Effect of gibberellic acid- and water-based pre-soaking treatments under different temperatures and photoperiods on the seed germination of Allium stracheyi Baker: An endangered alpine species of Central Himalaya,India

Allium stracheyi Baker (Alliaceae, 2600–3000 m asl), an endangered species of Central Himalaya, India, has low seed germination in its natural habitat. This study is an attempt to improve seed germination by determining the seed viability with a low mean germination time (MGT) and germination index (GI) under optimum temperature, light, and pre-soaking treatments. The seeds were pre-soaked in hot water (80°C), cold water (10°C), and gibberellic acid (GA₃ at 50 and 100 mg/l) for 24 h and subjected to light (12 h light and 12 h dark) and continuous dark (24 h) conditions with different temperature regimes (10, 15, 20, 25, and 30°C). The viability varied between 66.0% and 69.67% and declined rapidly after 12 months of storage. Our studies suggest that the 100 mg/l GA₃ treatment was beneficial for seed germination and seedling growth. Pre-soaking in a 100 mg/l GA₃ solution and incubation at 20°C under light conditions enhanced the germination significantly (p < 0.05) and resulted in the highest (97.3%) germination with the lowest MGT = 5.7 days, with GI = 8.11. The recommendations of this study support the conservation of alpine A. stracheyi via simple and cost-effective techniques for optimal seed germination.     

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.     

Response of Amaranthus retroflexus L. seeds to gibberellic acid, ethylene and abscisic acid depending on duration of stratification and burial

Freshly harvested, dormant seeds of Amaranthus retroflexus were unable to germinate at 25 and 35 °C. To release their dormancy at the above temperatures, the seeds were stratified at a constant temperature (4 °C) under laboratory conditions or at fluctuating temperatures in soil or by outdoor burial in soil. Fully dormant, or seeds stratified or buried (2006/2007 and 2007/2008) for various periods were treated with exogenous gibberellic acid (GA₃), ethephon and abscisic acid (ABA). Likewise, the effects of these regulators, applied during stratification, on seed germination were determined. The results indicate that A. retroflexus seed dormancy can be released either by stratification or by autumn–winter burial. The effect of GA₃ and ethylene, liberated from ethephon, applied after various periods of stratification or during stratification, depends on dormancy level. GA₃ did not affect or only slightly stimulated the germination of non-stratified, fully dormant seeds at 25 and 35 °C respectively. Ethylene increased germination at both temperatures. Seed response to GA₃ and ethylene at 25 °C was increased when dormancy was partially removed by stratification at constant or fluctuating temperatures or autumn–winter burial. The response to GA₃ and ethylene increased with increasing time of stratification. The presence of GA₃ and ethephon during stratification may stimulate germination at 35 °C. Thus, both GA₃ and ethylene can partially substitute the requirement for stratification or autumn–winter burial. Both hormones may also stimulate germination of secondary dormant seeds, exhumed in September. The response to ABA decreased in parallel with an increasing time of stratification and burial up to May 2007 or March 2008. Endogenous GA_n, ethylene and ABA may be involved in the control of dormancy state and germination of A. retroflexus. It is possible that releasing dormancy by stratification or partial burial is associated with changes in ABA/GA and ethylene balance and/or sensitivity to these hormones.     

Effects of temperature and light on germination and early seedling development of the pine pink orchid (Bletia purpurea)

Orchid seed physiology is a poorly understood phenomenon owing to an emphasis on production and the challenges associated with propagating orchids from minute seed. We investigated the role of simulated south Florida temperatures and illumination (dark and 12 h photoperiod) in regulating germination and seedling development using asymbiotic seed germination assays of Bletia purpurea. Our objectives were to determine whether in situ germination is limited by seasonal temperatures and to determine whether temperature alters responses to illumination. Bletia purpurea seeds were able to germinate to > 90% under all treatments. The greatest germination after 3 weeks was observed at 29/19°C under continual darkness and at 25°C under dark and illuminated conditions. The slowest germination was observed at simulated winter temperatures (22/11°C). Illumination initially inhibited germination and development, but resulted in equal or greater development by week six. Germination under 22/11°C was strongly inhibited by illumination, indicating an interaction between temperature and light sensing systems.     

Effect of temperature and light on seed germination of Erysimum naxense and Erysimum krendlii

Seed germination of two local Greek endemics was studied (Erysimum naxense, Erysimum krendlii). Seed viability was determined by using the tetrazolium method and germination was studied in synchronized cycles of five and four alternating temperatures [10/5 (for E. naxense only) and 15/10, 20/15, 25/20, and 30/25°C for both species, in cycles of 16 h day/8 h night], and in five light regimes (red, blue, green, white, and dark). Germination of E. naxense and E. krendlii seeds was determined daily for six and five weeks, respectively, with the data analyzed as viability adjusted accumulative seed germination at the end of each week. E. naxense’s seed viability was higher (90%) than that of E. krendlii (64%); seed germination (%) of both increased at low alternating temperatures (10/5°C, 15/10°C, 20/15°C). Germination of E. naxense seeds at low temperatures was light-independent, whereas at high temperatures it was increased with red light. Germination of E. krendlii seeds was inconsistently affected by light at the temperatures studied. Percentages of seed germination of both species were higher in experimental conditions similar to the ones of their natural habitats during autumn and/or spring (facilitated with Geographic Information Systems). These conclusions provide guidelines for species-specific propagation protocols and ex situ conservation.     

Comparative <Emphasis Type="Italic">in vitro</Emphasis> germination ecology of <Emphasis Type="Italic">Calopogon tuberosus</Emphasis> var. <Emphasis Type="Italic">tuberosus</Emphasis> (Orchidaceae) across its geographic range

Seed responses to temperature are often essential to the study of germination ecology, but the ecological role of temperature in orchid seed germination remains uncertain. The response of orchid seeds to cold stratification have been studied, but the exact physiological role remains unclear. No studies exist that compare the effects of either cold stratification or temperature on germination among distant populations of the same species. In two separate experiments, the role of temperature (25, 22/11, 27/15, 29/19, 33/24°C) and chilling at 10°C on in vitro seed germination were investigated using distant populations of Calopogon tuberosus var. tuberosus. Cooler temperatures promoted germination of Michigan seeds; warmer temperatures promoted germination of South Carolina and north central Florida seeds. South Florida seed germination was highest under both warm and cool temperatures. More advanced seedling development generally occurred at higher temperatures with the exception of south Florida seedlings, in which the warmest temperature suppressed development. Fluctuating diurnal temperatures were more beneficial for germination compared to constant temperatures. Cold stratification had a positive effect on germination among all populations, but South Carolina seeds required the longest chilling treatments to obtain maximum germination. Results from the cold stratification experiment indicate that a physiological dormancy is present, but the degree of dormancy varies across the species range. The variable responses among populations may indicate ecotypic differentiation.     

Viability of onion (Allium cega) seed as influenced by temperature during seed growth

The yield and performance of seeds from crops of winter-hardy, bolting-resistant onion grown at temperatures of 15–16, 18–19 and 22–23°C in 1979, 1980 and 1982 were compared. Yields of seed from crops grown at 22–23°C were lower than those from crops grown at lower temperatures but the seeds ripened between 11 and 32 days earlier. Seeds from crops grown at mean temperatures of above 18°C gave higher percentage germination when imbibed at 30°C than 20°C and they also gave higher percentage seedling emergence than those from crops grown at lower temperatures. Seedlings from seeds produced at mean temperatures above 18°C were heavier than those from seeds of a similar weight but produced at lower temperatures. None of these differences were associated with differences in seed weight, embryo weight or seed dormancy but were positively correlated with differences in seed N-concentration. The differences were also associated with the rate of imbibition of water as high germination, high N-content seeds had a slower rate of imbibition than low germination, low N-content seeds of the same weight.     

Variation in germination and seedling emergence of sugar beet at sub-optimal temperatures

Patterns of germination and seedling emergence at different temperatures were determined for a wide range of sugar beet accessions. There was good differentiation among seed lots and a close relationship between germination and emergence in temperature regimes of 20 h at 5 °C: 4 h at 7 °C, and constant 5 °C. Failure to germinate at temperatures between 5°C and 7 °C was more often associated with suppression of germination than with death of embryos. Results indicated that tests of germination at low temperatures would lead to more accurate predictions of field emergence than standard tests at 20 °C. Furthermore, tests at 20 °C may not give accurate estimates of potential viability.     

A seed respiration-based index of cold-sensitivity during imbibition in four macrothermal species

A research was carried out to evaluate the influence of temperature on seed respiration response of maize, cotton, grain sorghum and sunflower during imbibition, and to define reliable indices for a fast evaluation of cold-sensitivity at germination level in plants. The seed respiration activity was measured during seed imbibition at 25 °C (optimal) and 15 °C (suboptimal) constant temperatures, using a homemade respiration chamber adapted to an infrared gas analyzer. At 15 °C, sunflower and sorghum maintained high levels of seed germination (≥90 %), whilst this last dropped in cotton (36.7 %) and maize (27.8 %). With respect to this, cotton and maize seem to be cold sensitive during germination. Instantaneous seed respiration during imbibition versus temperature or thermal time could not be used as a good indicator for cold tolerance, since the levels of CO₂ recorded at 15 °C in cotton (higher than the other species) and maize (similar to that of sorghum and sunflower) did not correspond to adequate seed germination. Differently, the rates (b coefficient of linear regressions) of accumulation of CO₂ respired at optimal and suboptimal temperatures during the first hours of imbibition (up to approximately 24 h from the start of experiment), were significantly different in maize and cotton, whilst they did not differ in sorghum and sunflower. Therefore, the shift between slopes may represent a reliable index for seed cold-sensitivity assessment during early germination.     

Effect of temperature and cold stratification on seed germination of the Mediterranean wild aromatic Clinopodium sandalioticum (Lamiaceae)

A germination study was carried out on seeds of Clinopodium sandalioticum (Bacch. & Brullo) Bacch. & Brullo ex Peruzzi & Conti (Lamiaceae), a wild aromatic plant endemic to Sardinia. Seeds were incubated at a range of constant (5–25°C) and an alternating temperatures regime (25/10°C), with 12 hours of irradiance per day. The results achieved at 10°C were also compared with those obtained after a period of cold stratification at 5°C for three months. Final seed germination ranged from ca. 28% (5°C) to ca. 72% (25/10°C). A base temperature for germination (T_b) of ca. 5°C and a thermal constant for 50% germination (S) of 89.3°Cd were identified and an optimal temperature for germination (T_o) was estimated to be comprised between 20 and 25°C. Cold stratification negatively affected seed viability and germination at 10°C. Although a typical “Mediterranean germination syndrome”, could not be detected for C. sandalioticum seeds, these results were coherent with those previously reported for other Mediterranean Lamiaceae species.     

Tree fine root Ca/Al molar ratio – Indicator of Al and acidity stress

Many Aloe species are exploited as natural products. Generally, the leaves are unsustainably picked from wild plants to meet the market demand. Basic scientific information on seed biology and the ways of increasing levels of secondary metabolites in seedlings is still lacking for Aloe species. This study investigated seed germination requirements and evaluated levels of secondary metabolites in seedlings of Aloe arborescens, an important species in traditional medicine. The highest percentage germination (78%) and the fastest germination rate (GR) (10% d^? 1) with a mean germination time (MGT) of 9 days were achieved at 20°C under a 16-h photoperiod. At 25°C, maximum percentage germination (67%) (P < 0.05), higher GR (13% d^? 1) and shorter MGT (6 days) were obtained under constant light. These results indicate that temperature and light play a significant role in germination of A. arborescens seeds. Increasing osmotic pressure on seeds decreased percentage germination, whereas buffering the solution to a range of pH values (4–10) did not significantly affect germination. Smoke–water (1:500 v/v), smoke-isolated karrikinolide (10^? 8 and 10^? 9 M) and potassium nitrate (10^? 3 and 10^? 4 M) significantly promoted germination compared with the control at 25°C (supra-optimal temperature) under a 16-h photoperiod. These treatments were also effective in increasing secondary metabolite levels (flavonoids and phenolics) in A. arborescens seedlings.     

Temperature sensitive phases during the germination of lettuce (Lactuca sativa) seeds

Lettuce seeds cvs Hilde, Feltham King and Avoncrisp were subjected, at different phases during imbibition at 22°C, to a high temperature (33°C) inhibitory for germination, for periods ranging from 4 to 144 h, before returning them to 22°C. The results showed, that the first 4h of imbibition and also the phase between the commencement of mitosis and the onset of radicle emergence were more sensitive to the effects of high temperature than other phases in the germination process. Short exposures (8–24 h) to 33°C commencing at the latter phase delayed germination by up to 4 days, and at the earlier by up to 8 days. Percentage germination was unaffectd except after prolonged exposures (> 48 h) from the beginning of imbibition, which reduced it. Seedling emergence from moist sieved soil was both delayed and reduced when imbibing seeds were exposed for a short period from the beginning of imbibition to 33°C compared with seeds imbibing continuously at 19°C. Germination was delayed and not reduced when seed was exposed to 33°C at the phase between commencement of mitosis and the onset of radicle emergence.     

Energy metabolism in green cells

Background: Although fire is an important factor in determining cerrado vegetation, information about its effect on seed banks is sparse. Cerrado fires are rapidly moving surface fires with low residence time, producing only short-term heating of the uppermost centimetres of the soil. However, the reduction in vegetation cover and deposition of ashes increases the daily amplitude of soil temperature by as much as 35 °C.

Aims: To assess the effect of post-fire daily soil temperatures on the germination of one alien and nine native grasses.

Methods: Seeds were stored at alternating temperatures of 45 ºC/10 ºC (10 h/14 h) for 7 d or 30 d, simulating two different storage times in the soil seed bank before the onset of the rainy season. Germination was monitored over 30 d.

Results: The variation in temperature had a significant effect on the rate of seed germination in some species, either enhancing it (Aristida setifolia) or reducing it (Schizachyrium sanguineum). Increased storage time reduced the viability of S. sanguineum and Echinolaena inflexa. The invasive Melinis minutiflora had the highest germination rate and it also showed the best toleration of post-fire conditions (45 ºC/10 ºC) after 7 d, with significant reduction in the germination time after 30 d.

Conclusions: Fire seems to have a significant effect in the early life of cerrado grasses. Some native species responded positively to temperature oscillation, suggesting that they should be better prepared to compete with alien species after a fire, with more of their seeds germinating and/or at a more rapid rate.     

Hormonal and temperature regulation of seed dormancy and germination in <Emphasis Type="Italic">Leymus chinensis</Emphasis>

Fluctuating temperature plays a critical role in determining the timing of seed germination in many plant species. However, the physiological and biochemical mechanisms underlying such a response have been paid little attention. The present study investigated the effect of plant growth regulators and cold stratification in regulating Leymus chinensis seed germination and dormancy response to temperature. Results showed that seed germination was less than 2 % at all constant temperatures while fluctuating temperature significantly increased germination percentage. The highest germination was 71 % at 20/30 °C. Removal of the embryo enclosing material of L. chinensis seed germinated to 74 %, and replaced the requirement for fluctuating temperature to germinate, by increasing embryo growth potential. Applications of GA₄₊₇ significantly increased seed germination at constant temperature. Also, inhibition of GA biosynthesis significantly decreased seed germination at fluctuating temperatures depending upon paclobutrazol concentration. This implied GA was necessary for non-dormant seed germination and played an important role in regulating seed germination response to temperature. Inhibition of ABA biosynthesis during imbibition completely released seed dormancy at 20/30 °C, but showed no effect on seed germination at constant temperature, suggesting ABA biosynthesis was important for seed dormancy maintenance but may not involve in seed germination response to temperature. Cold stratification with water or GA₃ induced seed into secondary dormancy, but this effect was reversed by exogenous FL, suggesting ABA biosynthesis during cold stratification was involved in secondary dormancy. Also, cold stratification with FL entirely replaced the requirement of fluctuating temperature for germination with seeds having 73 % germination at constant temperature. This appears to be attributed to inhibition of ABA biosynthesis and an increase of GA biosynthesis during cold stratification, leading to an increased embryo growth potential. We suggest that fluctuating temperature promotes seed germination by increasing embryo growth potential, mainly attributed to GA biosynthesis during imbibitions. ABA is important for seed dormancy maintenance and induction but showed less effect on non-dormant seed germination response to temperature.     

Light,temperature, dry after‐ripening and salt stress effects on seed germination of Phleum sardoum (Hackel) Hackel

Phleum sardoum is an endemic psammophilous species of Sardinia, growing exclusively on coastal sandy dunes. The effect of glumes on seed germination, germination requirements at constant (5–25°C) and alternating (25/10°C) temperatures, both in the light (12/12 h) and in the dark were evaluated, as well as the effect of a dry after‐ripening period (90 days at 25°C), the salt stress effect (0–600 mmol NaCl) and its recovery on seed germination. The presence of glumes reduced final germination percentages. For fresh naked seeds, high germination percentages were observed at 10°C. Dry after‐ripening increased germination rate at low temperatures, but did not affect final germination percentages. NaCl determined a secondary salt‐induced dormancy which recovery interrupted only partially. Our results highlighted that this species has its optimum of germination during autumn–winter when, under a Mediterranean climate, water availability is highest and soil salinity levels are minimal.     

Effect of Temperature Regimes on Germination of Dimorphic Seeds of Atriplex prostrata

Dimorphic seeds of Atriplex prostrata were removed from cold dry storage monthly over a one year period to test for fluctuations in seed dormancy and germination rate. For each seed type, four replicates of 25 seeds were exposed to four alternating night/day temperature regimes mimicking seasonal fluctuations in Ohio: 5/15 °C; 5/25 °C; 15/25 °C and 20/35 °C with a corresponding 12-h photoperiod (20 μmol m⁻² s⁻¹; 400 – 700 nm). We found a significant three-way interaction of seed size, temperature and month for both percent germination and the rate of germination. Large seeds showed the greatest germination at the 20/35 °C and 5/25 °C temperature regimes and small seeds at the 5/25 °C regime. Large seeds had greater germination at all temperatures as compared to small seeds. Large seeds had the fastest germination rates at 20/35 °C followed by 5/25 °C whereas small seeds had the fastest rates at 5/25 °C followed by 20/35 °C. This revised version was published online in July 2006 with corrections to the Cover Date.     

Germination response of four pasture species to temperature,light, and post-harvest period

Abstract

The purpose of this research was to explore the effect of temperature, light, and post-harvest period, and their interactions, on seed germination ecology of four common pasture species in the Mediterranean environment. Mature seeds of Diplotaxis erucoides, Hirschfeldia incana, Hyoseris scabra (Mediterranean distribution) and Sonchus oleraceus (cosmopolitan distribution) were subjected to seven constant temperatures (10–40°C, at intervals of 5°C) under continuous darkness, or a 12 h/12 h light/dark photoperiod at 30, 150 and 270 days after harvest (DAH). Cumulative germination and germination speed were determined. In all the tested species, except S. oleraceus, light significantly enhanced germination. S. oleraceus seeds maintained germination values over 90%, in a wide range of temperatures (10–35°C), in the dark as well as in light. Seeds of H. incana germinated well soon after seed dispersal. In D. erucoides and H. scabra, germination increased with storage period, while in S. oleraceus there was no effect of seed age. In all the species, moreover, no germination was recorded at 40°C. Temperature, light, and post- harvest requirement may be regarded as an adaptation strategy to ensure optimal conditions for seedling development and survival in Mediterranean species, while the species with a cosmopolitan distribution germinates under almost all tested conditions.     

Germination response and viability of an endangered tropical conifer Widdringtonia whytei seeds to temperature and light

The tropical conifer Widdringtonia whytei Rendle is an endangered species endemic to Mulanje Mountain in Malawi. A study was conducted for the first time under controlled conditions in order to assess the effects of temperature and light on germination and viability of W. whytei seeds. Seeds incubated at a constant temperature of 20 °C attained the highest cumulative germination percentage (100%) followed by 87% germination under fluctuating temperatures of 15 °C night/25 °C day. No seed germination occurred at temperatures below 15 °C. Seeds that failed to germinate at temperatures below 15 °C showed the highest (> 90%) viability compared to the seeds incubated at 25 °C (60%). Across temperature regimes, germination was significantly higher under light (44.7%) than dark (35.6%) conditions. It is concluded that temperature is one of the critical factors for germination of W. whytei seed. The ability of W. whytei seeds to germinate both in light and darkness implies that the species would unlikely form a persistent soil seed bank, an attribute which is common in species that survive in habitats frequently disturbed by fires.     

Adaptation to habitat in Aquilegia species endemic to Sardinia (Italy): Seed dispersal,germination and persistence in the soil

Abstract

The autecology of the Sardinian endemics Aquilegia barbaricina Arrigoni et Nardi and A. nugorensis Arrigoni et Nardi were investigated. Peaks of anthesis and seed dispersal were recorded for five populations occurring in two distinct habitats, one riparian and one rupicolous. Germination tests were carried out on seed lots belonging to each population by sowing seeds at 10, 15, 20, 25 and 25/15°C. In addition, seeds were incubated for 2 months at either 25°C (summer), 5°C (winter) or 25°C for 2 months plus 2 months at 5°C (summer followed by winter–SW), and then moved to the germination temperatures. Embryo measurements were taken during pre-treatments and germination. Experimental seed burials were carried out for two populations of each species. Both species dispersed in summer. The population of A. nugorensis occurring on rocky outcrops differed in phenology from both the other A. nugorensis population from riparian vegetation and from A. barbaricina. Both species showed morphophysiological seed dormancy, with <50% germination under laboratory conditions. All riparian populations germinated only after the SW pre-treatment, while the rupicolous population germinated at 25°C, without any pre-treatment. Low germination percentages were observed in the experimental seed burials, suggesting the ability for both species to form a persistent soil seed bank.     

The impact of temperature on the production and fitness of microsclerotia of the fungal bioherbicide Mycoleptodiscus terrestris

The impact of growth temperature was evaluated for the fungal plant pathogen Mycoleptodiscus terrestris over a range of temperatures (20–36°C). The effect of temperature on biomass accumulation, colony forming units (cfu), and microsclerotia production was determined. Culture temperatures of 24–30°C produced significantly higher biomass accumulations and 20–24°C resulted in a significantly higher cfu. The growth of M. terrestris was greatly reduced at temperatures above 30°C and was absent at 36°C. The highest microsclerotia concentrations were produced over a wide range of temperatures (20–30°C). These data suggest that a growth temperature of 24°C would optimize the parameters evaluated in this study. In addition to growth parameters, we also evaluated the desiccation tolerance and storage stability of air-dried microsclerotial preparations from these cultures during storage at 4°C. During 5 months storage, there was no significant difference in viability for air-dried microsclerotial preparations from cultures grown at 20–30°C (>72% hyphal germination) or in conidia production (sporogenic germination) for air-dried preparations from cultures grown at 20–32°C. When the effect of temperature on germination by air-dried microsclerotial preparations was evaluated, data showed that temperatures of 22–30°C were optimal for hyphal and sporogenic germination. Air-dried microsclerotial preparations did not germinate hyphally at 36°C or sporogenically at 20, 32, 34, or 36°C. These data show that temperature does impact the growth and germination of M. terrestris and suggest that water temperature may be a critical environmental consideration for the application of air-dried M. terrestris preparations for use in controlling hydrilla.