Factors Influencing Seedling Establishment Rates in Zostera marina and Their Implications for Seagrass Restoration

Selection of strategies to efficiently utilize limited seed supplies in efforts to restore the seagrass Zostera marina (eelgrass) requires a better understanding of the processes that limit seedling establishment at potential restoration sites. We investigated the effect of seed distribution timing on seedling establishment and tested for interactive effects of seed burial and distribution timing. We also investigated the effect of habitat type on seedling establishment by distributing Z. marina seeds inside and outside of established Ruppia maritima (widgeongrass) patches and examined mechanisms causing habitat differences by manipulating seed position (buried or unburied) and vulnerability to seed predators (unprotected or protected in packets). Seeds distributed on the sediment surface in the summer (July or August) produced fewer seedlings than seeds distributed in fall (October) in five of six trials over 3 years. Seed burial increased success rates for seeds distributed in summer at one of two sites tested, eliminating the effect of season, but reduced success at the other site. Seeds placed in R. maritima generally produced fewer seedlings than seeds in bare sand, and although seed burial and protection in packets increased success in bare sand at three of four sites, the effect was less consistent in R. maritima. We conclude that seed predation and physical interactions were influential in reducing seedling establishment in R. maritima, contrary to hypotheses positing a nursery role for existing vegetation. Efficient restoration efforts with Z. marina seeds should target unvegetated areas after summertime sources of mortality have diminished. Direct seed burial may enhance seedling establishment rates.     

Innovative Techniques for Large‐scale Seagrass Restoration Using Zostera marina (eelgrass) Seeds

The use of Zostera marina (eelgrass) seeds for seagrass restoration is increasingly recognized as an alternative to transplanting shoots as losses of seagrass habitat generate interest in large‐scale restoration. We explored new techniques for efficient large‐scale restoration of Z. marina using seeds by addressing the factors limiting seed collection, processing, survival, and distribution. We tested an existing mechanical harvesting system for expanding the scale of seed collections, and developed and evaluated two new experimental systems. A seeding technique using buoys holding reproductive shoots at restoration sites to eliminate seed storage was tested along with new techniques for reducing seed‐processing labor. A series of experiments evaluated storage conditions that maintain viability of seeds during summer storage for fall planting. Finally, a new mechanical seed‐planting technique appropriate for large scales was developed and tested. Mechanical harvesting was an effective approach for collecting seeds, and impacts on donor beds were low. Deploying seed‐bearing shoots in buoys produced fewer seedlings and required more effort than isolating, storing, and hand‐broadcasting seeds in the fall. We show that viable seeds can be separated from grass wrack based on seed fall velocity and that seed survival during storage can be high (92–95% survival over 3 months). Mechanical seed‐planting did not enhance seedling establishment at our sites, but may be a useful tool for evaluating restoration sites. Our work demonstrates the potential for expanding the scale of seed‐based Z. marina restoration but the limiting factor remains the low rate of initial seedling establishment from broadcast seeds.     

An Effective Method for Collecting and Storing Seeds from Zostera marina (Eelgrass) in the Yellow Sea,China

The coast of the Yellow Sea in China, like many other temperate coastal zones, has been experiencing a dramatic decline in the abundance of seagrass. Intensive efforts have been made to restore seagrass communities along the coast to restore the function of the coastal ecosystem. Transplanting adult Zostera marina shoots is labor‐intensive, time‐consuming, expensive, and detrimental to donor beds; thus, restoring seagrass communities through the use of seeds is highly valued in current, large‐scale restoration trials. In this study, an effective method for collecting, processing, and storing Z. marina seeds was developed. From 2009 to 2013, respectively, 122,000, 421,000, 364,000, 1,041,000, and 1,091,000 seeds were successfully collected. Two‐way analysis of variance (ANOVA) showed the interaction between salinity and temperature significantly affected the cumulative germination rate (CGR) (p < 0.01) during the storage period and the viability (p < 0.01) of seeds after storage. The germination rate after storage was significantly affected by salinity and temperature (p < 0.01). The highest viability (89.8 ± 1.0%) and germination rate (75.6 ± 4.5%) were found among seeds stored at 4°C and a salinity of 44.5 psu for 7 months. The cost for planting 1 ha of sea bottom with Z. marina seeds ranged from $2,613 to $80,900 depending on the seeding density and seed loss during storage. The average cost per Z. marina seed in this study was $0.00586.     

Growing Zostera marina (eelgrass) from Seeds in Land‐Based Culture Systems for Use in Restoration Projects

The use of aquaculture systems to grow the seagrass Zostera marina (eelgrass) from seeds for restoration projects was evaluated through laboratory and mesocosm studies. Along the mid‐Atlantic coast of North America Z. marina seeds are shed from late spring through early summer, but seeds typically do not begin to germinate until the late fall. Fall is the optimal season to plant both seeds and shoots in this region. We conducted studies to determine if Z. marina seeds can be induced to germinate in the summer and seedlings grown in mesocosms to a size sufficiently large enough for out‐planting in the fall. Seeds in soil‐less culture germinated in the summer when held at 14°C, with percent germination increasing with lower salinities. Cold storage (4°C) of seeds prior to planting in sediments enhanced germination and seedling survival. Growth rates of seedlings were significantly higher in nutrient enriched estuarine sediments. Results from preliminary studies were used in designing a large‐scale culture project in which 15,000 shoots were grown and out‐planted into the Potomac River estuary in the Chesapeake Bay and compared with an equal number of transplanted shoots. These studies demonstrate that growing Z. marina from seeds is an alternative approach to harvesting plants from donor beds when vegetative shoots are required for restoration projects.     

Large‐Scale Zostera marina (eelgrass) Restoration in Chesapeake Bay,Maryland, USA. Part I: A Comparison of Techniques and Associated Costs

The Chesapeake Bay, like many other temperate estuaries, has exhibited dramatic declines in the abundance of submerged aquatic vegetation (SAV) during the later half of the twentieth century. Because of the functions SAV serve in maintaining a healthy estuarine ecosystem, SAV restoration has become an important component of Chesapeake Bay restoration. Specifically, recent water quality improvements in areas from which populations of Zostera marina (eelgrass) have been extirpated have suggested that Z. marina restoration could succeed. Early restoration efforts involved transplanting Z. marina plants from healthy source beds to restoration locations, but this was labor intensive, time consuming, expensive, and potentially detrimental to donor beds. This multi‐year project investigated new techniques for large‐scale Z. marina seed collection and processing and compared two seed dispersal methods to evaluate cost effectiveness. Tens of millions of mature Z. marina seeds were collected through snorkeling, SCUBA, or with a mechanical harvester. Seed storage conditions and processing techniques were manipulated in order to maximize seed yield. Seeds were dispersed using two methods: spring seed buoys and fall seed broadcasts. Our costs for planting 1 ha of bottom with Z. marina seeds ranged from $6,674 to $165,699 depending on seeding density and dispersal method used. The average cost per Z. marina seed was $0.17. Interannual variations in seed collection yield and seed viability after summer storage had great impact on final costs. Our results suggest that the use of seeds for large‐scale Z. marina restoration offers a competitive advantage to more traditional transplanting methods.     

The effect of extended exposure to hypersaline conditions on the germination of five inland halophyte species

In order to determine how salinity and exposure time affect seed viability and germination, seeds of five halophytes, Atriplex prostrata, Hordeum jubatum, Salicornia europaea, Spergularia marina, and Suaeda calceoliformis were exposed to 3.0, 5.0, and 10.0% NaCl solutions for 30, 60, 90, 365, and 730 d. Recovery experiments in distilled water indicated significantly different species responses to salinity over time. Percentage germination and rate of germination in H. jubatum were dramatically reduced following extended exposure and all seeds exposed to 10% NaCl for > 1 yr failed to germinate. Spergularia marina seeds were stimulated following short-term exposure to 3% NaCl; however, germination was delayed and overall germination was significantly reduced with exposure time in the two higher salinity levels. Percentage germination in A. prostrata decreased over time, but salinity level was not related to this reduction. Germination of S. europaea and S. calceoliformis, the most salt-tolerant species being tested, was stimulated by exposure to high salinity. Both species had a significant increase in percentage germination and in the germination rate when compared to seeds germinated in distilled water. Baseline germination data from seeds placed in 0, 1, 2, and 3% NaCl solutions indicated that S. europaea and S. calceoliformis were the only species to germinate in the 3% NaCl solution. Spergularia marina failed to germinate in the 2% NaCl treatment, and germination of A. prostrata and H. jubatum was significantly reduced at this salinity level. It is concluded that prolonged exposure to saline solutions can inhibit or stimulate germination in certain species, and the resulting germination and recovery responses are related to the duration and intensity of their exposure to salt in their natural habitats.     

Large‐Scale Zostera marina (eelgrass) Restoration in Chesapeake Bay,Maryland, USA. Part II: A Comparison of Restoration Methods in the Patuxent and Potomac Rivers

In response to systemic losses of submerged aquatic vegetation (SAV) in the Chesapeake Bay (east coast of North America), the U.S. Environmental Protection Agency's (EPA) Chesapeake Bay Program (CBP) and Maryland Department of Natural Resources (MD DNR) have considered SAV restoration a critical component in Bay restoration programs. In 2003, the CBP created the “Strategy to Accelerate the Protection and Restoration of Submerged Aquatic Vegetation in the Chesapeake Bay” in an effort to increase SAV area. As part of this strategy, large‐scale eelgrass (Zostera marina) restoration efforts were initiated in the Patuxent and Potomac Rivers in Maryland. From 2004 to 2007, nearly 4 million Z. marina seeds were dispersed over 10 ha on the Patuxent River and almost 9 million seeds over 16 ha on the Potomac River. Z. marina seedling establishment was consistent throughout the project (<4%); however, restored eelgrass survival was highly dependent on restoration site. Restoration locations on the Patuxent River experienced initial Z. marina seedling germination, but no long‐term plant survival. Restored Z. marina on the Potomac River has persisted and expanded, both vegetatively and sexually, beyond initial seeding areas. Healthy Z. marina beds now cover approximately five acres of the Potomac River bottom for the first time in decades. The differential success of Z. marina restoration efforts in the two rivers is evidence for the necessity of carefully considering site‐specific characteristics when using large‐scale seeding methods to achieve successful SAV restoration.     

威海天鹅湖大叶藻(Zostera marina)与日本鳗草(Zostera japonica)根际微生物群落结构及其驱动机制

[目的] 本文以威海天鹅湖大叶藻和日本鳗草根际沉积物为主要研究对象,探究不同生长时期的海草根际微生物群落结构多样性,并分析导致微生物群落结构差异的内在因素。[方法] 选取威海天鹅湖大叶藻和日本鳗草根际沉积物与非草区表层沉积物,采用高通量测序技术（Illumina MiSeq platform）解析海草不同生长时期下根际与非草区微生物群落特征。[结果] 微生物群落结构差异由海草生长时期以及海草是否定植共同驱动。在海草成熟期,丙酸菌属（Propionigenium）在大叶藻与日本鳗草根际有明显富集,其相对丰度分别为11.58%和14.26%;在海草幼苗期,脱硫球茎菌科（Desulfobulbaceae）在海草根际富集（大叶藻：2.299%,日本鳗草：4.092%）;在海草衰退期时,硫卵菌属（Sulfurovum）的相对丰度在根际较高（大叶藻：5.624%,日本鳗草：3.749%）。此外,海草生长时期对不同样品之间微生物群落结构差异的解释度最大（R²=0.20335,P=0.002）。PICRUSt2功能预测结果表明各功能基因在海草不同生长时期所呈现的趋势一致,但丰度上呈现出幼苗期 > 成熟期 > 衰退期的结果。[结论] 天鹅湖海草床沉积物微生物群落结构在海草不同生长时期呈现不同的多样性特征,具有明显的根际效应且不同种类海草的根际微生物群落无显著差异,不具有物种特异性。     

Suitable water temperature for seed storage of Zostera japonica for subtropical seagrass bed restoration

Restoration of Zostera japonica is needed. Laboratory culture experiments to know the germination characteristics might be helpful for implementation of actual restoration. As a part of germination experiments, we explored suitable water temperature for long-term storage of Z. japonica seeds. This work was based on earlier reports of Zostera marina, which presumably has similar physiological properties to Z. japonica. This study consisted of two experiments: (1) preservation experiments to investigate the fate of stored Z. japonica seeds and (2) germination experiments to investigate the germination potential of the stored seeds. The results of the preservation experiments suggested that seed condition, that is, germinated, degraded, unstable, stable, etc., showed variations between the seeds stored at 4 and 23 °C. The majority of the seeds stored at 4 °C were germinated, while those at 23 °C seemed to be degraded, presumably by bacteria and mold. The germination experiments suggested high germination potential of seeds stored at 4 °C even after 302 days had elapsed. In conclusion, including previously reported results on Z. marina, low temperature was suitable for the preservation of seeds to maintain germination potential.     

海草种子特性与海草床修复

海草床面积的急剧减少已引起人们对海草床生态修复的重视, 种子在海草床修复中具有较大的应用潜力。该文系统归纳了近年来有关海草种子的研究概况及其应用进展: 从种子生物学和生态学角度对海草种子的形态结构、发育、散布、休眠和萌发进行了归纳比较; 提出了在研究天然海草种子库的同时, 有必要建立人工海草种子库的观点, 并指出建立和完善人工海草种子库需基于种子生物学和生态学研究, 以指导种子的采集和保存; 对海草种子的播种方式和利用种子修复海草床的途径进行了阐述, 并指出了目前研究和应用中存在的一系列问题。最后对海草种子的研究和应用前景进行了展望, 提出海草种子生态学将成为今后的研究热点, 在应用方面, 仍然需要以提高种子萌发率和成苗率为研究重点, 并指出如果利用由种子获得的人工幼苗进行海草床修复, 需首先解决幼苗成活率低的难题。     

Seed germination and seedling growth of Zostera marina L. (eelgrass) in the chesapeake bay

Seed germination and seedling growth of Zostera marina L. were monitored in the Chesapeake Bay in 1979 and 1980. Harvested seeds were placed in small acrylic tubes at several sites representing the salinity range of Z. marina distribution. Seed germination was observed first in late September and continued through May, with peaks in the fall and spring. The majority of seeds that germinated (66%) did so between December and March when water temperatures ranged from 0–10°C. There was no correlation between sites (different salinity regimes) and frequency of germination rates, indicating that salinity was not a major factor in the germination process in this study. Additional information on seed germination was available for seeds collected in 1977 and 1980 and subsequently monitored for germination at only one site. These data were similar to germination frequency recorded in 1979–1980.Seedling growth was measured from individuals collected from an existing Zostera marina bed. Seedlings were collected from November through May, at which time we could no longer distinguish seedlings from existing vegetative stock. Growth was characterized by the increased length of the primary shoot, number of leaves per shoot and numbers of shoots per plant. Seedling growth was slow during the winter months (water temperature ? 10°C) but rapidly increased in the spring (temperatures > 10°C). The size range of the harvested seedlings indicated that seed germination in the field probably occurred from October through April, corroborating evidence from the seed germination experiments.     

Seed Dynamics of Early and Late Successional Tree Species in Tropical Abandoned Pastures: Seed Burial as a Way of Evading Predation

We explored different treatments to enhance the probability of sowed seeds of two early successional (ES, Cecropia obtusifolia and Ochroma pyramidale) and two late successional (LS, Brosimum costaricanum and Dialium guianense) species to escape predation and germinate in abandoned cattle‐raising pasture fields in Southeastern Mexico. ES species were sown in groups of 50 seeds under three treatments: invertebrate exclusion, burial, and exposition to seedeaters. LS species were sown in groups of 10 seeds under three treatments: vertebrate exclusion, burial, and exposition to seedeaters. We registered seed predation and germination 2, 4, 8, 16, 32, and 64 days after the initial sowing. Overall, ES showed higher predation rates (mean ± SE = 0.45 ± 0.07 seed seed^?1 day^?1; n = 3) than LS species (0.09 ± 0.02 seed seed^?1 day^?1). Cecropia obtusifolia was completely predated in all treatments after 8 days. Burial and exclusion treatments reduced final predation in circa 6% for O. pyramidale, relative to that of exposed seeds (85% after 8 days); most germination occurred in buried seeds (3.7%). In B. costaricanum, burial enabled germination by 10%; exposed and excluded seeds were removed 100%. Dialium guianense showed 12% germination in buried seeds and circa 20% of the seeds were not removed after 64 days. Direct sowing would be a recommended rainforest restoration practice for species with relatively large seeds if deposited in groups and buried. Studies which address variation across numerous sites are necessary in order to generate more consistent seed predation patterns and rainforest restoration principles in tropical pastures.     

Embryo growth and seed germination of four taxa of Narcissus (section Pseudonarcissi) conserved under non-recalcitrant seed conditions in germplasm banks

The aim of this study was to determine the germinative ability of the seeds of four Narcissus taxa belonging to Section Pseudonarcissi after they had been conserved under the conditions of non-recalcitrant seed storage protocols. For each taxon (N. alcaracensis, N. longispathus, N. radinganorum and N. pseudonarcissus subsp. munozii-garmendiae), one seed lot was desiccated to 4% moisture content (MC) and stored under laboratory conditions (22°C, 40–50% relative humidity (RH), whereas another was dehydrated to 3% MC and stored at −10°C. The latter treatment simulated standard conservation conditions for non-recalcitrant seeds. After 26 months, embryo growth and germination were evaluated. Seed responses were correlated with their MC upon dispersal. Seeds of N. alcaracensis, N. longispathus and N. radinganorum left to dry on the mother plant during maturation had 8–10% MC when dispersed, tolerated non-recalcitrant seed conservation and germinated to >90% under the most favorable incubation conditions. Narcissus pseudonarcissus subsp. munozii-garmendiae seeds did not undergo maturation drying and had 46.7% MC upon dispersal. They reached 100% germination after being desiccated to 4% and stored at 22°C, were not recalcitrant, but failed to germinate when stored at −10°C under non-recalcitrant seed conservation conditions. Therefore, N. alcaracensis, N. longispathus and N. radinganorum seeds can be conserved under non-recalcitrant seed conditions in germplasm banks, whereas those of N. pseudonarcissus subsp. munozii-garmendiae are moderately recalcitrant. Seed storage behavior is influenced primarily by the extent of maturation drying of the seeds on the mother plant.     

The role of after‐ripening in promoting germination of arid zone seeds: a study on six Australian species

The effects of after‐ripening (storage under warm, dry conditions) on seed germination was examined in six plant species from the arid zone of Western Australia with the aim of improving germination and germination rate for rehabilitation objectives. Study species (Acanthocarpus preissii, Anthocercis littorea, Dioscorea hastifolia, Eremophila oldfieldii, Thryptomene baeckeacea and Zygophyllum fruticulosum) were selected based on diverse plant habits, seed types and requirements for rehabilitation. After‐ripening was investigated by adjusting seed moisture content to 13 and 50 equilibrium relative humidity (eRH) at 23 °C and storing seeds at two temperatures (30 and 45 °C) from 1 to 18 months. Following storage, seeds were incubated in water, gibberellic acid (GA₃) or karrikinolide (KAR₁; the butenolide, 3‐methyl‐2H‐furo[2,3‐c]pyran‐2‐one). All after‐ripening conditions increased germination percentage and rate of A. littorea and D. hastifolia, with A. littorea only germinating when treated with GA₃ or KAR₁. The germination of Z. fruticulosum was dependent on after‐ripening temperature and seed moisture content. After‐ripening had a limited effect on the remaining three species. The restoration implications of the findings are discussed. © 2009 The Linnean Society of London, Botanical Journal of the Linnean Society, 2009, 161 , 411–421.     

Seagrass feeding choices and digestive strategies of the herbivorous fish Sarpa salpa

This is the first study investigating the plant–herbivore interaction between Sarpa salpa, which has overgrazed seagrass transplants in Portugal, and the seagrasses Cymodocea nodosa, Zostera marina and Zostera noltii, which have been considered for restoration. When offered the choice between the three seagrasses in outdoor tanks, adult S. salpa clearly preferred Z. noltii. Testing the seagrasses separately, mean ± s.d. feeding rates ranged from 21 ± 11 g seagrass fresh mass kg^?1 fish mass day^?1 for Z. marina to 32 ± 9 g seagrass fresh mass kg^?1 fish mass day^?1 for C. nodosa and 40 ± 11 g seagrass fresh mass kg^?1 fish mass day^?1 for Z. noltii (temperature = 16° C). Food‐processing rate in S. salpa did not differ between seagrasses, and there was no evidence of a regulation of processing rate according to food intake. Seagrasses differed substantially in nitrogen content and C:N, with C. nodosa containing the highest nitrogen content and lowest C:N (2·5 ± 0·1% and 14·0 ± 1·0), followed by Z. noltii (2·1 ± 0·1% and 17·0 ± 1·0) and Z. marina (1·4 ± 0·1% and 26·0 ± 2·0). Food‐processing rate in S. salpa and the nutritional value of the seagrasses were not correlated with the observed feeding preference and rate. The study suggests that C. nodosa and Z. marina are less at risk of overgrazing by S. salpa and might thus be preferable to Z. noltii for seagrass restoration in areas with noticeable abundances of this fish.     

Note on organic dormancy of estuarine Ruppia maritima L. seeds

Information on seed dormancy is one of the primary requirements for successful seedling propagation of submerged aquatics and seagrass. Studies on Ruppia maritima seed germination have been done, but the presence, requirements, and the types of dormancy have not been well understood. A laboratory study was conducted to understand presence and types of organic dormancy of the seeds of estuarine R. maritima collected from Lake Pontchartrain, Louisiana, USA. Our study results indicate that the brackish estuarine R. maritima population produces seeds that do not have any noticeable initial morphological, physical, and physiological dormancy. Although dry stratification reduced seed viability and final germination rates, drying seems to induce an earlier germination in R. maritima. Desiccation also appears to induce an environmental dormancy that can be disrupted by exposure to water. Further study on environmental dormancy is needed to provide information to develop methods for long-term seed storage that can be employed in greenhouse seedling propagation. Handling editor: P. Viaroli     

Effects of Moisture, Temperature, and Time on Seed Germination of Five Wetland Carices: Implications for Restoration

Successful restoration of sedge meadow wetlands is limited by lack of information regarding reintroduction of sedge (Carex) propagules. While restoration from seed is common for prairie restorations, little is known about the germination characteristics of many wetland plants, including sedges. We present the results of a 2.5-year study on seed germination and viability for five species of Carex common to sedge meadow and prairie pothole wetlands in temperate North America. Seed storage and germination conditions were investigated to determine the optimum combination for maintaining seed viability and stimulating germination rates over time. Seeds were germinated under seven different temperature and three moisture regimes after storage for 4, 10, and 14 months under one of four different storage regimes (dry-warm, dry-cold, moist-cold, and wet-cold). The efficacy of short-term wet-cold stratification to stimulate germination of 2.5-year-old seed after long-term dry storage was also investigated. Carex stricta, Carex comosa, and Carex lacustris showed the greatest germination response after wet-cold or moist-cold storage, while Carex lasiocarpa and Carex rostrata showed similar rates of germination after either wet-cold or dry-warm storage. Wet-cold long-term storage was associated with a high level of viability in all five species after 2.5 years. Viability and germination rates were reduced in Carex stricta, Carex comosa, and Carex lasiocarpa after long-term dry-cold storage. Germination rates of seeds stored dry for 2.5 years are not improved by short-term wet-cold treatment in any species tested. Carex seeds should be stored under wet-cold conditions to maintain seed viability over time, thus increasing the likelihood of seeding success for sedge meadow restoration.     

Seed- versus transplant-based eelgrass (Zostera marina L.) restoration success in a temperate marine lake

Despite active seagrass restoration gaining traction as a tool to halt and reverse worldwide seagrass losses, overall success remains limited. Restoration strategies, through seeding or transplantation, face different environmental bottlenecks that limit success. Choosing the most appropriate strategy of the two for a specific location, however, is hampered by lack of direct practical comparisons between strategies within a single system. To investigate potential life stage dependent bottlenecks, we compared seed-based and transplant-based restoration of Zostera marina in the subtidal saltwater Lake Grevelingen. Our results demonstrate that seedling recruitment was negatively impacted by bioturbation from the lugworm Arenicola marina and sediment movement due to hydrodynamic exposure. Transplant-based restoration was clearly more successful but surprisingly best predicted by leaf gluing by the ragworm Platynereis dumerilii. This previously undescribed interaction caused seagrass leaves to clump and reduce effective photosynthetic surface and leaf movement. We suggest that the observed behavior of these worms may result from a lack of foodweb interactions, illustrating the importance of  trophic control for seagrass restoration. Thus, in addition to recognizing life stage dependent environmental bottlenecks for restoration strategy selection, seagrass restoration may also require the active recovery of their associated food webs.     

Morphological characteristics of annual Zostera marina shoots at various germination temperatures

The timing of the transition from seed, seedlings and development into flowering is paramount importance in annual-type Zostera marina, because flowering is the first step of sexual reproduction. A majority of plants use environmental cues to regulate the transition to their developmental stages because plants must flower synchronously for successful outcrossing and must complete their sexual reproduction under favorable external conditions. The morphological characteristics (seeds and lateral shoot production, branch number, and inflorescence length) of reproductive shoots of Z. marina L. were examined in outdoor mesocosms to better understand the reproductive strategies of annual populations. Seeds in the germination experiment were divided into two groups: those exposed to cold (7 °C; vernalized group) and those left untreated (25-21 °C; non-vernalized group). All 600 seeds (300 from each group) were cultured for 2 months at 7, 10, 15, 20, and 25 °C in an indoor incubator. In the vernalized group, the germination rates were almost significantly higher than in the non-vernalized group. However, germination rates were not significantly affected by germination temperature. In outdoor mesocosms, production of vegetative shoots was observed in plants germinated at 15 and 20 °C in the vernalized group and at 10, 15 and 20 °C in the non-vernalized group. The highest number of vegetative shoots produced (35) was observed in plants germinated at 20 °C in the vernalized group, whereas seeds of either group failed to produce vegetative shoots when germinated at a low temperature (7 °C).In the flowering phase, the number of branches per shoot in the vernalized group was significantly higher than in the non-vernalized group. The total number of spadices on the 1st branches of plants in the vernalized group (germination at 20 °C) was significantly lower than that in the non-vernalized group at the same germination temperature. The total number of spadices per reproductive shoot in the vernalized group (germination at 10 °C) was also higher than in the non-vernalized group. Thus, both low temperature (vernalization) and seed germination temperature have implications for the sexual and asexual propagation of annual Z. marina populations.     

Effects of mucilage on seed germination of the desert ephemeral plant Plantago minuta Pall. under osmotic stress and cycles of wet and dry conditions

To test the role of the seed mucilage of Plantago minuta Pall. in regulating germination under osmotic stress and cycles of hydration and dehydration, two experiments were carried out using seeds with intact mucilage and mucilage‐free seeds. In Experiment 1 seeds were immersed in a range of iso‐osmotic polyethylene glycol solutions (?1.15 to 0 MPa) for 14 days; any ungerminated seeds were transferred to deionized water to investigate the recovery germination. In Experiment 2 seeds were immersed in deionized water for 24 h, and were then incubated on filter paper for an additional 13 days to ensure complete desiccation before reimbibition to test the germination recovery percentage. Under mild osmotic stress (?0.73 to 0 MPa), the intact seeds with mucilage were shown to have higher germination rates than the mucilage‐free seeds, indicating that the mucilage led to a “fast sprouting” germination strategy under mild osmotic stress. However, when seeds were exposed to high osmotic stress (?1.15 MPa), the mucilage apparently slowed the germination rate, resulting in a “risk‐balancing” germination strategy. Extreme drought induced by polyethylene glycol solution and the desiccation pretreatment accelerated germination rates compared to non‐pretreated seeds; both germination potential and recovery percentage of the mucilage seeds were significantly higher than that of the mucilage‐free seeds. Our results revealed that the seed mucilage of P. minuta plays a crucial role in regulating seed germination rates and the germination strategies adopted by controlling seed water absorption when the seeds experience different osmotic stresses or alternating wet and dry conditions.