Salt-Enhanced Reproductive Development of Suaeda salsa L. Coincided With Ion Transporter Gene Upregulation in Flowers and Increased Pollen K+ Content.
ABSTRACT: Halophytes are adapted to saline environments and demonstrate optimal reproductive growth under high salinity. To gain insight into the salt tolerance mechanism and effects of salinity in the halophyte Suaeda salsa, the number of flowers and seeds, seed size, anther development, ion content, and flower transcript profiles, as well as the relative expression levels of genes involved in ion transport, were analyzed in S. salsa plants treated with 0 or 200 mM NaCl. The seed size, flower number, seed number per leaf axil, and anther fertility were all significantly increased by 200 mM NaCl treatment. The Na+ and Cl- contents in the leaves, stems, and pollen of NaCl-treated plants were all markedly higher, and the K+ content in the leaves and stems was significantly lower, than those in untreated control plants. By contrast, the K+ content in pollen grains did not decrease, but rather increased, upon NaCl treatment. Genes related to Na+, K+ and, Cl- transport, such as SOS1, KEA, AKT1, NHX1, and CHX, showed increased expression in the flowers of NaCl-treated plants. These results suggest that ionic homeostasis in reproductive organs, especially in pollen grains under salt-treated conditions, involves increased expression of ion transport-related genes.
Project description:BACKGROUND:Halophytes show optimal reproduction under high-salinity conditions. However, the role of NaCl in reproduction and its possible mechanisms in the euhalophyte Suaeda salsa remain to be elucidated. RESULTS:We performed transcript profiling of S. salsa flowers and measured starch accumulation in ovules, sugar contents in flowers, and photosynthetic parameters in the leaves of plants supplied with 0 and 200?mM NaCl. Starch accumulation in ovules, sugar contents in flowers and ovules, and net photosynthetic rate and photochemical efficiency in leaves were significantly higher in NaCl-treated plants vs. the control. We identified 14,348 differentially expressed genes in flowers of NaCl-treated vs. control plants. Many of these genes were predicted to be associated with photosynthesis, carbon utilization, and sugar and starch metabolism. These genes are crucial for maintaining photosystem structure, regulating electron transport, and improving photosynthetic efficiency in NaCl-treated plants. In addition, genes encoding fructokinase and sucrose phosphate synthase were upregulated in flowers of NaCl-treated plants. CONCLUSIONS:The higher starch and sugar contents in the ovules and flowers of S. salsa in response to NaCl treatment are likely due to the upregulation of genes involved in photosynthesis and carbohydrate metabolism, which increase photosynthetic efficiency and accumulation of photosynthetic products under these conditions.
Project description:BACKGROUND AND AIMS:Secondary pollen presentation (SPP), a floral mechanism of reproductive adaptation, has been described for more than 200 years, with nine types SPP recorded. However, few studies have been done experimentally to link the floral mechanism of SPP to its functional roles in pollination process. This study aims to describe a new SPP mechanism from a wild ginger (Zingiber densissimum, Zingiberaceae) and explore how the pollen arrangement of SPP affects pollen removal during the interaction with different pollinators. METHODOLOGY/PRINCIPAL FINDINGS:Field observations and experiments revealed that flowers lasted for less than one day. The breeding system was partially self-incompatible. Two bee species, Macropis hedini (which carried pollen dorsally) and Amegilla zonata (which carried pollen ventrally) were the primary pollinators. About a third of pollen grains were relocated from the anther to the labellum staminode of flowers through the adherence of aggregated pollen chains, while other grains were presented on the anther. In a single visit, each bee species removed pollen grains from both the labellum staminode and the anther. Macropis hedini was more effective than Amegilla zonata. CONCLUSIONS/SIGNIFICANCE:Our study describes a new SPP mechanism in angiosperms. The new SPP mode enables pollen grains presented on the anther and the labellum staminode simultaneously via the adherence of aggregated pollen chains, thus promoting pollen to be taken away by different pollinators. This SPP mechanism plays a key role during pollen removal and may have evolved under the pressure to improve male fitness.
Project description:Irrigation with 200 mM NaCl significantly increases vegetative and reproductive growth of the extreme halophyte Suaeda salsa. However, little is known about how the progeny of S. salsa plants grown under a continuous NaCl supply behave in terms of growth and seed set parameters. We investigated various plant growth and reproductive parameters of the progeny that germinated from seeds harvested from mother plants grown under 0 or 200 mM NaCl over three generations. Seedling emergence, plant height, stem diameter, total branch length, flowering branch length, flowering branch ratio, and seed production were all significantly enhanced in the progeny produced by mother plants grown with 200 mM NaCl compared to progeny of mother plants grown on low salinity conditions. Therefore, irrigation with 200 mM of NaCl is beneficial to seed development in the halophyte S. salsa and possibly contributes to population establishment in high salinity environments. Likewise, the prolonged absence of NaCl in the growth environment inhibits seed development, results in lower seed quality, and thus limits seedling growth of the progeny, thereby restricting S. salsa to a high salinity ecological niche.
Project description:Successive stamen movement directly controls pollen presentation schedules through sequential stamen maturation and changes the extent of herkogamy by altering the positions of sexual organs. However, the implications of such movements in terms of pollination are not well understood. Pollen presentation theory predicts that staggered pollen presentation should be favoured when plants are subject to diminishing returns on pollen transfer. Herkogamy on the other hand, has been interpreted as an adaptive trait that reduces sexual interference in hermaphrodite flowers. In this study, we conducted floral manipulations to determine the function of successive stamen movement in pollen transfer. By artificially manipulating the flowers to present two anthers simultaneously in the floral centre, we attempted to investigate whether changes in the anther presentation strategy affect pollen removal, deposition and the efficiency of pollinators. Compared with the natural treatment, the pollen transfer efficiency of halictid bees decreased significantly when the flowers were manipulated to present two anthers simultaneously. Although the presentation of two anthers simultaneously led to a similar pollen removal rate, there was a significant reduction in pollen deposition on neighbouring stigmas. To evaluate the effect of movement herkogamy on pollen export and deposition and seed set, the flowers were manipulated with or without the movement of stamen bending out from the floral centre. Pollen export decreased significantly when the central anther was moved away from the pistil, and pollen deposition and seed set declined significantly when the five spent anthers were retained on the pistil. Our study provides good support for the pollen presentation theory and provides direct experimental evidence that successive stamen movement could increase pollen transfer efficiency by sequential stamen maturation. Moreover, movement herkogamy promotes pollen export, deposition and seed set, and could therefore be regarded as an effective mechanism to reduce interference between male and female functions.
Project description:Distyly is a widespread floral polymorphism characterized by the flowers within a population showing reciprocal placement of the anthers and stigma. Darwin hypothesizes that distyly evolves to promote precise pollen transfer between morphs. Primula chungensis exhibits two types of anther heights, and these two types of anthers show pollen of two different size classes. To understand whether the stigma could capture more pollen grains from the anthers of the pollen donor as the separation between the stigma of pollen receiver and the anther of pollen donor decreased, the present research assessed the source of the pollen load in a series of open-pollinated flowers with continuous variation of style lengths. Individuals with continuous variation of style length were tagged, and the selected flowers in the tagged plants were emasculated the day before dehiscence. The stigma of the emasculated flowers was fixed in fuchsin gel at the end of blooming. We assessed the pollen sources on each stigma by taking photos under a microscope and measured the diameter of each conspecific pollen grain with ImageJ. We found that a shorter distance from the stigmas to the anthers of a pollen donor gave the flower a higher capacity to receive pollen from those anthers. Our result provides a new evidence that distyly could promote the pollen transfer between morphs, which is consistent with Darwin's hypothesis of disassortative pollination. An alternative hypothesis for the evolution of distyly (e.g. selfing avoidance) might also be true, but less likely, because self-incompatibility would greatly avoid self-fertilization for many distylous species.
Project description:Gentiana leucomelaena manifests dramatic flower color polymorphism, with both blue- and white-flowered individuals (pollinated by flies and bees) both within a population and on an individual plant. Previous studies of this species have shown that pollinator preference and flower temperature change as a function of flower color throughout the flowering season. However, few if any studies have explored the effects of flower color on both pollen viability (mediated by anther temperature) and pollinator preference on reproductive success (seed set) in a population or on individual plants over the course of the entire flowering season. Based on prior observations, we hypothesized that flower color affects both pollen viability (as a function of anther temperature) and pollen deposition (as a function of pollinator preference) to synergistically determine reproductive success during the peak of the flowering season. This hypothesis was tested by field observations and hand pollination experiments in a Tibetan alpine meadow. Generalized linear model and path analyses showed that pollen viability was determined by flower color, flowering season, and anther temperature. Anther temperature correlated positively with pollen viability during the peak of the early flowering season, but negatively affected pollen viability during the peak of the mid- to late flowering season. Pollen deposition was determined by flower color, flowering season (early, or mid- to late season), and pollen viability. Pollen viability and pollen deposition were affected by flower color that in turn affected seed set across the peak of the flowering season (i.e., when the greatest number of flowers were being pollinated). Hand pollination experiments showed that pollen viability and pollen deposition directly influenced seed set. These data collectively indicate that the preference of pollinators for flower color and pollen viability changed during the flowering season in a manner that optimizes successful reproduction in G. leucomelaena. This study is one of a few that have simultaneously considered the effects of both pollen viability and pollen deposition on reproductive success in the same population and on individual plants.
Project description:The selective pressure imposed by maximizing male fitness (pollen dispersal) in shaping floral structures is increasingly recognized and emphasized in current plant sciences. To maximize male fitness, many flowers bear a group of stamens with temporally separated anther dehiscence that prolongs presentation of pollen grains. Such an advantage, however, may come with a cost resulting from interference of pollen removal by the dehisced anthers. This interference between dehisced and dehiscing anthers has received little attention and few experimental tests to date. Here, using one-by-one stamen movement in the generalist-pollinated Parnassia palustris, we test this hypothesis by manipulation experiments in two years. Under natural conditions, the five fertile stamens in P. palustris flowers elongate their filaments individually, and anthers dehisce successively one-by-one. More importantly, the anther-dehisced stamen bends out of the floral center by filament deflexion before the next stamen's anther dehiscence. Experimental manipulations show that flowers with dehisced anther remaining at the floral center experience shorter (1/3-1/2 less) visit durations by pollen-collecting insects (mainly hoverflies and wasps) because these 'hungry' insects are discouraged by the scant and non-fresh pollen in the dehisced anther. Furthermore, the dehisced anther blocks the dehiscing anther's access to floral visitors, resulting in a nearly one third decrease in their contact frequency. As a result, pollen removal of the dehiscing anther decreases dramatically. These results provide the first direct experimental evidence that anther-anther interference is possible in a flower, and that the selection to reduce such interferences can be a strong force in floral evolution. We also propose that some other floral traits, usually interpreted as pollen dispensing mechanisms, may function, at least partially, as mechanisms to promote pollen dispersal by reducing interferences between dehisced and dehiscing anthers.
Project description:Continuous exposure of tomato 'Trust' to high temperatures (day/night temperatures of 32/26 degrees C) markedly reduced the number of pollen grains per flower and decreased viability. The effect of heat stress on pollen viability was associated with alterations in carbohydrate metabolism in various parts of the anther during its development. Under control, favourable temperature conditions (28/22 degrees C), starch accumulated in the pollen grains, where it reached a maximum value 3 d before anthesis; it then diminished towards anthesis. During anther development, the concentration of total soluble sugars gradually increased in the anther walls and in the pollen grains (but not in the locular fluid), reaching a maximum at anthesis. Continuous exposure of the plants to high temperatures (32/26 degrees C) prevented the transient increase in starch concentration and led to decreases in the concentrations of soluble sugars in the anther walls and the pollen grains. In the locular fluid, however, a higher soluble sugar concentration was detected under the high-temperature regime throughout anther development. These results suggest that a major effect of heat stress on pollen development is a decrease in starch concentration 3 d before anthesis, which results in a decreased sugar concentration in the mature pollen grains. These events possibly contribute to the decreased pollen viability in tomato.
Project description:In habitats where resource availability declines during the growing season, selection may favor early-flowering individuals. Under such ephemerally favorable conditions, late-blooming species (and individuals) may be particularly vulnerable to resource limitation of seed production. In California, a region prone to seasonal drought, members of the annual genus Clarkia are among the last to flower in the spring. We compared pollen limitation (PL) of seed set and outcrossing rates between early- and late-flowering individuals in two mixed-mating Clarkia taxa to detect whether flowering time is associated with changes in seed set due to resource depletion, PL, or increased selfing. In 2008-2010, we hand-pollinated one flower on a total of 1855 individual plants either Early (near the onset of flowering) or Late (near the end of flowering) in the flowering season and compared seed set to adjacent, open-pollinated flowers on the same stem. To assess the contribution of pollen quality to reproduction, we first (2008) used allozymes to estimate outcrossing rates of seeds produced by Early and Late open-pollinated flowers. Second (2009), we conducted an anther-removal experiment to estimate self-pollen deposition. Seed set in Clarkia unguiculata was not pollen-limited. Clarkia xantiana ssp. xantiana was pollen-limited in 2008 and 2010, but not 2009. PL did not differ between Early and Late treatments. In both taxa, seed set of Early flowers was greater than Late flowers, but not due to PL in the latter. Reproduction was generally pollinator-dependent. Most pollen deposition was xenogamous, and outcrossing rates were >0.7 - and similar between Early and Late periods. These results suggest that pollen receipt and pollen quality remain seasonally consistent. By contrast, the resources necessary to provision seeds decline, reducing the fitness benefits associated with resource allocation to ovules.
Project description:Bird pollination systems are dominated by specialist nectarivores, such as hummingbirds in the Americas and sunbirds in Africa. Opportunistic (generalist) avian nectarivores such as orioles, weavers and bulbuls have also been implicated as plant pollinators, but their effectiveness as agents of pollen transfer is poorly known. Here, we compare the single-visit effectiveness of specialist and opportunistic avian nectarivores as pollinators of Aloe ferox, a plant that relies almost exclusively on birds for seed production. We found that the number of pollen grains on stigmas of flowers receiving single visits by opportunistic avian nectarivores was approximately threefold greater than on those receiving single visits by specialist sunbirds and about twofold greater than on those that received single visits by honeybees. The number of pollen grains on stigmas of flowers visited by sunbirds was similar to that on stigmas of unvisited flowers. These results show that opportunistic birds are highly effective pollinators of A. ferox, supporting the idea that some plants are specialized for pollination by opportunistic birds.