ca. 80% at 10 °C). Low germination occurred at warmer temperatures, and cold stratification (5 °C for 3 months) induced secondary dormancy. After radicle emergence, epicotyl emergence was delayed for ca. 2 months for seeds from three different populations. Mean time of epicotyl emergence was affected by GA(3) . Seeds of this species showed non-deep simple (root) - non-deep simple (epicotyl) morphophysiological dormancy, highlighting a high synchronisation with Mediterranean seasonality in all the investigated populations.

Thermal niche for in situ seed germination by Mediterranean mountain streams: model prediction and validation for Rhamnus persicifolia seeds

BACKGROUND AND AIMS Mediterranean mountain species face exacting ecological conditions of rainy, cold winters and arid, hot summers, which affect seed germination phenology. In this study, a soil heat sum model was used to predict field emergence of Rhamnus persicifolia, an endemic tree species living at the edge of mountain streams of central eastern Sardinia. METHODS Seeds were incubated in the light at a range of temperatures (10-25 and 25/10 °C) after different periods (up to 3 months) of cold stratification at 5 °C. Base temperatures (Tb), and thermal times for 50 % germination (θ50) were calculated. Seeds were also buried in the soil in two natural populations (Rio Correboi and Rio Olai), both underneath and outside the tree canopy, and exhumed at regular intervals. Soil temperatures were recorded using data loggers and soil heat sum (°Cd) was calculated on the basis of the estimated Tb and soil temperatures. KEY RESULTS Cold stratification released physiological dormancy (PD), increasing final germination and widening the range of germination temperatures, indicative of a Type 2 non-deep PD. Tb was reduced from 10·5 °C for non-stratified seeds to 2·7 °C for seeds cold stratified for 3 months. The best thermal time model was obtained by fitting probit germination against log °Cd. θ50 was 2·6 log °Cd for untreated seeds and 2·17-2·19 log °Cd for stratified seeds. When θ50 values were integrated with soil heat sum estimates, field emergence was predicted from March to April and confirmed through field observations. CONCLUSIONS Tb and θ50 values facilitated model development of the thermal niche for in situ germination of R. persicifolia. These experimental approaches may be applied to model the natural regeneration patterns of other species growing on Mediterranean mountain waterways and of physiologically dormant species, with overwintering cold stratification requirement and spring germination.

The Aichi Biodiversity Target 12 at regional level: an achievable goal?

The Aichi Biodiversity Target 12 aims to prevent the extinction risk of known threatened species and to improve their conservation status by 2020. We present the integrated strategy implemented in the last 10 years for the keystone plant species of Sardinia (Italy, W. Mediterranean Basin), which includes the following activities: conservation status assessment (following the IUCN protocol), ex situ conservation, in situ monitoring and active protection measures. To date, an average of 51.8% of keystone plant species have been subjected to the latter first three activities while, due to the higher costs, only few active conservation measures have been carried out. Considering the activities realised since 2004, we have also predicted the conservation effectiveness towards 2020 and have elaborated an index to evaluate it. Halfway through the strategic plan, we argue that more efforts are needed to guarantee the effective conservation of all threatened plants in Sardinia.

New findings on seed ecology of Ribes sardoum: can it provide a new opportunity to prevent the extinction of a threatened plant species?

Ribes sardoum, the most threatened endemic plant of Sardinia, is included in the Habitats Directive (92/43/EEC) and it was considered Critically Endangered in the global IUCN Red Lists. This species has been reported to have an extremely low fertility, scarce fruit production, low seed viability and a general inability to reproduce sexually. Fruits were collected for the first time from the remnant population, and the requirements for seed germination were investigated in the laboratory. Seeds were incubated at different temperatures (10, 15, and 20°C) and, in addition, they were exposed to a warm stratification (W) or a move-along treatment characterized by three cold temperature regimes (CCC). Seeds were also sown on the surface of 1% agar water with 250 mg·L−1 of GA3. At maturity, seeds have a linear underdeveloped embryo. Germination percentage between 35% and 65% were detected in the control and W groups. A low germination percentage occurred after CCC and during GA3 treatment. W treatment speeds up seed germination. Our results demonstrate that fruits of R. sardoum produce viable seeds, that are able to germinate under controlled conditions, with the assumption that the seeds have morphophysiological dormancy (MPD), and that propagation from the seeds is possible. Although the ability of seed germination was demonstrated, the lack of seedlings in the natural population seems to be a consequence of unfavourable climatic conditions for recruitment. However, our results indicate that seedlings obtained under controlled conditions could be useful for future translocation reducing and/or mitigating the extinction likelihood of this highly threatened plant.

Discovering the type of seed dormancy and temperature requirements for seed germination of Gentiana lutea L. subsp. lutea (Gentianaceae)

Aims There are a number of mechanisms that regulate germination; among these, seed dormancy, one of the most important, is an adaptative mechanism in plants to promote survival by dispersing germination in space and time until environmental conditions are favourable for germination. The main goals of this study were to determine the temperature requirements for seed dormancy release and germination of Gentiana lutea subsp. lutea, to identify the class and level of seed dormancy and to suggest an optimal germination protocol. Methods Seeds belonging to two different localities were subjected to various pre-treatments, including cold stratification (0 and 5°C), warm stratification (25/10°C) and different combinations of these, and then incubated at a range of constant temperatures (5–25°C) and 25/10°C. Embryo growth during pre-treatments and incubation conditions were assessed at different times by measuring the embryo to seed length ratio (E:S ratio). The final germination percentage (FGP) and the germination rate (t50) were calculated. Important Findings Fleshy mature seeds of G. lutea subsp. lutea have linear underdeveloped embryos. Cold stratification at 0°C was effective in overcoming the physiological dormancy (PD) and promoted embryo growth and subsequent germination. After cold stratification at 0°C, both the root and the shoot emerged readily under a wide range of temperatures. G. lutea subsp. lutea seeds showed an intermediate complex morphophysiological dormancy (MPD). As regards the optimal germination protocol for this taxon, we suggest a period of cold stratification at ca. 0°C followed by seed incubation at 10–20°C. The optimal germination temperatures found for seeds of this taxon, as well as its pre-chilling requirement at 0°C, suggest that it is well adapted to a temperate climate; this behavior highlights an increasing threat from global warming for G. lutea, which could reduce the level of natural emergence in the field, prejudicing also the long-term persistence of the natural populations in Sardinia.

The genetic structure and diversity of Gentiana lutea subsp. lutea (Gentianaceae) in Sardinia: further insights for its conservation planning

ABSTRACT Knowledge of the levels of genetic diversity and of the spatial genetic structure of plant species is important to ensure their effective management and conservation, especially in the case of endangered species. Gentiana lutea L. subsp. lutea is a long-lived plant which occurs in central and southern European mountains. It has a long-standing history of human exploitation, mainly in the liqueur and in the pharmaceutical industries and it is currently listed in the EU Habitats Directive 92/43/EEC Annex V. Mainly due to a prolonged root harvesting, its current distribution range in Sardinia consists of only a few groups of individuals limited to small areas of the Gennargentu massif (Central-Eastern area of the island). In this study, we investigated the levels of genetic diversity and the genetic structure of the species in Sardinia. We used AFLP (amplified fragment length polymorphism) markers to investigate the genetic variability of 182 samples from 13 subpopulations. A total of 433 fragments were detected, of which 75.5% were polymorphic. The levels of genetic diversity were generally high, but they tended to decrease in smaller subpopulations. Of the genetic variability 88% was found within subpopulations, while the genetic structure among them was fairly weak. In order to ensure the survival of these subpopulations, especially the smaller ones, ex situ and in situ management actions should be planned, such as the long term conservation of its seeds in germplasm repositories and their population reinforcements and monitoring.