Inbreeding and outbreeding reduces cocoon production in the earthworm Eisenia andrei

Earthworms are animals with reciprocal insemination. Eisenia andrei Bouché, 1972 is a simultaneous hermaphroditic earthworm that lives in manure heaps at high densities, with low opportunities of dispersal, thus very close inbreeding is expected. As the negative effects of inbreeding and outbreeding may be severe, we studied whether E. andrei adjusts its breeding effort according to the degree of mate relatedness. To test this, we performed laboratory experiments in which earthworms were mated with their sibs and with non-sibs from the same population and no-sibs from a geographically isolated population. Inbreeding and outbreeding matings caused a strong reduction of cocoon production, especially in genetic lines with high reproductive rates. As far as we know, this is the first study that indicates reproductive adjustment in earthworms according to the genetic divergence of their partners. Optimal outbreeding should be considered a crucial point in the management of breeding populations for applied purposes.     

Rodent control and food supplementation increase productivity of endangered San Clemente Loggerhead Shrikes (Lanius ludovicianus mearnsi)

Habitat loss and predation by exotic predators have contributed to the decline of many island populations of birds, including San Clemente Loggerhead Shrikes (Lanius ludovicianus mearnsi). This critically endangered subspecies has been the recipient of an extensive and expensive recovery effort, including a captive breeding and release program. We examined the effects of life-history characteristics, rainfall, and management, particularly food supplementation and rat (Rattus rattus) control, on the breeding productivity of 172 San Clemente Loggerhead Shrike pairs from 2000 to 2006. Annual fecundity and the number of fledglings reaching independence per pair varied with year and were closely correlated with prebreeding (November-March) precipitation levels. Annual fecundity increased as a result of management implemented between April and July each year. Shrike pairs receiving no management during this period produced a mean of 1.9 fledglings. Pairs receiving both food and rat control produced an average of 2.5 more fledglings than pairs receiving no management, while pairs receiving rat control alone produced an average of 1.1 more fledglings than pairs receiving no management during this period. Wild-origin females produced an average of 0.8 more fledglings per pair than captive-origin females, while older males produced an average of 0.9 more fledglings and independent young relative to inexperienced males. The effects of food supplementation and rat control on San Clemente Loggerhead Shrikes were most pronounced during dry years, suggesting such management should be emphasized when prebreeding precipitation levels are below historical averages. These techniques offer an effective means to increase breeding productivity of avian species in immediate threat of extinction.     

Genetics and extinction

The role of genetic factors in extinction has been a controversial issue, especially since Lande’s paper [Genetics and demography in biological conservation, Science 241 (1988) 1455-1460] paper in Science. Here I review the evidence on the contribution of genetic factors to extinction risk. Inbreeding depression, loss of genetic diversity and mutation accumulation have been hypothesised to increase extinction risk. There is now compelling evidence that inbreeding depression and loss of genetic diversity increase extinction risk in laboratory populations of naturally outbreeding species. There is now clear evidence for inbreeding depression in wild species of naturally outbreeding species and strong grounds from individual case studies and from computer projections for believing that this contributes to extinction risk. Further, most species are not driven to extinction before genetic factors have time to impact. The contributions of mutation accumulation to extinction risk in threatened taxa appear to be small and to require very many generations. Thus, there is now sufficient evidence to regard the controversies regarding the contribution of genetic factors to extinction risk as resolved. If genetic factors are ignored, extinction risk will be underestimated and inappropriate recovery strategies may be used.     

Effects of pollination distance on reproduction and offspring performance in Hypochoeris radicata: Experiments with plants from three European regions

Fragmentation of habitats has resulted in increased inbreeding for many plant species, while the introduction of foreign seed material for ecological restoration has resulted in crosses between plants from distant populations. Both processes may reduce plant fitness and increase the risk of extinction. Variation in the expression of inbreeding and outbreeding depression has been found among different genotypes and among populations, but little is known about large scale geographical patterns within species. We studied the effects of cross-proximity on seed production and offspring performance in the perennial meadow plant Hypochoeris radicata (Asteraceae) from five populations in each of three European regions (Bohemia in northwest Czechia, Northern Hesse in central Germany, and Salland in the central Netherlands). Five artificial cross types were conducted with varying proximity of mates: selfing (self), within family crosses (WF), within population crosses (WP), between population crosses (BP), and between region crosses (BR), and the offspring were grown in a common garden. Independent of the region of origin of the maternal plant, selfing, WF and BP crosses resulted in lower seed set and germination than WP crosses, indicating partial self-incompatibility, inbreeding depression and reduced performance in the F1 progeny resulting from outbreeding. However, crosses between regions resulted in similar seed set and germination as within population crosses. For late traits, the effects of inbreeding and interpopulation crosses differed among regions. WP crosses exhibited the highest survival, flowering and multiplicative fitness only in progeny from Czech maternal plants. Our results suggest that the sensitivity of populations to introgression may vary among regions and that outbreeding depression does not necessarily increase with interpopulation distance. However, the current study investigated only effects in the F1 in a common garden and outbreeding depression may be stronger in the F2 and in field populations.     

Influence of size and partner preference on the female function of the earthworm Eisenia andrei (Oligochaeta,Lumbricidae)

Many aspects of the reproduction and mating behaviour of earthworms remain poorly understood. In this study, we focused on body size as a possible trait that influences earthworm reproduction and mating processes. Eisenia andrei is a simultaneously hermaphroditic animal with reciprocal insemination and many hermaphrodites are expected to mate not primarily to get their own eggs fertilized, but rather to get the opportunity to fertilize the eggs of their partners. We investigated whether E. andrei has a size-dependent sex allocation, i.e. if larger earthworms are more biased toward female allocation and produce more egg mass and whether E. andrei has a size-related mate choice by studying the relationship between mating delay and cocoon production. To test this, we compared cocoon production between pairs of earthworms of equal and different size. Mature individuals of E. andrei were classified in two size classes (small and large) and we performed a two-factorial experiment with earthworm size and the size of the partner as factors. After copulation, earthworms were isolated and thereafter their mass and the number of cocoons they produced were recorded weekly for 18 weeks. We found no evidence of size-dependent sex allocation and we found no effect of size-assortative mating on cocoon production. With respect to the differences in the time to mate, the mating delay seems to indicate the existence of some kind of mate choice, independent of the earthworm size. Those earthworms that were matched sooner laid many cocoons, but those that waited a long time to mate laid fewer cocoons. This effect was stronger in those individuals paired with large partners, suggesting that some large partners are more desired ‘males’ than others; the reason for this remain as an open question. In general, our results confirm that earthworms are able to discriminate their partners and adjust their breeding effort accordingly.     

Decision analysis to guide recovery of the po‘ouli, a critically endangered Hawaiian honeycreeper

We used a probabilistic decision tree to help select among alternative recovery strategies for the Po‘ouli, a critically endangered forest bird endemic to the Hawaiian island of Maui. The Po‘ouli is one of the rarest birds in the world, with only three individuals known, and no breeding pairs. The most urgent conservation need for the species is to create a breeding pair and obtain eggs for captive propagation. Seven recovery strategies had been proposed, and there was disagreement among conservation workers about which strategy to pursue. In addition to lengthy discussions, a decision tree was introduced to provide an objective way of quantifying the chance of success under each alternative. All available information was used to attempt evaluation of each node in the decision tree. The overall chance of obtaining eggs was calculated by multiplying estimates at each node under each alternative. All options produced low estimates of potential success, but two options, removal of all birds to captivity and placement of the birds in a field aviary in an accessible location, produced similar estimates of success. The US Fish and Wildlife Service and the Hawaii Division of Forestry and Wildlife eventually agreed that removal to captivity was the preferred alternative because it could be implemented quickly, which was important considering the advanced age of the birds, and would make it easier to provide veterinary care and ensure their safety from predators, severe weather, and vandals. Decision trees and other structured models should not be relied on exclusively, but they can provide an objective method of helping to make difficult conservation decisions and provide a record of complex thought processes used in reaching a determination. The case of the Po‘ouli may serve as a template for navigation towards a decision to recover other species of extreme rarity.     

Polygyny and female breeding failure reduce effective population size in the lekking Gunnison sage-grouse

Populations with small effective sizes are at risk for inbreeding depression and loss of adaptive potential. Variance in reproductive success is one of several factors reducing effective population size (N_e) below the actual population size (N). Here, we investigate the effects of polygynous (skewed) mating and variation in female breeding success on the effective size of a small population of the Gunnison sage-grouse (Centrocercus minimus), a ground nesting bird with a lek mating system. During a two-year field study, we recorded attendance of marked birds at leks, male mating success, the reproductive success of radio-tagged females, and annual survival. We developed simulations to estimate the distribution of male reproductive success. Using these data, we estimated population size () and effective population size N_e for the study population. We also simulated the effects of population size, skewed vs. random mating, and female breeding failure on N_e. In our study population, the standardized variance in seasonal reproductive success was almost as high in females as in males, primarily due to a high rate of nest failure (73%). Estimated N_e (42) was 19% of in our population, below the level at which inbreeding depression is observed in captive breeding studies. A high hatching failure rate (28%) was also consistent with ongoing inbreeding depression. In the simulations, N_e was reduced by skewed male mating success, especially at larger population sizes, and by female breeding failure. Extrapolation of our results suggests that six of the seven extant populations of this species may have effective sizes low enough to induce inbreeding depression and hence that translocations may be needed to supplement genetic diversity.     

Evaluation of the extinction risk and of conservation alternatives for a very small insular population: the bearded vulture Gypaetus barbatus in Corsica

The bearded vulture Gypaetus barbatus is a large, long-lived osteophagus vulture whose abundance and breeding range have drastically declined during the last century, making it one of the most endangered European bird species. We evaluated the extinction risk of the bearded vulture population in Corsica (a small, isolated breeding population of 8-10 pairs), one of the last extant populations in Western Europe, and estimated its probability of extinction to be 0.165 over the next 50 years. A sensitivity analysis to assess the influence of uncertain demographic rates showed that it is critical to estimate precisely the values of pre-adult survival. Neither the type nor the parameters of density dependence acting on fecundity and survival rates influenced much the extinction risk of the Corsican population. We evaluated the effect of four realistic conservation actions that could be implemented on the Corsican bearded vulture population and rank them in terms of their respective decrease of the current extinction risk faced by this population. We found that the release of two juveniles every other year for 12 years and the increase of fecundity due to selective food provisioning would reduce by more than one-half the current extinction risk of Corsican bearded vulture population. In contrast, even substantial increases in the carrying capacity through large supplemental feeding produced very modest decreases in the extinction risk, thus calling into question the efficacy of one of the main pan-European conservation strategies for this species. Re-establishing a population network within the Mediterranean could be a potentially better strategy, though its efficacy depends on natal dispersal among populations that is currently unknown.     

What genetics tell us about the conservation of the critically endangered Balearic Shearwater?

The Balearic shearwater Puffinus mauretanicus is one of the most critically endangered seabirds in the world. The species is endemic to the Balearic archipelago, and conservation concerns are the low number of breeding pairs, the low adult survival, and the possible hybridization with a sibling species, the morphologically smaller Yelkouan shearwater (P. yelkouan). We sampled almost the entire breeding range of the species and analyzed the genetic variation at two mitochondrial DNA regions. No genetic evidence of population decline was found. Despite the observed philopatry, we detected a weak population structure mainly due to connectivity among colonies higher than expected, but also to a Pleistocene demographic expansion. Some colonies showed a high imbalance between immigration and emigration rates, suggesting spatial heterogeneity in patch quality. Genetic evidence of maternal introgression from the sibling species was reinforced, but almost only in a peripheral colony and not followed, at least to date, by the spread of the introgressed mtDNA lineages. Morphometric differences were not correlated with mtDNA haplotypes and introgression is probably due to a secondary contact between the two species several generations ago. Overall, results suggested that the very recent demographic decline in this critically endangered species has not yet decreased its genetic variability, and connectivity found among most colonies should help to reduce species extinction risk. Spreading of introgression should be monitored, but the species is not jeopardized at the moment by genetic factors and the major conservation actions should concentrate at enhancing adult survival.     

Contrasting measures of fitness to classify habitat quality for the black-throated sparrow (Amphispiza bilineata)

Habitat quality is an important consideration when identifying source and sink habitat and setting priority areas for avian conservation. The problem is that different measures may lead to different conclusions about habitat quality, and may also vary in the resources required to estimate them. Individual level measures, such as nest success, and fecundity, will often identify different high quality habitats than population level measures, such as abundance or the number of fledglings produced per unit area. We tested measures of fitness in the Black-throated Sparrow both at the individual and at the population level for six habitats in the northern Chihuahuan Desert, to explore their value as indicators of habitat quality. We compared clutch size, number of nestlings per nest, number of fledglings per successful nest, nest density, nest success, daily nest survival rate, season-long fecundity, number of fledglings produced per 100 ha, and adult abundance, in each habitat type. We also modeled source-sink dynamics to estimate the scale at which they operate, to infer survival rates, and to ascertain the relative source potential of each habitat. We found that fecundity is the best indicator of individual level habitat quality but a poor indicator of population level habitat quality. Nest success (or fecundity, if resources are available to adequately estimate it) plus nest density provide the most robust indicator of population level habitat quality, which is the level at which priority habitats for conservation should be identified. Mesa grassland and black grama grassland functioned as source habitats most consistently, and mesquite was consistently a sink but also probably a reservoir of individuals available to occupy other habitats.     

High levels of hatching failure in an insular population of the South Island robin: a consequence of food limitation?

Endangered birds in insular environments like New Zealand are often translocated to predator-free offshore islands for conservation purposes. Some translocated populations however exhibit reduced fecundity, and it has been suggested that either inbreeding (due to small number of founders) or resource shortages (due to high population density) may limit reproductive success. For example, the South Island robin (Petroica a. australis) population on Motuara Island (59 ha) was founded by only five birds but has increased to ∼600 individuals, or a density >10 times that of the mainland. Despite this apparent success, the rate of hatching failure in this population is three times higher than in mainland populations, with more than a third of eggs failing to hatch. To test if elevated hatching failure is the result of food limitation, we carried out a food supplementation experiment by providing females with mealworms equivalent to ∼50% of their daily energy requirements. Food supplementation had no effect on hatching success, clutch size, incubation attentiveness or nest size. Egg volume increased with food supplementation in one of the two years of this study, but both egg volume and incubation attentiveness were unrelated to rate of hatching failure. As previous genetic analyses confirmed that the bottlenecked population of robins on Motuara Island have significantly less genetic variation than their source population, we conclude that inbreeding depression, and not food limitation, is the most likely explanation for their high rate of hatching failure. We suggest that the experimental translocation of genetically dissimilar individuals be considered as a possible remedy for low productivity in island populations that were initiated with few founders.     

Reproduction of the African earthworm,Eudrilus eugeniae (Oligochaeta) — cocoons

Summary Although various authors have contributed to our knowledge of the biology of Eudrilus eugeniae some basic facts about the life cycle and reproduction are still not fully documented. Quantitative observations were made of the cocoons at 25 °C in different substrates. The incubation period for 166 cocoons was 16.89 days, based on cocoons produced by worms between the ages of 70 and 100 days. These cocoons produced a mean 2.12 hatchlings per cocoon after incubation in cattle manure, moist filter paper and distilled water. A smaller batch of cocoons incubated in cattle manure produced a mean of 2.7 hatchlings per cocoon. The hatching success of the cocoons was 84% in cattle manure, 50% in distilled water, and 48% on moist filter paper. The reproductive capabilities of E. eugeniae and Eisenia fetida were compared.     

Genetic variation of five species of Trifolium L. from south-west Turkey

Following a legume collection mission to south-west Turkey in 1996, five species of Trifolium were analysed for genetic variation within and between species in eleven morphological and flowering characters. The five species included two outcrossing species, T. michelianum and T. resupinatum, and three inbreeding species, T. clypeatum, T. glomeratum and T. tomentosum. The genetic diversity found was related to climate and edaphic factors. All five species showed significant amounts of genetic differentiation between sites and the species could be separated morphologically by principal components analysis and cluster analysis. The most significant source of genetic variation was found to be related to geographical distribution with those species which were widely distributed across south-west Turkey exhibiting much greater amounts of genetic variation between sites, than those which had a narrow distribution. The breeding system was found to be less important, but only the morphology of the outbreeding species showed any environmental clines in relation to climate. A multiple regression analysis was computed to estimate the effect of growing season on the days to flowering of each of the species.     

Novel microsatellite loci for the compost earthworm Eisenia fetida: A genetic comparison of three North American vermiculture stocks

Earthworms are important components of soil ecosystems worldwide, and have been used extensively as indicator species in ecotoxicology studies. Our understanding of mating systems, population structure, and genetic diversity in earthworms is limited by the current lack of available genetic tools. To address this gap, we developed 16 novel microsatellite markers for the compost earthworm Eisenia fetida, one of the most widely studied earthworm species. We tested the new markers in 3 putative populations of worms from commercial vermiculture operations in Canada and the U.S.A. All 16 loci were variable in at least one population, with the number of alleles per locus ranging from 3 to 10, observed heterozygosity from 0.000 to 0.783, and polymorphic information content from 0.032 to 0.587. One group had significantly reduced heterozygosity compared to the other 2, but overall there were only minor genetic differences among the tested suppliers, suggesting a possible bottleneck for this species in North America. The microsatellite loci we describe here will be extremely useful tools for future field and laboratory studies of E. fetida. Body size and condition in adult worms varied significantly by supplier, and breeding success and cocoon production were significantly higher in breeding pairs from one supplier compared to the other two. In the absence of evidence for genetic divergence among populations, our results suggest that the early rearing environment may be an important factor affecting adult breeding performance. We caution that the source of worms may affect the outcome of laboratory experiments involving growth and reproduction.     

Seed production, germinability and seedling growth for a bird-pollinated shrub in fragments of kwongan in south-west Australia

In this study we investigate the effect of population size on the proportion of flowers that produce a fruit (fruit set), the number of seeds per fruit (seed set), seed germinability, seedling mortality and growth in a range of population fragments for the bird-pollinated mixed mating system shrub Calothamnus quadrifidus R. Br. (Myrtaceae). We found no significant linear relationship (p < 0.05) between population fragment size and fruit set in any of the three years reproduction was studied. In contrast, we found a very strong positive correlation between the number of seeds produced per fruit and increasing population fragment size for each of the three years. We found no significant linear relationships between population fragment size and seed germination, or seedling growth and mortality. The most plausible explanation for the decline in seed set is increased inbreeding in smaller populations. Although a previous mating system analysis with allozymes did not reflect the above, we present evidence from other lines of inquiry to indicate that inbreeding does increase in smaller populations, but is masked by post-zygotic lethal systems that eliminate genetically incompetent homozygous embryos. We found no evidence that highly mobile pollinators transporting pollen among fragments rescue small fragments from inbreeding. We discuss the implications of our findings for the conservation of plant diversity in fragments of species rich Mediterranean climate shrublands.     

Agricultural intensification, rainfall patterns, and large waterbird breeding success in the extensively cultivated landscape of Uttar Pradesh, India

In countries with high human populations, using agricultural areas as multifunctional systems to produce food for humans and retain wildlife may be an efficient conservation strategy for many species. Inclusion of natural habitat and species requirements on agricultural landscapes explicitly into planning processes are precluded by lack of information on drivers of species persistence. Climate change is an additional emerging complexity, and adaptation plans for agricultural landscapes are biased towards intensification to secure long-range food production. I examine the conservation potential of an agricultural landscape in two districts of Uttar Pradesh, north India where agricultural intensification and altered rainfall patterns are predicted to occur. I assess stressors affecting breeding success over eight years of two large waterbirds of conservation concern – Sarus Cranes and Black-necked Storks. Both species had high breeding success that improved with total rainfall and more wetlands in breeding territories. Agricultural and township expansions deteriorated territory quality and reduced breeding success. Sarus Crane populations were predicted to decline relatively rapidly if development activities continued to displace breeding pairs. Black-necked Storks appeared resilient over the long-term notwithstanding reduced breeding success in low-rainfall years. Waterbird nesting habitats (wetlands and trees) were retained in Uttar Pradesh as community lands by villages and by state government via legal provisions suggesting the utility of multiple conservation approaches. Incorporating species requirements explicitly, alongside traditional land use practices conducive for habitat conservation, into adaptation planning and conservation policy will be necessary to retain long-term multifunctionality of such agricultural landscapes.     

Does reduced MHC diversity decrease viability of vertebrate populations?

Loss of genetic variation may render populations more vulnerable to pathogens due to inbreeding depression and depletion of variation in genes responsible for immunity against parasites. Here we review the evidence for the significance of variation in genes of the Major Histocompatibility Complex (MHC) for conservation efforts. MHC molecules present pathogen-derived antigens to the effector cells of the immune system and thus trigger the adaptive immune response. Some MHC genes are the most variable functional genes in the vertebrate genome. Their variation is clearly of adaptive significance and there is considerable evidence that its maintenance is mainly due to balancing selection imposed by pathogens. However, while the evidence for selection shaping MHC variation on the historical timescale is compelling, a correlation between levels of MHC variation and variation at neutral loci is often observed, indicating that on a shorter timescale drift also substantially affects MHC, leading to depletion of MHC diversity. The evidence that the loss of MHC variation negatively affects population survival is so far equivocal and difficult to separate from effects of general inbreeding. Some species with depleted MHC variation seem to be particularly susceptible to infection, but other species thrive and expand following severe bottlenecks that have drastically limited their MHC variation. However, while the latter demonstrate that MHC variation is not always critical for population survival, these species may in fact represent rare examples of survival despite of the loss of MHC variation. There is clearly a compelling need for data that would disclose the possible consequences of MHC diversity for population viability. In particular, we need more data on the impact of MHC allelic richness on the abundance of parasites or prevalence of disease in populations, while controlling for the role of general inbreeding. Before such evidence accumulates, captive breeding programs and other conservation measures aimed at inbreeding avoidance should be favoured over those protecting only MHC variation, especially since inbreeding avoidance programs would usually conserve both types of genetic diversity simultaneously.     

Inbreeding in red-cockaded woodpeckers: Effects of natal dispersal distance and territory location

Inbreeding depression constitutes a significant threat to the viability of small populations. In addition to small size and isolation of populations, short distance dispersal may elevate risk of inbreeding, but empirical evidence is scarce. Inbreeding depression has been demonstrated in the highly endangered red-cockaded woodpecker Picoides borealis. It has been suggested that conservation efforts to support extant populations should aim at spatially aggregating territories to enhance dispersal success. This however may aggravate inbreeding risk because distance between territories and hence dispersal distances become short. We analysed empirical data from a long-term study of the demography of the red-cockaded woodpecker and found that inbreeding risk varied inversely with natal dispersal distance of the mother. Using an individual-based, spatially explicit population model that incorporates simulations of environmental and demographic stochasticity and an empirically derived, species-specific estimate of inbreeding costs, we demonstrated that inbreeding depression significantly elevated extinction risk in this species. On the other hand, even though dispersal distances in populations with spatially aggregated territories were shorter and the proportion of inbred individuals was higher than in other populations of the same size, such populations were still more persistent. Despite the overall adverse effect of inbreeding depression on viability of red-cockaded woodpecker populations, lowering interterritorial distances can be viewed as a valuable conservation tool. Given the small size and isolated location of most extant red-cockaded woodpecker populations however, our findings suggest that inbreeding depression represents a significant threat to the survival of this species.     

The influence of food availability on breeding success of African penguins Spheniscus demersus at Robben Island, South Africa

From 1989 to 2004, the breeding success of African penguins Spheniscus demersus at Robben Island, South Africa was significantly related to estimates of the abundance of both their main prey species, anchovy Engraulis encrasicolus and sardine Sardinops sagax, and to the combined biomass of these species. When the combined spawner biomass of fish prey was less than 2 million ton, pairs fledged an average of 0.46 chicks annually. When it was above 2 million ton, annual breeding success had a mean value of 0.73 chicks per pair. Given previously estimated values of survival and age at first breeding, these levels of breeding success are inadequate to sustain the African penguin population. With the higher level of breeding success, an equilibrium situation might be attained if adult survival could be increased by 6-7% per annum. Attempts to reduce mortality of penguins have included the collection, cleaning and return to the wild of oiled birds, culling of Cape fur seals Arctocephalus pusillus pusillus seen preying on penguins around breeding localities and control of the spread of disease. Management of the purse-seine fishery should ensure adequate escapement of fish to maintain the combined biomass of anchovy and sardine above 2 million ton. The maintenance of suitable breeding habitat and removal of feral predators from breeding localities will also be important in improving breeding success.     

Reproductive biology and ecology of selected rare and endangered Oxalis L. (Oxalidaceae) plant species

About twenty-five percent of all southern African Oxalis species are rare/endangered and highly localized, making them especially vulnerable to extinction through inbreeding, low genetic variation, disrupted biological interactions and stochastic events; all consequences of small population sizes. Moreover, Oxalis displays tristyly, which is a rare and specialized sexual system that includes a strong self-incompatibility component between three floral morphs to promote out-crossing within populations. As tristyly requires the availability of plants with different floral morphs as well as effective pollinators for seed production, this breeding system can affect small populations when fully expressed. Factors that may have an effect on rarity in Oxalis were investigated by focusing on the expression of tristyly, levels of natural seed production, clonality and the ecology of eight rare/highly localized Oxalis species. Field experiments revealed that the reproductive success of some Oxalis species may be hampered by tristyly, resulting in extremely low levels of natural seed production. Other species display a more relaxed expression of self-incompatibility, which in combination with the possibility of cross-pollinations provides reproductive assurance regardless of population structure and pollinator availability. Others are rare and endangered, but appear not to be negatively affected by the tristylous breeding system. Most species are limited by their highly specific habitat requirements and are particularly vulnerable to variation in rainfall patterns.