Selfing Variants Research Articles

The genetic architecture of tristyly and its breakdown to self-fertilization.

The floral polymorphism tristyly involves three style morphs with a reciprocal arrangement of stigma and anther heights governed by two diallelic loci (S and M). Tristyly functions to promote cross-pollination, but modifications to stamen position commonly cause transitions to selfing. Here, we integrate whole-genome sequencing and genetic mapping to investigate the genetic architecture of the M locus and the genetic basis of independent transitions to selfing in tristylous Eichhornia paniculata. We crossed independently derived semi-homostylous selfing variants of the long- and mid-styled morph fixed for alternate alleles at the M locus (ssmm and ssMM, respectively), and backcrossed the F1 to the parental ssmm genotype. We phenotyped and genotyped 462 backcross progeny using 1450 genotyping-by-sequencing (GBS) markers and performed composite interval mapping to identify quantitative trait loci (QTL) governing style-length and anther-height variation. A QTL associated with the primary style-morph differences (style length and anther height) mapped to linkage group 5 and spanned ~13-27.5Mbp of assembled sequence. Bulk segregant analysis identified 334 genes containing SNPs potentially linked to the M locus. The stamen modifications characterizing each selfing variant were governed by loci on different linkage groups. Our results provide an important step towards identifying the M locus and demonstrate that transitions to selfing have originated by independent sets of mating-system modifier genes unlinked to the M locus, a pattern inconsistent with a recombinational origin of selfing variants at a putative supergene.

Histological and micro-CT evidence of stigmatic rostellum receptivity promoting auto-pollination in the madagascan orchid Bulbophyllum bicoloratum.

BackgroundThe rostellum, a projecting part of the gynostemium in orchid flowers, separates the anther(s) from the stigma and thus commonly prevents auto-pollination. Nonetheless, as a modified (usually distal) portion of the median stigma lobe, the rostellum has been frequently invoked of having re-gained a stigmatic function in rare cases of orchid auto-pollination. Here it is shown that a newly discovered selfing variant of Madagascan Bulbophyllum bicoloratum has evolved a modified rostellum allowing the penetration of pollen tubes from in situ pollinia.MethodsGynostemium micro-morphology and anatomy of selfing and outcrossing variants of B . bicoloratum was studied by using light and scanning electron microscopy and histological sections. Pollen tube growth in the selfing variant was further observed via X-ray computed microtomography (micro-CT), providing 3D reconstructions of floral tissues at a micron scale.FindingsSelfing variants possess a suberect (‘displaced’) rostellum rather than the conventional, erect type. Very early in anthesis, the pollinia of selfers are released from the anther and slide down onto the suberect rostellum, where pollen tube growth preferentially occurs through the non-vascularized, i.e. rear (adaxial) and (semi-) lateral parts. This penetrated tissue is comprised of a thin layer of elongate and loosely arranged cells, embedded in stigmatic exudates, as also observed in the stigmatic cavity of both selfing and outcrossing variants.ConclusionsOur results provide the first solid evidence of a stigmatic function for the rostellum in orchid flowers, thereby demonstrating for the first time the feasibility of the micro-CT technique for accurately visualizing pollen tube growth in flowering plants. Rostellum receptivity in B . bicoloratum probably uniquely evolved as an adaptation for reproductive assurance from an outcrossing ancestor possessing an erect (non-receptive) rostellum. These findings open up new avenues in the investigation of an organ that apparently re-gained its ‘primordial function’ of being penetrated by pollen tubes.

EFFECT OF RECURRENT SELFING ON INBREEDING DEPRESSION AND MATING SYSTEM EVOLUTION IN AN AUTOPOLYPLOID PLANT

Genome duplication resulting in polyploidy can have significant consequences for the evolution of mating systems. Most theory predicts that self-fertilization will be selectively favored in polyploids; however, many autopolyploids are outcrossing or mixed-mating. Here, we examine the hypothesis that the evolution of selfing is restricted in autopolyploids because the genetic cost of selfing (i.e., inbreeding depression) increases monotonically with successive generations of inbreeding. Using the herbaceous, autotetraploid plant Chamerion angustifolium, we generated populations with different inbreeding coefficients (F= 0, 0.17 and 0.36) through three consecutive generations of selfing and compared their magnitudes of inbreeding depression in a common environment. Mating system estimates for four natural populations confirmed that tetraploid selfing rates (s(m) = 0.25, SE = 0.02) are similar to those of diploids (s(m) = 0.12, SE = 0.12; F₁,₂ = 1.34, P= 0.37) indicating that both cytotypes are predominantly outcrossing. Compared to an outbred control line, mean inbreeding depression for seed production, survival, and height (vegetative and total) in the inbred line differed among generations (inbreeding coefficients). Across all stages, inbreeding depression (relative to control) was positively related to generation (inbreeding coefficient). Although the initial costs of inbreeding in extant and newly synthesized polyploids may be low compared to diploids, the monotonic increase in inbreeding depression with repeated inbreeding may limit the extent to which selfing variants are favored.

Evolutionary pathways to self‐fertilization in a tristylous plant species

Evolutionary transitions from outcrossing to selfing occur commonly in heterostylous genera. The morphological polymorphisms that characterize heterostyly provide opportunities for different pathways for selfing to evolve. Here, we investigate the origins and pathways by which selfing has evolved in tristylous Eichhornia paniculata by providing new evidence based on morphology, DNA sequences and genetic analysis. The primary pathway from outcrossing to selfing involves the stochastic loss of the short-styled morph (S-morph) from trimorphic populations, followed by the spread of selfing variants of the mid-styled morph (M-morph). However, the discovery of selfing variants of the long-styled morph (L-morph) in Central America indicates a secondary pathway and distinct origin for selfing. Comparisons of multi-locus nucleotide sequences from 27 populations sampled from throughout the geographical range suggest multiple transitions to selfing. Genetic analysis of selfing variants of the L- and M-morphs demonstrates recessive control of the loss of herkogamy, although the number of factors appears to differ between the forms. Early stages in the establishment of selfing involve developmental instability in the formation of flowers capable of autonomous self-pollination. The relatively simple genetic control of herkogamy reduction and frequent colonizing episodes may often create demographic conditions favouring transitions to selfing in E. paniculata.

Evaluation of the extent of among‐family variation in inbreeding depression in the perennial herb Scabiosa columbaria (Dipsacaceae)

Significantly different maternal line responses to inbreeding provide a mechanism for the invasion of a selfing variant into a population. The goal of this study was to examine the extent of family-level variation in inbreeding depression in the mixed-mating, perennial herb Scabiosa columbaria. Plants from one population were raised, and hand-pollinated to produce selfed and outcrossed progeny, and the effects of inbreeding depression on life-cycle traits were analyzed. Inbreeding depression significantly affected early life cycle traits. The pollination treatment by family interaction was significant for almost all traits, indicating a high family-level variation in inbreeding depression. The correlations between inbreeding depression values (e.g., percentage germination and flowering date, and flowering date and aboveground biomass) exhibited alternate signs, illustrating the type of association between inbreeding depression loci for different traits across the life cycle. Overall, it is concluded that the extent of among-family variation in inbreeding depression might allow a selfing variant of S. columbaria to invade an outcrossing population, though the pattern of correlations between inbreeding depression values might prevent effective purging of the deleterious genetic load.

EFFECTS OF DIFFERENT LEVELS OF INBREEDING ON PROGENY FITNESS IN PLANTAGO CORONOPUS.

Inbreeding depression (δ) is a major selective force favoring outcrossing in flowering plants. Many phenotypic and genetic models of the evolution of selfing conclude that complete outcrossing should evolve whenever inbreeding depression is greater than one-half, otherwise selfing should evolve. Recent theoretical work, however, has challenged this view and emphasized (1) the importance of variation in inbreeding depression among individuals within a population; and (2) the nature of gene action between deleterious mutations at different loci (epistasis) as important determinants for the evolution of plant mating systems. The focus of this study was to examine the maintenance of inbreeding depression and the relationship between inbreeding level and inbreeding depression at both the population and the individual level in one population of the partially self-fertilizing plant Plantago coronopus (L.). Maternal plants, randomly selected from an area of about 50 m2 in a natural population, were used to establish lines with expected inbreeding coefficients (f) of 0, 0.25, 0.50, 0.75, and 0.875. Inbreeding depression was estimated both in the greenhouse and at the site of origin of the maternal plants by comparing growth, survival, flowering, and seed production of the progeny with different inbreeding coefficients. No significant inbreeding depression for these fitness traits was detected in the greenhouse after 16 weeks. This was in strong contrast to the field, where the traits all displayed significant inbreeding depression and declined with increased inbreeding. The results were consistent with the view that mutation to mildly deleterious alleles is the primary cause of inbreeding depression. At the family level, significantly different maternal line responses (maternal parent × inbreeding level interaction) provide a mechanism for the invasion of a selfing variant into the population through any maternal line exhibiting purging of its genetic load. At the population level, evidence for synergistic epistasis was detected for the probability of flowering, but not for total seed production. At the family level, however, a significant interaction between inbreeding level and maternal families for both traits was observed, indicating that epistasis could play a role in the expression of inbreeding depression among maternal lines.

Stamen elongation, pollen size, and siring ability in tristylous elchhornia paniculata (Pontederiaceae)

Selfing variants in tristylous Eichhornia paniculata (Pontederiaceae) possess an elongated, short‐level stamen adjacent to mid‐level stigmas, which causes autonomous selfing. The variants commonly spread in dimorphic, but not trimorphic populations in northeast Brazil. We investigated the effect of stamen elongation on pollen size and siring ability. Competition experiments using controlled hand‐pollination and allozyme markers were used to compare different pollen types. Pollen from the elongated stamen was significantly larger in size than pollen from unmodified short‐level stamens. In mixed pollinations of mid‐level stigmas, pollen from the elongated stamen sired significantly more seed than pollen from unmodified short‐level stamens. Despite these differences, the size and compatibility of pollen from the elongated stamen were more similar to short‐ than mid‐level pollen, indicating that alterations to stamen level were not associated with major changes in pollen characteristics. The results suggest that the advantage of selfing variants in dimorphic populations is mainly due to efficient pollen transfer to mid‐level stigmas rather than increased postpollination siring success of pollen from modified stamens. In addition, the absence of major changes in pollen size and compatibility associated with stamen elongation support other lines of evidence indicating that selfing variants are not the result of recombination in the putative heterostyly supergene.

POLLEN DISCOUNTING AND THE SPREAD OF A SELFING VARIANT IN TRISTYLOUS EICHHORNIA PANICULATA: EVIDENCE FROM EXPERIMENTAL POPULATIONS.

Floral traits that increase self-fertilization are expected to spread unless countered by the effects of inbreeding depression, pollen discounting (reduced outcross pollen success by individuals with increased rates of self-fertilization), or both. Few studies have attempted to measure pollen discounting because to do so requires estimating the male outcrossing success of plants that differ in selfing rate. In natural populations of tristylous Eichhornia paniculata, selfing variants of the mid-styled morph are usually absent from populations containing all three style morphs but often predominate in nontrimorphic populations. We used experimental garden populations of genetically marked plants to investigate whether the effects of population morph structure on relative gamete transmission by unmodified (M) and selfing variants (M') of the mid-styled morph could explain their observed distribution. Transmission through ovules and self and outcross pollen by plants of the M and M' morphs were compared under trimorphic, dimorphic (S morph absent), and monomorphic (L and S morphs absent) population structures. Neither population structure nor floral morphology affected female reproductive success, but both had strong effects on the relative transmission of male gametes. The frequency of self-fertilization in the M' morph was consistently higher than that of the M morph under all morph structures, and the frequency of self-fertilization by both morphs increased as morph diversity of experimental populations declined. In trimorphic populations, total transmission by the M and M' morphs did not differ. The small, nonsignificant increase in selfing by the M' relative to the M morph was balanced by decreased outcross siring success, particularly on the S morph. In populations lacking the S morph, male gamete transmission by the M' morph was approximately 1.5 times greater than that by the M morph because of both increased selfing and increased success through outcross pollen donation. Therefore, gamete transmission strongly favored the M' morph only in the absence of the S morph, a result consistent with the distribution of the M' morph in nature. This study indicates that floral traits that alter the selfing rate can have large and context-dependent influences on outcross pollen donation.

Pollen Discounting and the Spread of a Selfing Variant in Tristylous Eichhornia paniculata: Evidence from Experimental Populations

Pollen Discounting and the Spread of a Selfing Variant in Tristylous Eichhornia paniculata: Evidence from Experimental Populations

Variation in Outcrossing Rates in Eichhornia paniculata: Temporal Changes in Populations of Contrasting Style Morph Structure

Abstract Surveys of mating‐system parameters in populations of the annual, self‐compatible, tristylous, emergent aquatic, Eichhornia paniculata (Pontederiaceae) from N.E. Brazil and Jamaica have indicated that the species exhibits a wide range of outcrossing rates. To investigate whether temporal variation in outcrossing rate was also a feature of populations, open‐pollinated families were sampled from five populations of contrasting style morph structure from N.E. Brazil over three consecutive years (1987–1989). Multilocus estimates of outcrossing rate (t) were obtained from assays of isozyme polymorphisms using starch gel electrophoresis. There was significant variation both among populations and between years in the frequency of outcrossing. Outcrossing in three tristylous populations was high (t > 0.80), with relatively small fluctuations occurring over the three‐year sampling period. In contrast, in a dimorphic and monomorphic population considerable self‐fertilization occurred and the frequency of outcrossing declined significantly from 1987 to 1989 in both populations. In the dimorphic population, increased selfing was associated with a marked reduction in population size and an increase in the frequency of selfing variants of the mid‐styled morph. The significance of temporal variation in outcrossing frequency in plant populations is discussed in relation to its effect on population genetic structure and recent models of mating‐system evolution.

Multiple origins of self‐fertilization in tristylous Eichhornia paniculata (Pontederiaceae): Inferences from style morph and isozyme variation

AbstractPopulations of Eichhornia paniculata (Pontederiaceae) exhibit a wide range of mating systems, from predominant outcrossing to high levels of self‐fertilization. The origin of self‐fertilization in this tristylous species is associated with the loss of style‐length morphs from populations and the spread of self‐pollinating, floral variants. We examined geographic variation in style morph and allozyme frequencies to determine whether the loss of style morphs and transition to selfing could have multiple origins in E. paniculata. Surveys of floral variation in 167 populations from six states in northeastern Brazil revealed that at least one style morph was absent from 29.3%. Non‐trimorphic populations occurred in all states and ranged in frequency from 9% in Ceará to 68% in Alagoas. Selfing variants occurred in 8.5% and 55% of trimorphic and non‐trimorphic populations, respectively, and were distributed among five of six states with primary concentrations in Alagoas and Pernambuco. A comparison of electrophoretic variation at 24 isozyme loci in 28 trimorphic, 13 dimorphic and 3 monomorphic populations indicated that non‐trimorphic populations contained 84% of the allelic variation present in trimorphic populations and were markedly differentiated from one another. Analyses of genetic distance and the distribution of rare alleles indicated that non‐trimorphic populations were often more similar to neighbouring trimorphic populations than to one another. Populations with selfing variants occurred at low frequency in three genetically distinct parts of the range. These results, in combination with genetic and morphological evidence suggest that style morphs are lost repeatedly from populations of E. paniculata and that selfing variants may have originated on at least three separate occasions in northeastern Brazil.

Variation in Outcrossing Rates in Eichhornia paniculata: The Role of Demographic and Reproductive Factors*

Abstract A survey of 32 populations of the self‐compatible, tristylous, aquatic, Eichhornia paniculata (Pontederiaceae), from N. E. Brazil and Jamaica, was undertaken to examine the role of demographic and reproductive factors on variation in outcrossing rate (t), and to investigate the association between t and population genetic structure. Multi‐locus outcrossing rates, estimated using isozyme techniques, varied widely (t̂= 0.002–0.960) among populations and were uniformly distributed across the entire range of t. Sixty percent of variation in t̂ was explained by style morph diversity (E) and the frequency of selfing variants within populations (M). Population size (N) and plant density (d) also accounted for a significant portion of variation in t̂, particularly in Jamaica, where variation in style morph diversity was low.Outcrossing rates were significantly correlated with the proportion of loci that were polymorphic (P), the average number of alleles per polymorphic loci (Na), the average observed heterozygosity, (Ho), and the inbreeding coefficient (f̂). A strong regional effect was evident in the association of f̂ with P and Na. This largely results from the confounding effect of genetic bottleneck(s) and high levels of self‐fertilization associated with the colonization of Jamaica. Comparisons of f̂ with the equilibrium inbreeding coefficient, (Feq), indicated that outcrossing populations had a significant deficiency of heterozygotes, while selfing populations had an excess. This demonstrates that levels of heterozygosity cannot be accounted for by the mating system alone. Outcrossing rate variation in E. paniculata is controlled by a complex interplay of local demographic and genetic factors. These influences operate within contrasting regional contexts as a result of the different evolutionary histories of populations in N. E. Brazil and Jamaica.

THE DISSOLUTION OF A COMPLEX GENETIC POLYMORPHISM: THE EVOLUTION OF SELF-FERTILIZATION IN TRISTYLOUS EICHHORNIA PANICULATA (PONTEDERIACEAE).

Eichhornia paniculata (Pontederiaceae) displays a wide range of outcrossing levels as a result of the dissolution of the tristylous genetic polymorphism and the evolution of semihomostyly. Population surveys, comparison of fitness components of the style morphs, and computer simulations were used to investigate the breakdown of tristyly and the selective mechanisms responsible for the evolution of self-fertilization. Of 110 populations surveyed in northeast Brazil and Jamaica, 53% were trimorphic, 25% were dimorphic, and 22% were monomorphic for style morph. The short (S) morph was underrepresented in trimorphic populations and absent from nontrimorphic populations. The mid (M) morph predominated in dimorphic populations and was the only morph in monomorphic populations. Stamen modifications promoting selfing, associated with semihomostyle evolution, were largely confined to the M morph. They were rare in trimorphic populations, common in dimorphic populations, and often fixed in monomorphic populations. Stochastic simulations and comparisons of fruit set in natural populations indicate that founder events, population bottlenecks, and lowered fertility of the S morph due to an absence of long-tongued pollinators can each account for loss of the S morph from trimorphic populations. A reduced level of disassortative mating can accentuate the rate at which the S morph is lost by both random and deterministic processes. Nontrimorphic populations occur at the geographical margins of the region surveyed and tend to be smaller and less dense than trimorphic populations. These observations and the higher fruit set of the M morph relative to the L morph in dimorphic populations suggest that reproductive assurance, favoring selfing variants of the M morph under conditions of low pollinator service, has been of primary importance in the origin of most monomorphic populations. Where pollinator service is reliable, however, automatic selection of selfing genes, aided by mating asymmetries between the morphs, can cause the M morph to spread to fixation in dimorphic populations.

RELATIVE FITNESSES OF SELFED AND OUTCROSSED PROGENY IN GILIA ACHILLEIFOLIA (POLEMONIACEAE).

The evolution of self-pollination is a recurrent theme in the history of the angiosperms, and has long been of interest to evolutionists (Darwin, 1876; Darlington, 1939; Mather, 1943; Baker, 1955; Stebbins, 1957; Allard et al., 1968; Jain, 1976; Maynard Smith, 1977; Lloyd, 1979). Yet despite the widespread attention that the topic has received, the factors important to the evolution of self-pollination (autogamy) are poorly understood. As noted by Jain (1976), there are many hypothesized advantages associated with autogamy, but few data bearing on these hypotheses. One of the commonly cited advantages of selfing, and the most general, has been referred to as the automatic selection advantage (Jain, 1976). This advantage arises from the fact that while an outcrossing individual contributes on the average, one haploid genome through its ovules and one through its pollen, a selfing variant contributes up to three haploid genomes on the average, two through its selfed offspring, and one by outcrossing as a pollen parent. Unless the inbred progeny have reduced fitness, mutations promoting selfing are expected to increase in frequency by selection (Fisher, 1941; Maynard Smith, 1977; Lloyd, 1979, 1980). Inbreeding depression in the progeny of selfed parents may impart a reduction in viability, fecundity or both, sufficient to impede the selection of genes for self-pollination. According to Lloyd (1979), in situations where a selfing mutant arises in an outcrossing population, the critical magnitude of inbreeding depression above which mutant genes promoting selfing are