Mixed Mating System Research Articles

Paternity Analysis of Tea (Camellia sinensis L. Kuntz) Hybrids Using Isozyme Marker

Tea plant has been categorized as self-incompatible crop. This is the reason behind the high genetic diversity. Natural pollination is possible to occur and the male parent is usually unknown, therefore, there is a need of method to identify male parent of hybrids through paternity analysis. Isozyme markers have been successfully used for paternity analysis due to their co-dominant polymorphism. This research aimed to predict male parents of hybrids by figuring out the mating system through isozyme banding patterns. In this experiment, seven enzyme systems were evaluated, of which only two of the enzyme systems i.e. esterase and shikimate dehydrogenase showing clear band pattern of Est-1, Est-2, and Shd-1 loci. The mating system of tea could be categorized as a mixed mating model, with high estimated out-crossing rate of 98.6 %. The pollen contributors were not always originated from the vicinity of the female parents. Key words: isozyme markers, paternity analysis, tea

Erratum to: Unexpectedly high outcrossing rate in both dense and sparse patches in self-compatible Pedicularis rex (Orobanchaceae)

Pedicularis rex, a self-compatible alpine herb, depends exclusively on pollinators for reproduction. Previous study has revealed that 74.7 % of genetic variability occurs among populations in Yunnan, indicating a mixed mating system for the species. The large floral display also indicates high potential geitonogamy. Therefore, plant density and display size may be important factors influencing the realized mating system of P. rex. In the study reported here we explored the effects of plant density and floral display size on pollinator movements and thus outcrossing rate in two patches of in P. rex at low and high plant densities. We examined foraging behavior of bumblebees, measured reproductive outputs under hand and natural pollination, and estimated the outcrossing rates using RAPD markers. Floral display size was significantly larger and bumblebees visited significantly more flowers in sequence on individual plants in the sparse patch than in the dense patch. We found a significant positive correlation between floral display size and successive visits in the sparse patch, but not in the dense patch. Regression analysis revealed a significant inverse correlation between fruit production and flower production per plant with a theoretical maximum of 63 fruits per plant, but no relationship between seed production and flower production per plant. Contrary to previous studies on genetic structure, however, we found an unexpectedly high outcrossing rate in both the sparse and the dense patch (tm = 1.151 and 0.924, respectively). Resource limitation may intensify seed competition and result in selected seed abortion, which may largely explain the unexpectedly high outcrossing rate.

Outcrossing rates in two self‐compatible, hybridising Rhinanthus species: implications for hybrid formation

The congeners Rhinanthus angustifolius and Rhinanthus minor, two annual hemiparasites pollinated by bumblebees, are known to hybridise in the wild. Both species are self-compatible, but the capacity for autonomous selfing is higher in R.minor. This suggests a difference in realized outcrossing rates, which have not been determined before in these species. Using microsatellites, both species turned out to have mixed mating systems, but with a much lower multilocus outcrossing rate (0.13) for R.minor compared to R.angustifolius (0.76). We hypothesised that a higher outcrossing rate should lead to a higher chance of heterospecific pollination, and we therefore determined the rate of hybrid formation on each species in an artificial mixed population. Hybrid seeds were produced at low frequency (4.5%), and no significant difference was found between the species. It is therefore likely that post-pollination processes influence hybrid seed formation to counteract the expected difference in heterospecific pollen deposition. We checked fruit set, seed set and the rate of autonomous selfing in controlled crosses in the greenhouse in 2 years, and found that fruit set (2003) or seed set (2010) were lower in R.angustifolius × R.minor crosses relative to the reciprocal cross. Hybrid seeds produced on R.angustifolius also had a much lower germination rate, so most of the established F1 hybrid plants have the R.minor cytoplasm. The formation of advanced hybrids depends on pollinator preference, which is biased towards R.angustifolius if present in sufficient numbers, because it offers more rewards.

Genetic diversity and relationships of the brown alga Undaria pinnatifida cultivated along the Dalian Coast as revealed by amplified fragment length polymorphism markers

In this study, the amplified fragment length polymorphism (AFLP) method was employed to estimate the genetic diversity of young sporophytes belonging to six Undaria pinnatifida populations collected from five seaweed cultivation farms along the Dalian Coast of China in 2008. A total of 397 loci were detected using ten combinations of selective primer pairs. Of these, 302 of which (76.07 %) were polymorphic, which was mainly caused by the 40 accessions of two populations collected from the LWT (44.08 %) and the JST (44.58 %) farms. According to the UPGMA dendrogram and biplot of principal component analysis, the 40 accessions represented high intra-population genetic diversity and far relationship from all of other populations. In contrast, the accessions from the other populations (HK, MS, LS, and QD) represented high intra-population similarity. The four populations may have originated from the same introduced U. pinnatifida strain. Analysis of molecular variance showed that a higher proportion of genetic variation resided among the populations (63.56 %) than that within populations (36.64 %). It is hypothesized that the differences in genetic traits, as well as the mixed mating system and reduced gene flow were the primary reasons that caused the high genetic variation among populations and the high similarity between accessions within the population. However, the genetic diversity of the partial U. pinnatifida cultivar populations in Dalian farms has been decreasing in the past years, which poses a potential risk of germplasm degradation in cultivation. In addition, the breeding techniques of cultivated U. pinnatifida in China and the application of AFLP in Undaria are discussed in this article. Based on the results, the identified polymorphic markers could be used for genetic improvement of the species through marker-assisted breeding and the results also provide excellent information for selecting indigenous gametophyte resources from Dalian farms using different breeding methods.

Epigenetics as a new avenue for the role of inbreeding depression in evolutionary ecology

Epigenetics as a new avenue for the role of inbreeding depression in evolutionary ecology

Sistema reprodutivo e polinização de Lepidagathis sessilifolia (Pohl) Kameyama ex Wassh. & J.R.I. Wood (Acanthaceae), em remanescente florestal da região sudoeste de Mato Grosso, Brasil

O gênero Lepidagathis apresenta distribuição pantropical com cerca de 100 espécies. No Brasil ocorrem 16 espécies, a maioria nas regiões Centro - Oeste e Sudeste. O estudo foi realizado em sub - bosque de remanescente florestal do município de Tangará da Serra - MT e teve como objetivo analisar a fenologia de floração, descrever a morfologia e biologia floral, verificar os visitantes florais e avaliar o sistema e o sucesso reprodutivo por meio de polinizações manuais. Lepidagathis sessilifolia apresenta inflorescências espiciformes, terminais, com cálice de cor rósea vistosa e corola de coloração branco-rósea. A floração ficou restrita aos meses de março a abril, durante a estação chuvosa. A senescência floral ocorreu após 24 ou 48 horas. A viabilidade dos grãos de pólen foi elevada (92,5%). O único polinizador observado visitando as flores de L. sessilifolia foi a abelha Partamona nhambiquara (Apidae - Meliponini). O sistema reprodutivo misto da espécie é caracterizado pela formação de frutos por meio de agamospermia, autopolinização e polinização cruzada. Esse sistema reprodutivo flexível é vantajoso, pois, garante a manutenção da espécie na área de estudo mesmo na ausência de polinizadores.

Morph-specific mating patterns in a low-density population of a heterodichogamous tree, Juglans ailantifolia

Heterodichogamy, including protandrous (PA) and protogynous (PG) morphs, is considered a mechanism to avoid selfing and promote disassortative mating. Although morphotypes are usually present in a population at a 1:1 ratio, this ratio may be biased in a low-density population by demographic stochasticity, resulting in a deficiency of mating partners in a neighbourhood dominated by a single morph. In this study, we determined morph ratio of the heterodichogamous tree species, Juglans ailantifolia by observing flowering in a low-density population during 2 years. The morph ratio (PG: PA) of 2.56:1 deviated significantly deviated from 1:1. We genotyped 59 reproductive trees and 405 offspring derived from eight PG-mother and three PA-mother trees with 11 microsatellite markers. Paternity analysis was conducted to clarify the effects of mother morph on the proportion of intra-morph mating. Then, we applied the Bayesian mixed effect mating model (MEMM) to clarify mating system, pollen dispersal, and individual fecundity of PG- and PA-mother trees. We found that the selfing rate and the distance of pollen dispersal were not clearly different between PG- and PA-mother trees. In contrast, the proportion of intra-morph mating was higher in the majority-morph (PG) mother trees than in the minor-morph (PA) mother trees. The MEMM indicated that mean dispersal distance of PG-mother trees was larger than that of PA-mother trees with large variance. Furthermore, we observed individuals with unusually high intra-morph fecundity for majority-morph (PG) trees. These findings indicate that intra-morph mating may occur when majority-morph mothers suffer a deficiency of potential inter-morph mates.

Nucleotide sequence diversity of floral pigment genes in Mexican populations of Ipomoea purpurea (morning glory) accord with a neutral model of evolution

The common morning glory (Ipomoea purpurea) is an annual vine native to Central and Southern Mexico. The genetics of flower color polymorphisms and interactions with the biotic environment have been extensively studied in I. purpurea and in its sister species I. nil. In this study, we examine nucleotide sequence polymorphism in 11 loci, 9 of which are known to participate in a pathway that produces floral pigments. A sample of 30 I. purpurea accessions from the native range of Central and Southern Mexico comprise the data, along with one accession from each of the two sister species I. alba and I. nil. We observe moderate levels of nucleotide sequence polymorphism of ~1%. The ratio of recombination to mutation parameter estimates (ρ/θ) of ~2.5 appears consistent with a mixed-mating system. Ipomoea resequencing data from these genic regions are noteworthy in providing a good fit to the standard neutral model of molecular evolution. The derived silent site frequency spectrum is very close to that predicted by coalescent simulations of a drift-mutation process, and Tajima's D values are not significantly different from expectations under neutrality.

The role of a mixed mating system in the reproduction of a Mediterranean subshrub (Fumana hispidula, Cistaceae)

Knowledge about mixed mating systems can improve our understanding of the evolutionary dynamics of reproductive systems. Here we report a study of the pollination system (floral and reproductive biology, pollen limitation and stigmatic pollen load, floral visitors and inbreeding depression) of Fumana hispidula, a Mediterranean subshrub belonging to a species group with a strong selfing tendency. Autonomous self-pollination, hand self- and hand cross-pollination, open pollination and controls yielded fruits (0.28, 0.65, 0.68, 0.38, and 0.67, respectively); however, autonomous self-pollination resulted in the lowest fruit set. All individuals were fully self-compatible but we found great inter-individual variation in autonomous self-pollination ability, which was probably related to a variable expression of herkogamy degree. Inbreeding depression was low (0.040 for fruit set, 0.015 for seed set and -0.026 for seed mass). The pollen supplementation experiment did not reveal pollen limitation and pollinators were seen visiting the flowers during the observation periods. These results support the idea that F. hispidula has a mixed mating strategy, which represents a successful reproduction mode in their patchy habitats.

Evolutionary Assurance vs. Mixed Mating

Reproductive assurance is a widely accepted explanation for the evolution of selfing, although theory suggests that an evolutionarily stable mixed mating strategy does not maximize seed production. We present a correlation analysis involving 28 species representing 23 families showing that selfing can evolve independently of inbreeding depression. We discuss the cost-benefit trade-off of selfing, in particular the incongruence of whether delayed selfing provides reproductive assurance in 22 species representing 14 families, in which pollen and seed discounting are minimized when pollinators or mates are scarce. Reproductive assurance, in response to frequent pollinator failure, can be reconciled with an evolutionarily stable mixed mating system contributed to by delayed selfing, which is still advantageous even if there is strong inbreeding depression.

Evidence for autonomous selfing in grassland Protea species (Proteaceae)

It has been assumed that species of the large African genus Protea have strong self-incompatibility systems. However, this assumption was based largely on studies conducted on a clade of bird-pollinated species that occur in the shrubby fynbos vegetation of the Cape region of southern Africa. To test whether self-incompatibility occurs in a grassland/savanna Protea clade, which is largely insect-pollinated, we performed controlled pollination experiments on four species, P. caffra, P. dracomontana, P. simplex and P. welwitschii. Although pollen–ovule ratios of all four species fall within the range for outcrossers, all four species are self-compatible and capable of autonomous seed production. Using fluorescence microscopy, we found that self-pollen tubes had the same probability of reaching ovules as cross-pollen tubes. In the small tree P. caffra, selfed progeny had rates of germination and survivorship that were identical to those of crossed progeny. The grassland Protea spp. studied are likely to have mixed mating systems on account of being both visited by insects and capable of autonomous selfing. If one assumes previous reports of self-incompatibility in Protea to be reliable, there have been at least five losses of self-incompatibility and two gains of autonomous selfing in this genus. However, earlier studies in the genus were often methodologically flawed and a thorough re-analysis of breeding systems in Protea is required. © 2012 The Linnean Society of London, Botanical Journal of the Linnean Society, 2012, 169, 433–446.

Microevolutionary Distribution of Isogenicity in a Self-fertilizing Fish (Kryptolebias marmoratus) in the Florida Keys

The mangrove rivulus Kryptolebias marmoratus and a closely related species are the world's only vertebrates that routinely self-fertilize. Such uniqueness presents a model for understanding why this reproductive mode, common in plants and invertebrates, is so rare in vertebrates. A survey of 32 highly polymorphic loci in >200 specimens of mangrove rivulus from multiple locales in the Florida Keys, USA, revealed extensive population-genetic structure on microspatial and micro-temporal scales. Observed heterozygosities were severely constrained, as expected for a hermaphroditic species with a mixed-mating system and low rates of outcrossing. Despite the pronounced population structure and the implied restrictions on effective gene flow, isogenicity (genetic identity across individuals) within and among local inbred populations was surprisingly low even after factoring out probable de novo mutations. Results indicate that neither frequent bottlenecks nor directional genetic adaptation to local environmental conditions were the primary driving forces impacting multilocus population-genetic architecture in this self-fertilizing vertebrate species. On the other hand, a high diversity of isogenic lineages within relatively small and isolated local populations is consistent with the action of diversifying selection driven by the extreme spatio-temporal environmental variability that is characteristic of mangrove habitats.

Reproductive Ecology and Severe Pollen Limitation in the Polychromic Tundra Plant, Parrya nudicaulis (Brassicaceae)

Pollen limitation is predicted to be particularly severe in tundra habitats. Numerous reproductive patterns associated with alpine and arctic species, particularly mechanisms associated with reproductive assurance, are suggested to be driven by high levels of pollen limitation. We studied the reproductive ecology of Parrya nudicaulis, a species with relatively large sexual reproductive investment and a wide range of floral pigmentation, in tundra habitats in interior montane Alaska to estimate the degree of pollen limitation. The plants are self-compatible and strongly protandrous, setting almost no seed in the absence of pollinators. Supplemental hand pollinations within pollinator exclusion cages indicated no cage effect on seed production. Floral visitation rates were low in both years of study and particularly infrequent in 2010. A diversity of insects visited P. nudicaulis, though syrphid and muscid flies composed the majority of all visits. Pollen-ovule ratios and levels of heterozygosity are consistent with a mixed mating system. Pollen limitation was severe; hand pollinations increased seed production per plant five-fold. Seed-to-ovule ratios remained low following hand pollinations, indicating resource limitation is likely to also be responsible for curtailing seed set. We suggest that pollen limitation in P. nudicaulis may be the result of selection favoring an overproduction of ovules as a bet-hedging strategy in this environmental context of highly variable pollen receipt.

Variability of individual genetic load: consequences for the detection of inbreeding depression

Inbreeding depression is a key factor affecting the persistence of natural populations, particularly when they are fragmented. In species with mixed mating systems, inbreeding depression can be estimated at the population level by regressing the average progeny fitness by the selfing rate of their mothers. We applied this method using simulated populations to investigate how population genetic parameters can affect the detection power of inbreeding depression. We simulated individual selfing rates and genetic loads from which we computed fitness values. The regression method yielded high statistical power, inbreeding depression being detected as significant (5% level) in 92% of the simulations. High individual variation in selfing rate and high mean genetic load led to better detection of inbreeding depression while high among-individual variation in genetic load made it more difficult to detect inbreeding depression. For a constant sampling effort, increasing the number of progenies while decreasing the number of individuals per progeny enhanced the detection power of inbreeding depression. We discuss the implication of among-mother variability of genetic load and selfing rate on inbreeding depression studies.

Higher fine-scale genetic structure in peripheral than core populations of a long-lived and mixed-mating conifer - eastern white cedar (Thuja occidentalis L.)

BackgroundFine-scale or spatial genetic structure (SGS) is one of the key genetic characteristics of plant populations. Several evolutionary and ecological processes and population characteristics influence the level of SGS within plant populations. Higher fine-scale genetic structure may be expected in peripheral than core populations of long-lived forest trees, owing to the differences in the magnitude of operating evolutionary and ecological forces such as gene flow, genetic drift, effective population size and founder effects. We addressed this question using eastern white cedar (Thuja occidentalis) as a model species for declining to endangered long-lived tree species with mixed-mating system.ResultsWe determined the SGS in two core and two peripheral populations of eastern white cedar from its Maritime Canadian eastern range using six nuclear microsatellite DNA markers. Significant SGS ranging from 15 m to 75 m distance classes was observed in the four studied populations. An analysis of combined four populations revealed significant positive SGS up to the 45 m distance class. The mean positive significant SGS observed in the peripheral populations was up to six times (up to 90 m) of that observed in the core populations (15 m). Spatial autocorrelation coefficients and correlograms of single and sub-sets of populations were statistically significant. The extent of within-population SGS was significantly negatively correlated with all genetic diversity parameters. Significant heterogeneity of within-population SGS was observed for 0-15 m and 61-90 m between core and peripheral populations. Average Sp, and gene flow distances were higher in peripheral (Sp = 0.023, σg = 135 m) than in core (Sp = 0.014, σg = 109 m) populations. However, the mean neighborhood size was higher in the core (Nb = 82) than in the peripheral (Nb = 48) populations.ConclusionEastern white cedar populations have significant fine-scale genetic structure at short distances. Peripheral populations have several-folds higher within-population fine-scale genetic structure than core populations. Anthropogenic disturbances and population fragmentation presumably have significant effects on fine-scale genetic structure in eastern white cedar. Core populations have higher neighborhood size than peripheral populations, whereas gene flow distances are higher in peripheral than in core populations. The results of our study contribute to the knowledge of poorly-understood spatial genetic structure of core versus peripheral populations in plants. As well, the information is of significance for conservation of genetic resources of eastern white cedar and perhaps of other long-lived forest trees with mixed-mating system.

Nucleotide Sequence Diversity of Floral Pigment Genes in Mexican Populations of Ipomoea purpurea (Morning Glory) Accord with a Neutral Model of Evolution

The common morning glory (Ipomoea purpurea) is an annual vine native to Central and Southern Mexico. The genetics of flower color polymorphisms and interactions with the biotic environment have been extensively studied in I. purpurea and in its sister species I. nil. In this study, we examine nucleotide sequence polymorphism in 11 loci, 9 of which are known to participate in a pathway that produces floral pigments. A sample of 30 I. purpurea accessions from the native range of Central and Southern Mexico comprise the data, along with one accession from each of the two sister species I. alba and I. nil. We observe moderate levels of nucleotide sequence polymorphism of ~1%. The ratio of recombination to mutation parameter estimates (ρ/θ) of ~2.5 appears consistent with a mixed-mating system. Ipomoea resequencing data from these genic regions are noteworthy in providing a good fit to the standard neutral model of molecular evolution. The derived silent site frequency spectrum is very close to that predicted by coalescent simulations of a drift-mutation process, and Tajima's D values are not significantly different from expectations under neutrality.

Genetics, evolution and conservation of Bromeliaceae

Bromeliaceae is a morphologically distinctive and ecologically diverse family originating in the New World. Three centers of diversity, 58 genera, and about 3,140 bromeliad species are currently recognized. We compiled all of the studies related to the reproductive biology, genetic diversity, and population structure of the Bromeliaceae, and discuss the evolution and conservation of this family. Bromeliads are preferentially pollinated by vertebrates and show marked variation in breeding systems, from predominant inbreeding to predominant outcrossing, as well as constancy in chromosome number (2n = 2x = 50). Autogamous or mixed mating system bromeliads have a high inbreeding coefficient (F(IS)), while outcrossing species show low F(IS). The degree of differentiation among populations (F(ST))of species ranges from 0.043 to 0.961, which can be influenced by pollen and seed dispersal effects, clonal growth, gene flow rates, and connectivity among populations. The evolutionary history of the Bromeliaceae is poorly known, although some studies have indicated that the family arose in the Guayana Shield roughly 100 Mya. We believe that genetic, cytogenetic, and reproductive data will be essential for diagnosing species status and for assisting conservation programs.

Inbreeding Depression in Solanum carolinense (Solanaceae) under Field Conditions and Implications for Mating System Evolution

The clonal weed Solanum carolinense exhibits plasticity in the strength of its self-incompatibility (SI) system and suffers low levels of inbreeding depression (δ) in the greenhouse. We planted one inbred and one outbred plant from each of eight maternal plants in a ring (replicated twice) and monitored clonal growth, herbivory, and reproduction over two years. Per ramet δ was estimated to be 0.63 in year one and 0.79 in year two, and outbred plants produced 2.5 times more ramets than inbred plants in the spring of year two. Inbred plants also suffered more herbivore damage than outbred plants in both fields, suggesting that inbreeding compromises herbivore resistance. Total per genet δ was 0.85 over the two years, indicating that S. carolinense is unlikely to become completely self-compatible, and suggesting that plasticity in the SI system is part of a stable mixed-mating system permitting self-fertilization when cross pollen limits seed production.

Pollination of the heterostylous Galápagos native, Cordia lutea (Boraginaceae)

The hypothesis of this study was that in the Galapagos Islands, fruit and seed set via nocturnal pollination would exceed that of diurnal pollination due to greater insect activity at night typical of hot, arid regions. Cordia lutea, a heterostylous member of the Galapagos flora, was submitted to pollination experiments, visitor observations, nectar sampling, pollen transfer studies, pollen-ovule ratio studies, and pollen measurements. Flowers set fruit and seed via open pollination, autonomous autogamy, facilitated autogamy, facilitated cross-pollination, diurnal pollination, and nocturnal pollination. There was a significant difference in fruit set between flowers experiencing legitimate cross-pollinations (pin × thrum) and those experiencing all other pollination treatments except facilitated autogamy. There was no significant difference in seed set among any of the treatments, but there was a trend toward greater seed set for flowers experiencing open-pollination, legitimate cross-pollination, and nocturnal pollination. There was no significant difference in fruit set or seed set between flowers experiencing diurnal pollination and nocturnal pollination, although there was a trend toward greater seed set resulting from nocturnal pollination. Carpenter bees were the most effective diurnal pollinators, whereas moths were the most effective nocturnal pollinators. Of the two, moths are more efficient at transporting pollen from plant to plant. Results indicate that an overall low productivity of this species is due to pollen limitation exacerbated by nectar robbing. Cordia lutea exhibits a mixed mating system, producing a relatively low level of fruits through a combination of self- and cross-pollination, facilitated by the relatively few insects that are available.

Molecular diversity, genetic structure and mating system of Calopogonium mucunoides Desv.

Calopogonium mucunoides Desv. is a species native of South and Central America that is used as green manure and a pasture crop. The molecular genetic diversity was characterized in 195 C. mucunoides accessions from a germplasm collection using 17 microsatellite markers. Outcrossing rate was estimated after the evaluation of six microsatellite loci in 200 genotypes originated from 10 open-pollinated progenies (20 genotypes per progeny). Six genetic clusters were identified in the germplasm collection by the STRUCTURE software analysis, neighbor-joining tree comparisons and principal component analysis, which highly correlated with the geographic locations where these accessions were originated or collected. These results were confirmed using AMOVA. The largest portion of the genetic variation was observed among clusters (64.38%). The results indicated that: multilocus outcrossing rate (tm) was 16.3%, suggesting a mixed mating system with a predominance of autogamy; single locus outcrossing rate (ts) was 11%; difference (tm−ts) was 0.054, indicating that only 5.4% of outcrossing occurred among related individuals; paternity correlation (rp) was 33% suggesting a low probability of finding full sibs among the progeny; parental coefficient of inbreeding (Fm) was 5.0%, indicating a low degree of inbreeding in each parent. A core collection for C. mucunoides was assembled to capture the allelic diversity found in this study. The complete allelic diversity was represented by only 15 accessions. These results should be useful for exploiting the genetic resources of C. mucunoides and could influence future conservation efforts and breeding programs.