Minor quantitative trait loci underlie floral traits associated with mating system divergence in Mimulus

The genetic basis of species differences provides insight into the mode and tempo of phenotypic divergence. We investigate the genetic basis of floral differences between two closely related plant taxa with highly divergent mating systems, Mimulus guttatus (large-flowered outcrosser) and M. nasutus (small-flowered selfer). We had previously constructed a framework genetic linkage map of the hybrid genome containing 174 markers spanning approximately 1800 cM on 14 linkage groups. In this study, we analyze the genetics of 16 floral, reproductive, and vegetative characters measured in a large segregating M. nasutus x M. guttatus F2 population (N = 526) and in replicates of the parental lines and F1 hybrids. Phenotypic analyses reveal strong genetic correlations among floral traits and epistatic breakdown of male and female fertility traits in the F2 hybrids. We use multitrait composite interval mapping to jointly locate and characterize quantitative trait loci (QTLs) underlying interspecific differences in seven floral traits. We identified 24 floral QTLs, most of which affected multiple traits. The large number of QTLs affecting each trait (mean = 13, range = 11-15) indicates a strikingly polygenic basis for floral divergence in this system. In general, QTL effects are small relative to both interspecific differences and environmental variation within genotypes, ruling out QTLs of major effect as contributors to floral divergence between M. guttatus and M. nasutus. QTLs show no pattern of directional dominance. Floral characters associated with pollinator attraction (corolla width) and self-pollen deposition (stigma-anther distance) share several pleiotropic or linked QTLs, but unshared QTLs may have allowed selfing to evolve independently from flower size. We discuss the polygenic nature of divergence between M. nasutus and M. guttatus in light of theoretical work on the evolution of selfing, genetics of adaptation, and maintenance of variation within populations.     

Inbreeding depression in two species of Mimulus (Scrophulariaceae) with contrasting mating systems

We examined the effect of self- and cross-pollination on germination success, flowering probability, pollen and ovule production, survivorship, and adult aboveground biomass in two species of Mimulus with contrasting mating systems: the highly seifing M. micranthus and an outcrossing population of M. guttatus. Cross-pollinations were performed both within and between populations in order to examine the scale at which the genetic load is distributed. We found significant inbreeding depression in M. guttatus in four of the six traits, with the highest inbreeding depression observed in biomass (68% and 69% based on within- and between-population crosses, respectively) and lowest in ovule production (21% based on between-population crosses only). M. micranthus displayed significant inbreeding depression in only two of the six traits examined. Again, we observed the highest inbreeding depression in biomass (47–60% based on within- and between-population crosses, respectively), but both traits showing significant differences between self and outcross progeny expressed lower inbreeding depression than in M. guttatus. We detected no significant inbreeding depression for either pollen or ovule production in M. micranthus. An estimate of total inbreeding depression based on the multiplicative effects of all traits was also lower in M. micanthus than∗∗∗ in M. guttatus. Our results are consistent with the expected purging of genetic load in populations with high selfing rates. The absence of inbreeding depression in M. micranthus pollen and ovule production, two traits with strong links to fitness in a selfing annual, further suggests the important role of directional selection in determining the population's genetic load. Comparison of cross-pollinations made within and between populations revealed little evidence of divergence of genetic load among the M. micranthus and M. guttatus populations examined.     

Modes of self-fertilization in Mimulus guttatus (Scrophulariaceae): a field experiment

We inferred Lloyd's modes of selfing in a natural population of the common monkeyflower, Mimulus guttatus. Estimates were obtained using floral manipulations combined with seed counts and isozyme analyses of selfing rates. Of the 25% selfing estimated from isozyme markers, about one-half was competing, about one-third was geitonogamous, and at least one-fifth (perhaps twice this) was due to biparental inbreeding. Estimates of prior and delayed selfing were small and did not significantly differ from zero. These results were obtained using plants with the characteristic pair of open flowers at an inflorescence node. The second-opening flower showed twice the rate of selfing, presumably because of protogynous-based geitonogamy differences. Solitary-flowered plants, which have smaller flowers but no geitonogamy, showed about 50% selfing, consisting of about equal components of competing selfing and biparental inbreeding. While geitonogamy and biparental inbreeding might be unavoidable by-products of adaptations for outcrossing, competing selfing is subject to more direct natural selection and warrants adaptive explanations.     

The relationship between mating system and penis morphology in ischnuran damselflies (Odonata: Goenagrionidae)

Zygopterans belonging to the genus Ischnura are unusual amongst damselflies because of the variety of mate guarding techniques employed by males of different species. The lack of post-copulatory guarding combined with lengthy copulations in one group of ischnuran species suggest that these males guard females in copula. An examination of the accessory penes of species in this group indicates that all but one species have considerable microspination on the distal end (the flagella) of their penes that can function in sperm displacement. The flagella of these species are long and thin compared to those of other ischnurans. This is likely an adaptation to gain access to the spermatheca of the female. Two species tandem guard their mates during ovipositing. These species are the only ischnurans missing a stout pair of basal spines on the penultimate segments of their penes. They have considerable microspination over much of their penes but their flagella are of only moderate length and stout. Ischnurans that do not mate guard have short, stout flagella and most species examined from this group (5 of 7) have litde microspination on their flagella tips. It is proposed that females of these species mate only once and therefore their males do not displace sperm.     

Inbreeding effects on mating system traits for two species of Lupinus (Leguminosae)

The consequences of inbreeding for reproductive traits were investigated for two closely related annual lupines that differ in their mating system. Lupinus bicolor (Leguminosae) is a primarily selling species while Lupinus nanus outcrosses at intermediate rates. A controlled crossing program was used for each species to produce selfed and outcrossed progeny. These progeny were then grown in a greenhouse and scored for the date of first flower, flower morphology, and autofertility. Selfed progeny of L. bicolor produced significantly smaller flowers but did not differ from outcrossed progeny for the remaining traits. Selfed progeny of L. nanus produced flowers that significantly differed in shape and had fewer ovules than the flowers of outcrossed progeny. Selfed progeny of L. nanus also had significantly lower rates of autofertility in comparison to outcrossed progeny. The significant effects of inbreeding on these mating system traits may indicate the presence of directional dominance at the loci underlying these characters. The consequences of these direct effects of inbreeding on reproductive traits for plants growing in natural populations may include nonadaptive changes in the outcrossing rate between generations.     

The genetic basis of floral traits associated with mating system evolution in Leptosiphon (Polemoniaceae): an analysis of quantitative trait loci

Mapping of quantitative trait loci (QTL) was used to investigate the genetic architecture of divergence in floral characters associated with the mating system, an important adaptive trait in angiosperms. Two species of Leptosiphon (Polemoniaceae), one strongly self-fertilizing (L. bicolor) and the other partially outcrossing (L. jepsonii), were crossed to produce F2 and both backcross progenies. For each crossing population, a linkage map was created using amplified fragment length polymorphism markers, and QTL were identified for several dimensions of floral size. For each of the five traits examined, three to seven QTL were detected, with independent datasets yielding congruent results in some but not all cases. The phenotypic effect of individual QTL was generally moderate. We estimated that many of the QTL were additive or showed dominance toward L. bicolor, whereas comparison of mean trait values for parental and cross progenies showed apparent overall dominance of L. jepsonii traits. Colocalization of QTL for different dimensions of floral size was consistent with high phenotypic correlations between floral traits. Substantial segregation distortion was observed in marker loci, the majority favoring alleles from the large-flowered parent. A low frequency of male sterility in the F2 population is consistent with the Dobzhansky-Muller model for the evolution of reproductive isolation.     

AFLP mapping and detection of quantitative trait loci (QTLs) for economically important traits in Pinus sylvestris: a preliminary study

We have applied a two-way pseudo-testcross strategy in an analysis of Pinus sylvestris for genetic mapping and detection of quantitative trait loci (QTLs) associated with economically important traits targeted in the Swedish tree-breeding programme. Based on 94 full-sib progeny of a cross between two plus-trees from northern Sweden we generated two parental maps using AFLP markers. The female map was comprised of 94 markers assigned to 15 linkage groups giving a size of 796 cM. On the male map 155 markers were assigned to 15 linkage groups, giving a total size of 1335 cM. The recombination frequency was found to be sex-dependent, being 29.3% higher in male than in female gametes. On the female map, 12 QTLs were detected (but none for branch diameter or wood density). Three QTLs for tree height accounted for 25.8% of the total phenotypic variation of this trait. When the QTLs detected for all the traits were taken independently, the percentages of phenotypic variance ranged from 9.3% to 22.7%. The highest value was observed for frost hardiness, an important trait in northern Sweden for which a major gene seemed to be involved. A cluster of QTLs for tree height, trunk diameter and volume was located on one linkage group. On the male map, four QTLs for trunk diameter and volume were detected. Due to the reduced number of individuals under study, the results are preliminary and have to be validated on more trees.     

The relationship between mating and oogenesis in monarch butterflies (Lepidoptera: Danainae)

We studied the relationship between the timing of mating and oogenesis in monarch butterflies (Danaus plexippus) to determine (1) the potential for male nutrient input into eggs and (2) whether mating stimulates egg development. Most females mated soon after they started maturing eggs. One and 2 days after mating, females contained the same number of mature oocytes as virgin females of the same age, while 3 days after mating they contained more mature oocytes than did virgins. These results confirm the potential for male-derived nutrients to augment oocyte production, but indicate that mating is not required for oocyte maturation to occur.     

Linkage between loci of quantitative traits and marker loci: multi-trait analysis with a single marker

An efficient approach to increase the resolution power of linkage analysis between a quantitative trait locus (QTL) and a marker is described in this paper. It is based on a counting of the correlations between the QTs of interest. Such correlations may be caused by the segregation of other genes, environmental effects and physiological limitations. Let a QT locus A/a affect two correlated traits, x and y. Then, within the framework of mixture models, the accuracy of the parameter estimates may be seriously increased, if bivariate densities f _aa(x, y), f _Aa(x, y) and f _AA(x, y) rather than the marginals are considered as the basis for mixture decomposition. The efficiency of the proposed method was demonstrated employing Monte-Carlo simulations. Several types of progeny were considered, including backcross, F₂ and recombinant inbred lines. It was shown that provided the correlation between the traits involved was high enough, a good resolution to the problem is possible even if the QTL groups are strongly overlapping for their marginal densities.     

Identification of several small main-effect QTLs and a large number of epistatic QTLs for drought tolerance related traits in groundnut (Arachis hypogaea L.)

Cultivated groundnut or peanut (Arachis hypogaea L.), an allotetraploid (2n = 4x = 40), is a self pollinated and widely grown crop in the semi-arid regions of the world. Improvement of drought tolerance is an important area of research for groundnut breeding programmes. Therefore, for the identification of candidate QTLs for drought tolerance, a comprehensive and refined genetic map containing 191 SSR loci based on a single mapping population (TAG 24 x ICGV 86031), segregating for drought and surrogate traits was developed. Genotyping data and phenotyping data collected for more than ten drought related traits in 2-3 seasons were analyzed in detail for identification of main effect QTLs (M-QTLs) and epistatic QTLs (E-QTLs) using QTL Cartographer, QTLNetwork and Genotype Matrix Mapping (GMM) programmes. A total of 105 M-QTLs with 3.48-33.36% phenotypic variation explained (PVE) were identified using QTL Cartographer, while only 65 M-QTLs with 1.3-15.01% PVE were identified using QTLNetwork. A total of 53 M-QTLs were such which were identified using both programmes. On the other hand, GMM identified 186 (8.54-44.72% PVE) and 63 (7.11-21.13% PVE), three and two loci interactions, whereas only 8 E-QTL interactions with 1.7-8.34% PVE were identified through QTLNetwork. Interestingly a number of co-localized QTLs controlling 2-9 traits were also identified. The identification of few major, many minor M-QTLs and QTL × QTL interactions during the present study confirmed the complex and quantitative nature of drought tolerance in groundnut. This study suggests deployment of modern approaches like marker-assisted recurrent selection or genomic selection instead of marker-assisted backcrossing approach for breeding for drought tolerance in groundnut.     

Association between loci with deleterious alleles and distorted sex ratios in an inbred line of tilapia (Oreochromis aureus)

Three microsatellite markers (UNH159, UNH231, and UNH216) were examined for association with both deleterious genes and sex-ratio distortions in a full-sib family of 222 progeny from the fourth generation of a meiogynogenetic tilapia line (Oreochromis aureus). The three markers were mapped previously to different linkage groups and were shown to be associated with genes with deleterious alleles in this line. A restricted maximum likelihood model was used for analysis of major effects and their interactions on sex ratio and viability. This model was based on selective mortality of genders, ignoring effects of possible sex-determining genes. The results showed that deleterious genes linked to UNH216 and UNH231 exert higher lethality in females than in males (P < .0005 and P < .05, respectively). UNH159 was not associated directly with sex ratio distortion, but acts strongly as a modifier of sex ratio in combination with UNH216 and UNH231. Each of the three loci was found to have a significant effect on viability (P < .05) in the maximum likelihood analysis. The deleterious single-locus effects act strongly against females, while most of the epistatic interactions exert higher lethality in males. This contradiction results in a close to 1:1 sex ratio at maturity. The genetic mechanism and significance of such a balance between genders are still unknown. A detailed analysis of sex-specific lethality may be applied by screening in appropriate series of matings and fine mapping with additional markers. Our data suggest that UNH216 and UNH231 are linked to sex ratio distortion genes and that UNH159 may be linked to a modifier of these genes.     

Evolution of novel morphological and reproductive traits in a clade containing Antirrhinum majus (Scrophulariaceae)

Phylogenetic analysis of DNA sequences of the chloroplast genes rcbL and ndhf revealed a highly supported clade composed of the families Plantaginaceae, Callitrichaceae, and Hippuridaceae in close association with the model organism Antirrhinum majus and other members of family Scrophulariaceae. Plantago has miniature actinomorphic wind-pollinated flowers that have evolved from zygomorphic animal-pollinated precursors. The aquatic Hippuridaceae have reduced windpollinated flowers with one reproductive organ per whorl, and three, rather than four, whorls. In monoecious aquatic Callitrichaceae, further reduction has occurred such that there is only one whorl per flower containing a single stamen or carpel. Optimization of character states showed that these families descended from an ancestor similar to Antirrhinum majus. Recent studies of plant developmental genetics have focused on distantly related species. Differences in the molecular mechanisms controlling floral development between model organisms are difficult to interpret due to phylogenetic distance. In order to understand evolutionary changes in floral morphology in terms of their underlying genetic processes, closely related species exhibiting morphological Variation should be examined. Studies of genes that regulate morphogenesis in the clade described here could aid in the elucidation of a general model tot such fundamental issues as how changes in floral symmetry, organ number, and whorl number are achieved, as well as providing insight on the evolution of dicliny and associated changes in pollination syndrome.     

The relationship between mating system and simultaneous hermaphroditism in the coral reef fish, Hypoplectrus nigricans (Serranidae)

The behaviour and reproductive ecology of the simultaneously hermaphroditic coral reef fish, Hypoplectrus nigricans (Serranidae), the black hamlet, was investigated in an effort to elucidate the relationship between simultaneous hermaphroditism and mating behaviour. The major features of the mating system are as follows. (1) Hamlets spawn only in pairs, one partner releasing eggs and the other fertilizing them. There is no selfing. (2) Eggs are planktonic. There is no parental care. (3) The major courtship display serves to advertise that an individual has ripe eggs. (4) A clutch is not released in a single spawn but is parcelled over several, usually with the same partner. (5) Partners generally alternate sex roles with each spawn: that is, they take turns giving up parcels to be fertilized. This active reciprocation of release of eggs, called egg trading, means that reproductive success as a male depends upon the ability to reproduce as a female. Since most of the reproductive effort of an individual is spent on female functions, egg trading provides a fecundity advantage to hermaphroditism, making it evolutionarily stable relative to dioecy (separate sexes). The advantage is analogous to that which parthenogenetic organisms have over sexual ones. The fecundity advantage under egg trading can account for the maintenance, but not the origin of simultaneous hermaphroditism, since hermaphroditism must be established before egg trading can evolve.     

Nectar traits in Nicotiana section Alatae (Solanaceae) in relation to floral traits, pollinators, and mating system

Nicotiana section Alatae exhibits great diversity among species in floral morphology, mating system, and predominant pollinators. As a first step towards estimating nectar's role in floral evolution, we studied nectar traits to determine whether they vary in association with predominant pollinators and mating system. Daily phenology determines when nectar becomes available to pollinators and differed between hummingbird- and moth-pollinated species. Nectar volume and concentration varied significantly among most species and pollinator groups, but were inversely correlated, so that total energy was similar among most species. In general, nectar volume was positively correlated with corolla length. The autogamous species, N. plumbaginifolia, had a nectar volume that matched expectations based on corolla length, but with lower concentration and total energy than predicted by corolla length, while nectar volume was lower than predicted by corolla length in the autogamous population of N. longiflora. Sugar and amino acid components (determined through HPLC) were similar among species, although differences did exist. The nectar of most species was sucrose-dominant, but the autogamous N. plumbaginifolia had nectar that contained similar proportions of sucrose, glucose, and fructose. Many nectar traits varied in association with the predominant pollinators and, in some cases, with the mating system.     

The joint evolution of mating system, floral traits and life history in Clarkia (Onagraceae): genetic constraints vs. independent evolution

Genetic correlations caused by pleiotropy or linkage disequilibrium may influence the joint evolution of multiple traits as populations or taxa diverge. The evolutionary transition from outcrossing to selfing has occurred numerous times and is often accompanied by phenotypic and genetic changes in multiple traits such as flower size, pollen-ovule ratio, stigma and anther maturity and the age of reproductive maturity. Determining whether the recurring patterns of multitrait change are because of selection on each trait independently and/or the result of genetic correlations among traits can shed light on the mechanism that accounts for such convergence. Here, we evaluate whether floral traits are genetically correlated with each other and/or with whole-plant traits within- and between-populations and taxa. We report results from a greenhouse study conducted on two pairs of sister taxa with contrasting mating systems: the autogamously selfing Clarkia exilis and its predominantly outcrossing progenitor C. unguiculata and the autogamous Clarkia xantiana ssp. parviflora and its outcrossing progenitor C. xantiana ssp. xantiana. We examined variation within and covariation among maternal families in three populations of each taxon with respect to the age at first flower, the rate of successive flower production and the number of days between bud break and anther dehiscence and stigma receptivity within individual flowers. Based on phenotypic divergence between sister taxa, bivariate regressions, correlations among maternal family means and analysis of covariance (ancova), we did not find unilateral support indicating that genetic constraints govern the joint distribution of floral and whole-plant traits.     

Quantitative Trait Loci (QTLs) mapping for growth traits in the mouse: A review

The attainment of a specific mature body size is one of the most fundamental differences among species of mammals. Moreover, body size seems to be the central factor underlying differences in traits such as growth rate, energy metabolism and body composition. An important proportion of this variability is of genetic origin. The goal of the genetic analysis of animal growth is to understand its "genetic architecture", that is the number and position of loci affecting the trait, the magnitude of their effects, allele frequencies and types of gene action. In this review, the different strategies developed to identify and characterize genes involved in the regulation of growth in the mouse are described, with emphasis on the methods developed to map loci contributing to the regulation of quantitative traits (QTLs).     

Many QTLs with minor additive effects are associated with a large difference in growth between two selection lines in chickens

Two growth-selected lines in chickens have been developed from a single founder population by divergent selection for body weight at 56 days of age. After more than 40 generations of selection they show a nine-fold difference in body weight at selection age and large differences in growth rate, appetite, fat deposition and metabolic characteristics. We have generated a large intercross between these lines comprising more than 800 F2 birds. QTL mapping revealed 13 loci affecting growth. The most striking observation was that the allele in the high weight line in all cases was associated with enhanced growth, but each locus explained only a small proportion of the phenotypic variance using a standard QTL model (1.3-3.1%). This result is in sharp contrast to our previous study where we reported that the two-fold difference in adult body size between the red junglefowl and White Leghorn domestic chickens is explained by a small number of QTLs with large additive effects. Furthermore, no QTLs for anorexia or antibody response were detected despite large differences for these traits between the founder lines. The result is an excellent example where a large phenotypic difference between populations occurs in the apparent absence of any single locus with large phenotypic effects. The study underscores the need for powerful experimental designs in genetic studies of multifactorial traits. No QTL at all would have reached genome-wide significance using a less powerful design (e.g. approx. 200 F2 individuals) regardless of the nine-fold phenotypic difference between the founder lines for the selected trait.