Search

The individual and combined effects of abiotic factors on pollinator-mediated selection on floral traits are not well documented. To examine potential interactive effects of water and nutrient availability on pollinator-mediated selection on three floral display traits of Primula tibetica, we manipulated pollination and nutrient availability in a factorial experiment, conducted at two common garden sites with different soil water content (natural vs addition). We found that both water and nutrient availability affected floral trait expression in P. tibetica and that hand pollination increased seed production most when both nutrient content and water content were high, indicating joint pollen and resource limitation. We documented selection on all floral traits, and pollinators contributed to directional and correlational selection on plant height and number of flowers. Soil water and nutrient availability interactively influenced the strength of both pollinator-mediated directional and correlational selection, with significant selection observed when nutrient or water availability was high, but not when none or both were added. The results suggest that resource limitation constrains the response of P. tibetica to among-individual variation in pollen receipt, that addition of nutrients or water leads to pollinator-mediated selection and that effects of the two abiotic factors are nonadditive., (© 2022 The Authors New Phytologist © 2022 New Phytologist Foundation.)

The trait-fitness relationship influences the strength and direction of floral evolution. To fully understand and predict the evolutionary trajectories of floral traits, it is critical to disentangle the direct and indirect effects of floral traits on plant fitness in natural populations. We experimentally quantified phenotypic selection on floral traits through female fitness and estimated the casual effects of nectar robbing with different nectar robbing intensities on trait-fitness relationships in both the L- (long-style and short-anther phenotype) and S-morph (short-style and long-anther phenotype) flowers among Primula secundiflora populations. A larger number of flowers and wider corolla tubes had both direct and indirect positive effects on female fitness in the P. secundiflora populations. The indirect effects of these two traits on female fitness were mediated by nectar robbers. The indirect effect of the number of flowers on female fitness increased with increasing nectar robbing intensity. In most populations, the direct and/or indirect effects of floral traits on female fitness were stronger in the S-morph flowers than in the L-morph flowers. In addition, nectar robbers had a direct positive effect on female fitness, but this effect varied between the L- and S-morph flowers. These results show the potential role of nectar robbers in influencing the trait-fitness relationships in this primrose species., (© 2019 German Society for Plant Sciences and The Royal Botanical Society of the Netherlands.)

Distyly is a mechanism promoting cross-pollination within a balanced polymorphism. Numerous studies show that the degree of inter-morph sexual organ reciprocity (SOR) within species relates to its pollen-mediated gene flow. Similarly, a lower interspecific SOR should promote interspecific isolation when congeners are sympatric, co-blooming and share pollinators. In this comparative study, we address the significance of SOR at both intra- and interspecific levels. Seventeen allopatric and eight sympatric populations representing four Primula species (P. anisodora, P. beesiana, P. bulleyana and P. poissonii) native to the Himalaya-Hengduan Mountains were measured for eight floral traits in both long- and short-styled morphs. GLMM and spatial overlap methods were used to compare intra- and interspecific SOR. While floral morphology differed among four Primula species, SOR within species was generally higher than between species, but in species pairs P. poissonii/P. anisodora and P. beesiana/P. bulleyana, the SOR was high at both intra- and interspecific levels. We did not detect a significant variation in intraspecific SOR or interspecific SOR when comparing allopatric versus sympatric populations for all species studied. As intraspecific SOR increased, disassortative mating may be promoted. As interspecific SOR decreased, interspecific isolation between co-flowering species pairs also may increase. Hybridisation between congeners occurred when interspecific SOR increased in sympatric populations, as confirmed in two species pairs, P. poissonii/P. anisodora and P. beesiana/P. bulleyana., (© 2018 German Society for Plant Sciences and The Royal Botanical Society of the Netherlands.)

Background and Aims: The outbreeding floral polymorphism heterostyly frequently breaks down, resulting in the evolution of self-fertilization as a result of homostyle formation. Here, the loss of floral polymorphism in distylous Primula oreodoxa, a sub-alpine species restricted to western Sichuan, China, was examined by investigating the ecological correlates and genetic consequences of mating system transitions. Several key questions were addressed. (1) What are the frequencies, geographical distribution and reproductive characteristics of floral morphs in distylous and homostylous populations? (2) Does increased elevation influence pollinator service and the likelihood of inbreeding in populations? (3) How often has homostyly originated and what are the consequences of the breakdown of distyly for the amounts and distribution of genetic diversity in populations?, Methods: Fourteen populations throughout the range of P. oreodoxa were sampled, and morph frequencies and floral characteristics were recorded. Polymorphism at microsatellite loci and chloroplast DNA (cpDNA) variation were used to quantify population genetic structure and genetic relationships among populations. Controlled pollinations and studies of pollen tube growth and fertility were conducted to determine the compatibility status of populations and their facility for autonomous self-pollination. Finally, visitation rates of long- and short-tongued pollinators to distylous and homostylous populations at different elevations were compared to determine if increased elevation was associated with deterioration in pollinator service., Key Results: In contrast to most heterostylous species, both distylous and homostylous morphs of P. oreodoxa are highly self-compatible, but only homostyles have the facility for autonomous self-pollination. Homostyles set significantly more fruit and seeds following open pollination than the distylous morphs. Visitation by long-tongued pollinators was significantly lower in homostylous populations, and overall rates of insect visitation decreased with elevation. Genetic diversity was significantly lower in homostylous populations, with evidence of increased inbreeding at higher elevation. Patterns of cpDNA variation were consistent with multiple transitions from distyly to homostyly and limited gene flow among populations., Conclusions: The results of this study support the hypothesis that the multiple loss of floral polymorphism in distylous P. oreodoxa is associated with unsatisfactory pollinator service, with homostyles benefiting from reproductive assurance as a result of autonomous self-pollination., (© The Author 2017. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com)

Darwin's studies on heterostyly in Primula described two floral morphs, pin and thrum, with reciprocal anther and stigma heights that promote insect-mediated cross-pollination. This key innovation evolved independently in several angiosperm families. Subsequent studies on heterostyly in Primula contributed to the foundation of modern genetic theory and the neo-Darwinian synthesis. The established genetic model for Primula heterostyly involves a diallelic S locus comprising several genes, with rare recombination events that result in self-fertile homostyle flowers with anthers and stigma at the same height. Here we reveal the S locus supergene as a tightly linked cluster of thrum-specific genes that are absent in pins. We show that thrums are hemizygous not heterozygous for the S locus, which suggests that homostyles do not arise by recombination between S locus haplotypes as previously proposed. Duplication of a floral homeotic gene 51.7 million years (Myr) ago, followed by its neofunctionalization, created the current S locus assemblage which led to floral heteromorphy in Primula. Our findings provide new insights into the structure, function and evolution of this archetypal supergene.

Heterostyly is a wide-spread floral adaptation to promote outbreeding, yet its genetic basis and evolutionary origin remain poorly understood. In Primula (primroses), heterostyly is controlled by the S-locus supergene that determines the reciprocal arrangement of reproductive organs and incompatibility between the two morphs. However, the identities of the component genes remain unknown. Here, we identify the Primula CYP734A50 gene, encoding a putative brassinosteroid-degrading enzyme, as the G locus that determines the style-length dimorphism. CYP734A50 is only present on the short-styled S-morph haplotype, it is specifically expressed in S-morph styles, and its loss or inactivation leads to long styles. The gene arose by a duplication specific to the Primulaceae lineage and shows an accelerated rate of molecular evolution. Thus, our results provide a mechanistic explanation for the Primula style-length dimorphism and begin to shed light on the evolution of the S-locus as a prime model for a complex plant supergene., Competing Interests: TE-L: The owner of Ebbing-Lohaus Gartenbau and an employee of Ebbing-Lohaus Vertriebsgesellschaft mbH. The other authors declare that no competing interests exist.

Primulina eburnea is a promising candidate for domestication and floriculture, since it is easy to culture and has beautiful flowers. An F₂ population of 189 individuals was established for the construction of first-generation linkage maps based on expressed sequence tags-derived single-nucleotide polymorphism markers using the massARRAY genotyping platform. Of the 232 screened markers, 215 were assigned to 18 LG according to the haploid number of chromosomes in the species. The linkage map spanned a total of 3774.7 cM with an average distance of 17.6 cM between adjacent markers. This linkage map provides a framework for identification of important genes in breeding programmes.

In 1862, Charles Darwin published his landmark study on the different forms of flower in Primula; he coined the term distyly and subsequently expanded his studies to other species, including those with tristyly. Darwin is widely recognized as the first to study pin and thrum flowers in Primula, and to provide an explanation for the functional significance of the two floral morphs. Our laboratory is pursuing the genes that underpin floral heteromorphy in Primula, work influenced by Darwin's observations. One day, while appreciating a print of Primula vulgaris from William Curtis' Flora Londinensis, I was struck by the fact that I was looking at images of dimorphic Primula flowers captured in a late-1700s copper-plate engraving that predated Darwin's observations by over 70 yr. This realization triggered a journey into archives of botanical texts, herbals and florilegia from the 16(th) to 19(th) Centuries, and correspondence archives, in search of earlier documents that could have influenced Darwin and the origins of an idea. Darwin was not the first to observe floral heteromorphy in Primula, but he was the first to realize the significance of the two floral morphs. Darwin's insight and exposition of purpose have underpinned all consequent work on the subject., (© 2015 The Author. New Phytologist © 2015 New Phytologist Trust.)

Background and Aims: Distyly is a floral polymorphism characterized by the presence of two discrete morphs with reciprocal positioning of anthers and stigmas in flowers on different plants within the same population. Although reciprocal herkogamy and associated floral traits are generally thought to be discrete and strict polymorphisms, little is known about variation in floral traits related to the distylous syndrome within and among populations of a single species. In this study, variation in floral morphology and reciprocal positioning of the sexual organs in the distylous Primula veris (cowslip) is quantified., Methods: Data were collected in ten populations occurring in two contrasting habitat types (grasslands and forests), and for each population the average level of reciprocity was assessed, the strength of the self-incompatibility system was determined, and seed production under natural conditions was quantified., Results: In grassland populations, flowers showed clear distyly with low and symmetric reciprocity indices at both the lower and upper level. In forests, P. veris produced larger flowers that showed strong deviations in stigma-anther separation, especially in the L-morph. This deviation was mainly driven by variation in stigma height, resulting in high and asymmetric reciprocity indices and the occurrence of several short-styled homostylous plants. Self-incompatibility was, however, strict in both habitats, and morph ratios did not differ significantly from isoplethy. The observed shift in reciprocity in forest populations was associated with a significant reduction in seed production in the L-morph., Conclusions: The results indicate that populations of P. veris show habitat-specific variation in flower morphology. Deviations from perfect reciprocal positioning of stigmas and anthers also translate into reduced seed production, suggesting that small changes in sexual organ reciprocity can have far-reaching ecological and evolutionary implications., (© The Author 2014. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

Background and Aims: Heterostyly is a floral polymorphism that has fascinated evolutionary biologists since Darwin's seminal studies on primroses. The main morphological characteristic of heterostyly is the reciprocal placement of anthers and stigmas in two distinct (distyly) floral morphs. Variation in the degree of intermorph sexual reciprocity is relatively common and known to affect patterns of pollen transfer within species. However, the partitioning of sexual organ reciprocity within and between closely related species remains unknown. This study aimed at testing whether intermorph sexual reciprocity differs within vs. between primrose species that hybridize in nature and whether the positions of sexual organs are correlated with other floral traits., Methods: Six floral traits were measured in both floral morphs of 15 allopatric populations of Primula elatior, P. veris and P. vulgaris, and anther-stigma reciprocity was estimated within and between species. A combination of univariate and multivariate approaches was used to test whether positions of reproductive organs were less reciprocal between than within species, to assess correlations between sexual organ positions and other corolla traits, and to quantify differences between morphs and species., Key Results: The three species were morphologically well differentiated in most floral traits, except that P. veris and P. vulgaris did not differ significantly in sexual organ positions. Overall, lower interspecific than intraspecific sexual organ reciprocity was detected. This decrease was marked between P. elatior and P. vulgaris, intermediate and variable between P. elatior and P. veris, but negligible between P. veris and P. vulgaris., Conclusions: Differences in anther and stigma heights between the analysed primrose species were of the same magnitude or larger than intraspecific differences that altered pollen flow within other heterostylous systems. Therefore, it is possible to suggest that considerable reductions of sexual organ reciprocity between species may lower interspecific pollen flow, with potential effects on reproductive isolation.

Background and Aims: Primula sieboldii is a perennial clonal herb that is distributed around the Sea of Japan and is endangered in Japan. Its breeding system is characterized by heteromorphic self-incompatibility, and the morph ratio within a population is very important for reproductive success. The aims of this study were to construct a linkage map, map the S locus as a qualitative trait and quantitative trait loci (QTLs) for floral morphological traits related to heterostyly, and predict the morph type in wild populations by using molecular markers for devising a conservation strategy., Methods: A linkage map was constructed with 126 markers. The QTLs for four floral traits and the S locus were mapped. Using the genotypes of loci that were located near both the S locus and the QTLs with large effects, morphs of 59 wild genets were predicted., Key Results: The linkage map consisted of 14 linkage groups (LGs). The S locus was mapped to LG 7. Major QTLs for stigma and anther heights were detected in the same region as the S locus. These QTLs exhibited high logarithm of the odds scores and explained a high percentage of the phenotypic variance (>85 %). By analysing these two traits within each morph, additional QTLs for each trait were detected. Using the four loci linked to the S locus, the morphs of 43 genets in three wild populations could be predicted., Conclusions: This is the first report of a linkage map and QTL analysis for floral morphology related to heterostyly in P. sieboldii. Floral morphologies related to heterostyly are controlled by the S locus in LG 7 and by several QTLs in other LGs. Additionally, this study showed that molecular markers are effective tools for investigating morph ratios in a population containing the non-flowering individuals or during the non-flowering seasons.

Darwin's focus on small quantitative variations as the raw material of evolution may have prevented him from discovering the laws of inheritance.

Background and Aims: Flower shapes are important visual cues for pollinators. However, the ability of pollinators to discriminate between flower shapes under natural conditions is poorly understood. This study focused on the diversity of flower shape in Primula sieboldii and investigated the ability of bumblebees to discriminate between flowers by combining computer graphics with a traditional behavioural experiment., Methods: Elliptic Fourier descriptors described shapes by transforming coordinate information for the contours into coefficients, and principal components analysis summarized these coefficients. Using these methods, artificial flowers were created based on the natural diversity of petal shape in P. sieboldii. Dual-choice tests were then performed to investigate the ability of the bumblebees to detect differences in the aspect ratio of petals and the depth of their head notch., Key Results: The insects showed no significant ability to detect differences in the aspect ratio of the petals under natural conditions unless the morphological distance increased to an unrealistic level. These results suggest the existence of a perception threshold for distances in this parameter. The bumblebees showed a significant preference for narrow petals even after training using flowers with wide petals. The bumblebees showed a significant ability to discriminate based on the depth of the petal head notch after training using artificial flowers with a deep head notch. However, they showed no discrimination in tests with training using extreme distances between flowers in this parameter., Conclusions: A new type of behavioural experiment was demonstrated using real variation in flower corolla shape in P. sieboldii. If the range in aspect ratios of petals expands much further, bumblebees may learn to exhibit selective behaviour. However, because discrimination by bumblebees under natural conditions was low, there may be no strong selective behaviour based on innate or learned preferences under natural conditions.

Grazing reduces litter thickness and vegetation height and may thereby indirectly affect reproductive success and selection on floral characters in plants with prostrate growth. Reductions in litter thickness and vegetation height should influence both the resource status of plants with leaves positioned close to the ground and the significance of inflorescence height for interactions with pollinators and seed predators. We experimentally examined how simulated grazing of surrounding vegetation affected pollen limitation, fruit predation and fecundity of short-scaped and long-scaped Primula farinosa, which differ markedly in floral display and therefore in expected attractiveness to pollinators. Litter removal and pruning of surrounding vegetation increased fruit and seed production per plant in the year of the treatment and the probability of flowering in the following year. Pollen limitation of fruit and seed production was stronger in the short-scaped morph than in the long-scaped morph, but was not significantly affected by litter removal and simulated grazing of surrounding vegetation. Supplemental hand-pollination reduced seed size in the year of the treatment and flowering probability in the second year, and these effects did not differ among scape morphs or grazing treatments. The results suggest that grazing indirectly favours seed production in P. farinosa, mainly because it increases the resource status of plants that escape damage. Contrary to expectation, there was no strong evidence that litter accumulation and tall vegetation increase the severity of pollen limitation or reduce the relative performance of the short-scaped morph.

Background and Aims: Petals are important for Primula sieboldii because of the commercial value of its flowers, and their form is a target characteristic for breeding. An appropriate understanding of petal form in terms of genetic mechanisms and environmental effects is necessary for improvement of this species. The aim of this study was to establish a quantitative evaluation method of petal shape by elliptic Fourier descriptors and principal component analysis (EF-PCA), and thus to investigate genotypic and environmental effects on petal morphology., Methods: EF-PCA describes an overall shape mathematically by transforming coordinate information concerning its contours into elliptic Fourier descriptors (EFDs) and summarizing the EFDs by principal component analysis. To examine varietal effects on principal component (PC) scores and petal area among commercial varieties, nested ANOVAs were performed (since the samples had a hierarchical structure with four sources, i.e. variety, plant, flower and petal)., Key Results: Petal shape variation could be evaluated successfully and the symmetrical and asymmetrical elements of the overall shape variation could be detected. The proportions of the variance component due to varietal differences were more than 70 % in the first five PCs of the symmetrical elements and petal area. By contrast, the proportions due to varietal effects of all PCs of the asymmetrical elements were less than 20 %, and the proportions of the variation within a flower were more than 75 %. It was also demonstrated that the yearly variance of petal shape was small, and that of petal area was large., Conclusions: Within a flower the major source of the symmetrical elements is genotypic and the asymmetrical elements are strongly affected by the environment. With respect to petal area, the contribution of genotypes is also large; it is, however, affected by the macro-environment more notably than is petal shape.

Primula flowers are heteromorphic with individual plants producing either pin-form or thrum-form flowers. We have used scanning electron microscopy to observe early development of wild-type flowers of primrose (Primula vulgaris), cowslip (P. veris), and the polyanthus hybrid (P. x tommasinii x P. vulgaris). Floral ontogeny in Primula is different from that observed in the well-studied models Antirrhinum majus and Arabidopsis thaliana and our studies reveal morphological landmark events that define the sequence of early floral development in Primula into specific stages. Pin-form and thrum-form flowers are indistinguishable during early development with differentiation of the two floral morphs occurring beyond the differentiation of floral organs. Early ontogeny of flowers with homeotic mutant phenotypes was also studied to determine the timing of developmental reprogramming in these mutants. Phenotypes studied included Hose in Hose and Jack in the Green that develop petaloid sepals and leafy sepals, respectively, and Jackanapes plants that carry both these dominant mutations. Recessive double and semi- double flowers that produce additional whorls of petals and/or stamens in place of carpels were also studied. We describe a previously undocumented recessive Primula mutant phenotype, sepaloid, that produces sepals in place of petals and stamens, and a new non-homeotic, dominant mutant phenotype Split Perianth, in which sepals and petals fail to fuse to form the typical calyx and corolla structures. The molecular basis of these mutant phenotypes in relation to the ABC model is discussed.