Herkogamy and mating patterns in the self-compatible daffodil Narcissus longispathus

BACKGROUND AND AIMS: Floral design in self-compatible plants can influence mating patterns. This study investigated Narcissus longispathus, a self-compatible bee-pollinated species with wide variation in anther-stigma separation (herkogamy), to determine the relationship between variation in this floral trait and the relative amounts of cross- and self-fertilization. METHODS: Anther-stigma separation was measured in the field in six populations of N. longispathus from south-eastern Spain. Variation in herkogamy during the life of individual flowers was also quantified. Multilocus outcrossing rates were estimated from plants differing in herkogamy using allozyme markers. KEY RESULTS: Anther-stigma separation varied considerably among flowers within the six populations studied (range = 1-10 mm). This variation was nearly one order of magnitude larger than the slight, statistically non-significant developmental variation during the lifespan of individual flowers. Estimates of multilocus outcrossing rate for different herkogamy classes (t(m) range = 0.49-0.76) failed to reveal a monotonic increase with increasing herkogamy. CONCLUSIONS: It is suggested that the lack of a positive relationship between herkogamy and outcrossing rate, a result that has not been previously documented for other species, could be mostly related to details of the foraging behaviour of pollinators.     

Variable herkogamy and autofertility in marginal populations ofGentianella germanica in the Netherlands

Gentianella germanica (Willd.) Börner, a rare plant species, occurs at the margin of its distribution area in the Netherlands. It is found only in isolated patches of chalk grassland surrounded by intensively-used agricultural land. In 1997 and 1998, we investigated the spatial and temporal separation of anthers and stigma in relation to autofertility. We performed our experiments in one large and two small populations to investigate the possible selection against herkogamy and for increased autofertility in marginal populations with pollination limitation. Dichogamy appeared to be very weak and varied within individuals. Homogamous and protandrous flowers were most frequent. Dichogamy is apparently no effective barrier against self-fertilization in this species. Herkogamy varied significantly between individuals and between populations, and plant and flower size were not correlated with anther-stigma separation. Both observations suggest a genetic basis for herkogamy. Autofertility was generally high in all three populations. Only in one small population was a correlation between autofertility and herkogamy found. In this population, most plants had the stigma positioned above the anthers (mean herkogamy +0.8 mm), while in the other two populations the stigma was positioned mainly in between or below the anthers (mean herkogamy ?0.55 mm). In comparison with earlier data from 1991 and 1992, plants in the large population were less herkogamous and more self-fertile in 1998. Since 1993 this population was mown each year one month earlier than usual, leading to dramatic reductions (bottlenecks) in population size. In addition, the early mowing time may have caused a further reduction of the already scarce visitation by pollinating insects. We hypothesize that both adverse conditions may have caused a selection of plants with smaller anther-stigma separation and higher autofertility and thus showing that human influence can have profound effects on the reproductive success and evolution of rare plant species, even if it occurs in the context of nature conservation.     

Anther-stigma separation is associated with inbreeding depression in Datura stramonium,a predominantly self-fertilizing annual

Abstract.— Genetically based variation in outcrossing rate generates lineages within populations that differ in their history of inbreeding. According to some models, mating-system modifiers in such populations will demonstrate both linkage and identity disequilibrium with fitness loci, resulting in lineage-specific inbreeding depression. Other models assert that differences among families in levels of inbreeding depression are mainly attributable to random accumulation of genetic load, unrelated to variation at mating-system loci. We measured female reproductive success of selfed and outcrossed progeny from naturally occurring lineages of Datura stramonium , a predominantly self-fertilizing annual weed that has heritable variation in stigma-anther separation, a trait that influences selfing rates. Progeny from inbred lineages (as identified by high degree of anther-stigma overlap) showed equal levels of seed production, regardless of cross type. Progeny from mixed lineages (as identified by relatively high separation between anthers and stigma) showed moderate levels of inbreeding depression. We found a significant correlation between anther-stigma separation and relative fitness of selfed and outcrossed progeny, suggesting that family-level inbreeding depression may be related to differences among lineages in inbreeding history in this population. Negative inbreeding depression in putatively inbred lineages may be due in part to additive effects or to epistatic interactions among loci.     

GENETIC AND ENVIRONMENTAL VARIATION IN FLORAL TRAITS AFFECTING OUTCROSSING RATE IN CLARKIA TEMBLORIENSIS (ONAGRACEAE)

Clarkia tembloriensis exhibits a wide range of variation among its natural populations in outcrossing rate and in separation of male and female function in space (anther-stigma separation or herkogamy) and in time (protandry). Here we show that outcrossing rate is highly correlated with protandry and anther-stigma separation. Both genetic and environmental variation contribute to inter- and intrapopulation variation in protandry and anther-stigma separation. Interpopulation differentiation for protandry and anther-stigma separation was found to be polygenic. Genetic variation for protandry and anther-stigma separation within populations was demonstrated by a significant among-family variance in two populations with contrasting breeding systems. Environmental effects on the expression of mating system traits were manifested in two ways. First, significant variation among lathhouse benches suggests that small-scale environmental heterogeneity may affect the development of floral traits. Second, protandry was shortened under hot summer conditions. Hence, hotter and drier habitats, typical of the more self-pollinating populations of C. tembloriensis, can promote self-pollination purely through environmental effects.     

Pollination adaptations of group-by-group stamen movement in a meadow plant with temporal floral closure

Floral sexual organ (stamen and pistil) movements are selective adaptations that have different functions in male-female reproduction and the evolution of flowering plants. However, the significance of stamen movements in the spatial–temporal function and separation of male and female organs has not been experimentally determined in species exhibiting floral temporal closure. The current study investigated the role of slow stamen (group-by-group) movement in male-female sexual function, and the effect of stamen movement on pollen removal, male-male and male-female interference, and mating patterns of Geranium pratense, a plant with temporal floral closure. This species uses stamen group-by-group movement and therefore anther-stigma spatial–temporal separation. Spatial separation (two whorls of stamen and pistil length) was shown to be stronger than temporal separation. We found that stamen movements to the center of the flower increase pollen removal, and the most common pollinators visited more frequently and for longer durations during the male floral stage than during the female floral stage. Petal movements increased both self-pollen deposition rate and sexual interference in G. pratense. The fruit and seed set of naturally and outcrossed pollinated flowers were more prolific than those of self-pollinated flowers. Group-by-group stamen movement, dehiscence of stamens, pistil movement, and male-female spatial–temporal functional separation of G. pratense before floral temporal closure may prevent male-female and stamen-stamen interference and pollen discounting, and may increase pollen removal and cross-pollination.     

Reproductive assurance through self-fertilization does not vary with population size in the alien invasive plant Datura stramonium

Autonomous self-fertilization is suggested to be associated with invasiveness in plants because it offers reproductive assurance when there is a shortage of suitable mates or pollinators. Given that shortages of mates and pollinators are a common cause of Allee effects in small plant populations, we predict that the benefits of self-fertilization in terms of reproductive assurance should be greatest in small populations. We tested this idea for the invasive herb Datura stramonium , a self-fertilizing species which is also cross-pollinated to some extent by insects (mainly hawkmoths and honeybees). During two consecutive years, we studied 20 and 55 populations, respectively, of different sizes. Untreated flowers of D. stramonium showed high levels of fruit and seed set in all populations studied. Although, fruit and seed set were generally reduced by about 90% in flowers in which self-fertilization was prevented through emasculation, this effect did not vary according to population size. By using a natural color (anthocyanin) dimorphism in 12 populations, we showed that the average outcrossing rate was low (1.3%) and that there was no relationship between outcrossing rate and population size. Pollen removal from flowers also did not vary according to population size, suggesting that the pollinator visitation rate is not lower in small populations. However, decreasing deviations of observed from expected fruit set with population size imply that small populations may have an increased chance of extinction due to demographic stochasticity. Overall, our results suggest that reproductive assurance through self-fertilization in invasive plants may be important for all stages of population establishment, and not just in the founder population.     

Effect of floral organ sizes on female reproductive success in<Emphasis Type="Italic">Erythronium japonicum</Emphasis> (Liliaceae)

We examined the effects of floral organ size on female reproductive success in self-incompatibleErythronium japonicum. We measured tepal size and anther-stigma separation and investigated the relationship between these measurements and fruiting and seeding success. We found that tepal length was positively correlated with fruiting success and the number of seeds per fruit This suggests that pollinator attraction is affected by tepal length in f.japonicum and that the number of pollinator visits affects female reproductive success. Anther-stigma separation was the most variable floral trait measured and was not correlated with either fruiting or seeding success in this species, suggesting that the proportion of outcross pollen deposited on stigmas by pollinators does not increase with anther-stigma separation. This is inconsistent with a previous report onEtyyhtonium. grandiflorum. Pollinator size might explain this interspecific difference in the effect of anther-stigma separation on female reproductive success.     

The role of breeding system and inbreeding depression in the maintenance of an outcrossing mating strategy in Silene virginica (Caryophyllaceae)

The goal of this study was to understand the interaction among breeding system, mating system, and expression of inbreeding depression in the hermaphroditic, primarily hummingbird-pollinated, iteroparous, short-lived perennial Silene virginica. We performed hand-selfed and hand-outcrossed pollinations in the field, conducted detailed floral observations within individual flowers and plants, and assayed adult tissue from flowering plants for a genetic estimate of population outcrossing rate. We quantified the opportunity for geitonogamy as the proportion of days each plant exhibited simultaneous male and female function, i.e., asynchronous expression of male- and female-phased flowers. Expression of cumulative inbreeding depression based on germination rate and total flower production in the glasshouse was ～40% and was congruent with the estimated high outcrossing rate of 0.89. Floral observations demonstrated strong temporal protandry within each flower (dichogamy) as well as complete spatial separation between male and female function within each flower (herkogamy). On average, 29% of the time there were both male- and female-phased flowers present on an individual plant. We conclude that our estimate of inbreeding depression is compatible with a largely outcrossing mating system and the amount of selfing observed, likely results from geitonogamy. This study illustrates the utility of examining both the causes and the consequences of inbreeding via selfing to provide additional insights into the evolution of plant mating systems.     

Predicting evolution of floral traits associated with mating system in a natural plant population

Evolution of floral traits requires that they are heritable, that they affect fitness, and that they are not constrained by genetic correlations. These prerequisites have only rarely been examined in natural populations. For Mimulus guttatus, we found by using the Riska-method that corolla width, anther length, ovary length and number of red dots on the corolla were heritable in a natural population. Seed production (maternal fitness) was directly positively affected by corolla width and anther size, and indirectly so by ovary length and number of red dots on the corolla. The siring success (paternal fitness), as estimated from allozyme data, was directly negatively affected by anther-stigma separation, and indirectly so by the corolla length-width ratio. Genetic correlations, estimated with the Lynch-method, were positive between floral size measures. We predict that larger flowers with larger reproductive organs, which generally favour outcrossing, will evolve in this natural population of M. guttatus.     

通泉草属3种植物柱头触敏性与花部特征之间的关系

选取3种具有不同传粉环境及花部结构差异明显的通泉草属(Mazus)植物,即毛果通泉草(M.spicatus Vaniot)、长蔓通泉草(M.longipes Bonati)和弹刀子菜(M.stachydifolius(Turcz.)Maxim.),观测其柱头触敏性行为,测量有关花部性状,对它们之间的相关性进行比较分析。结果显示,在相同强度的刺激下,毛果通泉草的柱头最为迟钝(闭合时间最长),弹刀子菜最为敏感,而长蔓通泉草介于两者之间。此外,柱头触敏性最强的弹刀子菜比柱头触敏性相对较弱的长蔓通泉草和毛果通泉草具有更深的花色、更大的单花展示、更长的药柱距以及更大的花粉胚珠比。研究结果表明通泉草属植物柱头触敏性的进化与花部特征密切相关,它们共同影响传粉过程中对传粉者的吸引、花粉落置、传递效率以及植物交配系统的构成式样。     

A model of the evolution of dichogamy incorporating sex-ratio selection, anther-stigma interference, and inbreeding depression

Historically, explanations for the evolution of floral traits that reduce self-fertilization have tended to focus on selection to avoid inbreeding depression. However, there is growing support for the hypothesis that such traits also play a role in promoting efficient pollen dispersal by reducing anther-stigma interference. The relative importance of these two selective pressures is currently a popular topic of investigation. To date, there has been no theoretical exploration of the relative contributions of selection to avoid the genetic costs of self-fertilization and selection to promote efficient pollen dispersal on the evolution of floral traits. We developed a population genetic model to examine the influence of these factors on the evolution of dichogamy: the temporal separation of anther maturation and stigma receptivity. Our analysis indicates that anther-stigma interference can favor dichogamy even in the absence of in-breeding depression. Although anther-stigma interference and inbreeding depression are the key forces driving the initial evolution of dichogamy, selection to match the timing of pollen dispersal to the availability of ovules at the population level becomes a more potent force opposing the further evolution of dichogamy as the extent of temporal separation increases. This result may help to explain otherwise puzzling phenomena such as why dichogamy is rarely complete in nature and why dichogamy tends to be associated with asynchronous flower presentation.     

Food or sex; pollinator–prey conflict in carnivorous plants

Carnivorous plants potentially trap their own pollinators and it has been argued that considerable spatial separation of flowers and traps has evolved to protect pollinators. We investigated flower-trap separation of Drosera and Utricularia . Short Drosera had a greater element of floral–trap separation than tall Drosera . Such a relationship is unexpected for plants whose peduncles were evolved to protect their pollinators. Utricularia can not trap pollinators but this genus still produces exceptionally long peduncles. We propose that flower-trap separation evolved because carnivorous plants are often short and need to project their flowers well above ground level to make them more attractive to pollinators.     

Sources of phenotypic variation in floral traits in wild radish, Raphanus raphanistrum (Brassicaceae)

Pollinator-mediated natural selection has been shown to act on phenotypic variation in floral morphology, and this variation has often been demonstrated to be heritable, but few details are available concerning the sources of floral variation. We examined phenotypic variation in seven floral traits in wild radish (Raphanus raphanistrum) at six levels: between two populations grown in a common garden, among plants within populations, among flowers measured on different weeks, between flowers on two flowering stalks measured on the same day, between adjacent flowers on a flowering stalk, and within individual flowers. There were no significant differences between plants derived from the two source populations, which were ～800 km apart. Most of the variance was within individual plants; repeatabilities were all <0.35. There were highly significant differences between flowers measured in different weeks and also highly significant plant by week interactions, indicating that the among-plant variation was not consistent over time. There was substantial variance among adjacent flowers on the same stalk, particularly in the gynoecium. This high within-plant variance is partly responsible for the low heritability of floral traits in the field and the weak selection on floral traits found in previous studies of wild radish.     

Drought alters the expression of mating system traits in two species of Clarkia

Variation in mating system traits can have important consequences for plant populations by affecting reproductive assurance, the expression of inbreeding depression, and the colonization of and persistence in new or altered habitats. Environmental stressors, such as drought, have been hypothesized to induce higher rates of self-fertilization, yet this hypothesis has rarely been tested. Here we measure the response of two sister species of self-compatible annual herbs from contrasting habitats, Clarkia breweri and C. concinna, to an experimentally imposed greenhouse drought treatment. We find that the species differ in their baseline per-flower autogamy rates and the degree of spatial and temporal separation of male and female function within their flowers. Both species show a reduction in temporal separation of anthesis and stigma receptivity with the drought treatment. However, the species from the more mesic habitat, C. concinna, increases its low autogamy rate under drought conditions, whereas the species from the more xeric habitat, C. breweri, decreases its high autogamy rate under drought conditions. Neither species showed a response to drought in flower size or anther-stigma distance. Our results demonstrate that the induction of selfing under environmental stress cannot be assumed and that, in this case, the developmental timing of flower maturation is more plastic than floral morphology.     

Impacts of floral gender and whole-plant gender on floral evolution in Ecballium elaterium (Cucurbitaceae)

Investigation of gender specialization in plants has led to several theories on the evolution of sexual dimorphism: reproductive compensation, based on enhanced reproductive efficiency with gender specialization (flowers should be larger on dioecious plants); Bateman's Principle, based on sex-specific selection (display for pollinator attraction in males and seed set in females); and intersexual floral mimicry, based on mimicry of a reward-providing gender by a non-reward providing gender (reduced dimorphism in dioecious plants due to increased spatial separation of male and female flowers). These theories were evaluated in Ecballium elaterium, which contains two subspecies, elaterium (monoecious) and dioicum (dioecious). Our results show that flowers of the dioecious subspecies are larger and allocate more to reproductive organs than do flowers of the monoecious subspecies. Both subspecies are sexually dimorphic (male flowers larger than female flowers). Variance in flower size among populations is greater in the dioecious subspecies. Finally, there is sufficient genetic variation to enable ongoing response to selection; genetic correlation constraints on independent response of female and male flowers may be stronger in the monoecious subspecies. Our findings provide support for aspects of all three theories, suggesting that the evolution of floral dimorphism is based on a complex interplay of factors.     

SEX EXPRESSION,BREEDING SYSTEM,AND POLLEN BIOLOGY OF RICINOCARPOS PINIFOLIUS: A CASE OF ANDRODIOECY?

A population of 54 Ricinocarpos pinifolius (Euphorbiaceae) plants contained male plants, which produced only staminate flowers, and hermaphrodites, which produced staminate and pistillate flowers. The fraction of pistillate flowers ranged continuously from 0 to 0.68. Insect pollination was effective and fruit set virtually complete except for losses to herbivores. Self pollen, outcross pollen from male plants, and outcross pollen from hermaphrodites were all equivalent in viability, germination, tube growth, ovule penetration, and fruit setting ability. Inbreeding depression was manifested as late abortion of some selfed seeds. Geitonogamous selfing is largely prevented by temporal separation of male and female functions within plants. This temporal separation, combined with population-wide synchrony of flowering, may create unusual conditions allowing male plants at low frequency to match hermaphrodites in reproductive success.     

ANDROMONOECY IN ZIGADENUS PANICULATUS (LILIACEAE): SPATIAL AND TEMPORAL PATTERNS OF SEX ALLOCATION

I describe patterns of sex allocation and gamete packaging in the andromonoecious lily Zigadenus paniculatus. In this species, pistil length was continuously, but bimodally, distributed within plants, and smaller pistils contained fewer mature ovules. In hermaphrodite flowers, ovule number per flower increased with blooming rank in small plants but decreased with blooming rank in large plants. Flowers with pistils less than three-fourths stamen length almost never produced fruits and were classified as males. The pedicel, tepals, stamens, and pistil of hermaphrodite flowers were all heavier than those of males. Hermaphrodite flowers were concentrated on the terminal raceme, males on the lower racemes. In combination with acropetal blooming, this spatial separation of flower types resulted in a seasonal decline in the proportion of open flowers that were hermaphrodite. However, individual flowers were protandrous, so that the population sex ratio, initially strongly male-biased, declined as the season progressed. Hand pollinations showed that plants were self-incompatible. Inflorescence size was positively correlated with bulb size, and plants with large inflorescences had a higher proportion of male flowers. Nutrient supplementation had no effect on inflorescence size, but increased the proportion of hermaphrodite flowers. Nutrient-supplemented plants also began blooming earlier than controls. I discuss these patterns in relation to the adaptive significance of andromonoecious breeding systems.     

Genetic engineering of parthenocarpic fruit development in tomato

Parthenocarpy was engineered in two genotypes of Lycopersicon esculentum Mill. by using the DefH9-iaaM chimeric gene. The parthenocarpic trait consists of fruit set and growth in the absence of fertilization. Seedless parthenocarpic fruits were obtained from emasculated flowers, and fruits with seeds from pollinated flowers. All parthenocarpic tomato plants analysed expressed the DefH9-iaaM gene during flower development. The fruit set percentage of emasculated transgenic flowers was similar to that of control plants. In 7 out of 8 independent transgenic plants, the fresh weight of fruits derived from pollinated or emasculated flowers did not significantly differ from that of fruits obtained by pollination of the control plants. The pH of the parthenocarpic fruit was generally unaffected and the soluble solid concentration was either unchanged or increased. Thus, the DefH9-iaaM gene is a genetic tool that might be used to improve tomato productivity.     

Tissue and subcellular distribution of the lectin from Datura stramonium (thorn apple).

Plants of Datura stramonium (thorn-apple) were dissected into their component tissues and examined for the presence of the Datura lectin. This lectin was easily detected in seeds and in various parts of the flowers of adult plants. Traces were also found in green (emerged) cotyledons and roots of seedlings. The specific lectin activity in seeds contained within the fruits increased as the seeds matured. Mature seeds were homogenized in sucrose and separated by differential centrifugation into four fractions, three of which were clearly of distinct composition. Most of the lectin activity sedimented with the low-speed (cell-wall/protein-body) pellet, but a similar specific activity was recovered from the other fractions. However, if EDTA was included in the homogenization medium, three or four times more lectin activity was recovered in the soluble fraction. Immunofluorescent staining of formaldehyde-fixed sections showed that the lectin was localized in the cytoplasm, with little associated with cell walls. The possible relevance of these results to the function of the lectin in plant cells is discussed.     

光照强度对曼陀罗和紫花曼陀罗生长发育的影响

以曼陀罗(Datura stramonium)及其变种紫花曼陀罗(Datura stramonium var. tatual)为研究材料, 采用光照培养箱对盆栽苗进行5个光照强度的处理, 研究其生长发育、叶片解剖结构、叶片及茎色素含量对不同光照强度的响应。结果表明: (1)在试验设置的光照强度范围内, 增强光照强度有利于曼陀罗和紫花曼陀罗的发育及茎、叶生物量的积累; 13 000和18 000 lx两个光照强度下两供试曼陀罗长势最好; 同等光照强度下紫花曼陀罗的株高、基茎粗、叶片数目、茎叶生物量大于曼陀罗, 但差异性不显著。(2)两供试曼陀罗叶片厚度、栅栏组织厚度、海绵组织厚度及气孔密度、气孔指数均随着试验光照强度的增强而增加; 紫花曼陀罗气孔密度及气孔指数明显高于曼陀罗。(3)两供试曼陀罗叶片中叶绿素含量均随试验设置光照强度的增强呈先升后降的趋势; 随试验设置光照强度的增强, 两供试曼陀罗茎中叶绿素a、b及类胡萝卜素含量变化趋势不一致; 茎中花色素苷、类黄酮及总酚含量的变化情况是: 两供试曼陀罗均与光照强度呈正相关, 但紫花曼陀罗比曼陀罗含量高, 这也是紫花曼陀罗花、茎等器官呈紫色的物质基础, 加上气孔密度和气孔指数较大等特性, 使紫花曼陀罗较曼陀罗适应性更强。