Efisio Mattana

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

BACKGROUND AND AIMSnMediterranean 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.nnnMETHODSnSeeds 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.nnnKEY RESULTSnCold 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.nnnCONCLUSIONSnTb 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.

Conservation genetics of two island endemic Ribes spp. (Grossulariaceae) of Sardinia: survival or extinction?

Measuring levels of population genetic diversity is an important step for assessing the conservation status of rare or endangered plant species and implementing appropriate conservation strategies. Populations of Ribes multiflorum subsp. sandalioticum and R. sardoum, two endangered endemic species from Sardinia, representing the whole genus on the island, were investigated using ISSR and SSR markers to determine levels and structure of genetic variability in their natural populations. Results indicated medium to low genetic diversity at the population level: Neis gene diversity for ISSR markers ranged from 0.0840 to 0.1316; the expected heterozygosity (HE ) for SSR ranged from 0.4281 to 0.7012. In addition, only one remnant population of R. sardoum showed a high level of inbreeding, in accordance with its very small size. Regarding the structure of the six R. sandalioticum populations, both principal coordinates analysis (PCoA) and STRUCTURE analysis of ISSR and SSR data highlighted low population structure, although two populations appeared to be clearly distinct from the others. The genetic pattern of the two taxa associated with their different ecological positions indicated resilience of R. sandalioticum populations in fresh and humid habitats and uncertain future resistance for the residual R. sardoum population in xeric calcareous stands. Hence, this study highlights the importance of an integrated conservation approach (genetic plus in situ and ex situ conservation studies/measures) for activating management programmes in these endemic and threatened taxa that can be considered as crop wild relatives of cultivated Ribes species.

Effects of NaCl stress on seed germination and seedling development of Brassica insularis Moris (Brassicaceae)

Brassica insularis is a protected plant that grows on both coastal and inland cliffs in the western Mediterranean Basin. The objective of this study was to test if any variability exists in the salt stress response during seed germination and seedling development in this species relative to its provenance habitat. Variability among three populations in the salt stress effects on seed germination and recovery under different temperatures was evaluated. The effect of nebulisation of a salt solution on seedling development was evaluated between populations growing at different distances from the sea. Seeds of B.xa0insularis could germinate at NaCl concentrations up to 200xa0mm. Seed viability was negatively affected by salt, and recovery ability decreased with increasing temperature or salinity. Inter-population variability was detected in salt response during the seed germination phase, as well as in seedling salt spray tolerance. The inland population seedlings had drastically decreased survival and life span and failed to survive to the end of the experiment. In contrast, at least 90% of the coastal seedlings survived, even when sprayed at the highest frequency with salt solution. This study allowed investigation of two natural factors, soil salinity and marine aerosols, widely present in the B.xa0insularis habitat, and provided the first insights into ecology of this protected species and its distribution in the Mediterranean. These results might be useful in understanding the actual distributions of other species with the same ecology that experience these same abiotic parameters.

Dependency of seed dormancy types on embryo traits and environmental conditions in Ribes species.

The hypothesis that seed dormancy may be dependent on environmental conditions and seed morphological traits was tested for six Ribes species, across an altitudinal gradient of 1300xa0m and a longitudinal separation of 120°. Embryo measurements and seed germination experiments were conducted for R. alpinum L., R. hudsonianum Richardson var. petiolare (Douglas) Jancz., R. nevadaense Kellogg, R. roezlii Regel var. cruentum (Greene) Rehder and R. speciosum Pursh, and data taken from the literature for R. multiflorum Kit. ex Schult. ssp. sandalioticum Arrigoni. Germination was compared with seed viability to reveal proportional seed dormancy, which was then correlated to seed/embryo morphological traits and these traits related to the seed provenance environment. The embryos of all the investigated species are linear underdeveloped and all had a morphological component of seed dormancy (MD). Seeds of R.xa0roezlii, R.xa0hudsonianum and R.xa0nevadaense required a temperature and/or hormone pre-treatment in order to germinate, highlighting morphophysiological seed dormancy (MPD). Seed dormancy was found to be strongly negatively correlated with embryo length, but not with embryo to seed (E:S) ratio or seed mass. Initial embryo length was positively related to mean annual temperature. Seed dormancy in the investigated Ribes species could be quantified and predicted by the interaction of embryo traits and environmental conditions. This approach may be helpful in assessing and predicting seed dormancy in the Ribes genus and in other genera and families with underdeveloped embryos.

Conserving seeds of useful wild plants in Mexico: main issues and recommendations

The efficient storage and germination of seeds underpin the effective use of plants for livelihoods and sustainable development. A total of 204 wild species useful for local communities of the Tehuacán–Cuicatlán Valley were collected and stored in seed banks in country for long term conservation, and 66xa0% (i.e., 134) duplicated in the U.K., as an effective means of ex situ conservation. Of the 204 species, 147 (122 of which also duplicated in the U.K.) were previously listed as useful plants in the ethnofloristic inventory of the Valley. Based on literature surveys, we found that one of the major impediments to the use of stored seeds of wild species is the lack of knowledge of how to germinate the seed. In detailed studies, we found that seeds of 18 useful plant species from 10 different families germinated readily and could be propagated. In contrast, four species (Actinocheita filicina, Bursera submoniliformis, Karwinskia mollis and Lippia graveolens) produced dormant seeds and therefore further studies are needed before their use can be maximised in large scale propagation programmes in support of conservation and livelihoods. Overall, this large-scale study on useful wild plant species in Mexico confirms that conventional seed banking can effectively support sustainable development and livelihood programmes.

Effects of pre-treatments and temperature on seed viability and germination of Juniperus macrocarpa Sm.

The effects of collecting season, collection site, laboratory pre-treatments and temperatures on seed viability and germination of Juniperus macrocarpa were investigated. Ripe cones were collected in four Sardinian dune systems, in two seasons, from plant and soil. Warm (W) and cold (C) stratifications, two combinations of them (W+C, C+W), and no pre-treatment (0) were applied. Seeds were incubated in a range of constant (10-25°C) and an alternating (25/10°C) temperature regime. Seed viability was low (ca. 40%) and varied significantly according to the collecting season. Seed germination was also low (ca. 10%), the 0 and W were the most effective pre-treatments on stimulating germination. The best germination temperature, without any pre-treatment, was 15°C (ca. 20%). J.xa0macrocarpa seeds are dormant and the achieved results suggested that the presence of secondary dormancy is induced by cold stratification. Spring appeared to be the best season for seed collecting, whereas autumn was the best for sowing. These results give new findings for restoration activities on this species.

Dissecting seed dormancy and germination in Aquilegia barbaricina, through thermal kinetics of embryo growth

Threshold-based thermal time models provide insight into the physiological switch from the dormant to the non-dormant germinating seed. This approach was used to quantify the different growth responses of the embryo of seeds purported to have morphophysiological dormancy (MPD) through the complex phases of dormancy release and germination. Aquilegia barbaricina seeds were incubated at constant temperatures (10-25xa0°C) and 25/10xa0°C, without pre-treatment, after warm+cold stratification (W+C) and GA3 treatment. Embryo growth was assessed and the time of testa and endosperm rupture scored. Base temperatures (Tb ) and thermal times for 50% (θ50 ) of embryo growth and seed germination were calculated. W+C enabled slow embryo growth. W+C and GA3 promoted rapid embryo growth and subsequent radicle emergence. The embryo internal growth base temperature (Tbe ) was ca. 5xa0°C for W+C and GA3 -treated seeds. GA3 treatment also resulted in similar Tb estimates for radicle emergence. The thermal times for embryo growth (θe50 ) and germination (θg50 ) were four- to six-fold longer in the presence of GA3 compared to W+C. A.xa0barbaricina is characterised by a multi-step seed germination. The slow embryo growth during W+C reflects continuation of the maternal programme of development, whilst the thermal kinetics of both embryo and radicle growth after the removal of physiological dormancy are distinctly different. The effects of W+C on the multiphasic germination response in MPD seeds are only partially mimicked by 250xa0mg·l-1 GA3 . The thermal time approach could be a valid tool to model thermal kinetics of embryo growth and radicle protrusion.

Conservation of indigenous plants to support community livelihoods: the MGU – Useful Plants Project

Through the MGU – Useful Plants Project (2007–2015) led by the Royal Botanic Gardens Kew, high-quality seed collections and research information have been gathered on useful indigenous plants selected by communities in Botswana, Kenya, Mali, South Africa, and Mexico. Local communities were trained in seed conservation, plant propagation, and planting activities, while revenue generation was promoted directly through the sustainable use of plants and plant products and indirectly through wider environmental and cultural services. The success of this project was determined by its participative approach, involving local communities in plant conservation activities and using plant research to support it. However, the promotion of sustainable use and income generation highlighted issues that confirmed the importance of using a ‘holistic approach’ to address the objective of poverty reduction and contribute to improved livelihoods in the communities.

Integration of genetic and seed fitness data to the conservation of isolated subpopulations of the Mediterranean plant Malcolmia littorea

Plant autoecology and population genetics provide a perspective on the likelihood of natural regeneration, which is critical when designing conservation strategies for endangered species. The threatened coastal plant Malcolmia littorea (Brassicaceae) was sampled across its European distribution and studied for genetic diversity and seed fitness, with the aim of providing information for the conservation of isolated and declining populations. Nine microsatellite markers (five chloroplast and four nuclear) were analysed to assess population genetic diversity and structure and to conduct a spatial analysis using the software DIVA-GIS. Germination percentages and rates were assessed by incubating the seeds under eight constant temperatures (0-27xa0°C). The genetic diversity was found to be similar among subpopulations (chloroplast Hxa0=xa00.04-0.17; nuclear Ho xa0=xa00.20-0.37), with no correlation between subpopulation diversity and the area of occupancy (AOO). The subpopulations were found to be clustered in three genetic groups, and three of them were identified as conservation priorities due to their unique genetic composition. The germination study revealed a significant influence of the maternal environment and AOO on seed germination, with the smaller subpopulations showing lower germination percentages (Pxa0<xa00.05). These results highlight the importance of obtaining information on isolated subpopulations through different experimental approaches (e.g. seed germination plus population genetics) to enable planning of effective conservation actions. For M.xa0littorea, seed collection for both inxa0situ and ex situ conservation should take into account the local adaptation of the subpopulation and the genetic structure of the species.

Morphological and functional seed traits of the wild medicinal plant Dioscorea strydomiana, the most threatened yam in the world

Morphological and functional seed traits have important roles in characterising the species regeneration niche and help to understand the reproductive biology of rare and threatened plants, which can thus support appropriate plant conservation measures. Seed morphometric and dispersal kinetics of the critically endangered Dioscorea strydomiana were measured and compared with those of four other Dioscorea species, and seed germination response under constant temperatures (5-35xa0°C) was compared with that of the congeneric and widespread D.xa0sylvatica. Seed mass of D.xa0strydomiana (ca. 14xa0mg) was twice that of D.xa0sylvatica, but similar to or smaller than the other species examined. Seeds of D.xa0strydomiana have the lowest speed of descent and lowest variability in most of the morphological traits considered, suggesting lower phenotypic plasticity but higher variance in the wing-loading value. Seeds of D.xa0strydomiana reached maximum germination at 15xa0°C (ca. 47%), which decreased slightly to ca. 37% at 25xa0°C and was completely inhibited at 35xa0°C. D.xa0sylvatica seeds started to germinate at 10xa0°C (ca. 3%), reached 75-80% germination at 15-20xa0°C and maximum (ca. 90%) at 25-30xa0°C. Base temperatures for germination (Tb ) were 9.3 and 5.7xa0°C, for D.xa0strydomiana and D.xa0sylvatica, respectively. Due to the higher germination percentages of D.xa0sylvatica, ceiling and optimum temperatures could also be modelled for this species, suggesting higher sensitivity to high temperature for seeds of D.xa0strydomiana. The detected poor seed lot quality of D.xa0strydomiana suggests difficulties in reproduction from seed, highlighting the need for further investigation and conservation actions for this threatened yam species.