Quantitative and qualitative consequences of reduced pollen loads in a mixed‐mating plant

Greater pollination intensity can enhance maternal plant fitness by increasing seed set and seed quality as a result of more intense pollen competition or enhanced genetic sampling. We tested experimentally these effects by varying the pollen load from a single pollen donor on stigmas of female flowers of Dalechampia scandens (Euphorbiaceae) and measuring the effects on seed number and seed mass. Seed set increased rapidly with pollen number at low to moderate pollen loads, and a maximum set of three seeds occurred with a mean pollen load of 19 pollen grains. We did not detect a trade‐off between the number of seeds and seed mass within a fruit. Seed mass increased with increasing pollen load, supporting the hypothesis of enhanced seed quality via increased pollen‐competition intensity or genetic sampling. These results suggest that maternal fitness increases with larger pollen loads, even when the fertilization success is already high. Our results further highlight the importance of high rates of pollen arrival onto stigmas, as mediated by reliable pollinators. Comparing the pollen‐to‐seed response curve obtained in this experiment with those observed in natural populations suggests that pollen limitation may be more severe in natural populations than predicted from greenhouse studies. These results also indicate that declines in pollinator abundance may decrease plant fitness through lowered seed quality before an effect on seed set is detected.     

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.     

Heat and smoke pre‐treatment of seeds to improve restoration of an endangered Mediterranean climate vegetation type

Invasive alien plants impact ecosystems, which often necessitates their removal. Where indigenous species recovery fails following removal alone, an active intervention involving reintroduction of seed of native species may be needed. This study investigated the potential for a combination of the fire cues of smoke and heat as a pre‐treatment of seeds in breaking dormancy and facilitating increased germination. Species were selected to represent different functional types within Cape Flats Sand Fynbos; a fire‐prone, critically endangered vegetation type in South Africa. Seeds were exposed to either a heat pulse (temperatures between 60 and 300°C for durations of between 30 s and 20 min) or dry after‐ripening (1 or 2 months at milder temperatures of 45°C or less). Thereafter, seeds were soaked in smoke solution for 18 h and subsequently placed on agar at 10/20°C for germination. Most species fell into one of two main groups: Seed germination in the first group was greatest following a lower temperature (60°C) heat pulse, an extended period of mild temperature (20/40°C or 45°C) exposure, or no pre‐treatment with heat. Seed germination in the second group was promoted after brief exposure to higher (100°C) temperatures. No germination occurred in any species following heat treatments of 150°C or higher. Species which responded better to higher temperatures were mainly those possessing physical dormancy, but seed morphology did not correlate with germination success. This study showed that heat stimulation of seeds is more widespread in fynbos plant families than previously known and will enable the development of better seed pre‐treatment protocols before large‐scale sowing as an active restoration treatment after alien plant clearing.     

Investigating seed dormancy in cotton (Gossypium hirsutum L.): understanding the physiological changes in embryo during after‐ripening and germination

• The dormancy of seeds of upland cotton can be broken during dry after‐ripening, but the mechanism of its dormancy release remains unclear.
• Freshly harvested cotton seeds were subjected to after‐ripening for 180 days. Cotton seeds from different days of after‐ripening (DAR) were sampled for dynamic physiological determination and germination tests. The intact seeds and isolated embryos were germinated to assess effects of the seed coat on embryo germination. Content of H₂O₂ and phytohormones and activities of antioxidant enzymes and glucose‐6‐phosphate dehydrogenase were measured during after‐ripening and germination.
• Germination of intact seeds increased from 7% upon harvest to 96% at 30 DAR, while embryo germination improved from an initial rate of 82% to 100% after 14 DAR. Based on T₅₀ (time when 50% of seeds germinate) and germination index, the intact seed and isolated embryo needed 30 and 21 DAR, respectively, to acquire relatively stable germination. The content of H₂O₂ increased during after‐ripening and continued to increase within the first few hours of imbibition, along with a decrease in abscisic acid (ABA) content. A noticeable increase was observed in gibberellic acid content during germination when ABA content decreased to a lower level. Coat removal treatment accelerated embryo absorption of water, which further improved the accumulation of H₂O₂ and changed peroxidase content during germination.
• For cotton seed, the alleviation of coat‐imposed dormancy required 30 days of after‐ripening, accompanied by rapid dormancy release (within 21 DAR) in naked embryos. H₂O₂ acted as a core link between the response to environmental changes and induction of other physiological changes for breaking seed dormancy.

     

Microbial responses to warming enhance soil carbon loss following translocation across a tropical forest elevation gradient

Tropical soils contain huge carbon stocks, which climate warming is projected to reduce by stimulating organic matter decomposition, creating a positive feedback that will promote further warming. Models predict that the loss of carbon from warming soils will be mediated by microbial physiology, but no empirical data are available on the response of soil carbon and microbial physiology to warming in tropical forests, which dominate the terrestrial carbon cycle. Here we show that warming caused a considerable loss of soil carbon that was enhanced by associated changes in microbial physiology. By translocating soils across a 3000 m elevation gradient in tropical forest, equivalent to a temperature change of ± 15 °C, we found that soil carbon declined over 5 years by 4% in response to each 1 °C increase in temperature. The total loss of carbon was related to its original quantity and lability, and was enhanced by changes in microbial physiology including increased microbial carbon‐use‐efficiency, shifts in community composition towards microbial taxa associated with warmer temperatures, and increased activity of hydrolytic enzymes. These findings suggest that microbial feedbacks will cause considerable loss of carbon from tropical forest soils in response to predicted climatic warming this century.     

RopGEF2 is involved in ABA‐suppression of seed germination and post‐germination growth of Arabidopsis

The involvement of Rho of Plants (ROP) GTPases in abscisic acid (ABA) signalling in Arabidopsis has been demonstrated in many studies. However, the roles of RopGEFs (Rop guanine nucleotide exchange factors), which modulate ROP activities in ABA signalling, are poorly understood. Here, we demonstrate that RopGEF2 may play a negative role in ABA‐suppressed seed germination and post‐germination growth. We show that disruption of RopGEF2 enhances sensitivity to exogenous ABA in seed germination assays and that RopGEF2pro‐GUS is mainly expressed in developing embryos and germinating seeds. Interestingly, YFP‐RopGEF2 is located in both the cytoplasmic region and in mitochondria. Notably, the PRONE2 (plant‐specific ROP nucleotide exchanger 2) domain of RopGEF2 is detected in mitochondria, whereas the N‐terminus of RopGEF2 is shown to be in the cytosol. After ABA treatment, degradation of RopGEF2 is triggered in the cytosol through the ubiquitin‐26S proteasome system. The binding of RopGEF2 to ROP2, ROP6 or ROP10, which has been demonstrated to be involved in ABA signalling, not only alters the localization of RopGEF2 but also enables RopGEF2 to escape degradation in the cell. Thus, in this study, we deduce a sophisticated mechanism of ABA‐mediated RopGEF2‐ROP signalling, which potentially implicates the inactivation of ROPs in responsiveness to ABA.     

Tree seed rain and seed removal,but not the seed bank,impede forest recovery in bracken (Pteridium aquilinum (L.) Kuhn)‐dominated clearings in the African highlands

Considerable areas dominated by bracken Pteridium aquilinum (L.) Kuhn occur worldwide and are associated with arrested forest recovery. How forest recovery is impeded in these areas remains poorly understood, especially in the African highlands. The component processes that can lead to recruitment limitation—including low seed arrival, availability and persistence—are important determinants of plant communities and offer a potential explanation for bracken persistence. We investigated key processes that can contribute to recruitment limitation in bracken‐dominated clearings in the Bwindi Impenetrable National Park, Uganda. We examined if differences in seed rain (dispersal limitation), soil seed bank, or seed removal (seed viability and persistence) can, individually or in combination, explain the differences in tree regeneration found between bracken‐dominated areas and the neighboring forest. These processes were assessed along ten 50‐m transects crossing the forest–bracken boundary. When compared to the neighboring forest, bracken clearings had fewer seedlings (bracken 11,557 ± 5482 vs. forest 34,515 ± 6066 seedlings/ha), lower seed rain (949 ± 582 vs. 1605 ± 335 tree seeds m^?2 year^?1), comparable but sparse soil seed bank (304 ± 236 vs. 264 ± 99 viable tree seeds/m²), higher seed removal (70.1% ± 2.4% vs. 40.6% ± 2.4% over a 3‐day interval), and markedly higher rodent densities (25.7 ± 5.4 vs. 5.0 ± 1.6 rodents per 100 trapping sessions). Camera traps revealed that rodents were the dominant animals visiting the seeds in our seed removal study. Synthesis: Recruitment limitation contributes to both the slow recovery of forest in bracken‐dominated areas, and to the composition of the tree species that occur. Low seed arrival and low persistence of unburied seeds can both explain the reduced density of seedlings found in bracken versus neighboring forest. Seed removal, likely due to rodents, in particular appears sufficient to constrain forest recovery and impacts some species more severely than others.     

The effects of rainfall on different components of seasonal fecundity in a tropical forest passerine

Seasonal fecundity is a composite metric that is determined by component parameters such as clutch size, nest survival and re‐nesting probability. Many of these component parameters are known to vary with environmental conditions, in particular rainfall prior to or during the breeding season. In some species, seasonal fecundity is positively related to rainfall, but little is known about which component parameters of seasonal fecundity respond most strongly to rainfall. We used intensive nest monitoring of a multi‐brooded tropical forest passerine, the Montserrat Oriole Icterus oberi, to examine the effects of rainfall during the pre‐breeding season on component parameters of annual fecundity. We monitored all nests of a total of 42 pairs over 5 years in which rainfall varied substantially. We then related clutch size, nest survival, onset and length of the breeding season, re‐nesting probability and re‐nesting interval to pre‐breeding season rainfall using generalized linear mixed models that accounted for random variation across sites and individual pairs, and incorporated other variables known to affect the response. Higher pre‐breeding season rainfall led to an increase in clutch size and a decrease in re‐nesting interval, but nest survival, re‐nesting probability and length of the breeding season were not affected by variation in rainfall. The onset of the breeding season was delayed in very dry years. We conclude that higher rainfall is likely to increase food availability and thus body condition of female Montserrat Orioles, leading to an increase in fecundity due to larger clutch sizes.     

Assessing seed priming,sowing date,and mulch film to improve the germination and survival of direct‐sown Miscanthus sinensis in the United Kingdom

Direct sowing of Miscanthus seed could lower crop establishment costs, and increase the rate of grower uptake and biomass supply for the emerging bio‐economy. A replicated field trial was conducted at two contrasting UK sites: Aberystwyth (ABR) in mid‐Wales and Blankney (BLK) in Lincolnshire. These sites encompass the west–east meteorological gradient in the United Kingdom where the growing season at ABR is cooler and wetter while BLK is warmer and drier. Primed and unprimed Miscanthus sinensis seeds were sown directly onto the soil surface with and without a clear biodegradable mulch film, at nine dates interspersed from May to October. Average daily mean soil surface temperatures measured over the first 2 months after sowing under the mulch film were higher than control plots (2.7°C ABR and 4.2°C BLK). At both sites, the film covering also affected soil volumetric moisture relative to uncovered control plots (?3% ABR and 8% BLK), demonstrating the negative impact of mulch film when sowing on dry soil. Over nine sowings, seed germination at ABR under film varied between ?28% and +18% of germination under control conditions. Seedlings from the first three sowings at both sites under film had sufficient physiological maturity to survive the first winter period. At BLK, mulch film significantly increased tiller count and height in both the first and second years after sowing. At ABR, where temperatures were lower, film covering significantly increased tiller height but not count. Water priming had no significant effect on seed viability or germination in the field tests. Base temperatures for germination of primed and unprimed seeds on a thermal gradient plate were 7.0°C and 5.7°C, respectively, with a ± 1.7°C confidence interval. Based on our results for M. sinensis in the United Kingdom, we recommend the sowing of unprimed seed in May under film and only when the soil is moist.     

Smoke and heat effects on soil seed bank germination for the re‐establishment of a native forest community in New South Wales

The effects of plant‐derived smoke and of heat on the emergence of seedlings from seeds were assessed. Seeds had been stored in forest topsoil used for mine site rehabilitation. The study was carried out in a dry sclerophyll, spotted gum (Corymbia maculata), forest community at the Mount Owen open‐cut coal mine in the Hunter Valley region of New South Wales. Samples of the surface 2.5 cm of topsoil were either exposed to cool smoke from eucalypt foliage for 60 min, heated to 80°C, or left untreated. Seedling emergence from the seed bank in this soil was then monitored in a glasshouse. Within the first month, smoke alone promoted a 4.3‐fold increase in the density of seedlings relative to control. There were 540 emergents per m² in the control and 2309 per m² in the smoke treated topsoil. Many annual and perennial herbs emerged but grasses responded most strongly to smoke. Germination in seven of the 20 grass species was promoted by smoke. Smoke promoted the germination of some introduced species as well as native species, and accelerated the rate at which seedlings emerged, although these differences sometimes declined with time. Heat also stimulated germination but smoke and heat stimuli appeared to be complementary in their promotion of seedling emergence from the topsoil seed bank. Each treatment increased the density of different species, enhanced the species richness of different components of the seed bank, and had different effects on the rate of emergence. The results suggest that increased seed germination in the field immediately following a moderate intensity fire may sometimes be the result of smoke stimulation and sometimes the result of heat stimulation of the soil seed bank. These findings may have important implications for minesite revegetation programs where topsoils are replaced after mining and rapid germination of seeds stored in these soils is required during short periods when conditions are favourable for germination.     

Effect of maturation conditions on light and temperature requirements during seed germination of Citrullus colocynthis from the Arabian Desert

• Seed germination of Citrullus colocynthis, as in many other species of Cucurbitaceae, is inhibited by light, particularly at low temperatures. Germination response to light and temperature has been attributed to day length and temperature during seed maturation. This study assessed the effects of these factors on the germination response of C. colocynthis to temperature and light quality.
• Ripe fruits were collected from natural habitats during December and February and germinated at three temperatures (15/25, 20/30 and 25/35 °C) in five light treatments (dark, white light and Red:Far Red (R:FR) ratios of 0.30, 0.87 and 1.19). Additionally, unripe fruits were also collected from natural habitats and completed their maturation in growth chambers under different day lengths (6, 16 and 24 h of darkness) at 10/20 °C, and in darkness at both 10/20 °C and 25/35 °C. Mature seeds of the different treatments were germinated in the same five light treatments at 15/25 °C.
• Germination was significantly higher in the dark than that in any light treatment. Seeds matured at higher temperatures (i.e. seeds from the December collection and those matured at 25/35 °C) had significantly higher germination than those matured at lower temperatures (i.e. seeds from the February collection and those matured at 10/20 °C). Dark germination was significantly higher for the December collection than for the February collection. Seeds of the two collections germinated in the dark only at 15/25 °C. However, seeds matured in a growth chamber at 10/20 °C in darkness germinated at 15/25 °C in all light treatments, except for the R:FR ratio 0.30. Seeds of the different treatments failed to germinate in FR‐rich light.
• This study demonstrates that both temperature and day length during seed maturation play significant roles in the germination response of C. colocynthis. Additionally, the dark requirement for germination is likely beneficial for species with the larger seeds, such as C. colocynthis, which produce bigger seedlings that are able to emerge from deep soils and are competitively superior under dense vegetation and resource‐limited conditions.

     

Contrasting reproductive traits in two species of mangrove‐dwelling littorinid snails in a seasonal tropical habitat

Previous studies on the reproductive biology of littorinid snails have focused on rocky shore species, investigating how these gastropods can achieve maximal reproductive success, as well as on processes of sexual selection. This study documented differences in the reproductive traits of two mangrove‐dwelling littorinids, Littoraria ardouiniana and L. melanostoma, in Hong Kong. Reproductive activity of both species was most intense during the summer months. Mating pairs of the two species generally occurred in the tree canopies. Few false mating pairs (same sex or heterospecific pairs: <10%) were recorded, and members of both species showed size‐assortative mating. Littoraria ardouiniana had a shorter reproductive season but a higher intensity of mating and higher seasonal fecundity, than did L. melanostoma. Members of both species showed bi‐lunar periodicities of egg or larval release, synchronized with spring tides. Fecundity showed a strong positive relationship with body size in L. ardouiniana, but not in L. melanostoma. Females of L. ardouiniana released entire broods of larvae in a single brief event, whereas females of L. melanostoma released fewer eggs over 1–8 d. Release of larvae in L. ardouiniana involved a series of short bursts and was much faster than the trickle release of eggs in L. melanostoma. The contrasting reproductive traits in these two species represent different strategies to optimize reproductive success in mangrove habitats.     

Recovery of tree and mammal communities during large‐scale forest regeneration in Kibale National Park,Uganda

Tropical landscapes are changing rapidly as a result of human modifications; however, despite increasing deforestation, human population growth, and the need for more agricultural land, deforestation rates have exceeded the rate at which land is converted to cropland or pasture. For deforested lands to have conservation value requires an understanding of regeneration rates of vegetation, the rates at which animals colonize and grow in regenerating areas, and the nature of interactions between plants and animals in the specific region. Here, we present data on forest regeneration and animal abundance at four regenerating sites that had reached the stage of closed canopy forest where the average dbh of the trees was 17 cm. Overall, 20.3 percent of stems were wind‐dispersed species and 79.7 percent were animal‐dispersed species, while in the old‐growth forest 17.3 percent of the stems were wind‐dispersed species. The regenerating forest supported a substantial primate population and encounter rate (groups per km walked) in the regenerating sites was high compared to the neighboring old‐growth forests. By monitoring elephant tracks for 10 yr, we demonstrated that elephant numbers increased steadily over time, but they increased dramatically since 2004. In general, the richness of the mammal community detected by sight, tracks, feces, and/or camera traps, was high in regenerating forests compared to that documented for the national park. We conclude that in Africa, a continent that has seen dramatic declines in the area of old‐growth forest, there is ample opportunity to reclaim degraded areas and quickly restore substantial animal populations.