Mycorrhizal associations and reproductive isolation in three closely related Orchis species

Background and Aims

The maintenance of species boundaries in sympatric populations of closely related species requires some kind of reproductive isolation that limits gene flow among species and/or prevents the production of viable progeny. Because in orchids mycorrhizal fungi are needed for seed germination and subsequent seedling establishment, orchid–mycorrhizal associations may be involved in acting as a post-mating barrier.

Methods

We investigated the strength of post-mating barriers up to the seed germination stage acting between three closely related Orchis species (Orchis anthropophora, O. militaris and O. purpurea) and studied the role of mycorrhizal fungi in hybridization by burying seed packets of pure and hybrid seeds. After retrieval and assessment of seed germination, the fungi associating with protocorms originating from hybrid and pure seeds were determined and compared with those associating with adult individuals using DNA array technology.

Results

Whereas pre-zygotic post-mating barriers were rather weak in most crosses, post-zygotic post-mating barriers were stronger, particularly when O. purpurea was crossed with O. anthropophora. Germination trials in the field showed that seed germination percentages of hybrid seeds were in most cases lower than those originating from pure crosses. In all species pair combinations, total post-mating reproductive isolation was asymmetric. Protocorms associated with a smaller range of fungal symbionts than adult plants, but there was considerable overlap in mycorrhizal associations between protocorms and their respective parents.

Conclusions

Our results suggest that mycorrhizal associations contribute little to reproductive isolation. Pre-mating barriers are probably the main factors determining hybridization rates between the investigated species.     

Behavioral asymmetries in a moving hybrid zone

Three narrow hybrid zones exist between hermit and Townsend'swarblers in the Pacific Northwest. Character transition curvessuggest that these zones are moving and that Townsend's warblershave a selective advantage over hermit warblers. This studycompares year-to-year return rates, male persistence on territories,pairing success, pairing patterns, and territory quality of hermitand Townsend's warblers and their hybrids in the WashingtonCascades hybrid zone. There was no difference in the year-to-yearreturn rate between the parental species. Townsend's males weremore successful in maintaining territories and attracting matesthan hermit males. Among mated pairs there were few hermit malescompared to females, independently supporting the inferiorityof hermit males in competition for territories or female mate choicefor Townsend's-like males. I found no difference in the qualityof hermit and Townsend's territories; however, in poor habitatsTownsend's males were more successful at attracting mates, suggestingfemale preference for Townsend's-like males. In high-qualityhabitats, there was no difference in pairing success betweenthe parental species. The fitness of hybrids relative to parentalsaffects the width and movement of the zone. Hybrids were intermediatein their ability to maintain territories and to attract mates, whichshould increase the width of the zone and accelerate its movement.Fewer hybrid males returned to from one year to the next, andamong mated pairs there were few hybrid females relative tomales, suggesting hybrid inferiority, which should narrow thehybrid zone and slow its movement.     

Response of three arid zone floodplain plant species to inundation

For species to persist on floodplains and in temporary wetlands in arid climates, where large and unpredictable water level fluctuations are common, at least one life history stage must be able to survive inundation. We investigated the survival and performance (RGR, total biomass and above-to-belowground biomass (A:B)) of three common and often coexisting arid zone floodplain species: Xanthium strumarium, Cyperus gymnocaulos and Ludwigia peploides. Observations suggested the species had different responses to inundation, which was tested in a controlled pond experiment. Plants were held at three elevations (+ 10 cm, ? 20 and ? 70 cm) and subjected to three hydrological regimes (static 90 cm, 1 and 5 cm day^?1 inundation) for 16 weeks. Xanthium strumarium died when completely inundated for longer than 4 weeks but when partially flooded survived, showed lower growth rates, increased A:B and produced adventitious roots. C. gymnocaulos showed reduced growth rates when partially flooded and senesced to rhizomes when completely inundated for longer than 4 weeks, which re-sprouted after inundation pressure was removed. L. peploides responded positively to flooding with increased A:B and the production of adventitious roots. The species exhibited three contrasting responses to inundation, which do not necessarily fit neatly within existing water regime functional classification frameworks.     

Gene‐flow across the European crow hybrid zone – a spatial simulation

In hybrid zones populations that are otherwise allopatric meet and produce hybrids. One of the most well‐known hybrid zones occurs throughout much of Europe between the hooded and carrion crows. Even though these species (or subspecies) of crows look very different, genetic differentiation is weak, and introgression seems to be common. In a spatial simulation that was based on empirically estimated values, we investigated how introgressing alleles that would confer some fitness advantage would flow across the zone. The advantage was assumed to be some unknown factor that enhanced survival for carriers relative to non‐carriers. We varied the yearly survival advantage between 0 to 10% and recorded how this would affect zone shape and position. In the simulation we cycled ‘yearly’ events such as reproduction, mortality and juvenile dispersal. We started the simulation by equipping all individuals of one crow type outside the zone with a homozygotic set of the beneficial allele. At all levels of advantage the allele first rapidly became fixed in the donator crow type, then transgressed into the receiving type more slowly and finally became fixed in all individuals. The time until fixation varied from around 176 yr for a 10% advantage to around 20 000 yr for 0% advantage. An exciting discovery was that the position of the zone would move during the introgression but then stop. The reason is that the beneficial allele would be common in the donating type but not in the receiving type during the introgression event. At large the crow hybrid zone appears to have been stable for a long time but there have been small recent northward movements of carrion crows in both Scotland and Denmark. We suggest that introgression by itself is a factor that should be considered as an explanation for why hybrid zones move temporarily.     

Trophic dynamics within a hybrid zone – interactions between an abundant cyprinid hybrid and sympatric parental species

1. Recent proliferation of hybridisation in response to anthropogenic ecosystem change, coupled with increasing evidence of the importance of ancient hybridisation events in the formation of many species, has moved hybridisation to the forefront of evolutionary theory. 2. In spite of this, the mechanisms (e.g. differences in trophic ecology) by which hybrids co‐exist with parental taxa are poorly understood. A unique hybrid zone exists in Irish freshwater systems, whereby hybrid offspring off two non‐native cyprinid fishes often outnumber both parental species. 3. Using stable isotope and gut content analyses, we determined the trophic interactions between sympatric populations of roach (Rutilus rutilus), bream (Abramis brama) and their hybrid in lacustrine habitats. 4. The diet of all three groups displayed little variation across the study systems, and dietary overlap was observed between both parental species and hybrids. Hybrids displayed diet, niche breadth and trophic position that were intermediate between the two parental species while also exhibiting greater flexibility in diet across systems.     

Cytonuclear disequilibria in a spatially structured hybrid zone

A hybrid zone model was developed that uses a stepping-stone model of population structure along with both nuclear and cytoplasmic genotypes to evaluate the effect that migration has on random mating organisms when no selection is present. Numerical simulations indicate that a number of different allele frequency and cytonuclear disequilibrium patterns can be found across the hybrid zone when it has reached equilibrium, and these results are discussed in the context of relative migration rates of the two sexes and the two source populations. The importance of numbers of subpopulations sampled, census time, and the persistence of pure species individuals into the hybrid zone are also discussed. Although this model does not consider selection or assortative mating in the hybrid zone, it should provide a useful baseline for evaluating joint cytonuclear genetic data from a structurally complex hybrid zone.     

Nonrandom distribution of chromosomal aberrations induced by three chemicals

The G-band locations of 3244 breakpoints induced by cis-platinum (II) diamminedichloride (PDD), 1460 breakpoints induced by cytosine arabinoside (ara-C), and 1257 breakpoints induced by triethylenemelamine (TEM) in human lymphocyte chromosomes were identified. The breakpoints induced by each of these chemicals demonstrated a significantly nonrandom distribution within the human karyotype. The overall pattern of the interarm distribution was dependent upon the chemical used, but certain chromosomes arms exhibited similar responses to all 3 chemicals. Comparison of the frequencies of breakpoints within individual G-bands indicated that (1) certain bands were susceptible to damage induced by all 3 chemicals; (2) certain bands were resistant to damage by all 3 chemicals; (3) certain bands demonstrated variable susceptibility to induced damage dependent upon the chemical agent; and (4) other bands demonstrated near expected frequencies of damage (by length) to all 3 agents.     

Multi-scale spatial patterns of three seagrass species with different growth dynamics

Knowledge of landscape spatial patterns of seagrasses and their rates of loss and natural colonization is critical for understanding the ecology of this group of submerged aquatic plants. Seagrasses form extensive meadows that occupy sheltered coastal seas of the world. In this paper, we examine the multi-scale variability of three seagrass species over a large near-shore region (42 km²) in Western Australia. Geostatistical non-parametric methods were used to explore spatial variation in presence of Amphibolis griffithii , Posidonia coriacea and P. sinuosa , and to identify the spatial scales at which distinct patterns in the species distributions occur: <50, 50–610 and >610 m. Each species showed unique variance structure across local (<50 and 50–610 m) and regional scales (>610 m), suggesting differences in species biology, environmental requirements, inter-species interactions, and their ability to modify their environment. These observations reflect that 1) seagrass landscapes are created by processes that independently act on each seagrass species at different spatial scales; 2) the species' distributions differ in their hydrodynamic forcing, and; 3) seagrass species distributions reflect colonization history such that related species are separated in space because they have different places in the successional sequence. This cross-scale study demonstrates that shoot studies only partly address the spatial structure of seagrass landscapes and further large-scale spatially-explicit research is required before we can interpret the driving processes.     

Genetic diversity in three colonizing orchids with contrasting mating systems

Comparative studies on the genetic attributes of colonizing orchids with diverse mating systems are lacking in the literature. Eulophia sinensis, Spiranthes hongkongensis, and Zeuxine strateumatica are colonizing orchids that frequently occupy newly created habitats in Hong Kong. Mating system studies showed that E. sinensis is a self-compatible but pollinator-dependent outcrossing species, S. hongkongensis is a self-pollinating taxon, and Z srateumatica is apomictic. Population genetic attributes of these orchid species were investigated. Despite their contrasting mating systems, electrophoretic surveys revealed a striking lack of allozyme variation, both within and among populations, in all three species. However, gene duplications were evident in these species, due to their likely polyploid origins, The percentage of duplicate loci exhibiting "fixed heterozygosity" was 10, 21.4, and 20% in E. sinensis, S. hongkongensis, and Z. strateumatica, respectively. The genetic attributes of these orchids are compared to those found in other colonizing plant species.     

Genetic structure in a dynamic baboon hybrid zone corroborates behavioural observations in a hybrid population

Behaviour and genetic structure are intimately related: mating patterns and patterns of movement between groups or populations influence the movement of genetic variation across the landscape and from one generation to the next. In hybrid zones, the behaviour of the hybridizing taxa can also impact the incidence and outcome of hybridization events. Hybridization between yellow baboons and anubis baboons has been well documented in the Amboseli basin of Kenya, where more anubis-like individuals tend to experience maturational and reproductive advantages. However, it is unknown whether these advantages are reflected in the genetic structure of populations surrounding this area. Here, we used microsatellite genotype data to evaluate the structure and composition of baboon populations in southern Kenya. Our results indicate that, unlike for mitochondrial DNA, microsatellite-based measures of genetic structure concord with phenotypically based taxonomic distinctions and that the currently active hybrid zone is relatively narrow. Isolation with migration analysis revealed asymmetric gene flow in this region from anubis populations into yellow populations, in support of the anubis-biased phenotypic advantages observed in Amboseli. Populations that are primarily yellow but that receive anubis gene flow exhibit higher levels of genetic diversity than yellow populations far from the introgression front. Our results support previous work that indicates a long history of hybridization and introgression among East African baboons. Specifically, it suggests that anubis baboons are in the process of gradual range expansion into the range of yellow baboons, a pattern potentially explained by behavioural and life history advantages that correlate with anubis ancestry.     

Asymmetrical gene flow in a hybrid zone of Hawaiian Schiedea (Caryophyllaceae) species with contrasting mating systems

Asymmetrical gene flow, which has frequently been documented in naturally occurring hybrid zones, can result from various genetic and demographic factors. Understanding these factors is important for determining the ecological conditions that permitted hybridization and the evolutionary potential inherent in hybrids. Here, we characterized morphological, nuclear, and chloroplast variation in a putative hybrid zone between Schiedea menziesii and S. salicaria, endemic Hawaiian species with contrasting breeding systems. Schiedea menziesii is hermaphroditic with moderate selfing; S. salicaria is gynodioecious and wind-pollinated, with partially selfing hermaphrodites and largely outcrossed females. We tested three hypotheses: 1) putative hybrids were derived from natural crosses between S. menziesii and S. salicaria, 2) gene flow via pollen is unidirectional from S. salicaria to S. menziesii and 3) in the hybrid zone, traits associated with wind pollination would be favored as a result of pollen-swamping by S. salicaria. Schiedea menziesii and S. salicaria have distinct morphologies and chloroplast genomes but are less differentiated at the nuclear loci. Hybrids are most similar to S. menziesii at chloroplast loci, exhibit nuclear allele frequencies in common with both parental species, and resemble S. salicaria in pollen production and pollen size, traits important to wind pollination. Additionally, unlike S. menziesii, the hybrid zone contains many females, suggesting that the nuclear gene responsible for male sterility in S. salicaria has been transferred to hybrid plants. Continued selection of nuclear genes in the hybrid zone may result in a population that resembles S. salicaria, but retains chloroplast lineage(s) of S. menziesii.     

Dispersion movements in ants: spatial structuring and density-dependent effects

This paper examines whether the characteristics of individual dispersion movements in ants are changed when workers are moving solitarily or in a group. We analyzed the trajectories of workers of the species Messor sancta moving solitarily or in groups of different size (5, 10, 15 individuals), tested for density-dependent effects on their trajectory characteristics and investigated through resampling techniques whether ants are able to spatially structure their movements through direct (e.g. contact) or indirect (pheromone deposited on the ground) interactions. In addition to group size, the effects of the nutritional state of the colony and of the state of the area on which ants were dispersing were also examined. Solitary ants moved faster and had more sinuous trajectories than ants moving in a group. We found however no significant differences in trajectory characteristics between groups of different size. Whatever the group size, ants from starved colonies moved more slowly and had more direct trajectories than their counterpart coming from fed colonies. On the other hand, the state of the area on which ants were moving had no direct significant effect on dispersion movement. Ants dispersing in a group moved independently and did not coordinate their movements through direct or indirect interactions. However, the geometry of their path was changed not only through the effect of random encounters with other workers but also through an active modification of their movement when they perceived directly or indirectly the presence of nearby workers.     

A narrow hybrid zone between two Cottus species in Wills Creek, Potomac drainage

We describe a narrow hybrid zone between the mottled sculpin (Cottus b. bairdi) and the Blue Ridge sculpin (C. caeruleomentum). Seven characters (dorsal fin rays, pectoral fin rays, caudal base band condition, male spawning coloration, and one frequency and two fixed allozyme differences) distinguish the two taxa in the hybrid zone. C. caeruleomentum and C. b. bairdi diverged in these characters in allopatry as indicated by their distribution on opposite sides of the Atlantic-Ohio divide. However, a stream capture placed these two taxa in secondary contact in Wills Creek, Potomac drainage (Atlantic slope). Allozyme data indicate the presence of post-F(1) hybrids in the zone of secondary contact. Changes in allozymes, morphology, and spawning coloration along a transect in Wills Creek reveal the hybrid zone is less than 20 river kilometers in length. Estimates of root mean square dispersal and gene flow tentatively suggest that selection is operating in the Wills Creek hybrid zone. C. b. bairdi and C. caeruleomentum are maintaining their identity in seven distinguishing characters on opposite ends of the hybrid zone revealing these two taxa are independent evolutionary lineages.     

Environment-dependent admixture dynamics in a tiger salamander hybrid zone

After an estimated five million years of independent evolution, the barred tiger salamander (Ambystoma tigrinum mavortium) was introduced by bait dealers into the native range of the California tiger salamander (A. californiense). Hybridization and backcrossing have been occurring in central California for 50-60 years, or an estimated 15-30 generations. We studied genetic and ecological factors influencing admixture of these two divergent gene pools by analyzing frequencies of hybrid genotypes in three kinds of breeding habitats: natural vernal pools, ephemeral man-made cattle ponds, and perennial man-made ponds. Perennial ponds tended to have higher frequencies of nonnative alleles than either type of seasonal pond, even in cases where perennial and seasonal ponds are within a few hundred meters. Thus, the hybrid zone has a mosaic structure that depends on pond hydrology or ecology. The presence of some broadly acting constraints on admixture is suggested by linkage disequilibria between physically unlinked molecular markers within ponds. In addition, we found several marker-specific deviations from Hardy-Weinberg equilibrium. One marker showed a consistent deficit of heterozygotes across pond types. Another showed heterozygote deficits only in vernal pools. A third was more likely to have heterozygote excess in ephemeral cattle ponds. These patterns indicate that admixture is influenced by complex genotype-by-environment interactions.     

Risk assessment by crow phenotypes in a hybrid zone

Predation is one of the most selective forces in evolution and, thus, predation may select against hybrids in narrow hybrid zones. It may be possible that parental phenotypes and hybrids differ in their responses towards predators or humans. As predation is difficult to observe I used flight-initiation distance (FID) as a metric of risk assessment. FID is a measurable outcome of the trade-off between fleeing and remaining. Here, I tested whether hybrid and parent crow phenotypes (Corvus corone, Corvus cornix) from the hybrid zone in Eastern Germany differ in their FID. Further, I measured many environmental and social variables to control statistically for their influence on FID. I sampled 154 individuals (53 hooded crows, 54 carrion crows, and 48 hybrids) in the hybrid zone in eastern Germany. I calculated a general linear model using a stepwise backward procedure to establish a minimum model containing only significant variables that explained FID in crows. The variable phenotype (hooded, carrion, hybrid) was then added to the model. There were no differences in FID between hybrids and both parental phenotypes types, suggesting similar risk assessment. This suggests that hybrids may behave similarly in their decision to flee as their parent phenotypes, which, in turn, provides no evidence for a selective disadvantage. An additional analysis focusing on pure phenotypic flocks showed that hybrids in pure hybrid flocks had a lower FID than both parental species in pure flocks. This suggests that hybrids in pure hybrid flocks may be at a disadvantage.     

Fine scale spatial structuring of sex and mitochondria in Silene vulgaris

Fine scale spatial structure (FSSS) of cytoplasmic genes in plants is thought to be generated via founder events and can be amplified when seeds germinate close to their mother. In gynodioecious species these processes are expected to generate FSSS in sex ratio because maternally inherited cytoplasmic male sterility genes partially influence sex expression. Here we document a striking example of FSSS in both mitochondrial genetic markers and sex in roadside populations of Silene vulgaris. We show that in one population FSSS of sexes influences relative fruit production of females compared to hermaphrodites. Furthermore, FSSS in sex ratio is expected to persist into future generations because offspring sex ratios from females are female-biased whereas offspring sex ratios from hermaphrodites are hermaphrodite-biased. Earlier studies indicated that pollen limitation is the most likely mechanism underlying negative frequency dependent fitness of females. Our results support the theoretical predictions that FSSS in sex ratio can reduce female fitness by decreasing the frequency at which females experience hermaphrodites. We argue that the influence of FSSS on female fitness is complementary to the influence of larger scale population structure on female fitness, and that population structure at both scales will act to decrease female frequencies in gynodioecious species. Better comprehension of the spatial structure of genders and genes controlling sex expression at a local scale is required for future progress toward understanding sex ratio evolution in gynodioecious plants.