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Background and Aims: Pollinator-limited seed-set in some terrestrial orchids is compensated for by the presence of long-lived flowers. This study tests the hypothesis that pollen from these insect-pollinated orchids should be desiccation tolerant and relatively long lived using four closely related UK terrestrial species; Anacamptis morio, Dactylorhiza fuchsii, D. maculata and Orchis mascula., Methods: Pollen from the four species was harvested from inflorescences and germinated in vitro, both immediately and also after drying to simulate interflower transit. Their tolerance to desiccation and short-term survival was additionally assessed after 3 d equilibration at a range of relative humidities (RHs), and related to constructed sorption isotherms (RH vs. moisture content, MC). Ageing of D. fuchsii pollen was further tested over 2 months against temperature and RH, and the resultant survival curves were subjected to probit analysis, and the distribution of pollen death in time (σ) was determined. The viability and siring ability, following artificial pollinations, were determined in D. fuchsii pollen following storage for 6 years at -20 °C., Key Results: The pollen from all four species exhibited systematic increases in germinability and desiccation tolerance as anthesis approached, and pollen from open flowers generally retained high germinability. Short-term storage revealed sensitivity to low RH, whilst optimum survival occurred at comparable RHs in all species. Similarly, estimated pollen life spans (σ) at differing temperatures were longest under the dry conditions. Despite a reduction in germination and seeds per capsule, long-term storage of D. fuchsii pollen did not impact on subsequent seed germination in vitro., Conclusions: Substantial pollen desiccation tolerance and life span of the four entomophilous orchids reflects a resilient survival strategy in response to unpredictable pollinator visitation, and presents an alternative approach to germplasm conservation., (© The Author 2014. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Seed deterioration is detrimental to plant germplasm conservation, and predicting seed germination and vigor with reliability and sensitivity means is urgently needed for practical problems. We investigated the link between hydrogen peroxide (H2O2) flux, oxygen influx and seed vigor of Caragana korshinskii by the non-invasive micro-test technique (NMT). Some related physiological and biochemical changes in seeds were also determined to further explain the changes in the molecular fluxes. The results showed that there was a good linear relationship between germination and H2O2 flux, and that O2 influx was more suitable for assessing seed vigor. H2O2 flux changed relatively little initially, mainly affected by antioxidants (APX, CAT and GSH) and H2O2 content; afterward, the efflux increased more and more rapidly due to high membrane permeability. With the damage of mitochondrial respiration and membrane integrity, O2 influx was gradually reduced. We propose that monitoring H2O2 and O2 fluxes by NMT may be a reliable and sensitive method to evaluate seed germination and vigor.

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.

Background and Aims: The importance of thermal thresholds for predicting seed dormancy release and germination timing under the present climate conditions and simulated climate change scenarios was investigated. In particular, Vitis vinifera subsp. sylvestris was investigated in four Sardinian populations over the full altitudinal range of the species (from approx. 100 to 800 m a.s.l)., Methods: Dried and fresh seeds from each population were incubated in the light at a range of temperatures (10-25 and 25/10 °C), without any pre-treatment and after a warm (3 months at 25 °C) or a cold (3 months at 5 °C) stratification. A thermal time approach was then applied to the germination results for dried seeds and the seed responses were modelled according to the present climate conditions and two simulated scenarios of the Intergovernmental Panel on Climate Change (IPCC): B1 (+1·8 °C) and A2 (+3·4 °C)., Key Results: Cold stratification released physiological dormancy, while very few seeds germinated without treatments or after warm stratification. Fresh, cold-stratified seeds germinated significantly better (>80 %) at temperatures ≥20 °C than at lower temperatures. A base temperature for germination (T(b)) of 9·0-11·3 °C and a thermal time requirement for 50 % of germination (θ(50)) ranging from 33·6 °Cd to 68·6 °Cd were identified for non-dormant cold-stratified seeds, depending on the populations. This complex combination of thermal requirements for dormancy release and germination allowed prediction of field emergence from March to May under the present climatic conditions for the investigated populations., Conclusions: The thermal thresholds for seed germination identified in this study (T(b) and θ(50)) explained the differences in seed germination detected among populations. Under the two simulated IPCC scenarios, an altitude-related risk from climate warming is identified, with lowland populations being more threatened due to a compromised seed dormancy release and a narrowed seed germination window.

The maximal potential desiccation tolerance (MPDT) of tea (Camellia sinensis) seeds has been a matter of debate for decades. Here we assessed the ability of tea seeds from three sites in China to germinate after desiccation. Desiccation tolerance was greatest in Kunming, followed by Puer and Lincang, with Kunming seeds tolerating drying to 8% moisture content (MC), or ∼0.5 water activity (a(w)). Such tolerance was observed in Lincang seeds only when hydrogen peroxide (H₂O₂) at 0.5 or 1M was applied to seeds, indicating a stimulatory role for H₂O₂ in post-desiccation germination. Puer seeds exhibited MPDT of 16% MC (∼0.7 a(w)). Therefore, seeds from all three sites were not recalcitrant. The length of the dry season after dispersal and the high ratio of seed coat to seed mass (>0.3) support the observation of non-recalcitrant behaviour. The seeds were not immature, as the lipid signal in embryonic axes mirrored that of the cotyledons (30% oil). Even after high survival [>60% total germination (TG)] on drying to 10-13% MC, no Kunming seeds tolerated 1 month storage at -20 °C coinciding with lipid transitional changes at this temperature. The results indicate that tea seeds from China are neither recalcitrant nor storable at -20 °C., (Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.)

Ex situ seed storage underpins global agriculture and food supplies and enables the conservation of thousands of wild species of plants within national and international facilities. As an insurance policy against extinction, ex situ seed conservation is estimated to cost as little as 1% of in situ conservation. The assumptions, costs, risks and scientific challenges associated with ex situ plant conservation depend on the species, the methods employed and the desired storage time. Recent, relatively widespread evidence of less than expected longevity at conventional seed bank temperatures, innovations in the cryopreservation of recalcitrant-seeded species and economic comparators provide compelling evidence that ultra-cold storage should be adopted for the long-term conservation of plants. Policy instruments, such as the Global Strategy for Plant Conservation (2011-2020), should respond to the evidence base and promote the implementation of cryopreservation for both tropical and temperate plants.

The effects of fruit maturity, at the time of natural dispersal, on subsequent desiccation tolerance and sub-zero storage was investigated in three lots of Acer pseudoplatanus (sycamore) collected from northern to southern Europe. Fruits from the native plant distribution range in Italy had significantly higher desiccation tolerance (0.16 g water per g DW) than those from England (0.30) and Norway (0.50), confirming that the maximum potential desiccation tolerance in sycamore exceeds that of the recalcitrant type. In contrast, the unfrozen water content varied only slightly between seedlots, but systematically reduced with development (0.35 to 0.27 g water per g DW). Maximum survival (60 percent fruit germination) of seven days sub-zero temperature storage coincided with drying the Italian fruit lot to c. 0.2 g water per g DW followed by holding at -20 degree C, above the onset temperature for freezing, or at -196 degree C (liquid nitrogen). Fruit survival was much lower in the Italian fruits when held at this water content and -70 degree C, and in all other combinations of water content, temperature and fruit lot provenance. As the risk of nucleation in partially dried fruits held at -20 degree C is high, we recommend sycamore fruits are cryopreserved for long-term conservation.

In the tropics, species with recalcitrant or desiccation-sensitive, Type III seeds are largely restricted to regions with comparatively high rainfall, because desiccation-induced seed death will be minimal in these environments. However, species with recalcitrant seeds do occur in drylands, although little is known about ecological adaptations to minimize seed death in these environments. Here we present data for the seed desiccation tolerance of 10 African dryland species and examine the relationships between seed size, rainfall at the time of seed shed, and desiccation tolerance for these and a further 70 species from the scientific literature. The combined data set encompasses species from 33 families. Three species (Syzygium cumini, Trichilia emetica, and Vitellaria paradoxa) had desiccation-sensitive seeds, and the remaining seven species investigated were desiccation-tolerant. The desiccation-sensitive species had large (>0.5 g) seeds, germinated rapidly, and had comparatively small investments in seed physical defenses. Furthermore, seed was shed in months of high rainfall (>60 mm). In comparison, for species with desiccation-tolerant seeds, seed mass varied across five orders of magnitude, and seed was shed in wet and dry months. Although infrequent in dryland environments (approximately 11% of the species examined here), species with desiccation-sensitive seeds do occur; large size, rapid germination, and the timing of dispersal all reduce the likelihood of seed drying. Furthermore, desiccation-sensitivity may be advantageous for large-seeded species by increasing the efficiency of resource use in seed provisioning.

The effects of desiccation and storage temperature on the viability and longevity of willow seeds was investigated using two hybrids, Salix rehderiana x (Salix x capreola) [cross 458] and Salix x sericans x Salix viminalis [cross 512]. Freshly harvested seed of both crosses survived silica gel drying down to c. 3 to 5% moisture content. Hybrid 458 seed stored in liquid nitrogen (-196C) for 3 d retained viability when equilibrated to < or = 45% RH (pre-storage), showed slightly reduced survival at 65% RH and exhibited no survival at > or =82% RH. The level of survival after 68 d for seeds pre-equilibrated to either c. 10 or 65% (5 or 10% moisture) and stored at four temperatures was -196C > -20C > 2C > 16C. At all temperatures, drier seed stored better than wetter seed. For hybrid 512, seed longevity at 20C > 40C > 60C, and a 10% fall in pre-storage seed RH resulted in a c. 2-fold increase in longevity at each storage temperature. The response of hybrid willow seeds to desiccation and cooling raises possibilities for the long-term seed conservation of Salix species by cryopreservation.