AtDSEL, an Arabidopsis cytosolic DAD1-like acylhydrolase, is involved in negative regulation of storage oil mobilization during seedling establishment

Mobilization of seed storage reserves is essential for seed germination and seedling establishment. Here, we report that AtDSEL, an Arabidopsis thalianaDAD1-like Seedling Establishment-related Lipase, is involved in the mobilization of storage oils for early seedling establishment. AtDSEL is a cytosolic member of the DAD1-like acylhydrolase family encoded by At4g18550. Bacterially expressed AtDSEL preferentially hydrolyzed 1,3-diacylglycerol and 1-monoacylglycerol, suggesting that AtDSEL is an sn-1-specific lipase. AtDSEL-overexpressing transgenic Arabidopsis plants (35S:AtDSEL) were defective in post-germinative seedling growth in medium without an exogenous carbon source. This phenotype was rescued by the addition of sucrose to the growth medium. In contrast, loss-of-function mutant plants (atdsel-1 and atdsel-2) had a mildly fast-growing phenotype regardless of the presence of an exogenous carbon source. Electron microscopy revealed that 5-day-old 35S:AtDSEL cotyledons retained numerous peroxisomes and oil bodies, which were exhausted in wild-type and mutant cotyledons. The impaired seedling establishment of 35S:AtDSEL was not rescued by the addition of an exogenous fatty acid source, and 35S:AtDSEL seedling growth was insensitive to 2,4-dichlorophenoxybutyric acid, indicating that β-oxidation was blocked in AtDSEL-overexpressers. These results suggest that AtDSEL is involved in the negative regulation of seedling establishment by inhibiting the breakdown of storage oils.     

Relative effects of Justicia americana litter on germination,seedlings and established plants of Polygonum lapathifolium

Justicia americana (L.) Vahl is one of the most common species associated with seasonally variable aquatic habitats in the south-central U.S.A. When water levels decline, litter of Justicia decomposes on the exposed mudflats where most emergent aquatic species typically germinate and establish. In this study, we have tested the effects of aqueous leachates of Justicia shoots on the germination, seedling growth and established plant growth of Polygonum lapathifolium L., a common annual colonizer of exposed mudflats. Maximal leachate concentrations were prepared using quantities of litter comparable to those estimated for natural stands of Justicia. At these concentrations, seed germination and transplant growth were unaffected, but seedling growth was inhibited. When Polygonum was tested with lower concentrations of Justicia leachate, germination was still unaffected, but seedling growth was found to be enhanced, especially at the lowest concentrations. Using mannitol as an inert osmotic agent, Polygonum seedling growth was found to be inhibited at osmotic potentials equivalent to those of all leachate concentrations. Thus, the effects of leachate on seedling growth include both osmotic and non-osmotic components. Overall, the results support the hypothesis that the seedling stage of the plant life cycle is the stage most susceptible to permanent effects by the chemicals released from decomposing litter.     

The Response of Citrus limon Seedlings to a Symbiont,Glomus etunicatus,and a Pathogen,Radopholus similis

The influences of a vesicular-arbuscular mycorrhiza (Glomus etunicatus) and burrowing nematode (Radophohts similis), alone and in combination, on the growth of rough lemon (Citrus limon) seedlings were studied in the greenhouse. Growth of mycorrhizal seedlings was significantly greater than that of nonmycorrhizal seedlings or seedlings inoculated with R. sindlis. Mycorrhizal stimulation of seedling growth was inhibited by nematode infection. When seedlings were inoculated with G. etunicatus arid R. similis, suppression of seedling growth by R. similis was less on VAM seedlings than on nonmycorrhizal seedlings, Nonmycorrhizal seedlings infected with R. similis were significantly smaller than nonmycorrhizal seedlings free of R. similis. Vesicle formation and mycelia growth were less in nematode-infected roots.     

Genetic variation of piperidine alkaloids in Pinus ponderosa: a common garden study

Background and Aims

Previous measurements of conifer alkaloids have revealed significant variation attributable to many sources, environmental and genetic. The present study takes a complementary and intensive, common garden approach to examine genetic variation in Pinus ponderosa var. ponderosa alkaloid production. Additionally, this study investigates the potential trade-off between seedling growth and alkaloid production, and associations between topographic/climatic variables and alkaloid production.

Methods

Piperidine alkaloids were quantified in foliage of 501 nursery seedlings grown from seed sources in west-central Washington, Oregon and California, roughly covering the western half of the native range of ponderosa pine. A nested mixed model was used to test differences among broad-scale regions and among families within regions. Alkaloid concentrations were regressed on seedling growth measurements to test metabolite allocation theory. Likewise, climate characteristics at the seed sources were also considered as explanatory variables.

Key Results

Quantitative variation from seedling to seedling was high, and regional variation exceeded variation among families. Regions along the western margin of the species range exhibited the highest alkaloid concentrations, while those further east had relatively low alkaloid levels. Qualitative variation in alkaloid profiles was low. All measures of seedling growth related negatively to alkaloid concentrations on a natural log scale; however, coefficients of determination were low. At best, annual height increment explained 19·4 % of the variation in ln(total alkaloids). Among the climate variables, temperature range showed a negative, linear association that explained 41·8 % of the variation.

Conclusions

Given the wide geographic scope of the seed sources and the uniformity of resources in the seedlings'' environment, observed differences in alkaloid concentrations are evidence for genetic regulation of alkaloid secondary metabolism in ponderosa pine. The theoretical trade-off with seedling growth appeared to be real, however slight. The climate variables provided little evidence for adaptive alkaloid variation, especially within regions.Key words: Pinus ponderosa var. ponderosa, Pinaceae, 2,6-disubstituted piperidine alkaloids, secondary products, geographic variation, progeny study, plant defense, Growth–Differentiation Balance Hypothesis, PRISM     

Effect of salinity on germination,seedling emergence,seedling growth and ion accumulation of a euhalophyte Suaeda salsa in an intertidal zone and on saline inland

The effect of salinity on germination, seedling emergence, seedling growth and ion accumulation of a euhalophyte Suaeda salsa L. in an intertidal zone and on saline inland were investigated. Brown seeds of S. salsa were heavier and better developed than black seeds in both the intertidal zone and on saline inland. The brown seeds/black seeds ratio for S. salsa in the intertidal zone was much higher than that for S. salsa on saline inland. More germinated seeds grew as seedlings under high salinity for S. salsa from the intertidal zone than S. salsa on saline inland; high salinity decreased the shoot length more severely for S. salsa from saline inland than for S. salsa from the intertidal zone; the seedling growth at a range of NaCl, measured either as shoot length or shoot dry weight, for S. salsa from the intertidal zone was lower than that of S. salsa from saline inland. In conclusion, for S. salsa from the intertidal zone there appears to be selection for slower growth and producing more brown seeds. The establishment of populations of S. salsa in different saline environments depends on the responses of seed germination, seedling emergence and seedling growth to salinity. These characteristics may determine the natural distributions of S. salsa populations in different saline environments.     

How significant are endophytic fungi in bromeliad seeds and seedlings? Effects on germination,survival and performance of two epiphytic plant species

In bromeliads, nothing is known about the associations fungi form with seeds and seedling roots. We investigated whether fungal associations occur in the seeds and seedling roots of two epiphytic Aechmea species, and we explored whether substrate and fungal associations contribute to seed germination, and seedling survival and performance after the first month of growth. We found a total of 21 genera and 77 species of endophytic fungi in the seeds and seedlings for both Aechmea species by Illumina MiSeq sequencing. The fungal associations in seeds were found in the majority of corresponding seedlings, suggesting that fungi are transmitted vertically. Substrate quality modulated the germination and growth of seedlings, and beneficial endophytic fungi were not particularly crucial for germination but contributed positively to survival and growth. Overall, this study provides the first evidence of an endophytic fungal community in both the seeds and seedlings of two epiphytic bromeliads species that subsequently benefit plant growth.     

Comparative Population Biology of Critically Endangered Dracocephalum austriacum (Lamiaceae) in Two Distant Regions

Our study aims to compare the population dynamics of critically endangered species, Dracocephalum austriacum, in two distant regions (Czech and Slovak Karst) with similar habitat conditions, and to evaluate if conclusions concerning factors affecting population performance in one region could help in understanding species dynamics in the other region. Transition matrix models were used to examine population dynamics of the species. Results show that populations in the Slovak Karst are performing better than populations in the Czech Karst. The transitions contributing most to the population growth rates were those of stasis in the small and large adult plant stage. These transitions are, however, stable between populations and years, suggesting that they are not very sensitive to the ongoing changes. Transitions that contribute most to variations in population growth rate include seed and seedling production by large adult plants, seed germination and stasis and growth of small adult plants. These transitions seem to be affected by habitat conditions (soil depth and bare rock cover), genetic parameters of the populations, climate fluctuations (especially severe droughts), and shrub expansion. The transitions contributing most to the population growth rates are very similar between the regions except for contributions of seedling growth and stasis of large adults. These differences need to be considered before using information about the most important life cycle transitions in other regions. We suggest that similar results would be found for other species with narrow habitat requirements occupying very similar habitats, even in very distant regions.     

Germination,survival, and growth of grass and forb seedlings: Effects of soil moisture variability

Seed germination and seedling growth, survivorship, and final biomass and their responses to watering interval were studied in two grass and six forb species to assess germination and seedling growth responses to increased soil moisture variability as might occur with future increases in precipitation variability. Seeds were planted in prairie soil and watered at 1, 2, 4, or 7 d intervals (I). Seed germination peaked at I = 4 d whereas leaf growth in grasses and forbs, and final biomass in grasses peaked at I = 7 d, suggesting that growth and biomass were favored at greater soil moisture variability than seed germination. Biomass responses to I were stronger than the germination responses, suggesting that soil moisture variability more strongly influenced post germination growth. Individual species responses to I fell into three groups; those with responses to I for: (1) seed germination and seedling survival, (2) biomass, or (3) both germination and biomass production. These species groups may be more useful than life form (i.e., grass/forb) for understanding seed germination and seedling dynamics in grasslands during periods of soil moisture variability. Seed germination and early growth may assume more importance in grassland plant community dynamics under more variable precipitation patterns.     

Forest Structure,Stand Composition,and Climate-Growth Response in Montane Forests of Jiuzhaigou National Nature Reserve,China

Montane forests of western China provide an opportunity to establish baseline studies for climate change. The region is being impacted by climate change, air pollution, and significant human impacts from tourism. We analyzed forest stand structure and climate-growth relationships from Jiuzhaigou National Nature Reserve in northwestern Sichuan province, along the eastern edge of the Tibetan plateau. We conducted a survey to characterize forest stand diversity and structure in plots occurring between 2050 and 3350 m in elevation. We also evaluated seedling and sapling recruitment and tree-ring data from four conifer species to assess: 1) whether the forest appears in transition toward increased hardwood composition; 2) if conifers appear stressed by recent climate change relative to hardwoods; and 3) how growth of four dominant species responds to recent climate. Our study is complicated by clear evidence of 20^th century timber extraction. Focusing on regions lacking evidence of logging, we found a diverse suite of conifers (Pinus, Abies, Juniperus, Picea, and Larix) strongly dominate the forest overstory. We found population size structures for most conifer tree species to be consistent with self-replacement and not providing evidence of shifting composition toward hardwoods. Climate-growth analyses indicate increased growth with cool temperatures in summer and fall. Warmer temperatures during the growing season could negatively impact conifer growth, indicating possible seasonal climate water deficit as a constraint on growth. In contrast, however, we found little relationship to seasonal precipitation. Projected warming does not yet have a discernible signal on trends in tree growth rates, but slower growth with warmer growing season climates suggests reduced potential future forest growth.     

Adaptive Differentiation in Seedling Traits in a Hybrid Pine Species Complex,Pinus densata and Its Parental Species,on the Tibetan Plateau

Evidence from molecular genetics demonstrates that Pinus densata is a natural homoploid hybrid originating from the parent species Pinus tabuliformis and Pinus yunnanensis, and ecological selection may have played a role in the speciation of P. densata. However, data on differentiation in adaptive traits in the species complex are scarce. In this study, we performed a common garden test on 16 seedling traits to examine the differences between P. densata and its parental species in a high altitude environment. We found that among the 16 analyzed traits, 15 were significantly different among the species. Pinus tabuliformis had much earlier bud set and a relatively higher bud set ratio but poorer seedling growth, and P. yunnanensis had opposite responses for the same traits. P. densata had the greatest fitness with higher viability and growth rates than the parents. The relatively high genetic contribution of seedling traits among populations suggested that within each species the evolutionary background is complex. The correlations between the seedling traits of a population within a species and the environmental factors indicated different impacts of the environment on species evolution. The winter temperature is among the most important climate factors that affected the fitness of the three pine species. Our investigation provides empirical evidence on adaptive differentiation among this pine species complex at seedling stages.     

On the role of a Lipid-Transfer Protein. Arabidopsis ltp3 mutant is compromised in germination and seedling growth.

Plant Lipid-Transfer Proteins (LTPs) exhibit the ability to reversibly bind/transport lipids in vitro. LTPs have been involved in diverse physiological processes but conclusive evidence on their role has only been presented for a few members, none of them related to seed physiology. Arabidopsis seeds rely on storage oil breakdown to supply carbon skeletons and energy for seedling growth. Here, Arabidopsis ltp3 mutant was analyzed for its ability to germinate and for seedling establishment. Ltp3 showed delayed germination and reduced germination frequency. Seedling growth appeared reduced in the mutant but this growth restriction was rescued by the addition of an exogenous carbon supply, suggesting a defective oil mobilization. Lipid breakdown analysis during seedling growth revealed a differential profile in the mutant compared to the wild type. The involvement of LTP3 in germination and seedling growth and its relationship with the lipid transfer ability of this protein is discussed.     

Seedling growth and biomass allocation of endemic and threatened shrubs of rupestrian fields

The increasing anthropogenic pressure in the rare rupestrian fields in southeastern Brazil has led to the expansion of degraded areas on the extremely nutrient-deficient quartzitic soils. On the other hand, the use of rupestrian field native species in reclamation programmes has been hampered by the lack of studies involving seedling physiological ecology. The present study evaluated biomass allocation and seedling growth rate during early seedling growth of four Fabaceae shrubs: Collaea cipoensis, Calliandra fasciculata, Chamaecrista ramosa, and Mimosa foliolosa. The following hypotheses were tested: (i) species proportionally allocate higher biomass to the roots, presenting a high root/shoot ratio; and (ii) species exhibit low phenotypic variation because they have adapted to poor nutritional environments. A 12-month greenhouse experiment was carried out to evaluate seedling growth and biomass allocation performance in substrates with contrasting levels of soil fertility. The four species studied presented values of root/shoot ratio lower than one in both fertility conditions of the substrate. Growth parameters for Collaea and Calliandra increased with increasing soil fertility, while no differences were observed for Mimosa and Chamaecrista. Although the four species are naturally adapted to low nutritional quality soils, seedling development was not hindered by high fertility substrate conditions. Despite the remarkable differences in fertility between the substrates, the responsiveness in growth and allocation in Chamaecrista and Mimosa was lower than that expected if the species would exhibit high phenotypic variation. The implications for rupestrian field restoration are discussed.     

Growth Reduction of Apple Seedlings by Pratylenchus penetrans as Influenced by Seedling Age at Inoculation

Apple seedlings of different ages (1, 3, and 5 weeks) were inoculated with 6,900 Pratylenchus penetrans per seedling in 10-cm-diam pots in a growth chamber. Rate of growth suppression and total growth suppression of seedlings by P. penetrans were inversely proportional to seedling age at time of nematode inoculation. Younger seedlings were found to contain a higher number of nematodes per gram root.     

Phylogenetically diverse AM fungi from Ecuador strongly improve seedling growth of native potential crop trees

In many deforested regions of the tropics, afforestation with native tree species could valorize a growing reservoir of degraded, previously overused and abandoned land. The inoculation of tropical tree seedlings with arbuscular mycorrhizal fungi (AM fungi) can improve tree growth and viability, but efficiency may depend on plant and AM fungal genotype. To study such effects, seven phylogenetically diverse AM fungi, native to Ecuador, from seven genera and a non-native AM fungus (Rhizophagus irregularis DAOM197198) were used to inoculate the tropical potential crop tree (PCT) species Handroanthus chrysanthus (synonym Tabebuia chrysantha), Cedrela montana, and Heliocarpus americanus. Twenty-four plant-fungus combinations were studied in five different fertilization and AMF inoculation treatments. Numerous plant growth parameters and mycorrhizal root colonization were assessed. The inoculation with any of the tested AM fungi improved seedling growth significantly and in most cases reduced plant mortality. Plants produced up to threefold higher biomass, when compared to the standard nursery practice. AM fungal inoculation alone or in combination with low fertilization both outperformed full fertilization in terms of plant growth promotion. Interestingly, root colonization levels for individual fungi strongly depended on the host tree species, but surprisingly the colonization strength did not correlate with plant growth promotion. The combination of AM fungal inoculation with a low dosage of slow release fertilizer improved PCT seedling performance strongest, but also AM fungal treatments without any fertilization were highly efficient. The AM fungi tested are promising candidates to improve management practices in tropical tree seedling production.     

Sod cutting and soil biota effects on seedling performance

Sod cutting (i.e. top soil removal) is a restoration management option for enhancing seedling establishment and for lowering the nutrient concentration in eutrophicated soils of nutrient-poor species-rich grasslands. Removal of the upper soil changes not only abiotic soil properties but may also affect the resident soil community. We investigated the effects of sod cutting on the establishment and performance of two endangered plant species (Cirsium dissectum and Succisa pratensis) while simultaneously manipulating the interaction between seedlings and soil biota. In intact grassland and sod-cut areas at two localities, seedlings were grown in plastic tubes. Half of the tubes had a filter that excluded roots but allowed entry of fungal hyphae and soil microorganisms. The other tubes were closed (i.e. no contact with the surrounding soil). In a greenhouse experiment we studied the effect of soil solutions (with or without fungal tissue) from three grasslands and three sod-cut areas on seedling growth. Sod cutting had a positive net effect on seedling growth for S. pratensis. Access to (mycorrhizal) fungi and other soil biota resulted in a negative impact on seedling growth of both plant species, both in grassland and sod-cut areas. The greenhouse experiment confirmed that the soil biota in these meadows reduced seedling growth. Although sod cutting did not mitigate negative plant-soil feedback, it enhanced seedling growth, presumably by decreasing competition for light. Sod cutting is therefore very useful when seedling establishment needs to be stimulated.     

The effects of shrub patch sizes on the colonization of pioneer plants on the volcano Mount Koma,northern Japan

Vegetation recovery on Mount Koma, Hokkaido, Japan, has been slow after the catastrophic eruption in 1929, due to undeveloped soil and limited plant colonization. Nowadays, the seedling establishment is supported mostly by a nurse plant, Salix reinii forming shrub patches, facilitates the plant colonization. Although the effects of shrub patches should differ with patch sizes, the size effects have not been examined well. To examine the size effects, seed-sowing experiments were conducted on two common pioneer herbaceous species, Miscanthus sinensis and Polygonum sachalinense, in the field. The seed germination and seedling survival were monitored by the seeds sown into S. reinii patches (0.97 m²–4.12 m² in area) for 4 months during snow-free periods. Microenvironments altered by the patches were measured. Lab-experiments were performed to characterize the seed germination and seedling growth.Larger patches decreased light intensity and temperature more and increased litter and water content. The large patches promoted the seed germination of the two species. Interspecific interactions, examined by a seed mixture experiment, showed that the interaction increased the seed germination on M. sinensis and decreased that on P. sachalinense. On the lab-experiments at three temperatures (15, 20 and 25 °C), M. sinensis seeds germinated more at higher temperatures and obtained higher seedlings biomass. P. sachalinense germinated the seeds more at 20 °C and grew faster at lower temperatures. The total biomass of the two species was reduced by shade that intercepted 50% of light intensity. The seed germination and seedling growth of these two species became higher on litter with 2 cm in depth than on no litter. Soil water supported seed germination when the seeds of these two species were mixed while the water reduced the growth of P. sachalinense seedlings. Therefore, the dry soils were suitable for their growths. In all the treatments, P. sachalinense seedlings showed higher mortalities than M. sinensis.In conclusion, the large patches facilitated more to the colonization of pioneer plants via seed germination and growth. Large patches acted as a nursery supporting the natural regeneration in the disturbed area by improving litter accumulation, maintaining soil water, reducing strong light and/or protecting from heat.     

Effects of sea level rise on mangrove Avicennia population growth,colonization and establishment: Evidence from a field survey and greenhouse manipulation experiment

Mangroves will either face longer tidal inundation or retreat landwards in response to on-going accelerated sea level rise. However, little is known about the growth, regeneration or colonization of mangrove seedlings under the different tidal inundation regimes associated with accelerated sea level rise. In the present study, a field survey and a greenhouse mesocosm experiment were conducted to evaluate possible effects of accelerated sea level rise on colonization, establishment and seedling growth of a mangrove pioneer species, Avicennia marina. Avicennia populations at different elevations of the intertidal zone on Xiamen Island in Fujian Province, China showed distinctive patterns in both seedling density and plant age. The seedlings at lower elevations had less annual biomass accumulation and population productivity, but higher shoot to root ratios, suggesting that elevation has positive effects on seedling growth. The greenhouse mesocosm experiment with 1-year-old A. marina seedlings utilized five inundation periods (0, 2, 4, 6 and 12 h in a semidiurnal tidal cycle) and two inundation depths (root immersion and canopy immersion). Both inundation period and depth exerted significant and negative effects on biomass accumulation, photosynthetic rate, leaf electron transportation and water use efficiency. However, the negative effects of canopy immersion were more profound than root immersion. Canopy immersion exacerbated the effects of prolonged inundation, with no seedlings surviving under canopy immersion at the 12-hour treatment. These results suggest that at lower elevations with higher sea level, canopy immersion will have greater negative effects on seedling colonization, establishment and early growth of A. marina. This finding is instrumental in predicting the future dynamics of mangrove forests under increasing sea levels.     

The effects of gap disturbance on the seedling emergence,survival and growth of two different native species in Inner Mongolia

A field study was conducted to investigate the effects of gap disturbance on the seedling establishment process of two native species. Seeds of Agropyron cristatum and Stipa krylovii were reseeded to artificially created gaps in a degraded steppe in North China. There were seven treatments: shoot gaps and root gaps (10 cm, 20 cm and 40 cm in diameters), no gaps (control). Shoot gaps were formed by removing above ground vegetation and below ground biomass without restricting the re-growth of neighbor roots back into the gap. The root gaps were accomplished by using polyvinyl chloride pipes sunk in the soil of shoot gaps to exclude neighboring roots. Seedling emergence, survival and growth performance after 90 days of growing were recorded for both species. Gap significantly increased soil moisture, especially for root gaps. Emergence increased significantly for both species as gap size increased. Seedling emergence and survivorship of both species were greater in gaps than in controls. However, the gap size showed a significantly negative effect on Agropyron cristatum's survivorship. Growth performance of Agropyron cristatum and Stipa krylovii differ in their response to gap disturbance. Gap had positive effects on seedling growth (including seedling height, dry weight, and numbers of tillers and leaves) of Stipa krylovii, but had negative effects on seedling growth of Agropyron cristatum. The two species have significantly different responses to gap disturbance. All results suggest that Stipa krylovii is a gap-enhanced species, and Agropyron cristatum is not. Predation by insects may be one of the key reasons to explain the stand dominance in this grassland.     

Soil microbial communities from an elevational cline differ in their effect on conifer seedling growth

Sub-alpine environments consist of altitudinal gradients associated with dramatic changes in plant growth and community composition, but the role of soil feedbacks and microbe interactions is largely unknown. Here, we examine the influence of the overall soil microbial community, with a focus on ectomycorrhizal and dark septate endophytic root colonizing fungi, from low, mid, and high elevations on the growth of Pinus contorta and Picea glauca × engelmannii. The influence of the soil microbial community was tested on seedlings from the same three elevations in order to determine ‘home’ versus ‘away’ effects on conspecifics of differing elevations. The low elevation soil was the most fertile and harbored a soil microbial community with an overall negative effect on seedling growth. In contrast, the high elevation soil was the least fertile and had a microbial community that enhanced seedling growth. However, only the soil microbial community in the highest elevation soil resulted in a stronger influence on the native P. contorta seedlings than seedlings originating from lower elevations. Despite the overall influence of the soil microbial community, ectomycorrhizal colonization was significantly correlated with P. glauca × engelmannii growth rates, but colonization by dark septate endophytes showed no relationship with seedling growth. The results provide evidence that plant—soil microbial community relationships are dependent on soil environment. Moreover, our results provide further support for the importance of soil microbes in facilitating seedling growth toward the edge of their elevational range.