Gender-specific inbreeding depression in a gynodioecious plant, Geranium maculatum (Geraniaceae)

In gynodioecious species, females coexist with hermaphrodites in natural populations even though hermaphrodites attract more pollinators, are capable of reproducing through pollen, and can self-fertilize. This study tests the hypothesis that inbreeding depression helps to maintain females in natural populations. It also examines whether gender lineages that differ in selfing rates might experience different levels of inbreeding depression. Female and hermaphroditic lineages of the gynodioecious species Geranium maculatum were used in self, sib-cross and outcross experiments to examine inbreeding depression levels and to determine whether these levels differ between hermaphroditic and female lineages. Six fitness correlates were measured in the greenhouse and compared among pollination types and between genders. Severe inbreeding depression was found for both individual fitness traits and cumulative fitness in early life history stages. Inbreeding depression levels were slightly higher in hermaphroditic than in female lineages, but this difference was not statistically significant. Because females are unable to self-pollinate and are less likely to experience inbreeding than hermaphrodites under natural conditions, these results suggest that severe inbreeding depression could confer a selective advantage for females that could help to maintain females in natural populations.     

Female frequency and relative fitness of females and hermaphrodites in gynodioecious Geranium sylvaticum (Geraniaceae)

We determined female frequency of 23 populations of the gynodioecious Geranium sylvaticum (Geraniaceae) in Finland. We compared our results to previous results on this species from the 1960s in order to reveal putative changes in female frequencies. Because females may be maintained in gynodioecious populations if their seed production or offspring quality is higher than that of hermaphrodites, we explored reproductive success of females and hermaphrodites in detail in 11 populations for two consecutive years. Female frequencies varied from 0.4 to 27.2%; this variation is similar to that observed in the 1960s. Contrary to previous results that indicated lower seed production in females, females produced 1.2 and 1.7 times more seeds per flower than hermaphrodites in 2000 and 2001, respectively. Females also had higher fruit set than hermaphrodites. Thus, higher seed production of females partly explains the maintenance of gynodioecy in this species. Furthermore, female frequency correlated negatively with relative seed fitness of hermaphrodites suggesting that relative seed fitness is related to population sex ratio. Female frequency and the distance of the population from the most southern population also tended to correlate positively, suggesting that harsher environmental conditions in the north may benefit female plants. Given the observed yearly variation, our results also highlight the importance of temporal variation for the relative seed fitness of females and hermaphrodites.     

Frequency-dependent pollinator discrimination acts against female plants in the gynodioecious Geranium maculatum

MethodsTo test the severity and consequences of this type of pollinator discrimination in Geranium maculatum, experimental populations with the range of sex ratios observed in nature were created, ranging from 13 % to 42 % females. Pollinators were observed in order to measure the strength of discrimination, and pollen deposition and seed production of both sexes were measured to determine the fitness consequences of this discrimination. Additionally a comparison was made across the sex ratios to determine whether discrimination was frequency-dependent.ConclusionsThe results suggest that pollinator discrimination negatively affects females'' relative fitness when they are rare. Thus, the initial spread of females in a population, the first step in the evolution of gynodioecy, may be made more difficult due to pollinator discrimination.     

Mating system contributes only slightly to female maintenance in gynodioecious Geranium maculatum (Geraniaceae)

Gynodioecy, the co-occurrence of female and hermaphroditic individuals within a population, is an important intermediate in the evolution of separate sexes. The first step, female maintenance, requires females to have higher seed fitness compared with hermaphrodites. A common mechanism thought to increase relative female fitness is inbreeding depression avoidance, the magnitude of which depends on hermaphroditic selfing rates and the strength of inbreeding depression. Less well studied is the effect of biparental inbreeding on female fitness. Biparental inbreeding can affect relative female fitness only if its consequence or frequency differs between sexes, which could occur if sex structure and genetic structure both occur within populations. To determine whether inbreeding avoidance and/or biparental inbreeding can account for female persistence in Geranium maculatum, we measured selfing and biparental inbreeding rates in four populations and the spatial genetic structure in six populations. Selfing rates of hermaphrodites were low and did not differ significantly from zero in any population, leading to females gaining at most a 1–14% increase in seed fitness from inbreeding avoidance. Additionally, although significant spatial genetic structure was found in all populations, biparental inbreeding rates were low and only differed between sexes in one population, thereby having little influence on female fitness. A review of the literature revealed few sexual differences in biparental inbreeding among other gynodioecious species. Our results show that mating system differences may not fully account for female maintenance in this species, suggesting other mechanisms may be involved.     

Floral dimorphism, pollination, and self-fertilization in gynodioecious GERANIUM RICHARDSONII (Geraniaceae)

The selective maintenance of gynodioecy depends on the relative fitness of the male-sterile (female) and hermaphroditic morphs. Females may compensate for their loss of male fitness by reallocating resources from male function (pollen production and pollinator attraction) to female function (seeds and fruits), thus increasing seed production. Females may also benefit from their inability to self-fertilize if selfing and inbreeding depression reduce seed quality in hermaphrodites. We investigated how differences in floral resource allocation (flower size) between female and hermaphroditic plants affect two measures of female reproductive success, pollinator visitation and pollen receipt, in gynodioecious populations of Geranium richardsonii in Colorado. Using emasculation treatments in natural populations, we further examined whether selfing by autogamy and geitonogamy comprises a significant proportion of pollen receipt by hermaphrodites. Flowers of female plants are significantly smaller than those of hermaphrodites. The reduction in allocation to pollinator-attracting structures (petals) is correlated with a significant reduction in pollinator visitation to female flowers in artificial arrays. The reduction in attractiveness is further manifested in significantly less pollen being deposited on the stigmas of female flowers in natural populations. Autogamy is rare in these protandrous flowers, and geitonogamy accounts for most of the difference in pollen receipt between hermaphrodites and females. Female success at receiving pollen was negatively frequency dependent on the relative frequency of females in populations. Thus, two of the prerequisites for the maintenance of females in gynodioecious populations, differences in resource allocation between floral morphs and high selfing rates in hermaphrodites, occur in G. richardsonii.     

Taxonomic revision of Geranium sect. Gracilia (Geraniaceae)

Geranium sect.Gracilia (Geraniaceae) comprises nine species in the Andes of Venezuela and Colombia.Geranium lindenianum andG. gracilipes are synonymized withG. holosericeum. The identity ofG. multiceps is clarified. Its differences from the two Colombian species,G. santanderiense andG. lainzii, are discussed.Geranium lignosum, G. sebosum, G. stoloniferum, G. subnudicaule, andG., velutinum are now considered to be members of sect.Gracilia. The identity ofG. pilgerianum could not be clarified because the type was destroyed and other collections have not been found. Micromorphological leaf and seed characters are discussed. Six lectotypes and one neotype are designated.

---

Resumen  Se revisaGeranium sect.Gracilia (Geraniaceae), un grupo formado por nueve especies distribuidas por los Andes de Venezuela y Colombia. Se sinonimizanG. lindenianum y G. gracilipes aG. holosericeum. Se aclara la identidad deG. multiceps y se establecen sus diferencias conG. santanderiense y conG. lainzii, ambas esencialmente colombianas. Se aceptan en esta secciónG. lignosum, G. stoloniferum yG. sebosum, propuestos tras la descripción de la sect.Gracilia, yG. subnudicaule yG. velutinum, ya tratados por otros autores en como pertenecientes a este grupo. Por el contrario, la identidad deG. pilgerianum no ha podido ser aclarada tras la destrucción del tipo y ante la falta de otros materiales de la especie. Se estudian diversos caractere micromorfológicos, especialmente algunos referibles a la anatomía foliar y la de la semilla. Se designan seis lectótipos y un neótipo.

     

Morphology, ontogeny and histochemistry of secretory trichomes of Geranium robertianum (Geraniaceae)

Geranium robertianum bears three types of glandular uniseriate trichomes which originate from a single protodermal cell and develop through periclinal divisions. Type I trichomes are procumbent and have an oval apical cell, two stalk cells and a basal cell. Type II trichomes are erect and have a pear shaped apical cell, two stalk cells and a basal cell. Type III trichomes are much longer than the other two types and have an elongated apical cell, five long stalk cells and a basal cell. Type I and type II trichomes are common on leaves while III trichomes are more abundant on flower structures.
Type I and type II trichomes secrete terpenoids and phenols. Type III trichomes are characterized by the accumulation of anthocyanins in the apical cell and secrete flavonoids.     

The influence of plant and corolla size on pollen deposition and seed set in Geranium sanguineum (Geraniaceae)

In a coastal grassland area the influence of plant and corolla size on insect visitation rate, pollen deposition on stigmas and seed set was measured in a natural population of Geranium sanguineum. Visits by bumble bees, the primal pollinators, were significantly affected by plant size measured as the number of flowers, and differences in visit rates among plants were explained by size differences among these. On average, larger plants had bigger corollas. The mean corolla area affected pollen deposition positively whereas deposition did neither affect seed set (number per flower) nor seed weight. Insects reaction to corolla size was dependent on plant size, and thus we found the effect of corolla size on pollen deposition to decline with plant size. Instead pollen deposition per plant seemed to be correlated to overall floral display. We argued that resource status of the individual plants could explain corolla size and seed set variation. The finding of some plant setting seeds while other plant with comparable size did not set any seeds could not be explained satisfactory by resource limitation.     

Development of nuclear microsatellite markers for the threatened wetland plant Geranium soboliferum var. kiusianum (Geraniaceae)

Nuclear microsatellite markers were developed for the threatened plant Geranium soboliferum var. kiusianum, which has decreased its population size as a result of loss of its wetland habitat in Kyushu, Japan. Utilizing RNA‐seq data obtained by next‐generation sequencing techniques, 10 polymorphic microsatellite markers with 3–16 alleles in a nuclear genome were developed and characterized. Two to 15 alleles were observed in G. soboliferum. These markers will be used to investigate the genetic circumstance of remnant populations of G. soboliferum var. kiusianum and their phylogenetic relationship with G. soboliferum.     

Geranium hanaense (Geraniaceae), a new species from Maui,Hawaiian Islands

Geranium hanaense, a new species endemic to two small montane bogs (20 ha total) on northeast Haleakala, Maui, Hawaiian Islands, is described. This new species is postulated to have evolved from the high elevation xeromorphic species,G. cuneatum, in a fashion similar to that of the two other bogGeranium species. The ecology and evolution of HawaiianGeranium species, includingG. hanaense, are discussed.     

Geranium kikianum sp. nov. (Geraniaceae) from the southern Peloponnese,Greece

Geranium kikianum Kit Tan & G. Vold sp. nov. (Geraniaceae) is illustrated and described as a new species endemic to Greece. It occurs beside streams and other wet places in open Pinus nigra forest at the foothills of Mt Taigetos in the southern Peloponnese. It is closely related to G. macrorrhizum, a species more widely distributed in southern and central Europe, ranging from the southern Alps to the Balkan Peninsula. It differs from the latter by its smaller and narrower deflexed petal limb, slender claw and occurrence in wet habitats. The genome sizes of G. kikianum and of Greek populations of G. macrorrhizum are presented here for the first time. The nuclear DNA content (2C values) of 2.84 pg for G. kikianum and 2.83 and 2.87 pg for two G. macrorrhizum populations from Greece probably corresponds to a 2n=92 cytotype while the smaller genome size of 1.45 pg in a population of G. macrorrhizum from Mt Smolikas, Greece may correspond to a 2n=46 cytotype.     

Volatiles of Geranium purpureum Vill. and Geranium phaeum L.: Chemotaxonomy of Balkan Geranium and Erodium Species (Geraniaceae)

The essential oils obtained by hydrodistillation of Geranium purpureum and G. phaeum were characterized by GC‐FID and GC/MS analyses (the former for the first time in general). In total, 154 constituents were identified, accounting for 89.0–95.8% of the detected GC peak areas. The investigated essential oils consisted mainly of fatty acids and fatty‐acid‐derived compounds (45.4–81.3%), with hexadecanoic acid and (E)‐phytol as the major components. The chemotaxonomic significance of the variations in the essential‐oil composition/production of the presently and previously investigated Geranium and highly related Erodium taxa from Serbia and Macedonia was assessed by multivariate statistical analyses. The main conclusions drawn from the high chemical similarity of the two genera, visible from the obtained dendrograms and biplots, confirm the close phylogenetic relationship between the investigated Geranium and Erodium taxa, i.e., that there is no great intergeneric oil‐composition variability. Changes in the composition and production of essential oils of the herein investigated taxa and 60 other randomly chosen species belonging to different plant genera were also statistically analyzed. The results put forward pro arguments for the oil‐yield–oil‐composition correlation hypothesis.     

A cladistic analysis of Geranium subgen. Erodioidea (Picard) Yeo (Geraniaceae)

Subgenus Erodioidea , comprising 19 species, is diagnosed exclusively by the type of fruit discharge and associated morphological characters, all of which are of widespread occurrence within the Geraniaceae. We have no evidence to suggest that the three included sections ( Erodioidea, Aculeolata and Subacaulia) form a monophyletic group. Cladistic analysis of a data set containing 30 morphological characters has confirmed our doubts of monophyly of the subgenus by placing part of the outgroup within the ingroup. Subsequent analyses of the two included non-monotypic sections, Erodioidea and Subacaulia , give different results. For both sections, the monophyly assumption is reasonably well-supported by characters. However, while analysis of sect. Erodioidea results in an explicit hypothesis of relationships and a fully resolved cladogram, resolution within sect. Subacaulia is low. For analysis of the latter, seven different outgroups were used and, in each one, the successive approximations weighting procedure (available in the program Hennig 86) was followed to improve resolution. Despite this, lower nodes in the cladogram remained unresolved, and strict consensus trees of all the analyses resulted in only a few replicated subterminal clades. These results are discussed in connection with biogeographical data and with the hypothesis that representatives of sect. Subacaulia are schizoendemics.     

Fruit-discharge-type in Geranium (Geraniaceae): its use in classification and its evolutionary implications

Three main types of seed-discharge in Geranium are made the basis of its division into subgenera: Geranium subgenus Geranium , with a ballistic expulsion of the seed from the mericarp, termed 'seed-ejection'; Geranium subgenus Robertium , with forcible discharge of the mericarp with the seed in it, separately from the awn, termed 'carpel-projection', and Geranium subgenus Erodioideae , with the seed-containing mericarp being thrown off with the attached awn, which becomes helically coiled, called the 'Erodium-type'. Variants of the seed-ejecting type permit the division of Geranium subgenus Geranium into three sections. Other criteria are used to divide Geranium subgenus Robertium into eight sections and Geranium subgenus Erodioideae into two. Species are fully enumerated except for Geranium section Geranium , which comprises the bulk of the genus, and for which some tentative subgroups are given in an Appendix. Diversity of fruit-type in Geranium is greatest in the Mediterranean Region. Characters of the fruit in other genera of Geraniaceae are surveyed. Geographical distributions, chromosome numbers, pollen morphology and phytochemistry are reviewed. It is suggested that Erodium-type fruit discharge, shared with the four remaining genera of the family, is primitive, and that carpel-projection and seed-ejection arose from it separately, the latter probably more than once. The very large, mainly perennial, Geranium subgenus Geranium is contrasted with Geranium subgenus Robertium , half of which is hapaxanthic, and which occupies marginal habitats and shows greater morphological and chromosomal variation despite its being only one tenth the size. Geranium subgenus Erodioideae is smaller still and probably relictual.     

Germination of up to 129-year old, dry-stored seeds of Geranium bohemicum (Geraniaceae)

Old dry-stored seeds of the annual pyrophyte Geranium bohemicum from seed collections and herbaria were tested to determine their germinability. Ten samples of seed, ranging in age from 3 to 200 years, were used. The oldest seeds to germinate (three out of ten) were from a 129-year old collection. All three seedlings developed into healthy plants. This finding supports the assumption that G bohemicum can emerge from a bank of long-lived seeds after forest fires.     

Preferences of pollinators and herbivores in gynodioecious Geranium sylvaticum

BACKGROUND AND AIMS: For the maintenance of gynodioecy (i.e. the coexistence of female and hermaphroditic plants), females need to compensate for the lack of pollen production through higher seed production or better progeny quality compared to hermaphrodites. In Geranium sylvaticum, females produce more seeds per flower than hermaphrodites. This difference in seed production might be modified by biological interactions with pollinators and herbivores that may favour one sex and thus affect the maintenance of gynodioecy. METHODS: Sexual dimorphism in flower size and flowering phenology, and in attractiveness to pollinators, pre-dispersal seed predators and floral herbivores were examined in natural populations of G. sylvaticum. KEY RESULTS: Pollinators preferred hermaphrodites 25 % more often than females in two of the three study populations, and floral herbivores attacked hermaphrodites 15 % more often than females in two of the six study populations. These preferences might be explained by the larger flower size of hermaphrodites. In contrast, seed predators did not prefer either sex. CONCLUSIONS: The data suggest that pollinator preference does not benefit females, whereas the higher floral herbivory of hermaphrodites might enhance the maintenance of females in G. sylvaticum. Thus, although the data support the view that ecological factors may contribute to the maintenance of gynodioecy, they also suggest that these contributions may vary across populations and that they may function in opposite directions.