Resource reallocation patterns within Sagittaria trifolia inflorescences following differential pollination.

PREMISE OF THE STUDY: Understanding resource allocation to reproduction, a key factor in life history tradeoffs, has long intrigued plant ecologists. Despite the recognized importance of understanding the movement of resources among flowers following variable pollination, the patterns of resource reallocation to plant reproductive organs have not been thoroughly addressed. In this study, we aimed to empirically explore how resources redistribute within inflorescences in response to differential pollination intensities. METHODS: Using a common herb, Sagittaria trifolia, we conducted supplemental and controlled pollination for single, some, or all flowers in simple and complex inflorescences, and compared their resulting fruiting probabilities, seed production, and average seed masses. KEY RESULTS: Pollen supplementation of a single flower significantly increased its fruiting probability; however, the same manipulation of an inflorescence did not increase its overall reproduction. Single pollen-supplemented flowers had a higher percentage fruit set than inflorescences receiving supplemental pollination. In complex inflorescences, supplemental pollination had no effect on the reproductive success of flowers on the lateral or main branches. CONCLUSIONS: We provided evidence of resource reallocation from controlled to pollen-supplemented flowers in simple inflorescences; however, resources were unlikely to be reallocated between the main and lateral branches in the complex inflorescences, suggesting that flowering branches represent integrated physiological units in S. trifolia. The results also demonstrated that single-flower supplemental pollination would exaggerate pollen limitation and lead to a biased understanding of a plant's reproductive status.

Unique growth paths of heterospecific pollen tubes result in late entry into ovules in the gynoecium of Sagittaria (Alismataceae).

Pollen-pistil interactions are a fundamental process in the reproductive biology of angiosperms and play a particularly important role in maintaining incipient species that exist in sympatry. However, the majority of previous studies have focused on species with syncarpous gynoecia (fused carpels) and not those with apocarpous gynoecia (unfused carpels). In the present study, we investigated the growth of conspecific pollen tubes compared to heterospecific pollen tubes in Sagittaria species, which have apocarpous gynoecia. We conducted controlled pollinations between S. pygmaea and S. trifolia and observed the growth of conspecific and heterospecific pollen tubes under a fluorescence microscope. Heterospecific and conspecific pollen tubes arrived at locules within the ovaries near simultaneously. However, conspecific pollen tubes entered into the ovules directly, whereas heterospecific tubes passed through the carpel base and adjacent receptacle tissue, to ultimately fertilize other unfertilized ovules. This longer route taken by heterospecific pollen tubes therefore caused a delay in the time required to enter into the ovules. Furthermore, heterospecific pollen tubes displayed similar growth patterns at early and peak pollination. The growth pattern of heterospecific pollen tubes at late pollination was similar to that of conspecific pollen tubes at peak pollination. Heterospecific and conspecific pollen tubes took different routes to fertilize ovules. A delayed entry of heterospecific pollen into ovules may be a novel mechanism of conspecific pollen advantage (CPA) for apocarpous species.

Water level-dependent morphological plasticity in Sagittaria montevidensis Cham. and Schl. (Alismataceae). / Plasticidade morfológica dependente do nível da água em Sagittaria montevidensis Cham. And Schl. (Alismataceae).

Aquatic plants are able to alter their morphology in response to environmental condition variation, such as water level fluctuations. The aim of this study was to evaluate the effect of water level on Sagittaria montevidensis morphology through measures of vegetative structures formed in drought and flood periods. We hypothesised that the plant height and the biomass of S. montevidensis leaves will increase during flood periods, while the biomass and diameter of petioles, and the basal plant area will increase during dry periods. We sampled a total amount of 270 individuals in nine sediment banks per visit, totalling 1080 plants. In order to compare plant morphology between dry and flood periods, we measured the water level in each bank and took the following variables for each plant: diameter, height and diameter of the biggest petiole. In order to compare biomass allocation between dry and flood periods, we sampled a total amount of 90 individuals in nine sediment banks per visit, totalling 360 plants. Plants were dried and weighed in the laboratory. All measured morphologic traits, as well as the biomass of leaf blades and petioles, were higher during flood periods, indicating that water level highly influences the morphology of S. montevidensis individuals. Our results suggest that these morphological responses allow survival and maintenance of S. montevidensis populations under environmental stress. These results can be linked to the invasive potential of S. montevidensis and sheds light on basic management practices that may be applied in the future.

As plantas aquáticas são capazes de alterar a sua morfologia em resposta a variações nas condições ambientais, tais como mudanças no nível da água. Nosso objetivo foi avaliar o efeito do nível da água na morfologia de Sagittaria montevidensis através de medidas de estruturas vegetativas formadas em períodos de seca e de cheia. Nós hipotetizamos que a altura dos indivíduos e a biomassa das folhas de S. montevidensis aumentarão durante períodos de cheia, enquanto a biomassa e diâmetro dos pecíolos, além da área basal da planta, aumentarão durante períodos de seca. Nós amostramos um total de 270 indivíduos, distribuídos em nove bancos de sedimento, por visita, totalizando 1080 plantas. Para comparar a morfologia das plantas entre os períodos de cheia e seca nós medimos o nível de água em cada banco e tomamos as seguintes medidas para cada planta: diâmetro, altura e diâmetro do maior pecíolo. Para comparar a alocação de biomassa entre os períodos de cheia e seca nós amostramos um total de 90 indivíduos em nove bancos de sedimento por visita, totalizando 360 plantas. As plantas foram secas em estufa e pesadas em laboratório. As plantas foram maiores no período de cheia e também apresentaram maior número e biomassa de folhas, maior diâmetro e biomassa de pecíolos e maiores áreas basais das rosetas. Nós concluímos que o nível da água influencia muito na morfologia de S. montevidensis. Nossos resultados sugerem que essas respostas morfológicas podem permitir a sobrevivência e manutenção de populações de S. montevidensis em estresse ambiental. Esses resultados podem ser ligados ao potencial invasivo de S. montevidensis e lançam luzes sobre práticas de manejo que poderão ser aplicadas no futuro.

Water level-dependent morphological plasticity in Sagittaria montevidensis Cham. and Schl. (Alismataceae).

Aquatic plants are able to alter their morphology in response to environmental condition variation, such as water level fluctuations. The aim of this study was to evaluate the effect of water level on Sagittaria montevidensis morphology through measures of vegetative structures formed in drought and flood periods. We hypothesised that the plant height and the biomass of S. montevidensis leaves will increase during flood periods, while the biomass and diameter of petioles, and the basal plant area will increase during dry periods. We sampled a total amount of 270 individuals in nine sediment banks per visit, totalling 1080 plants. In order to compare plant morphology between dry and flood periods, we measured the water level in each bank and took the following variables for each plant: diameter, height and diameter of the biggest petiole. In order to compare biomass allocation between dry and flood periods, we sampled a total amount of 90 individuals in nine sediment banks per visit, totalling 360 plants. Plants were dried and weighed in the laboratory. All measured morphologic traits, as well as the biomass of leaf blades and petioles, were higher during flood periods, indicating that water level highly influences the morphology of S. montevidensis individuals. Our results suggest that these morphological responses allow survival and maintenance of S. montevidensis populations under environmental stress. These results can be linked to the invasive potential of S. montevidensis and sheds light on basic management practices that may be applied in the future.

Identification of differentially expressed genes relevant to corm formation in Sagittaria trifolia.

Sagittaria trifolia is a good model of wetland plants to elucidate the formation of corm. However, few studies have been conducted to uncover the complexity of gene expression involved in corm formation. In this study, high-throughput tag-sequencing based on Solexa Genome Analyzer Platform was applied to monitor the changes in gene expression with three libraries of differentially expressed genes (DEGs) (C1 library: stolon stage, C2 library: initial swelling stage and C3 library: swelling stage) during corm formation in Sagittaria trifolia. Approximately 6.0 million tags were sequenced, and 5854021, 5983454, and 5761079 clean tags including 138319, 116804, and 101739 distinct tags were obtained after removal of low quality tags from each library, respectively. About 46% distinct tags were unambiguous tags mapping to the reference genes, and 33% were unambiguous tag-mapped genes. Totally, 20575, 19807, and 18438 were annotated in C1, C2, and C3 libraries, respectively, after mapping their functions in existing databases. In addition, we found that profiling of gene expression in C1/C2 and C2/C3 libraries were different among most of the selected 20 DEGs. Most DEGs in C1/C2 libraries were relevant to hormone synthesis and response; energy metabolism and stress response, while most of the genes in C2/C3 libraries were involved in carbohydrate metabolism. All up-regulated transcriptional factors and 16 important genes relevant to corm formation in three libraries were also identified. To further analyze the expression of 9 genes, from the results of tag-sequencing, qRT-PCR was applied. In summary, this study provides a comprehensive understanding of gene expression, during the formation of corm in Sagittaria trifolia.

Effects of population size on synchronous display of female and male flowers and reproductive output in two monoecious Sagittaria species.

BACKGROUND: Flowering synchrony and floral sex ratio have the potential to influence the mating opportunities and reproductive success through female function. Here, we examine the variances in synchronous display of female and male function, ratio of male to female flowers per day and subsequently reproductive output in small populations of two monoecious plants, Sagittaria trifolia and Sagittaria graminea. METHODOLOGY/PRINCIPAL FINDING: We created plant populations of size 2, 4, 10 and 20 and recorded the daily number of blooming male and female flowers per plant to determine daily floral display, flowering synchrony index and ratio of male to female flowers per day. We also harvested the fruits, counted the seeds and calculated the number of fruits and seeds per flower to measure reproductive success through female function. There is less overlap in flowering time of female and male function in smaller populations than in larger populations. Most importantly, we found that male-biased floral sex ratio and imbalanced display period of female and male function for individual plant can lead to a population-size-dependent ratio of male to female flowers per day. Increasing ratio of male to female flowers per day was generally associated with a greater percentage of fruit production. CONCLUSIONS/SIGNIFICANCE: Our results highlight the importance of flowering synchrony of female and male function and population-size-dependent ratio of male to female flowers per day for female reproductive success. This finding improves our understanding of a mechanism that reduces reproductive success in small populations.

Trade-offs between clonal and sexual reproduction in Sagittaria latifolia (Alismataceae) scale up to affect the fitness of entire clones.

Many plants combine sexual reproduction with vegetative propagation, but how trade-offs between these reproductive modes affect fitness is poorly understood. Although such trade-offs have been demonstrated at the level of individual shoots (ramets), there is little evidence that they scale up to affect genet fitness. For hermaphrodites, reproductive investment is further divided between female and male sexual functions. Female function should generally incur greater carbon costs than male function, which might involve greater nitrogen (N) costs. Using a common garden experiment with diclinous, clonal Sagittaria latifolia we manipulated investment in reproduction through female and male sex functions of 412 plants from monoecious and dioecious populations. We detected a 1:1 trade-off between biomass investment in female function and clonal reproduction. For male function, there was no apparent trade-off between clonal and sexual reproduction in terms of biomass investment. Instead, male function incurred a substantially higher N cost. Our results indicate that: trade-offs between investment in clonal propagation and sexual reproduction occur at the genet level in S. latifolia; and sexual reproduction interferes with clonal expansion, with investment in female function limiting the quantity of clonal propagules produced, and investment in male function limiting the nutrient content of clonal propagules.

Extra-gynoecial pollen-tube growth in apocarpous angiosperms is phylogenetically widespread and probably adaptive.

• Fusion of floral carpels (syncarpy) in angiosperms is thought to have allowed for significant improvements in offspring quantity and quality in syncarpous species over gymnosperms and apocarpous (free-carpelled) angiosperms. Given the disadvantages of apocarpy, it remains an evolutionary puzzle why many angiosperm lineages with free carpels (apocarpy) have been so successful and why some lineages show reversals to apocarpy. • To investigate whether some advantages of syncarpy may accrue in other ways to apocarpous species, we reviewed previous studies of pollen-tube growth in apocarpous species and also documented pollen-tube growth in nine additional apocarpous species in six families. • Anatomical studies of a scattering of apocarpous paleodicots, monocots, and eudicots show that, after transiting the style, 'extra' pollen tubes exit fully fertilized carpels and grow to other carpels with unfertilized ovules. In many species this occurs via openings in the simple carpels, as we report here for Sagittaria potamogetifolia, Sagittaria pygmaea, Sedum lineare, and Schisandra sphenanthera. • The finding that extra-gynoecial pollen-tube growth is widespread in apocarpous species eliminates the possibility of a major fitness cost of apocarpy relative to syncarpy and may help to explain the persistence of, and multiple reversals to, apocarpy in the evolutionary history of angiosperms.

Short-term accumulation of atrazine by three plants from a wetland model system.

This work describes the accumulation and distribution of the herbicide atrazine in soil, water, and roots from three wetland model systems using the monocots Typha domingensis, Sagittaria lancifolia, and Echinochloa pyramidalis. Results were analyzed from a 3(3) full factorial experimental design, in order to describe the effect of accumulation of atrazine and times of exposure in the species evaluated. We found that accumulation depends on the species and the herbicide concentration; about 30% was accumulated in soil, 40% in roots, and 10-20% in water. By the end of the experiment, E. pyramidalis accumulated 8.47 mg/l of atrazine and 14.39 mg/l T. domingensis; in all cases, adsorption accounted for 1.4%, fitting a Langmuir model with a k(d) of 14.47.

Life-history differentiation and the maintenance of monoecy and dioecy in Sagittaria latifolia (Alismataceae).

The existence of monoecious and dioecious populations within plant species is rare. This limits opportunities to investigate the ecological mechanisms responsible for the evolution and maintenance of these contrasting sexual systems. In Sagittaria latifolia, an aquatic flowering plant, monoecious and dioecious populations exist in close geographic proximity but occupy distinct wetland habitats differing in the relative importance of disturbance and competition, respectively. Life-history theory predicts contrasting evolutionary responses to these environmental conditions. We propose that the maintenance of monoecy and dioecy in S. latifolia is governed by ecological selection of divergent life-history strategies in contrasting habitats. Here we evaluate this hypothesis by comparing components of growth and reproduction between monoecious and dioecious populations under four conditions: natural populations, a uniform glasshouse environment, a common garden in which monoecious and dioecious populations and their F1 progeny were compared, and a transplant experiment using shaded and unshaded plots in a freshwater marsh. Plants from dioecious populations were larger in size and produced heavier corms in comparison with monoecious populations. Monoecious populations flowered earlier and produced more flowers, clonal ramets, and corms than dioecious populations. The life-history differences between the sexual systems were shown to have a quantitative genetic basis, with F1 progeny generally exhibiting intermediate trait values. Survival was highest for each sexual system in field plots that most closely resembled the habitats in which monoecious (unshaded) and dioecious (shaded) populations grow. These results demonstrate that monoecious and dioecious populations exhibit contrasting patterns of investment in traits involved with growth and reproduction. Selection for divergent life histories between monoecious and dioecious populations of S. latifolia appears to be the principal mechanism maintaining the integrity of the two sexual systems in areas of geographic overlap.

Metal bioavailability and speciation in a wetland tailings repository amended with biosolids compost, wood ash, and sulfate.

Lead poisoning of waterfowl from direct ingestion of wetland mine tailings has been reported at the Coeur d'Alene River basin in Idaho. A greenhouse study was conducted to evaluate the effects of surface applications of amendments on lead bioavailability in the tailings. Treatments included sediment only, and sediment with three different surface amendments: (i) biosolids compost plus wood ash, (ii) compost + wood ash + a low SO4(2-) addition as K2SO4, and (iii) compost + wood ash + a high SO4(2-) addition. Measured variables included growth and tissue Pb, Zn, and Cd concentration of arrowhead (Sagittaria latifolia Willd.) and cattail (Typha latifolia L.) and soil pH, redox potential (Eh), pore water Pb, Pb speciation by X-ray absorption spectroscopy, and in vitro Pb bioavailability. The compost + ash amendment alleviated phytotoxicity for both plant species. Bioavailability of Pb as measured by a rapid in vitro extract decreased by 24 to 34% (over control) in the tailings directly below the amendment layer in the compost + SO4 treatments. The ratio of acid volatile sulfide (AVS) to simultaneously extracted metals (SEM) also indicated a reduction in Pb bioavailability (1:40 control, 1:20 compost, 1:8 compost + low SO4, and 1:3 compost + high SO4). Extended X-ray adsorption fine structure (EXAFS) and X-ray absorption near edge structure (XANES) spectroscopy data indicated that lead sulfide was greater after 99 d in the treatments that included additions of sulfate. These results indicated that, under reducing conditions, surface amendments of compost + wood ash (with or without sulfate) decreased the bioavailability of Pb in metal-contaminated mine tailings.

Gender variation of sequential inflorescences in a monoecious plant Sagittaria trifolia (Alismataceae).

In protogynous plants, female flowers of early blooming plants are at a reproductive disadvantage because they cannot set fruit due to the lack of available pollen. To study this phenomenon, gender expression of the monoecious herb Sagittaria trifolia was investigated over the entire flowering season in two field and two cultivated populations in Hubei and Hunan Provinces, China. In racemes of S. trifolia, flowers open sequentially from bottom to top, with female flowers opening first followed by male flowers. This creates a temporal separation of sexes in the species. Under field conditions small plants are often male, with production of both male and female flowers increasing with plant size. Femaleness increased among sequential inflorescences since female flower production increased whereas male flower production did not. Seed production was greater in large inflorescences because they contain more female flowers, and the number of ovules increased in female flowers at basal positions within the raceme. A consistent pattern of high seed set was observed in flowers from both field and cultivated populations. About 1 % of unfertilized ovules resulted from no pollination and 2 % of the seeds produced were only partly developed due to resource limitation. In the first inflorescence of the six experimental populations, 6.7-40.0 % of individuals produced only male flowers, and female flowers of 1.9-6.5 % individuals were aborted. The occurrence of male flowers in early blooming inflorescences could be an adaptive strategy to conserve resources and enhance pollination of female flowers in protogynous S. trifolia.