Soil seed bank and seed dormancy in wild populations of lima bean (Fabaceae): considerations for in situ and ex situ conservation

Seed dormancy and its impact on the soil seed bank for wild Lima bean (Phaseolus lunatus) populations were studied in the Central Valley of Costa Rica. Five populations were selected in contrasted environments. In all cases, distribution of seeds in the soil was limited to 3 cm depth. No innate dormancy was observed but combination of hard seed coat and hilum opening controlled by environmental conditions were responsible for an induced dormancy and the constitution of a persistent seed bank. Breaking of this dormancy was obtained by a brief elevation of temperature from 25° to 45°C. Impacts of this phenomenon concern both genetic and demographic aspects of in situ conservation of the species. Consequences on ex situ conservation are mainly related with the regeneration of the seed collection.     

Dormancy,germination and seed bank storage: a study in support of ex situ conservation of macrophytes of southwest Australian temporary pools

1. Vernal pools and rock pools (gnammas) in the Southwest Australian Floristic Region are forms of temporary wetlands that are under threat. Some of their aquatic macrophytes are rare and/or endemic, and there is a need to develop off‐site seed banks to assure their conservation. Here, we report results of the first comprehensive study of the seed germination, dormancy and seed storage behaviour of nine indigenous macrophyte species. 2. Seeds of Glossostigma drummondii, Myriophyllum balladoniense, M. lapidicola, M. petreaum and Triglochin linearis were non‐dormant, whereas those of Damasonium minus, Glossostigma sp. (currently undescribed), G. trichodes and Myriophyllum crispatum were dormant. Non‐dormant seeds germinated over a range of temperatures (5–20 °C) but temperatures at which highest germination occurred were species specific. All species demonstrated a germination preference for the light. Warm stratification substantially increased germination of dormant Glossostigma trichodes seeds and all dormancy‐breaking treatments partially overcame dormancy in Glossostigma spp. 3. Seeds possessed orthodox storage behaviour (tolerating drying to 5% moisture content and storage at ?18 °C) and are thus amenable to seed banking as a means of ex situ conservation. 4. It appears that seeds of most species are able to germinate upon inundation as long as they are situated at the soil surface. Thus, species are opportunistic and respond to the first rains of the season providing prompt ecological cuing in an environment vulnerable to rapid drying events. Maintaining the integrity of the soil crust may be an important first step for on‐site conservation if seeds are in the superficial layers.     

Restoration of ditch bank plant species richness: The potential of the soil seed bank

Abstract. Small‐scale landscape elements, such as ditch banks, play an important role in preserving plant species richness in agricultural landscapes. In this study, we investigate whether the seed bank might be useful for restoring the above‐ground plant species richness. We studied the vegetation and seed bank composition at six species‐rich and six species‐poor ditch banks, where agri‐environment schemes are running to maintain and enhance ditch bank plant diversity. We show that the number of species in the seed bank was low, regardless of the number of species in the established vegetation. Moreover, the seed bank was always dissimilar to the established vegetation. Target species for nature conservation were occasionally present in the seed bank at both species‐poor and species‐rich sites, but rarely so if the species was absent from the established vegetation. We conclude that the potential use of the seed bank for restoration of ditch banks is minimal. At present, plant species richness seems to be largely controlled by germination opportunities; high biomass and competition appear to hamper germination at species‐poor sites. We recommend continued nutrient reduction at such sites. Soil disturbance measures and deliberate sowing should also be considered.     

Systematic overestimation of Salicaceae seed survival using radicle emergence in response to drying and storage: implications for ex situ seed banking

Biodiversity conservation programmes are underpinned by seed banking following drying to low water contents (WC), and supported by both the assessment and prediction of seed viability over time. The means of judging viability is thus crucial to the comprehension of seed vigour. We selected seeds of three species and one hybrid in the Salicaceae likely to have variation in tolerance to drying, processing and storage, including in relation to cryobanking, and compared survival growth as radicle emergence (germination) and normal seedling production. With three seed lots of Salix gracilistyla, air-drying to 8–10 % WC enhanced seed survival after 40 days’ storage at 5 °C as compared with non-treated seeds at 14–20 % WC. Four seed lots of Populus alba × P. glandulosa showed equally high germination (88–100 %) and proportions of normal seedlings (81–99 %) when stored at 5 °C for 7–10 weeks. Among seven seed lots of S. gracilistyla, two groups with different storage behaviour could be statistically distinguished with normal seedling production ranging from 0 to 45 % after storage at 5 °C for 13 weeks. Seed tolerance to WC manipulation and cryopreservation was very variable among species and seed lots. Seed lots of S. hallaisanensis and S. gracilistyla with ~80 % germination survived cryopreservation at 10 % WC, but were sensitive to lower WCs. In contrast, Populus seeds had greater desiccation tolerance combined with cryopreservation capability. With seed lots of all species and hybrids, cryopreservation had little effect on viability unless the high moisture freezing limit had been exceeded (~10–20 % WC, depending on seed lot). However, under all conditions of handling (drying, rehydration, storage at 5 °C or cryopreservation) using germination as the only indicator of viability over-estimated survival compared with normal seedling production.     

Ecological correlates of ex situ seed longevity: a comparative study on 195 species

Background and Aims

Extended seed longevity in the dry state is the basis for the ex situ conservation of ‘orthodox’ seeds. However, even under identical storage conditions there is wide variation in seed life-span between species. Here, the effects of seed traits and environmental conditions at the site of collection on seed longevity is explored for195 wild species from 71 families from environments ranging from cold deserts to tropical forests.

Methods

Seeds were rapidly aged at elevated temperature and relative humidity (either 45°C and 60% RH or 60°C and 60% RH) and regularly sampled for germination. The time taken in storage for viability to fall to 50% (p₅₀) was determined using Probit analysis and used as a measure of relative seed longevity between species.

Key Results

Across species, p₅₀ at 45°C and 60% RH varied from 0·1 d to 771 d. Endospermic seeds were, in general, shorter lived than non-endospermic seeds and seeds from hot, dry environments were longer lived than those from cool, wet conditions. These relationships remained significant when controlling for the effects of phylogenetic relatedness using phylogenetically independent contrasts. Seed mass and oil content were not correlated with p₅₀.

Conclusions

The data suggest that the endospermic seeds of early angiosperms which evolved in forest understorey habitats are short-lived. Extended longevity presumably evolved as a response to climatic change or the invasion of drier areas. The apparent short-lived nature of endospermic seeds from cool wet environments may have implications for re-collection and re-testing strategies in ex situ conservation.Key words: Gene banks, seed ageing, seed longevity, taxonomic trends, climate, seed structure     

Potential for seed bank formation in seven Great Lakes sand dune species

Seeds of seven dune species were collected from sand dunes of Lakes Erie and Huron and buried to various depths in a natural sand dune habitat along Lake Huron. The seed samples were then retrieved after varying lengths of time and examined for their germinability and dormancy. Results showed that buried seeds remained viable for at least 2.5 years and had the potential to form a persistent seed bank. Seed banks were larger and longer lasting at greater depths of burial than those at shallow burial depths. The results suggested that failure to verify the existence of effective seed banks in previous studies may be due to insufficient number of samples, shallow sample depth, local population variations, and fruiting events. Several species also possessed a temporary, aboveground seed reserve formed by retention of a small proportion of viable seeds on the previous year's inflorescences. In some species, seeds retained aboveground were dormant and thus capable of forming a persistent seed bank when they entered the soil.     

Soil nematode assemblages indicate the potential for biological regulation of pest species

In concept, regulation or suppression of target nematode pest species should be enhanced when an abundance of predator species is supported by ample availability of bacterial- fungal- and non-damaging plant-feeding prey species. We selected soils from natural and managed environments that represented different levels of resource availability and disturbance. In microcosm chambers of each soil, in its natural state or after heat defaunation, we introduced test prey species not already resident in the soils (Meloidogyne incognita and Steinernema feltiae). Survival of the test prey was determined after a 5-day bioassay exposure. Across the soils tested, predator abundance and biomass were greater in undisturbed soils with plentiful resources and lower in soils from agricultural sites. Suppressiveness to the two introduced species increased with both numerical abundance and metabolic footprint of the predator assemblages. The magnitude of the increase in suppressiveness was greater at low numbers of predators then dampened to an asymptotic level at greater predator abundance, possibly determined by temporal and spatial aspects of the bioassay system and/or satiation of the predators. The more resource-limited the predators were and the higher the metabolic predator footprint, the greater the suppressiveness. The applied implications of this study are that soil suppressiveness to pest species may be enhanced by increasing resources to predators, removing chemical and physical constraints to their survival and increase, and altering management practices so that predators and target prey are co-located in time and space.     

Physiological aspects of Taxus brevifolia seeds in relation to seed storage characteristics

Water relations, desiccation tolerance and longevity of Taxus brevifolia (Nutt.) seeds were studied to determine the optimal stage of development and storage conditions for seeds of this species. Seeds equilibrated to a range of relative humidities (RHs) had unusually low water contents which can be accounted for by the high lipid content of gametophyte tissues (71% of the dry mass). Water relations of embryonic tissue were more typical of those reported for other seed species. The water content below which freezing transitions were not observable in the embryo was ca 0.24 g H₂O (g dry weight)⁻¹ (g g⁻¹) for all maturity classes studied. Embryos did not achieve significant levels of desiccation tolerance (survival to water contents less than 0.5 g g⁻¹) until the latter stages of development when dry matter was maximal. Mature embryos could be dried to 0.025 g g⁻¹ (seed water content of 0.010 g g⁻¹) with no loss of viability. Thus, at the latter stages of development, embryo water content could be optimized to avoid both desiccation and freezing damage. Survival of mature seeds declined over a 2-year period when seeds were stored at temperatures between 5 and 35°C and RHs between 14 and 75%, corresponding to seed water contents between 0.015 and 0.07 g g⁻¹. The deterioration rate was slowest for seeds stored at the lowest RH and temperature. Our data indicate that seeds of Taxus brevifolia show orthodox rather than recalcitrant storage characteristics, but that the optimum water content for storage was extremely low. The results suggest that even if stored at optimal water contents and low temperatures, T. brevifolia seeds will be relatively short lived. The high quantity of lipids or reducing sugars may be contributing factors in the poor storage characteristics.     

Effectiveness of dry heat as a seed sterilisation technique: Implications for ex situ conservation

Seed-borne pathogens are a daily issue for ex situ collection managers who try to solve it using various chemicals more or less harmful to the staff and the plant material stored. The most common physical method for seed sterilisation is moist heat, in contrast to dry heat which is used much less frequently. Consequently, the efficiency of dry heat as disinfection method and the behaviour of seeds undergoing this treatment are currently poorly known, especially for wild species. In this study, seed viability was estimated by performing germination tests according to standard procedures. Germination tests were conducted on 13,200 seeds from 66 wild species of temperate regions belonging to 22 different families. Results indicate that dry seeds exposed to 60°C during 1hour were less infected by seed-borne pathogens in 14% of the cases, whereas no change has been registered in the other cases. For all 66 studied species, no decrease in germination percentage was detected after the heat treatment. Given its positive effect on infection control without affecting seed viability, dry heat treatment as proposed here opens opportunities for seed bank managers, but also for the disinfection of herbarium collections.     

Effective seed harvesting strategies for the ex situ genetic diversity conservation of rare tropical tree populations

Tropical rain forests harbor a high diversity of tree species, a large portion of which are considered rare and threatened by anthropogenic factors such as land-use change. Addressing the global need for ex situ rescue populations of such species, we investigated whether certain methods of wild-source seed collection harvest greater amounts of genetic variation from the source population while minimizing the costs expended. We used an individual-based computer program (NEWGARDEN) to conduct simulation trials examining which aspects of seed procurement provided the most cost-effective methods for maximal rare allele capture for single isolated populations of rare tropical tree species. Aspects examined included the geometric pattern of harvesting, location of seed collections within the spatial distribution of the population (central vs. peripheral), geometric partitioning of sampling regions (contiguous vs. subdivided), gene dispersal distance, linear travelling costs, and ratio of seeds collected to plants sampled. Results demonstrated that ratio of seeds collected to plants sampled had the highest effect on genetic diversity harvested relative to sampling location or geometric pattern of collection, explaining 82 and 79% of the variance in alleles harvested, respectively. Collecting from subdivided peripheral sampling regions and populations having long distance gene dispersal led to increases of up to 39 and 51% in the amount of genetic variation conserved respectively. These results suggest that ex situ conservation efforts for rare tropical trees can benefit by collecting fewer seeds from each of a larger number of trees, sampling from peripheral subdivided regions of the population rather than in a contiguous centrally located one under the transect or nearest neighbor strategies, and adjusting harvesting based on the dispersal biology of the source population. This study underscores the need for more information regarding the dispersal characteristics of the target species and demonstrates the value of predictive modeling for planning seed collection projects.     

Metal species indicate the potential of constructed wetlands for long-term treatment of metal mine drainage

Experimental peat-based wetlands at the former Bell Copper Mine (Smithers, B.C., Canada) removed copper from low strength (0.3–1.0 ppm, pH 6–8) and high strength (35–50 ppm, pH 3) copper-contaminated mine drainage. Copper species retained in wetlands sediments were quantified as a means to assess the long-term potential of these wetlands for treating mine drainage. Previous microbial and chemical analyses had indicated that sulphide was biologically generated in these wetlands, and mineralogical analysis confirmed that copper sulphides were present in wetland sediments. However, sequential leaching of these sediments showed that copper was predominantly in organically-bound and oxide-bound phases, with a smaller proportion present as sulphides. The available data suggest that much of the copper was retained as sulphides when low strength mine drainage was applied to the wetlands.     

Reproductive status of endemic felid species in Latin American zoos and implications for ex situ conservation

Reproductive evaluations were conducted on 185 male cats representing eight endemic Latin American species that were maintained in 44 zoos and private facilities in 12 Latin American countries. Reproductive assessments (testicular measures, ejaculate quality, and blood testosterone/cortisol concentration) were used to establish normative values for large‐ and small‐sized cats in Latin American collections. Data also were analyzed using multiple regression to study the impact of proven breeder status, diet, and various animal housing combinations. Most felids (>95%) in the survey were of wild‐born origin, and <20% had produced offspring in captivity. Larger felids had bigger testes and produced more semen, but tended to produce low‐sperm‐density ejaculates. The ejaculates of small felids were more sperm‐concentrated, but contained fewer total spermatozoa. Sperm motility was unrelated to species size, and certain species (puma, margay, tigrina, and jaguarundi) consistently produced few (<40%) normal sperm forms. Across species, >50% of males had low sperm counts (<1 million total sperm per ejaculate). Among large cats (jaguars and pumas), proven breeders had larger (P<0.05) testes, greater semen volume, and more normal sperm than nonbreeders. Males on adequate diets had higher (P<0.05) circulating cortisol. Among small‐sized felids, proven breeders had higher (P<0.05) testosterone, and males housed alone or paired with a conspecific female had more (P<0.05) total sperm per ejaculate and greater (P<0.05) seminal and testicular volumes. Fifty‐nine ejaculates (potentially representing ～100 artificial insemination (AI) or 26,000 in vitro fertilization (IVF) procedures) were cryopreserved for a felid genome resource bank. In conclusion, breeding success and reproductive traits for many endemic felids in Latin American zoos appear to be suboptimal, and likely would benefit from improvements in diet and exhibitry. Technology transfer and continued training of zoo staff and scientists in Latin American countries are essential if these zoos are to achieve their tremendous conservation potential for felids and other threatened endemic species. Zoo Biol 22:421–441, 2003. © 2003 Wiley‐Liss, Inc.     

Restoration of high species density in calcareous grassland: the role of seed rain and soil seed bank

Abstract. A small-scale restoration experiment concerning a species-rich calcareous grassland was conducted over a 7-yr period. 20 yr of abandonment (1971-1990) resulted in both a strong reduction of the number of species and an absolute dominance of the shrub Crataegus monogyna and the grass Brachypodium pinnatum - which contributed 98% of the above-ground herb phytomass. The plot (3 m x 6 m) was cleared in September 1990 and thereafter yearly mown in autumn. Small soil seed bank samples were taken immediately after clearing and again in the autumn of 1996. The inner part of the plot showed a reduced soil seed bank compared to the plot underneath the vegetation outside the plot in 1990. Six years later an increased number of seeds in the soil in this inside area could be recorded mainly in the upper soil layer, although the amount was only 40% compared to the outside or control area. Ca. 20% of the species present in the restored vegetation were recorded in seed traps during the main seed dispersal period in 1997. Restoration of high species richness in the plot was a fast process during the first few years: from seven species in 1990 towards 39 three years later. This was significantly faster than the restoration process of the calcareous grassland at the same slope 20 yr earlier. This was probably due to the increased size of the community species pool at the site as a result of appropriate management during the last decades. Relative species number increase was highest at the largest scale during this experiment, viz. at the total plot area. The success of restoration of species-rich, dry grassland vegetation largely depends on the interaction between several conditions, e.g. seed availability, either originating from the seed rain or soil seed bank, favourable germination conditions and a successful development to a mature, seed producing individual. Germination and establishment strongly depend on suitable vegetation structure as a result of sustainable management.     

Seasonal and spatial variance of seed bank species composition in an oligotrophic wet meadow

Seasonal and small-scale spatial variation of the seed bank in an oligotrophic wet meadow (Molinion) was assessed by two methods: (1) direct determination of seeds extracted from soil samples (6 cm in diameter, 10 cm depth) and (2) germination of seeds according to Ter Heerdt et al. [1996. An improved method for seed bank analysis: seedling emergence after removing the soil by sieving. Funct. Ecol. 10, 144–151]. Samples were taken three times during the year (July and November 2001, March 2002) from three distinct microhabitats (moss layer, gaps, Molinia caerulea tussocks). Using the extraction method, more seeds were found in the upper 0–3 cm soil layer than in the deeper 3–7 cm layer, significant differences in the number of seeds were found among microhabitats (with the lowest seed bank densities in Molinia tussocks). The highest number of seeds was found in the spring census, but the test was not significant (P<0.10). However, seasonal variation was found when damaged seeds were included in the analysis. Changes in damaged/undamaged and light/heavy seed ratios, and in numbers of grass/herbs/sedges seeds are also discussed. The spatial distribution of seeds is probably influenced to a great degree by abiotic factors, such as microtopography (round and heavy seeds of sedges prevail in gaps and in deeper soil layers). The species composition of the seed bank varied among microhabitats, depth, and throughout the year. No differences were found by the seed germination method.     

Comparative study of storage compound breakdown in germinating seeds of three lupine species

Storage lipid and protein breakdown in germinating seeds of yellow (Lupinus luteus L.), white (L. albus L.), and Andean lupine (L. mutabilis Sweet) and regulatory function of sucrose were investigated. Less oil bodies were detected in organs of yellow lupine seeds, whereas the highest content of oil bodies was noticed in the Andean lupine seeds. Mature, air-dried yellow, white and Andean lupine seeds do not contain starch. Starch grains appear the earliest in white lupine seeds during imbibition. Sucrose deficiency in tissues enhances breakdown of storage lipid, protein and temporary starch in cotyledons. In sucrose starved embryo axes of all investigated lupine species, an increased level of vacuolization was noted. Interconnections between catabolism of storage protein and storage lipid in germinating lupine seeds were identified by applying ¹⁴C-acetate. To assess the importance of key processes in storage lipid breakdown NaF (inhibitor of glycolysis and gluconeogenesis), KCN, NaN₃ and SHAM (inhibitors of mitochondrial electron transport chain) and MSO (inhibitor of glutamine synthetase) were used. Radioactivity coming from ¹⁴C-acetate was released as ¹⁴CO₂ but mostly was incorporated into ethanol-soluble fraction of embryo axes and cotyledons. Respiratory inhibitors caused a significant decrease in ¹⁴CO₂ and ethanol fractions in all three lupine species studied. MSO stimulated release of ¹⁴CO₂ and radioactivity of ethanol fractions in yellow lupine organs fed with sucrose, but in Andean lupine MSO enhanced the production of ¹⁴CO₂ and radioactivity of ethanol fractions both in organs fed and not fed with sucrose. Different strategies of storage compound breakdown are proposed, depending on relative proportion in storage protein and lipid content in lupine seeds.     

Effects of an antisense napin gene on seed storage compounds in transgenic Brassica napus seeds

To manipulate the quantity and quality of storage components in Brassica napus seeds, we have constructed an antisense gene for the storage protein napin. The antisense gene was driven by the 5-flanking region of the B. napus napin gene to express antisense RNA in a seed-specific manner. Seeds of transgenic plants with antisense genes often contained reduced amounts of napin. In some transgenic plants, no accumulation of napin was observed. However, the total protein content of transgenic and wild-type seeds did not differ significantly. Seeds lacking napin accumulated 1.4 to 1.5 times more cruciferin than untransformed seeds, although the oleosin content was not affected. Fatty acid content and composition in the seeds of transgenic plants were also analyzed by gas chromatography. Though the total fatty acid content of the transformants was the same as that of non-transformants, there was a reduction in 18:1 contents and a concomitant increase of 18:2 in seeds with reduced napin levels. This observed change in fatty acid composition was inherited in the next generation.     

岷江干旱河谷灌丛土壤种子库及其自然更新潜力评估

为了解岷江干旱河谷区灌丛植被土壤种子库的特征及预测灌丛植被自然更新的潜力,利用土壤种子库"萌发法"(每个地段15个2m×2m样方内分土表层、0-5cm土层和5-10cm土层采集土样)及野外植被调查方法(每个地段15个1m×1m的调查样方),研究了岷江干旱河谷3个地段(河谷下游边缘汶川地段、核心茂县地段以及上游边缘松潘地段)阳坡灌丛植被土壤种子库及其与地上植物群落的关系。发现:土壤种子库发芽试验共观察到4274株幼苗,分属于113个物种。土壤种子库密度在102—103ind./m2,物种数在12—23种/0.06m2,边缘地段的土壤种子库平均种子密度(ind./m2)和平均物种数(种/0.06m2)均比中心地段(茂县)高。3个地段土壤种子库和地上植被的组成物种均多为多年生植物,地上灌丛与其土壤种子库的密度及物种数均呈不显著相关性;物种组成的Sorensen相似性指数较低,不足30%,但土壤种子库均比地上植被具有更高的物种丰富度。综合分析表明,岷江干旱河谷灌丛具有依靠土壤种子库实现自然更新的潜力,但由于物种组成相似性较低,现存灌丛植被一旦遭到破坏后,单依靠土壤种子库自然恢复到现存植被是困难的,需要在自然恢复潜力评估基础上积极充分采取人工促进自然恢复的策略,同时也应充分考虑干旱河谷植被及其自然更新潜力的空间异质性。探明岷江干旱河谷区灌丛植被的土壤种子库特征,是认识灌丛植被自然更新潜力的基础,是合理制订该区植被恢复与管理措施的重要依据。     

Digging deep for diversity: riparian seed bank abundance and species richness in relation to burial depth

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