Balance between inbreeding and outcrossing in a nannandrous species,the moss Homalothecium lutescens

Epiphytic dwarf males on the females present a possible solution to the problem of short fertilization distances in mosses. However, leptokurtic spore dispersal makes dwarf males likely to be closely related to their host shoot, with an accompanying risk of inbreeding. The capacity of a female to harbour a high number of different dwarf males suggests that there may be mechanisms in place that counteract inbreeding, such as polyandry and post-fertilization selection. We have genotyped sporophytes, female host shoots and dwarf males in four populations of the moss Homalothecium lutescens. We found no evidence of selective sporophyte abortion based on level of heterozygosity. The occurrence of entirely homozygous sporophytes together with significantly positive inbreeding coefficients in three of the populations (mean F_IS between 0.48 and 0.64) suggest frequent mother–son mating events. However, 23% of all sampled sporophytes had a higher level of heterozygosity compared with the mean expected heterozygosity at the population level. Polyandry was frequent, on average 59% of the sporophytes on a female shoot were sired by distinct fathers. In conclusion, sporadic fertilizations by dwarf males originating from nonhost female shoots appear to counteract strong inbreeding.     

Costs of sporophyte production in the moss,Dicranum polysetum

We investigated the cost of sporophyte production in the moss Dicranum polysetum both by examining patterns of growth and reproduction in unmanipulated shoots and by experimentally manipulating sexual reproduction.The estimated proportion of total carbon investment allocated to sexual reproduction in sporophyte-producing shoots over the study period was 74.8%. Unmanipulated shoots that aborted all sporophytes had a significantly higher growth in the top shoots than shoots that produced sporophytes. In sporophyte-producing shoots, total apical growth decreased proportionately with the number of sporophytes.Experimental prevention of sporophyte development resulted in significantly higher total apical growth of the gametophytes. Shoots where current perichaetia were lacking when marked had a mass increase in the top shoots similar to manipulated sporophytic shoots whereas sporophytic control shoots grew significantly less than these two categories.The difference between control shoots and manipulated shoots in the mass of vegetative apical growth was mainly because of different length increments whereas mass per unit length was similar between groups. The probability to reproduce sexually in the year after the manipulation, and the biomass allocated to this reproduction, were not affected by the experimental treatment.     

Reproductive traits in the tropical moss Octoblepharum albidum Hedw. differ between rainforest and coastal sites

Abstract

Octoblepharum albidum Hedw. is an autoicous moss commonly occurring in tropical savannas, dry forests, rainforests and coastal habitats. It frequently reproduces by spores and asexual structures (gemmae and protonemata or buds at leaf tips), making it a good model for understanding how reproductive traits change with respect to habitat type. Our aims were to characterize the different life-history traits in O. albidum relative to sexual and asexual cycles and to detect variations in reproductive performance among the different habitats, trade-offs among these traits, and relationships among reproductive traits and plant length. We studied colonies from two Atlantic rainforests and two coastal sites in north-eastern Brazil. Shoots in the coastal sites, compared to those of the forest sites, had higher numbers of sporophytes, male and female branches per shoot, male gametangia per sexual branch, and longer setae. Numbers of female gametangia per sexual branch did not differ between forest and coastal sites. A male-biased sex ratio of branches and gametangia occurred in all sites. Compared to gemmae, sporophytes and protonemata or buds were more likely to be found on longer shoots than on shorter ones, but this relationship was only applicable to forest sites. The abundant production of gemmae and protonemata or buds at leaf tips, and sporophytes (spores) in O. albidum are important components in explaining colonization success and maintenance in this tropical moss. Longer sporophytic setae in addition to a higher reproductive performance (especially for number of male gametangia and sporophytes per shoot) may favour spore dispersal and colonization in plants of coastal sites.     

Restriction of sexual reproduction in the moss Racomitrium lanuginosum along an elevational gradient

1. Terrestrial plant populations located at the margins of species’ distributions often display reduced sexual reproduction and an increased reliance on asexual reproduction. One hypothesis to explain this phenomenon is that the decline is associated with environmental effects on the energetic costs to produce reproductive organs.
2. In order to clarify the changing processes of sexual reproduction along an elevational gradient, we investigated the sexual reproductive parameters, such as the number of sporophytes and gametangia, in Racomitrium lanuginosum, a dioicous moss found on Mt. Fuji.
3. Matured sporophytes were present only below 3,000 m, and the number of sporophytes per shoot tended to be lower at higher elevation habitats. The numbers of male inflorescences per shoot and antheridia per inflorescence and shoot significantly decreased with increasing elevation. In contrast, the numbers of female inflorescences per shoot and archegonia per inflorescence and shoot varied little across elevations.
4. Synthesis. Our results suggest that the reasons for this limitation are assumed to be limitations in sporophyte development that result in abortion, and the spatial segregation between males and females. Possible reasons for the abortion of sporophytes are the inhibitory effects of low air temperature, a shortened growth period, and winter environmental conditions at higher elevations. Remarkable differences between male and female on various reproductive parameters found in this study are thought to affect the mode of sexual reproduction under the harsh environment. These differences between males and females may be caused by differences in the costs of production and development of gametangia, sensitivity to environmental stressors, and phenological patterns.

     

Selective spore germination on shoots of Homalothecium lutescens,a moss with dwarf males

Spores from three bryophyte species with dwarf males (Homalothecium lutescens, Homalothecium sericeum and Isothecium alopecuroides) were sown on shoots of H. lutescens in vitro. After 10 months, presence and fertility of dwarf plants were scored. Spores of the more distantly related I. alopecuroides were unable to develop into dwarf plants on H. lutescens. Spores of both H. lutescens and H. sericeum developed into dwarf plants. In fact, dwarf plants of H. sericeum were both more abundant and more often fertile than those of H. lutescens. The ability of H. sericeum spores to develop into dwarf males on shoots of H. lutescens suggests a possible pathway for hybridization between the two species. On the other hand, the inability of I. alopecuroides to develop into dwarf males on shoots of H. lutescens suggests that regulation of spore germination and dwarf male development on host shoots is associated with the degree of relatedness between species.     

Molecular biodiversity in the moss <Emphasis Type="Italic">Leptodon smithii</Emphasis> (Neckeraceae) in relation to habitat disturbance and fragmentation

Bryophytes seem particularly suitable to investigate genetic diversity in relation to habitat disturbance due to their large employment as bioindicators and to the recent application of molecular markers to moss population studies. Genetic variation and structure were analysed in seven urban, extraurban and remote populations of Leptodon smithii, an epiphytic moss of Quercus ilex, a phanerogamic species of Mediterranean climax vegetation. A total of 210 individual shoots were DNA extracted and amplified with internal simple sequence repeat (ISSR) primers, and 54 haplotypes were identified. An uneven distribution of haplotype number and frequencies was observed among sites, with a higher number of haplotypes and more homogeneous haplotype frequencies in the extraurban/remote populations. Molecular diversity indices were overall higher in the extraurban sites than in the urban ones. Multilocus linkage disequilibrium values were in line with the occurrence of sexual/asexual reproduction in the seven populations. The isolation-by-distance model was not supported by Mantel test among sites; however, within-population fixation index (F_ST) highlighted a clear relation between genetic and physic distances among trees, suggesting a limited dispersal range for L. smithii’s spores. The genetic structure was mainly affected by population size, wood structure and extent, and genetic drift consequent to habitat fragmentation and human-induced disturbance.     

CONTROL OF SEX RATIOS IN HAPLOID POPULATIONS OF THE MOSS,CERATODON PURPUREUS

Female biased sex ratios occur in a number of unrelated mosses. Such ratios refer to the relative numbers of male and female gametophytes in moss populations and are therefore more comparable to the numbers of pollen grains and ovules in populations of seed plants than to the numbers of male (microsporangiate) and female (megasporangiate) sporophytes. A survey of 11 populations of the moss, Ceratodon purpureus, showed that sex ratios are heterogeneous, but that female biases occur in more than half the populations. One hundred and sixty single spore isolates representing 40 sporophytes from one population demonstrated that female gametophytes outnumbered males by a ratio of 3:2 at the time of germination. Female gametophytic clones formed significantly more biomass than male clones, and individual female shoots were more robust. Male clones, however, produced more numerous stems. These sexually dimorphic traits may be related to life history differences between male and female gametophytes since females must provide nutritional support to the “parasitic” sporophyte generation, a burden that males do not share.     

An experimental demonstration of the cost of sex and a potential resource limitation on reproduction in the moss Pterygoneurum (Pottiaceae)

The cost of sexual reproduction is incurred when the current reproductive episode contributes to a a decline in future plant performance. To test the hypotheses that a trade-off exists between current sexual reproduction and subsequent clonal regeneration and that resources limit reproduction and regeneration, plants of the widespread moss Pterygoneurum ovatum were subjected to induced sporophytic abortion, upper leaf removal, and nutrient amendment treatments. Sexually reproducing plants were slower or less likely to produce regenerative structures (protonemata or shoots) and produced fewer regenerative tissue areas or structures. The ability and the timeline to reproduce sexually and regenerate clonally were unaffected by an inorganic nutrient amendment. However, when leaves subtending the sporophyte were removed, the sporophytes were less likely to mature, tended to take a longer time to mature, and were smaller compared to sporophytes from shoots with a full complement of upper leaves. Our findings indicate that plants investing in sexual reproduction suffer a cost of decreased clonal regeneration and indicate that sporophyte maturation is resource-limited, with upper leaves contributing to the nutrition of the sporophyte. This study represents only the second explicit experimental demonstration of a trade-off between sexual and asexual reproduction in bryophytes.     

Do males time their mate-guarding effort with the fertile phase in order to secure fertilisation in Cayo Santiago rhesus macaques?

In contrast to most mammalian species, female sexual activity is not limited to the fertile phase of the ovarian cycle in anthropoid primates, which has long been proposed to conceal the timing of ovulation to males. It is now generally believed that females are still most attractive during the fertile phase, leading to high-ranking males successfully mate-guarding them specifically during this period. While studies conducted in species exhibiting exaggerated sexual swellings (probabilistic signal of the fertile phase) have generally supported this hypothesis, mixed support comes from others. Here, we investigated whether high-ranking males timed mate-guarding effort towards female fertile phases in rhesus macaques (Macaca mulatta). In this species, adult females do not exhibit sexual swellings, but undergo facial skin colour variation, an alternative oestrogen-dependent graded-signal of female reproductive status. We collected behavioural, hormonal and genetic paternity data during two mating seasons for one group of the free-ranging population of Cayo Santiago. Our results show that mate-guarding by top-ranking males did not completely cover the entire female fertile phase and that this tactic accounted for only 30-40% of all fertilisations observed. Males tended to prolong mate-guarding into the luteal phase (null probability of fertilisation), which mirrors the pattern of male attraction to female facial colour reported in an earlier study. These findings suggest that males may have limited knowledge regarding the exact timing of females' fertile phase in rhesus macaques, which presumably allows females to gain more control over reproduction relative to other anthropoid primate species.     

Clonal distribution,fertility and sex ratios of the moss Plagiomnium affine (Bland.) T.Kop. in forests of contrasting age

Abstract

Six populations of the clonal forest floor moss Plagiomnium affine from forests of different age were screened for genetic variation at 23 allozyme loci, of which nine were polymorphic. Samples consisting of two adjacent unconnected shoots were taken at regular intervals along one transect from each population. A total of 602 shoots was analysed. Almost 80% of the shoots were sterile (i.e. not expressing male or female gender). Sex remained unknown for only 10% of shoots after identification of genets based on electrophoretic data. We identified a mean number of 3.7 fertile clones per population. The mean length of clones along transects in each population ranged between 2 and 3 m. The size distribution within populations was bimodal, with a few dominant clones and a varying number of much smaller clones. The overall sex ratio was slightly female biased at the ramet level, but balanced at the genet level. Forest age was negatively correlated with percentage of sterile shoots and positively correlated with frequency of sporophytes. In both cases correlations were significant only if population 1, which was subject to extreme soil disturbance by badgers, was excluded. We conclude that the effective population size is larger, and the susceptibility to genetic drift is lower, in old forests.     

Regeneration strategies of dominant boreal forest dwarf shrubs in response to selective removal of understorey layers

Abstract. The deciduous Vaccinium myrtillus and the evergreen Vaccinium vitis‐idaea were subjected to five removal treatments of understorey layers: control, removal of the moss layer, removal of the field layer, removal of both moss and field layers and removal of moss, field and humus layers. A second factor, sowing, was included to investigate sexual reproduction after disturbance. Density of new ramets and seedlings and growth of annual shoots were studied for the first two growing seasons, whereas cover was measured for five growing seasons after disturbance treatment. Initially, vegetative production of new ramets and species cover increased rapidly in all disturbed plots, except for the most severe treatment, in which production of new ramets was virtually absent throughout the study. Full recovery following removal of the field layer only or both field and moss layers was reached after four years for V. myrtillus. V. vitis‐idaea recovered more quickly, after one year (removal of field layer only) and four years (removal of field and moss layers). The relative growth of V. myrtillus and V. vitis‐idaea increased in the latter treatment in terms of production of annual shoots and length of annual shoots, respectively. Seedling density increased after sowing in the most severe treatment. The results underscore the importance of vegetative growth for recovery of these species at moderate‐level disturbances. The high rate of sexual reproduction in the most severe treatment implies that strong mechanical disturbance is needed to enhance the establishment of new genotypes in these species.     

The cost of realized sexual reproduction: assessing patterns of reproductive allocation and sporophyte abortion in a desert moss

The desert moss Syntrichia caninervis exhibits one of the most skewed sex ratios in the plant kingdom, with female individuals far outnumbering male individuals (exceeding 14♀:1♂). The "cost of sex hypothesis" derives from allocational theory and predicts that the sex which is most expensive should be the rarer sex. This hypothesis, which, as considered here represents the realized cost of sexual reproduction, is contingent upon two assumptions that are explored: (1) that male sex expression is more expensive than female sex expression, and (2) that sexual reproduction is resource limited. Using inflorescence biomass and discounting sperm, male sex expression was found to be in the neighborhood of one order of magnitude more expensive than female sex expression, and this difference is reflected in higher numbers of gametangia per male inflorescence, presence of paraphyses in male inflorescences, and a much longer developmental time for male inflorescences. The realized cost of female reproduction from two communities dominated by S. caninervis was found to be lower than the realized cost of male sexual reproduction. Resource-limited reproduction was assessed by determining the frequency of sporophyte abortion, the age distribution of sporophyte abortions, and patterns of sporophyte abortion that may be density dependent. Among ten sexually reproducing populations, abortive sporophytes occurred at a frequency of 0.64. Abortive sporophytes averaged 8% the mass of mature sporophytes, and cohort sporophytes from the same individual female were found to abort in a density-dependent pattern. We conclude that the two assumptions, upon which the cost of sex hypothesis depends, are supported.     

Arbuscular mycorrhiza and water and nutrient supply differently impact seedling performance of dry woodland species with different acquisition strategies

Background: We recently demonstrated that dwarf male plants are much more common among mosses than hitherto thought. Dwarf plants, producing functional male sexual organs occur in 10–20% of the moss species worldwide.

Aims: We investigated how an inadvertent omission of dwarf males affects estimates of male sex expression rates and sex ratios.

Methods: We recorded frequencies of specimens with female-expressing plants, male-expressing normal-sized and/or dwarf plants, and sporophytes in herbarium material of five model species from Macaronesia. We analysed how frequency estimates of male occurrence and sex ratios were affected by excluding dwarf males.

Results: Occurrence of dwarf males was positively associated with sporophyte formation. When dwarf males were excluded, male mate availability was missed in 51–61% of specimens (in 74–76% of sporophyte-containing specimens). Specimen-level sex ratio was balanced in material with sporophytes in the three species exhibiting male dwarfism, but strongly female biased if dwarf males were omitted.

Conclusions: We provide the first quantitative evidence that sex-expressing dwarf males are essential for sexual reproduction and sporophyte production in mosses that form such males. Considering its abundance, male dwarfism therefore deserves more attention among plant biologists. Innovative approaches are required to further study the minute male plants.     

Sexual reproduction, clonal diversity and genetic differentiation in patchily distributed populations of the temperate forest herb Paris quadrifolia (Trilliaceae)

Clonal plant species have been shown to adopt different strategies to persist in heterogeneous environments by changing relative investments in sexual reproduction and clonal propagation. As a result, clonal diversity and genetic variation may be different along environmental gradients. We examined the regional and local population structure of the clonal rhizomatous forest herb Paris quadrifolia in a complex of forest fragments in Voeren (Belgium). Relationships between population size (the number of shoots), shoot density (the number of shoots per m²) and local growth conditions were investigated for 47 populations. Clonal diversity and genetic variation within and among 19 populations were investigated using amplified fragment length polymorphism markers. To assess the importance of sexual reproduction, seed set, seed weight and germination success were determined in 18 populations. As predicted, local growth conditions largely affected population distribution, size and density of P. quadrifolia. Populations occurring in moist and relatively productive sites contained significantly more shoots. Here, shoots were also much more sparsely distributed compared to populations occurring in dry and relatively unproductive sites, where shoots showed a strongly aggregated distribution pattern. Clonal diversity was relatively high, compared with other clonal species (G/N ratio = 0.43 and Simpson’s D=0.81). Clonal diversity significantly (P<0.01) decreased with increasing shoot density while molecular genetic variation was significantly (P<0.01) affected by population size and local environmental conditions. Lack of recruitment and out-competition of less-adapted genotypes may explain the decreased genetic variation in dry sites. Analysis of molecular variance revealed significant genetic variation among populations (Φ _ST=0.42, P<0.001), whereas pairwise genetic distances were not correlated to geographic distances, suggesting that gene flow among populations is limited. Finally, the number of generative shoots, the number of seeds per fruit and seed weight were significantly and positively related to population size and local growth conditions. We conclude that under stressful conditions populations of clonal forest plant species can slowly evolve into remnant populations characterized by low levels of genetic variation and limited sexual reproduction. Conservation of suitable habitat conditions is therefore a prerequisite for effective long-term conservation of clonal forest plant species.     

Sex ratio,sex‐specific pattern in vegetative growth and gemma production in an aquatic liverwort,Scapania undulata (Marchantiophyta: Scapaniaceae)

Scapania undulata is an aquatic dioicous liverwort growing in shallow streams in boreal to subtropical zones. We studied the expressed sex ratio, sex‐specific differences in shoot architecture and possible trade‐off between sexual and asexual reproduction in ten populations of S. undulata by surveying 100 plots in ten streams in southern Finland. The expressed sex ratio was male biased, in contrast with the sex ratio in most dioicous bryophytes. It was also highly variable between the streams, but individual plots frequently comprised shoots from only one sex. The overproduction of males might be a strategy to overcome sperm dilution and ensure fertilization over longer distances in water. No size differences between females and males were detected, but they differed in branching patterns. Evidence for a higher cost of sexual reproduction in females than males can be seen from the following: the male‐biased sex ratio; low number of sex‐expressing female shoots in female‐only plots; no co‐occurrence of gemmae and female sex organs on a single branch, and no more than one sexual branch per female shoot. In contrast, high gemma production of male and female sex‐expressing shoots indicates a minimal trade‐off between sexual and asexual reproduction. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 175 , 229–241.     

Reproductive effort and costs of reproduction do not explain female-biased sex ratios in the moss Pseudocalliergon trifarium (Amblystegiaceae)

A fundamental assumption in life-history theory is that reproduction is costly. Higher reproductive investment for fruits than for flowers may result in larger costs of reproduction in females than in males, which is often used to explain male-skewed sex ratios in unisexual seed plants. In contrast, bryophytes have predominantly female-biased sex ratios, suggested to be a product of a higher average cost of sexual reproduction in males. Empirical evidence to support this notion is largely lacking. We investigated sex-specific reproductive effort and costs in the unisexual moss Pseudocalliergon trifarium that has a female-dominated expressed sex ratio and rarely produces sporophytes. Annual vegetative segment mass did not differ among male, female, and non-expressing individuals, indicating that there was no threshold-size for sex expression. Mean and annual mass of sexual branches were higher in females than in males, but branch number per segment did not differ between sexes. Prefertilization reproductive effort for females was significantly greater (11.2%) than for males (8.6%). No cost for sexual branch production in terms of reduced relative vegetative growth or decreased investment in reproductive structures in consecutive years was detected. A higher realized reproductive cost in males cannot explain the unbalanced sex ratio in the study species.     

Reproductive biology of Syzygiella rubricaulis (Nees) Steph. (Adelanthaceae,Marchantiophyta), a liverwort disjunctly distributed in high‐altitude Neotropical mountains

Syzygiella rubricaulis is a dioecious leafy liverwort disjunctly distributed and restricted to high‐altitude mountains in the Neotropics and the Azores. This study is part of a larger project examining the phylogeography of S. rubricaulis in the Neotropics, and our main goals were to understand its reproductive biology, where sex expression occurs, if vegetative propagules are frequently found, how the sexes are distributed in populations, how frequently sporophytes are formed and what environmental conditions influence sexual expression. S. rubricaulis patches are mostly female, but all patches also contain non sex‐expressing shoots. Out of 42 patches examined, 29 (69%) were sex‐expressing: 25 were unisexual (21 female and four male) and four of mixed sex (two male‐biased and two unbiased). At shoot level, out of 4200 shoots 18% were female and 7% male; among sex‐expressing shoots, 73% were female, representing a sex ratio of 0.8 (female‐biased). We encountered a total of 33 sporophytes in six patches (in Brazil, Venezuela and Ecuador). Leaf regenerants were found in one patch in Mexico. Low rates of sporophytes were likely related to low frequencies of male shoots and large distances between the sexes. As 25% of S. rubricaulis shoots expressed sex (occasionally producing sporophytes), we suggest that short‐distance (and rarely long‐distance) spore dispersal events occur in mountainous areas on a short‐term basis. On a long‐term basis, however, these events likely contribute to dynamic exchanges among populations in the Neotropics.     

Male sexual ornament size is positively associated with reproductive morphology and enhanced fertility in the stalk-eyed fly <Emphasis Type="Italic">Teleopsis dalmanni</Emphasis>

Background  

Exaggerated male ornaments and displays often evolve in species where males only provide females with ejaculates during reproduction. Although "good genes" arguments are typically invoked to explain this phenomenon, a simpler alternative is possible if variation in male reproductive quality (e.g. sperm number, ejaculate content, mating rate) is an important determinant of female reproductive success. The "phenotype-linked fertility hypothesis" states that female preference for male ornaments or displays has been selected to ensure higher levels of fertility and has driven the evolution of exaggerated male traits. Females of the stalk-eyed fly Teleopsis dalmanni must mate frequently to maintain high levels of fertility and prefer to mate with males exhibiting large eyespan, a condition-dependent sexual ornament. If eyespan indicates male reproductive quality, females could directly increase their reproductive success by mating with males with large eyespan. Here we investigate whether male eyespan indicates accessory gland and testis length, and then ask whether mating with large eyespan males affects female fertility.     

Can the meiotic sex ratio explain the sex ratio bias in adult populations in the dioicous moss Drepanocladus lycopodioides?

Sex ratio variation is commonly observed in natural populations of many organisms with separate sexes and genetic sex determination, including bryophytes. Most bryophyte populations exhibit female-skewed expressed adult sex ratios, generally inferred from counts of sexually mature plants. For the rarely sexually reproducing perennial dioicous moss Drepanocladus lycopodioides, we showed that a female bias also exists in the genetic adult sex ratio, using a specifically designed molecular sex-associated marker. Here, we investigated whether the meiotic spore sex ratio contributes to the observed bias in genetic adult sex ratio in natural populations. Earlier attempts to study meiotic sex ratios have involved commonly cultivated ruderals that rapidly express sex in the laboratory. We established single-spore cultures from field-collected sporophytes from these populations and used the marker to assess the sex of individual sporelings. Spore germinability was (near) complete, and mortality among sporelings was virtually absent. The true meiotic sex ratio did not differ from equality, but strongly differed both from the observed genetic sex ratios in the natural adult populations, and from the European scale genetic sex ratio. We conclude that the biased population sex ratios in this species arise at life cycle stages after spore germination. Sexual dimorphism may selectively favour female proliferation during some phase of gametophyte development. Based on methodological progress, we successfully used a perennial study species with rare sexual reproduction, which significantly broadens the life history spectrum investigated in bryophyte sex ratio studies.     

Polymorphic microsatellite loci in the bush‐cricket Metrioptera roeseli

Metrioptera roeseli is a common bush‐cricket in Europe which prefers grassland as habitat. We developed microsatellite loci in order to study the population genetic structure and the effects of different habitat types on the genetic structure of the bush‐cricket in a rural landscape. In this paper, we report on six polymorphic microsatellite loci with seven to 16 alleles per locus in 20 individuals from the Lahn‐Dill‐Bergland in Hesse, Germany.