Seed bank of Littorella uniflora (L.) Asch. in the Czech Republic,Central Europe: does burial depth and sediment type influence seed germination?

Littorella uniflora (Plantaginaceae) is a scarce and declining aquatic plant in Europe. Its population has been strongly reduced by changes in fishpond management (eutrophication) in the Czech Republic since the 1950s. We studied its seed bank in both recent (n = 8) and historical localities (the last found from 1972 to 2000; n = 10) and tested the effects of sediment type and burial depth on seed germination using extracted seeds from two recent populations. The seeds were found in 60% of the historical localities, mostly in low densities (≤6 seeds per 3.75 l of sediment), and also in 100% of the recent localities in various densities (8–1390 seeds per 3.75 l of sediment); however, low germination rates (0–13.3%) were estimated. The seeds germinated best on wet filter paper, followed by nutrient-rich fishpond sediment, but poorly on sand mixed with different substrates. Burial depth significantly affected seed germination. The seeds germinated only on the soil surface and at the depth of 1 cm but no seed germinated at the depth of 3 and 5 cm. Besides low water transparency, both high rate of sediment accumulation and the absence of summer drainage may endanger L. uniflora populations in fishponds.

     

In vitro mature embryo culture protocol of einkorn (Triticum monococcum ssp. monococcum) and bread (Triticum aestivum L.) wheat under boron stress

Mature embryos of einkorn (Triticum monococcum ssp. monococcum) and bread (Triticum aestivum L.) wheat were used for callus induction on media containing four different doses (0, 1, 2 and 4 mg L^?1) of 2,4-D and dicamba supplemented with five different boron concentrations (0, 6.2, 12.4, 24.8, and 37.2 mg L^?1). The obtained callus was transferred to culture media with three (0, 0.5, and 2 mg L^?1) different BAP doses with five boron concentrations for further regeneration. The maximum callus weight in einkorn wheat was in culture media with 1 mg L^?1 dicamba and 6.2 mg L^?1 (3.71?±?0.13 g). Bread wheat had the maximum callus weight on culture media with 4 mg L^?1 dicamba and 12.4 mg L^?1 (3.46?±?0.40 g). The highest plantlet numbers were in only 2 mg L^?1 BAP (2.92?±?0.88) for einkorn wheat and 0.5 mg L^?1 BAP supplemented with 6.2 mg L^?1 boron (3.71?±?1.12) for bread wheat. This indirect regeneration protocol using mature embryos of einkorn and bread wheat under boron stresses expected to be useful for future wheat breeding studies.

     

Osmoconditioning and Ageing of Pepper Seeds During Storage

The effects of osmoconditioning on the germination at 15 and25 °C of pepper (Capsicum annuum L.) seeds were studiedover a 3-year period with respect to temperature of storage.Untreated seeds stored at 5 °C showed high germinabilitythroughout the entire storage period, whereas untreated seedsstored at 25 °C showed a progressive decline in germinability,especially when assayed at 15 °C. Seeds that had been osmoconditionedprior to storage retained a high level of germinability irrespectiveof either storage or germination temperatures. When seeds thathad been stored at 25 °C were osmoconditioned after storage,there was a significantly higher germinability (assayed at 15 °C) in comparison with the corresponding untreated seeds.Seeds that were osmoconditioned twice (prior to and after storage)germinated in a similar way to those that had been osmoconditionedonce only Lactuca saliva L., lettuce, Hordeum oulgare L., barley, seed storage, moisture content, relative humidity, water potential, temperature, oxygen     

Comparison of seed and seedling functional traits in native Helianthus species and the crop H. annuus (sunflower)

• Seed functional traits of native Helianthus species contribute towards ecosystem services but limitations to their use in managed programmes exist. Many perennial Helianthus possess seed dormancy. The ability for germination to occur under different temperature and drought conditions, as well as the capacity of germinated seeds to convert into normal seedlings is rarely considered. Our aim was to identify and quantify these constraints through functional trait analyses.
• In five seed lots of native Helianthus (four perennial and one annual) and five genotypes of sunflower (H. annuus) for comparison, dormancy, thermal and hydro thresholds and times, morphology, mass, oil content and conversion into normal seedlings were quantified. The influence of the seed collection site environment on these traits was also explored.
• Seed dormancy of the perennial species was overcome by scarification followed by germination in 5 mm GA₃. Thermal and hydro‐time analyses revealed slower germination for the native seed lots (>1350 °Ch) in comparison to the sunflower genotypes (<829.9 °Ch). However, native seed lots had a higher capacity to convert into normal seedlings at high temperatures and low water potentials than sunflower genotypes. For the native seed lots, the average monthly temperature of the collection site was negatively correlated with thermal time.
• Variability in seed functional traits of native Helianthus and greater capacity for germinated seeds to convert into normal seedlings suggests they are better equipped to cope with high temperature and drought scenarios than sunflower. Effective dormancy alleviation is required to facilitate the use of native Helianthus species.

     

Morpho‐colorimetric analysis and seed germination of Brassica insularis Moris (Brassicaceae) populations

Brassica insularis is a perennial plant growing on both coastal and inland cliffs. Three seed lots from Sardinia were analysed using an image analysis system to detect differences in seed morphology, both within and among populations. Germination requirements at constant (5–25 °C) and alternating temperatures (25/10 °C), both in light and in darkness, were evaluated for all populations. In addition, the effect of a dry after‐ripening period (90 days at 25 °C) was also investigated. Morpho‐colorimetric analysis clearly identified seeds from different populations and discriminated three chromatic categories for seeds belonging to the Isola dei Cavoli coastal population, but not for the inland Masùa and the coastal Planu Sartu. Inter‐population variability was also observed in germination behaviour. B. insularis seeds germinated, with percentages up to 60%, in a wide range of temperatures (5–25 °C), and neither light nor dry after‐ripening affected final germination percentages. Moisture content measurements were made for seeds of each colour, but there were no particular differences among colours. Inter‐populational variability in germination behaviour may be a survival strategy for species growing under unpredictable environmental conditions, such as under Mediterranean climate, while heteromorphy may be due to independent evolutionary divergence processes of the Isola dei Cavoli population.     

Effects of salinity and nitrate on production and germination of dimorphic seeds applied both through the mother plant and exogenously during germination in Suaeda salsa

Salinity and nitrogen are two important environmental factors that affect the distribution of halophytes in their natural saline habitats. Seeds of the euhalophyte Suaeda salsa L. were harvested from plants that had been treated with 1 or 500 mm NaCl combined with 0.5 or 5 mm NO₃^?‐N (nitrate) for 115 days in a glasshouse. Germination was evaluated under different concentrations of NaCl and nitrate. Plants exposed to high salinity (500 mm ) and low nitrate (0.5 mm ) tended to produce heavy seeds. Either high salinity (500 mm ) or high nitrate (5 mm ) increased the brown/black seed ratio. The concentrations of Na⁺, K⁺, and Cl^? were higher in brown than in black seeds, and NO₃^? concentrations were higher in black than in brown seeds, regardless of NaCl and nitrate treatments during plant culture. Regardless of NaCl and nitrate concentrations during germination, seeds from plants grown with 0.5 mm nitrate generally germinated more rapidly than seeds from plants grown with 5 mm nitrate, and the difference was greater for black than for brown seeds. Exogenous nitrate during germination enhanced the germination of brown seeds less than that of black seeds. Producing more brown seeds and heavy black or brown seeds appears to be an adaptation of S. suaeda to saline environments. Producing more black seeds, which tend to remain dormant, should reduce competition for nitrogen and appears to be an adaptation to nitrogen‐limited environments. In conclusion, nitrate provided exogenously or by mother plants to black seeds may act as a signal molecule that enhances the germination of black S. suaeda seeds.     

Selection of somaclonal variants with low-temperature germinability in melon (Cucumis melo L.)

Summary Plants were regenerated from adventitious buds and somatic embryos (R₀) of melon (Cucumis melo L.), the cultivar Andes. Somaclonal variants of melon with low temperature germinability were selected from the progenies (R₁) of R₀ plants. Among 5,618 R₁ seeds harvested from 23 R₀ plants that were regenerated from adventitious buds 4 seeds germinated after 5 days of culture at 15 °C (selection rate; 0.07%). However, among 374 R₂ seeds harvested from 2 R₁ plants no seed germinated after 7 days of culture at 14 °C. Among 9,181 R₁ seeds harvested from 50 R₀ plants regenerated from somatic embryos 110 seeds germinated after 5 days of culture at 15 °C (selection rate; 1.20%). Among 3,717 R₂ seeds harvested from 17 R₁ plants 113 seeds germinated after 7 days of culture at 14 °C (selection rate; 3.04%). R₃ seeds were collected from these R₂ plants following self-pollination. Forty-five of the 47 lines (R₃) originated from 10 R₀ plants showed higher germination rates than that of the original cultivar. Selected lines with low-temperature germinability showed greater fruit growth rate than the original cultivar during the middle stage when they were cultivated in a greenhouse under low-temperature conditions. Of fruits harvested from 31 lines, 15 lines showed greater fruit volume than the original cultivar.     

Cryopreservation and low-temperature storage of seeds of Phaius tankervilleae

In this study we established reliable methods for conservation of seeds of Phaius tankervilleae as an orchid genetic resource. The seeds, which were dehydrated to 5% water content and preserved at 4°C, showed no decrease in viability and germinability after three months. After storage for six months, however, the seeds showed a drastic decrease in germinability, even though survival rate was high. For long-term preservation of seeds of P. tankervilleae, cryopreservation is applied to the freshly harvested seeds. When the seeds were cryopreserved by the vitrification method for up to 12 months there was no apparent deterioration effect of storage time. These results indicate that cryopreservation by the vitrification method is useful for long-term conservation of P. tankervilleae seeds, which are difficult to preserve for more than three months under dry and low-temperature conditions.     

Ecology of seed germination of eight non-pioneer tree species from a tropical seasonal rain forest in southwest China

We compared various aspects of the seed biology of eight non-pioneer tree species from a tropical seasonal rain forest in Xishuangbanna, SW China, that differ in time of dispersal, size and fresh seed moisture content (MC). Seeds were tested for germination under laboratory conditions after dehydration to different moisture levels and under 3.5, 10 and 30% solar irradiances in neutral-shade houses. For six species, germination was also compared in forest understory (3.5% light) and center of a forest gap (32.5% light). Under continuous dehydration over activated silica gel, 100% of seeds of four species had lost the ability to germinate after 48 h, and those of all species except Castanopsis hystrix (decreased from >90 to 30% germination) had lost the ability to germinate after 120 h. Four species did not differ in final germination percentages at the three irradiances (i.e. uniform germination). However, final germination percentages of Horsfieldia pandurifolia and Litsea pierrei var. szemaois were significantly lower in 30% than in 10 or 3.5% light, and seeds of Antiaris toxicaria and C. hystrix germinated to higher percentages in 30 and 10% than in 3.5% light. Mean time to germination (MTG) of the eight species (forest and shade house data combined) ranged from 5–5 days for Pometia tomentosa to 72–207days for L. pierrei; MTG for four species was ≤21 days. There was no obvious relationship between relative desiccation resistance and either time of dispersal, MTG or uniformity of germination at the three light levels, or between seed size and MC or MTG. However, the relationship between seed MC at maturity (25–60% fresh mass basis) and MC at 50% loss of seed viability (12.4–42.5%) was significant. Seven of the species fit Garwood’s (Ecol Monogr 53:159–181, 1983) rapid-rainy germination syndrome and one, L. pierrei, either her delayed-rainy or intermediate-dry germination syndrome. However, fresh, non-dehydrated seeds of all eight species germinated in ≤30 days at constant 30°C in light.     

Survival of Schismus arabicus seedlings exposed to desiccation depends on annual periodicity

Schismus arabicus, a desert annual grass, is one of the most common pasture annuals in the deserts of Israel and Asia. S. arabicus exhibits a unique set of adaptations and survival strategies, which enable it to germinate, develop and produce seeds even in years with annual rainfall of less than 100 mm. The current study examined whether an annual rhythm exists in the survival ability of S. arabicus seedlings exposed to desiccation. Our results indicate that survival of S. arabicus seedlings after six different periods of 7 to 42 days of desiccation depended on the month of germination of the caryopses (seeds). Seed germination was 80–100% in all experiments, regardless the month of germination; however, seedlings that germinated in different months varied in their root and shoot elongation rates. None of about 2,500 seedlings that germinated in July (in each of the 4 years) survived the desiccation treatment. The percentages of surviving seedlings in each month of June from 2002 to 2005 were less than 40%. In contrast, over 80% of the seedlings that germinated in each of the months of December and January survived after the desiccation periods of 7–42 days. Seedlings that survived were transferred to 5 L soil pots in which the seedlings developed into mature plants, completed their life cycle and produced seeds that germinated well. The current study demonstrated a novel phenomenon indicating that seedling survival in plants may depend on an annual periodicity according to the date of germination.     

Seed germination characteristics of Maianthemum dilatatum (Wood) Nels. et Macbr. (Asparagaceae)

The seed germination characteristics of Maianthemum dilatatum were investigated in a laboratory experiment and the results compared with those of other species in the subfamily Asparagoideae, LILIACEAE (Engler's system). M. dilatatum seeds mature in late September to October in montane to subalpine areas across Japan. Germination percentages and rates were low for fresh seeds at 10 to 30°C. Seeds cold stratified for 4 months or longer showed increased germination percentages and rates. The seeds lost germinability with decreasing moisture content. The seeds germinated well in dark conditions. The process of germination from the cotyledonary sheath/petiole breaking through the seed coat to the appearance of the first and second leaves was examined. After emergence of the cotyledonary sheath/petiole, a root emerged from it, and additional roots appeared after 1–2 months. The plumule emerged from the cotyledonary sheath/petiole after the seedling had three roots. Seeds dispersed in autumn, and germinate slowly in the next spring after exposure to low temperature even though dark condition as buried seed. We compared the seed germination characteristics among species in the Asparagoideae or with other recent taxonomy, and found that seedlings of Paris verticillate and Trillium apetalon, which belong to Melanthiaceae, and Streptopus streptopoides Var. japonica and Clintonia udensis, which belong to Liliaceae (linear cotyledon), were different from Asparagaceae, showing a globose cotyledon in the APG II.     

Germination ofRubus palmatus var.coptophyllus andR. microphyllus seeds buried in soil for 7.5 years

Seeds of twoRubus species,R. palmatus var.coptophyllus andR. microphyllus, buried for 7.5 years in soil were subjected to germination tests to investigate their germinability and germination traits. Most of the retrieved seeds were viable, and germinated at the alternating temperatures of 20/30°C in both light and dark. The twoRubus species showed similar responses of germination to temperature and light, although the final percentages of germination were slightly higher inR. palmatus var.coptophyllus. These characteristics of seed dormancy and germination would be involved in the species' utilization of ephemeral habitats created by unpredictable and infrequent disturbances.     

Diversity in seed germination behavior in relation to heterocarpy inSalsola komarovii Iljin

Seed germination behavior in two types of dimorphic fruits inSalsola komarovii Iljin (Chenopodiaceae) was studied. The germination percentage was much higher in the long-winged type than in the short-winged type when the fruits were tested for germinability. If lignified perianths were removed from the fruits, seeds of both types germinated at higher percentages and the difference in the germination percentage between two types decreased. The extract of the perianths inhibited the seed germination. Abscisic acid was detected in the extract of the perianths by enzyme-linked immunosorbent assay and its level was higher in the short-winged type than in the long-winged type. Germinability in both types was lost one year after harvest when the fruits were stored at room temperature and the germination was tested with fruits. When the seeds were tested, however, the short-winged type germinated even two years after the harvest; the germinability of the short-winged type lasted one year longer than that of the long-winged type. Light irradiation was not required for the seed germination in either type regardless of presence or absence of the perianths. No clear difference between the two types was observed either in optimal temperature for the germination or in strength of the resistance to salinity.     

Seed germination during floatation and seedling growth of Carapa guianensis, a tree from flood-prone forests of the Amazon

Carapa guianensis Aubl. (Meliaceae), a hard wood tree from the Brazilian Amazon, has large recalcitrant seeds that can germinate and establish in both flood-free (terra-firme) and flood-prone (várzea) forests. These seeds, although large, can float. This study was designed to experimentally examine seed longevity under floating conditions ex-situ and its effects on subsequent germination and seedling growth. Many seeds germinated while floating, and radicle protrusion occurred from 3 to 42 d after the start of the floating treatment (tap water, room temperature 20–30 °C). Shoots of newly germinated floating seedlings may elongate up to 37.0 cm in 20 d without loss of viability. Epicotyl and first leaf emergence were delayed by floating. Seeds that did not germinate while floating were then placed on vermiculite and watered daily, where many seeds resumed germination. Germination during and after floating was affected by the length of the floating treatment: 88% germinated after 1 mo, 82% germinated after 2 mo and 70% germinated after 2.5 mo. These results indicate that Carapa guianensis has physiological variation regarding dormancy in response to seed floatation. The fact that floatation induces dormancy in recalcitrant seeds of this economically important species can be relevant to initiatives of ex situ storage of seeds.     

Seed dispersal in the mycoheterotrophic orchid Yoania japonica: Further evidence for endozoochory by camel crickets

• Although orthopterans are rarely considered to be effective seed dispersal agents, the large flightless crickets known as ‘weta’ have been suggested to function as ecological replacements for small mammals in New Zealand, where such mammals are absent. In addition, a recent study reported that camel crickets mediate seed dispersal of several heterotrophic plants, including Yoania amagiensis in Japan.
• I investigated the seed dispersal mechanism of Yoania japonica because the fruit morphology is similar to Y. amagiensis. Specifically, I aimed to determine whether Y. japonica fruits are consumed by camel crickets and, if so, whether the seeds defecated by camel crickets remains intact, by checking seed viability with TTC staining, and whether germination rate is different between seeds collected directly from fruits and defecated seeds by comparing in situ seed germinability.
• The present study provides evidence that camel crickets function as seed dispersal agents of Y. japonica. Camel crickets were important consumers of Y. japonica fruits, and a substantial portion of the consumed seeds remained viable after passing through the digestive tract. In situ seed germination experiments revealed that the seeds defecated by camel crickets actually germinated in the field. In addition, the germination rate of defecated seeds was even higher than that of intact seeds, although the difference was not significant.
• Taken together with recent reports of insect‐mediated endozoochory, such a seed dispersal system may be common in plants with fleshy indehiscent fruits and small seeds, even in locations where other seed dispersal agents are present.

     

Overexpression of SoACLA-1 Gene Confers Drought Tolerance Improvement in Sugarcane

Drought is one of the most severe stresses which limit sugarcane production in China. ATP citrate lyase (ACL) is a major enzyme responsible for the production of acetyl-CoA in cytoplasm and plays an important role in plant metabolism and stress response. In this study, sugarcane ACL gene SoACLA-1 was cloned. The plant overexpression vector of SoACLA-1 was built and transformed into sugarcane calli by Agrobacterium-mediated transformation, and PCR analysis confirmed that SoACLA-1 gene had been stably present in the T0, T1, and T2 generations of the transgenic sugarcane. In order to evaluate the drought resistance of the transgenic lines and verify the function of SoACLA-1 gene in the transgenic sugarcane, T1 generation of the SoACLA-1 transgenic sugarcane lines was used as the material to investigate the physiological and biochemical characteristics at 0 day, 3 days, 6 days, and 9 days after water stress and rewatering for 3 days. Comprehensive evaluation of four indicators (chlorophyll, malondialdehyde, proline, soluble sugar) related to drought resistance was done with membership fuzzy function method. The results showed that the drought resistance of five transgenic sugarcane lines from strong to weak, in turn, was RT2?>?RT4?>?RT3?>?RT1?>?WT, and the recovery ability after drought, in turn, was RT1?>?RT2?>?RT4?>?RT3?>?WT. The T2 generation of the SoACLA-1 transgenic sugarcane lines was used to analyze the physiological and biochemical changes and the expression of drought-related genes under water stress. The results showed that the transgenic sugarcane lines were more tolerant to drought as compared with the wild-type plants. Our findings indicated that SoACLA-1 gene plays an important role as a positive factor in response to water stress, and overexpression of SoACLA-1 can enhance drought tolerance in transgenic sugarcane plants.

     

The impact of flooding and drought on seeds of Cnidium dubium, Gratiola officinalis, and Juncus atratus, three endangered perennial river corridor plants of Central European lowlands

The effects of flooding and drought on the post-flooding survival and germinability of seeds in the soil were investigated for three endangered river corridor angiosperms Cnidium dubium (Schkuhr) Thell., Gratiola officinalis L. and Juncus atratus Krocker. Freshly matured seeds were placed in small nylon bags and were buried in pots filled with soil collected from a wetland along the Lower Havel River. The pots were subjected to different winter/spring flooding treatments (30, 60, 120, and 180 days) and to drought for up to 3 years. Every year in spring, after drawdown, the seeds were exhumed, counted and tested for germination. Seed mortality of flooded seeds was low, except for C. dubium. Flooding did not affect germination of seeds of J. atratus. Flooding of seeds of C. dubium and G. officinalis had inconsistent, but no beneficial effects on germination of either. The year of harvest and the interannual variability of weather are found to be the main factors affecting germinability of C. dubium and G. officinalis in the course of the three consecutive years.The experiment showed that the capacity of the flooded seed bank to maintain germination ability and viability for extended periods of time could be directly related to the flooding duration of natural habitats of the three species. The three species are resilient to variable flooding. Seeds appeared desiccation tolerant, and single flooding events do not deplete their seed bank.     

The role of natural agents in the removal of coat-imposed dormancy in Dichrostachys cinerea (L.) Wight et Arn. seeds

Dichrostachys cinerea seeds are impermeable to water and do not germinate readily. The plant, however, contributes to a bush encroachment problem, minimising visibility in wildlife reserves and reducing the area available for grazing. Natural dormancy-breaking conditions must therefore exist, promoting the germinability of these normally dormant seeds. Diurnal temperature fluctuations (50°C/15°C), especially when combined with moisture were found to break dormancy. High temperatures, such as those generated by fire, destroyed seeds. Seeds ingested by herbivorous browsers such as nyala, became permeable. Seed burial for 48 weeks, was a more effective treatment than storage in promoting permeability. Distribution of D. cinerea will therefore be restricted to areas in which these conditions or factors occur. Seeds germinated optimally at 30C in the laboratory.     

Seed viability and seed dormancy of non-native Phragmites australis in suburbanized and forested watersheds of the Chesapeake Bay, USA

The non-native, invasive haplotype of Phragmites australis is rapidly invading tidal and non-tidal wetlands across North America. Phragmites has the potential to spread by seeds and rhizomes. Seed viability and dormancy differences were quantified among 18 patches of non-native Phragmites in subestuarine wetlands in developed (i.e., suburbanized) vs. forested watersheds of the Chesapeake Bay. We used tetrazolium and germination assays to assess seed viability and compared germination percentages and rate of germination among fresh seeds, cold–moist treated seeds, and warm–dry treated seeds to evaluate seed dormancy. Seed viability was <1% in most patches but a few patches produced abundant viable seeds (5–21%). Seed viability, however, did not differ significantly between wetlands in forested vs. developed watersheds. Contrary to studies of Phragmites seed dormancy in European populations, some Phragmites seeds were dormant at maturity; cold–moist treated seeds germinated faster and to higher percentages than fresh seeds or warm–dry treated seeds.     

Germinating seeds or bulbils in 87 of 113 tested Arctic species indicate potential for ex situ seed bank storage

Arctic plant species are expected to lose range due to climate change. One approach to preserve the genetic and species diversity for the future is to store propagules in seed vaults. However, germinability of seeds is assumed to be low for Arctic species. We evaluated ex situ storage potential of 113 of the 161 native angiosperms of Svalbard by studying seed ripening and germination. Seeds or bulbils were collected, and germinability was tested after one winter of storage in the Svalbard Global Seed Vault. Twenty-six of the species did not produce ripe propagules, 8 produced bulbils, and 79 produced seeds. Bulbils sprouted to high percentages. Seeds of 10 species did not germinate, 22 had low germination (<20 %), 34 had germination of 21–70 %, and 13 had high germination percentages (>70 %). More than 70 % of the species belonging to Asteraceae, Brassicaceae, Caryophyllaceae, Juncaceae, Rosaceae, and Saxifragaceae germinated. Cold tolerant, common species had higher germination percentages than relatively thermophilous, rare species. Germination percentages were six times higher than observed in 1969 (n = 51) and 0.7 times that observed in 2008 (n = 22), indicating that recent climate warming improves germination in the Arctic. While in situ conservation is of vital importance, ex situ conservation in seed banks is a potential complementary conservation strategy for the majority of Arctic vascular plant species. For species that did not germinate, other methods for ex situ conservation should be sought, for example, growing in botanical gardens.