Fitness-related selection on floral traits in the distylous plant Primula poissonii (Primulaceae)

Aims Reproductive fitness of different floral phenotypes varies within and/or among populations.These variations are important to understand the process of natural selection and the evolution of floral traits.In this study,we focused on a distylous,self-incompat-ible species,Primula poissonii,to investigate fitness-related selec-tion on floral traits.Our aim was to determine how traits vary as targets of natural selection and whether morph-specific selection occurs.Methods This study was conducted at two sites(Yushuizhai at 2700 m and Haligu at 3200 m)in the Lijiang Alpine Botanical Garden,northwest-ern Yunnan,southwestern China.Insects visiting flowers of P.pois-sonii were observed,captured and identified.Randomly selected plants of long-and short-styled morphs were labeled.Five floral/inflorescence traits were measured including floral display,corolla width(CW),floral tube length(FTL),tube opening width(TOW)and floral scape height.Fruit and seed set were recorded.The total num-ber of seeds per individual plant(plant fitness)and seed production per capsule(flower fitness)were calculated.Multiple regression analyses were used to quantify selection gradients.Important Findings The frequencies of the two morphs did not deviate from the expected 1:1 ratio at both sites.Except for FTL,the four other traits did not dif-fer significantly between the long-and short-styled morphs.Floral scape height,floral display and FTL differed between two sites.The selection regimes differed between two morphs and between two sites.At the Yushuizhai site,linear selection for shorter floral tubes was stronger in the short-styled morph.However,nonlinear selec-tion on the floral display was stronger in the long-styled morph than selection on the short-styled morph.At the Haligu site,linear selec-tion for a smaller corolla was stronger in the long-styled morph.A morph-specific nonlinear selection on CW and floral display was also detected.Morph-specific selections were detected through the estimation of flower fitness only in Haligu population.In this site,morph-specific linear selection was also detected for CW and floral display.Morph-specific nonlinear selection on traits was detected only in CW.We found that butterflies and sphingid moths dominated at Yushuizhai,while long-tongued bees dominated at Haligu.The difference in pollinator fauna suggested that selection on floral tubes may be due to differences in pollinator assemblages.Overall,variation of floral and/or inflorescence traits in P.poissonii was probably driven by pollinator selection.Selection regime dif-ferences between two morphs,in part,due the inter-morph diver-gences of sexual functions in distylous plant.

Patterns and drivers of plant sexual systems in the dry-hot valley region of southwestern China

Sexual systems play important roles in angiosperm evolution and exhibit substantial variations among different floras. Thus, studying their evolution in a whole flora is crucial for understanding the formation and maintenance of plant biodiversity and predicting its responses to environmental change. In this study, we determined the patterns of plant sexual systems and their associations with geographic elements and various life-history traits in dry-hot valley region of southwestern China, an extremely vulnerable ecosystem. Of the 3166 angiosperm species recorded in this area, 74.5% were hermaphroditic,13.5% were monoecious and 12% were dioecious, showing a high incidence of diclinous species. Diclinous species were strongly associated with tropical elements, whereas hermaphroditic species were strongly associated with temperate and cosmopolitan elements. We also found that hermaphroditism was strongly associated with showy floral displays, specialist entomophily, dry fruits and herbaceous plants.Dioecy was strongly associated with inconspicuous, pale-colored flowers, generalist entomophily, fleshy fruits, and woody plants, whereas monoecy was strongly associated with inconspicuous, pale-colored flowers, anemophily, dry fruits, and herbaceous plants. In addition, hermaphroditic species with generalist entomophily tended to flower in the dry season, whereas diclinous species with specialist entomophily tended to flower in the rainy season. However, independent of sexual systems, plants that produce dry fruits tended to flower in the rainy season and set fruits in the dry season, but the opposite pattern was found for fleshy fruit-producing plants. Our results suggest that in the dry-hot valleys, plant sexual systems are associated with geographic elements as well as various life-history traits that are sensitive to environmental change.

Covariance of Floral and Vegetative Traits in Four Species of Ranunculaceae:A Comparison between Specialized and Generalized Pollination Systems

Theory predicts that tighter correlation between floral traits and weaker relationship between floral and vegetative traits more likely occur in specialized flowers than generalized flowers, favoring by precise fit with pollinators. However, traits and trait correlations frequently vary under different environments. Through detecting spatiotemporal variation in phenotypic traits （floral organ size and vegetative size） and trait correlations in four Ranunculaceae species, we examined four predictions. Overall, our results supported these predictions to a certain degree. The mean coefficient of variation （CV） of floral traits in two specialized species （Delphinium kamaonense and Aconitum gymnandrum） was marginally significantly lower than that of another two generalized species （Trollius ranunculoides and Anemone obtusiloba）. The two specialized species also showed marginally significantly smaller CV in floral traits than vegetative size across the two species. The absolute mean correlation between floral and vegetative traits, or that between floral traits in species with specialized flowers was not significantly lower, or higher than that in generalized plants, weakly supporting the predictions. Furthermore, we documented a large variation in trait correlations of four species among different seasons and populations. Study of covariance of floral and vegetative traits will benefit from the contrast of results obtained from generalized and specialized pollination systems.

New Temperature Sensitive Genic Male Sterile Lines with Better Outcrossing Ability for Production of Two-Line Hybrid Rice

An investigation was carried out with three newly developed temperature sensitive genic male sterile （TGMS） lines for their floral traits, seed production potential and outcrossing ability in ten cross combinations. In the TGMS lines, fertile pollens had an average diameter of 0.89 mm while the sterile pollens was with 0.02 mm diameter.TS-29-150GY produced the biggest fertile pollens with 0.92 mm and other two lines produced relatively smaller pollens with 0.91 and 0.85 ram. Pollen fertility during the fertility reversion period was an average of 60.7%. TS-29-150GY had the maximum of 66.9% spikelet fertility whereas other two lines （TNAU18S and TNAU60S） had relatively lower spikelet fertility of 27.8% and 26.7%, respectively. Average of 17.00 g of seed yield was obtained in the TGMS lines during the fertility reversion period. TS-29- 150GY had the highest value of 21.20 g of seed yield while TNAU18S and TNAU60S produced 16.6 g and 13.2 g of seed yield, respectively. The low seed production ability of these three TGMS lines was attributed only to the environmental conditions prevailing during the period. All three TGMS lines had considerable outcrossing potential of 41.2%, 24.6% and 25.0%, respectively. The cross combinations viz. TNAU18S/IET21508 （36 g/plant）, TNAU18S/IET21044 （13 g/plant）, TNAU18S/IET21009 （26.5 g/plant）, TNAU60S/CB-09-106 （26.2 g/plant）, TNAU60S/IET21009 （14 g/plant） and TS29-150-GY/DRR 3306 （39.2 g/plant） showed perfect synchronization with acceptable hybrid seed yield, indicating suitability of TGMS system under Indian condition. Based on the outcrossing related traits viz. panicle exertion, angle of glume opening, stigma length and pollen size, TNAU18S was identified as the best, followed by TS-29-150GY.

Quantitative Variation in Water-Use Efficiency across Water Regimes and Its Relationship with Circadian, Vegetative, Reproductive, and Leaf Gas-Exchange Traits

Drought limits light harvesting, resulting in lower plant growth and reproduction. One trait important for plant drought response is water-use efficiency （WUE）. We investigated （1） how the joint genetic architecture of WUE, reproductive characters, and vegetative traits changed across drought and well-watered conditions, （2） whether traits with distinct developmental bases （e.g. leaf gas exchange versus reproduction） differed in the environmental sensitivity of their genetic architecture, and （3） whether quantitative variation in circadian period was related to drought response in Brassica rapa. Overall, WUE increased in drought, primarily because stomatal conductance, and thus water loss, declined more than carbon fixation. Genotypes with the highest WUE in drought expressed the lowest WUE in well-watered conditions, and had the largest vegetative and floral organs in both treatments. Thus, large changes in WUE enabled some genotypes to approach vegetative and reproductive trait optima across environments. The genetic architecture differed for gas-exchange and vegetative traits across drought and well-watered conditions, but not for floral traits. Correlations between circadian and leaf gas-exchange traits were significant but did not vary across treatments, indicating that circadian period affects physiological function regardless of water availability. These results suggest that WUE is important for drought tolerance in Brassica rapa and that artificial selection for increased WUE in drought will not result in maladaptive expression of other traits that are correlated with WUE.

Unravelling host-mediated effects on hemiparasitic Mexican mistletoe Psittacanthus calyculatus(DC.)G.Don traits linked to mutualisms with pollinators and seed dispersers

Aims While the influence of mistletoe-host interactions on each other’s evolution is well-recognized,the role of interactions between mis-tletoes and its mutualistic pollinators and seed dispersers mediated by host species is relatively unexplored.Methods Here,we examine the effects of host species(Mexican hawthorn Crataegus mexicana DC,black cherry Prunus serotina Ehrh.,leather-leaf Mexican oak Quercus crassipes Bonpl.)on flower morphology,nectar production,pollinator visitation rate and female reproduct-ive fitness in hemiparasitic Mexican mistletoe Psittacanthus caly-culatus(DC.)G.Don(Loranthaceae)populations at three different locations.We first measured the lengths of corolla,style,stamen,exerted stamen and anther,the length and width of the ovary and nectar availability for mistletoe flowers in natural populations.Then,we evaluated flower visitation and measured(length and width)and weighed the fruits and its seeds of mistletoes growing on each of the three host species.Finally,we evaluated the effects of host species,location and flower traits on fruit or seed size variation(both as proxies of reproductive fitness).Important Findings We found mistletoes growing on natural hosts P.serotina and Q.crassipes produced larger flowers,fruits and seeds than mistle-toes on cultivated C.mexicana.However,these differences varied across space.The amount of available nectar and hummingbird vis-itation rates in flowers of mistletoes on C.mexicana was higher at the three sites than in flowers of mistletoes on P.serotina or Q.cras-sipes.The effects of host species,study site and floral trait covariates affected significantly all fitness measures,indicating that mistletoes’reproductive fitness is affected differently depending on the host species and their site of occurrence.These host-associated differ-ences in reproduction might have implications for interactions with mutualistic vectors.

Genome-wide Association Analyses Reveal the Genetic Basis of Stigma Exsertion in Rice

Stigma exsertion, a key determinant of the rice mating system, greatly contributes to the application of heterosis in rice. Although a few quantitative trait loci associated with stigma exsertion have been fine map- ped or cloned, the underlying genetic architecture remains unclear. We performed a genome-wide associ- ation study on stigma exsertion and related floral traits using 6.5 million SNPs characterized in 533 diverse accessions of Oryza sativa. We identified 23 genomic loci that are significantly associated with stigma exsertion and related traits, three of which are co-localized with three major grain size genes GS3, GW5, and GW2. Further analyses indicated that these three genes affected the stigma exsertion by controlling the size and shape of the spikelet and stigma. Combinations of GS3 and GW5 largely defined the levels of stigma exsertion and related traits. Selections of these two genes resulted in specific distributions of floral traits among subpopulations of O. sativa. The low stigma exsertion combination gw5GS3 existed in half of the cultivated rice varieties; therefore, introducing the GW5gs3 combination into male sterile lines is of high potential for improving the seed production of hybrid rice.

Discovery of Gynoecium Color Polymorphism in an Aquatic Plant

Flower color polymorphism exhibited by natural populations provides an opportunity for understanding the evolutionary mechanisms contributing to the diversity of floral morphology. However, little is known about the color polymorphism of female organs in flowering plants. Here we report gynoecium color polymorphism in Butornus urnbellatus （Butomaceae）, an emergent, aquatic monocot. Populations from Mishan, northeastern China comprised two morphs; gynoecia are either pink, as observed in other areas, or white. We measured floral traits and female fecundity in the two gynoecium color morphs in the field. There was no significant difference in plant height, pedicel length, and flower size including petal, sepal and gynoecium between the two morphs, but plants with pink gynoecia had wider inflorescence stalks, larger inner whorl anthers and produced more pollen and ovules than those with white gynoecia. Correspondingly, we found that seed production was significantly higher in the pink than in the white morph. This new finding suggested selection against white gynoecia in part because of low fecundity, consistent with the rarity of the white gynoecium morph in this species.

Does persistence of showy calyces in Limonium leptolobum enhance pollinator attraction?

Aims The calyx,the outermost whorl of a flower(usually green),has been considered to function to protect flowers.in some species,how-ever,calyces are colorful and retained during seed development.Limonium species have been exploited as cut flower crops because the calyces persist for several months after the corolla has closed.To explore the adaptive significance of the persistent calyx in a desert plant Limonium leptolobum,we ask whether persistence of caly-ces can enhance pollinator attraction by enlarging floral displays,increasing reproductive success in this self-incompatible species.Methods The yellow flower of L.leptolobum lasted 1-2 days but its white,membranous calyx extended fully after the corolla closed,and per-sisted for over 2 months in the field,making hundreds of‘showy flowers’on one individual.To examine the ecological function of calyces,we test the pollinator attraction hypothesis.in an experi-mental population,we compared the difference in visit frequency and visitor behavior between intact inflorescences and inflores-cences with their calyces removed on the same individual plants.Important findingsin four experimental plots four types of floral visitors were observed including bees,butterflies,syrphid flies and day-flying moths.No significant preference was observed between calyx-free and intact inflorescences for both first arrivals and total visit frequency of all types of floral visitors,indicating that the persistence of calyces did not make plants more attractive to potential pollinators.The pollina-tor attraction hypothesis for the showy calyces was not supported by the current data.Whether the calyx in this desert plant helps seed development or has other functions needs further study.

Discovery of androecium color polymorphism in Epimedium pubescens with habitat preference of anther/pollen color in the genus

Aims Interspecific and intraspecific variation in flower color in natural populations provides an opportunity for us to understand the evolu-tion and maintenance of diversity of floral traits.Compared to cor-olla color,little is known about the color polymorphism of sexual organs in flowering plants.to explore evolutionary transitions of androecium color and polymorphism within species,interspecific and intraspecific variation in androecium(anther and pollen)color in the genus Epimedium(Berberidaceae)was investigated.Methods to explore the geographical patterns of anther/pollen color variation in Epimedium species,data of 45 species were collected and their phyl-ogeny was constructed based on available DNA sequences.to investi-gate whether intraspecific variation in androecium color relates to habitat preference,three environmental factors were measured in the field popu-lation of Epimedium pubescens in northeastern Sichuan,China,which plants had green or yellow androecia.Vegetative and reproductive traits of this species were compared between the two color morphs.Important Findings Androecium(anther and pollen)color polymorphism in field populations of Epimedium pubescens is reported here where nine populations are monomorphic with a green androecium but three populations are dimorphic with individuals having either a green or a yellow androecium.Inflorescence stalk height,stalk diameter,leaf number,flower number and spur length(as well as spur and nectar volume)were not significantly different between two morphs.Compared to the yellow morph,the green morph had relatively larger leaves and anthers,but smaller sepals.the green morph produced more pollen and larger seeds,but the same number of ovules.Seed set was not significantly different between green and yellow morph.Investigations of environmental factors in the color dimorphic popu-lations of E.pubescens indicated that the green morph was more likely to occur in habitats with relatively lower light intensity.the distribution survey of 45 Epimedium species showed that species with a green androecium tended to appear at lower elevations.Comparative phylogenetic analysis showed that transitions from yellow to a green androecium or to androecial color dimorphism occurred at least seven times.this genus,characterized by anther color diversity and containing some species with anther color polymorphism,provides a model system in which to study the evolution and maintenance of colorful sexual organs in flowering plants.

How do belowground organisms influence plant-pollinator interactions?

Aims The majority of angiosperms are pollinated by animals,and this interaction is of enormous importance in both agricultural and natural systems.Pollinator behavior is influenced by plants’floral traits,and these traits may be modified by interactions with other community members.In recent years,knowledge of ecological linkages between above-and belowground organ-isms has grown tremendously.Soil communities are extremely diverse,and when their interactions with plants influence floral characteristics,they have the potential to alter pollinator attrac-tion and visitation,but plant-pollinator interactions have been neglected in studies of the direct and indirect effects of soil organism-root interactions.Here,we review these belowground interactions,focusing on the effects of nitrogen-fixing bacteria,arbuscular mycorrhizal fungi and root-feeding herbivores,and their effects on floral traits and pollinators.Further,we identify gaps in our knowledge of these indirect effects and recommend promising directions and topics that should be addressed by future research.Important Findings Belowground organisms can influence a wide variety of floral traits that are important mediators of pollinator attraction,including the number and size of flowers and nectar and pollen production.Other traits that are known to influence pollinators in some plant species,such as floral volatiles,color and nectar composition,have rarely or never been examined in the context of belowground plant interac-tions.Despite clear effects on flowers,relatively few studies have measured pollinator responses to belowground interactions.When these indirect effects have been studied,both arbuscular mycorrhi-zal fungi and root herbivores were found to shift pollinator visitation patterns.Depending on the interaction,these changes may either increase or decrease pollinator attraction.Finally,we discuss future directions for ecological studies that will more fully integrate below-ground ecology with pollination biology.We advocate a multilevel approach to these questions to not only document indirect effect pathways between soil interactions and pollination but also iden-tify the mechanisms driving changes in pollinator impacts and the resultant effects on plant fitness.A more thorough understanding of these indirect interactions will advance ecological theory and may inform management strategies in agriculture and conservation biology.

The role of the pollination niche in community assembly of Erica species in a biodiversity hotspot

Aims Closely related species frequently co-occur in plant communities.The role of niche partitioning for allowing their coexistence has been called into question,but most studies on community ecol-ogy neglect pollination as a niche component,and thus poten-tially underestimate niche partitioning.Here,we investigate the importance of the pollination niche for determining coexistence in a biodiversity hotspot.We specifically test whether co-flowering insect-pollinated species of Erica,the most species-rich plant genus of the South African Cape Floristic Region,are characterized by dif-ferent functional floral traits and different specialized pollination systems.Methods Communities of co-flowering Erica species were studied at three sites in the fynbos biome in the Western Cape of South Africa.We focused on five species with flowers that conform to an insect pol-lination syndrome.We assessed whether floral traits important for pollinator attraction and mechanical fit overlap by comparison of scent(gas chromatography coupled with mass spectrometry),col-our(spectrophotometry and models of bee vision),nectar volume and concentration and corolla morphology.Pollinator partitioning was quantified by comparison of floral visitor profiles and pollinator importance,calculated as the product of visitation rate,number of flowers probed per visit and pollen load for each functional visitor group among species within communities.Potential effects of com-petition were assessed by comparing visitation rates both directly and using anther ring disturbance as a proxy for lifetime visitation.Important Findings Floral phenotypes differed among species within communities in all functional traits.Visitor assemblages were diverse,including ants,beetles,flies,honeybees,moths and solitary bees.However,the Cape honeybee,Apis mellifera capensis,was the main pollinator of all but one Erica species.Insect visitation rates were significantly lower for E.imbricata and E.calycina relative to congeners,but high rates of anther ring disturbance,which differed significantly in only two out of seven comparisons,suggest that long-term visitation rates are mostly similar among species that share pollinators.This study shows that coexistence of co-flowering insect-pollinated Erica species is not determined by specialization for different pollination systems.Pollinator sharing might rather lead to facilitation among co-flowering species,and floral constancy mediated by extensive interspecific floral trait variation might mitigate its potentially nega-tive effects.

Shortened anther-stigma distance reduces compatible pollination in two distylous Primula species

Aims Distyly has been regarded as an adaptation to improve compatible pollination between two floral morphs with reciprocal herkogamy.The hypothesis that the different positions of anthers and stigmas within flowers as well as their reciprocal position between morphs,reduce the probability of self pollination raised by Darwin has been rarely tested.in this study,we measured stigmatic pollen loads in response to reduced reciprocal herkogamy in two Primula species.Methods To see whether reciprocal herkogamy can increase compatible and/or reduce incompatible pollen deposition,thus promoting compat-ible pollination,we shortened the distance between anthers and stigmas within the flowers by changing the position of the corolla tube,to which the anthers were fused,i.e.reduced herkogamy in natural populations of Primula secundiflora and P.poissonii and quantified stigmatic pollen loads in the field over 2 years.Important Findingsin both species,stigmatic pollen loads were significantly higher in the long-styled(l-morph)than in the short-styled morph(s-morph)in both control and manipulated flowers,but percentage of compatible pollen in s-morph were higher.Flowers manipulated to halve the anther-stigma distance showed a similar pattern for 2 years:total pollen grain counts on stigmas did not differ significantly but compatible pollen grains in l-and s-morphs were significantly decreased in both species.The percentage of compatible pollen loads was decreased by 68.7%in P.secundiflora and 65.3%in P.poissonii in l-morphs,while it decreased by 30.6%and 2.9%in s-morphs,respectively.our manipulation of the relative position of anthers and stigmas in the two distylous species indicated that a lower degree of herkogamy reduced compatible but incompatible pollen transfer was likely to increase.The higher proportion of compatible pollen in the s-morph than in the l-morph in the two Primula species could be attributed to the accessibility of two-level sexual organs,floral orientations and pollinator behaviors.This is a first attempt to manipulate intraflower herkogamy for understanding adaptation of heterostyly,shedding insights into how the reciprocal herkogamy promotes compatible pollination.