From species to individuals: combining barcoding and microsatellite analyses from non-invasive samples in plant ecology studies

By means of DNA barcoding and microsatellite analyses, we studied the species and individuals of legitimate seed dispersers of the Mediterranean shrub Pistacia lentiscus, a keystone species that represents the main source of food in winter for frugivorous birds. We collected dropping of birds containing seeds, and after DNA extraction we amplified and sequenced a fragment of the mitochondrial COI gene. Through BLASTN queries of the sequenced fragments against registered sequences in the GenBank database we identified the bird species that are currently dispersing P. lentiscus seeds. Further, through the amplification of specific nuclear microsatellite loci we calculated standard genetic diversity parameters of the population of birds from the genus Sylvia (the blackcap and Sardinian warbler), the most important dispersers of P. lentiscus. Five bird species were identified as seed dispersers through their barcode match. Further, we found that S. melanocephala displayed slightly lower levels of genetic diversity than S. atricapilla. In this study we show how the genetic analyses of environmental faecal samples can be a useful and convenient tool for the study of plant-frugivore interactions through the ascertainment of the identity of the species involved and through the analyses of genetic variability of their populations.     

Non-redundancy in seed dispersal and germination by native and introduced frugivorous birds: implications of invasive bird impact on native plant communities

Seed dispersal by vertebrate animals is important for the establishment of many fleshy-fruited plant species. Different frugivorous species can provide different seed dispersal services according to their specific dietary preferences as well as behaviour and body traits (e.g. body size and beak size of birds). Our aim was to study redundancies and complementarities in seed dispersal and germination between the two main native seed disperser birds and the introduced silver pheasant Lophura nycthemera in the temperate Patagonian forests. For this, we collected fresh droppings from the studied species and analyzed seed content. We conducted germination trials for four plant species common in bird droppings; two native species (Aristotelia chilensis and Rhaphithamnus spinosus) and two invasive non-native species (Rubus ulmifolius and Rosa rubiginosa). Both native frugivorous birds and the silver pheasant dispersed fruits of non- native fleshy-fruited plants, but their roles were non-redundant in terms of species dispersed and effect on seed germination. The silver pheasant dispersed a proportionally high number of non-native seeds, while native birds dispersed a high number of native seeds. In addition, the effect of gut treatment in seed germination differed between seed dispersers. Native birds promoted the germination for the two native plant species studied, while the silver pheasant promoted the germination of one non-native plant. This suggests that seed dispersal by the silver pheasant may contribute to the spread of some invasive fleshy-fruited plants in the ecosystems that otherwise would not be dispersed by any other bird. The understanding of redundancies and complementarities on seed dispersal and germination between native and introduced birds will allow improving the management of fleshy-fruited non-native plants.     

Famine Foods of Brazil’s Seasonal Dry Forests: Ethnobotanical and Nutritional Aspects

Famine Foods of Brazil’s Seasonal Dry Forests: Ethnobotanical and Nutritional Aspects. Famine foods are used by people in times of food scarcity. Although the northeast of Brazil regularly faces periods of drought that cause a shortage of traditional foods, the use of famine foods is insufficiently recorded. The purpose of this study was to record the knowledge of famine food in two rural Caatinga communities, Carão and Cachoeira, in the states of Pernambuco and Paraiba, respectively, and to determine the chemical composition of the major famine foods utilized. The ethnobotanical aspect was divided into two stages: free lists and semistructured interviews. Based on the interviews, the main famine foods known by communities were selected for nutritional analysis. The study revealed that certain foods are used by communities only in times of shortages. Carão showed a higher species richness than Cachoeira, but the food knowledge was more widespread among the interviewees in Cachoeira. In Carão, the main species mentioned were Dioclea grandiflora and Manihot dichotoma, whereas in Cachoeira, Encholirium spectabile and Pilosocereus gounellei were the major species indicated. The species examined had a high carbohydrate content, especially D. grandiflora, Manihot glaziovii, and M. dichotoma, and could be used for complementary feeding of the population if adverse effects could be minimized. Mandevilla tenuiflora is indicated for future studies of potential agro-industrial applications and the beneficial aspects of its root, which could represent an alternative source of income for the population.     

Differences in reward removal efficiency and its consequences for seed germination in the ant-dispersed sedge <Emphasis Type="Italic">Carex tristachya</Emphasis> (Cyperaceae)

Reward removal is an essential step for seed dispersal mutualism because residual rewards inhibit germination. Nevertheless, variation in the reward removal efficiency (RRE) among dispersers and its consequences for germination have rarely been reported. In this study, we compared the RREs of two sympatric seed-dispersing ants, Formica japonica and Pheidole noda, using seeds of the ant-dispersed sedge Carex tristachya. Then, we conducted seed sowing experiments in a non-heated glasshouse to evaluate the effect of RRE on the percentage and speed of germination. The majority (85%) of seeds handled by F. japonica had residual elaiosomes, while elaiosomes were completely removed from all seeds handled by P. noda, demonstrating that P. noda has much higher RRE than F. japonica. The seed sowing experiments revealed that RRE, defined by the presence or absence of residual elaiosomes, was not associated with the percentage germination within a year. However, high RRE seeds with no residual elaiosomes germinated significantly faster than low RRE seeds with residual elaiosomes. Similarly, artificial removal of elaiosomes from C. tristachya seeds accelerated germination speed without affecting germination percentage. These results suggest that RRE is one of the most important parameters determining the effectiveness of a seed dispersal agent.     

Dispersal of remnant endangered trees in a fragmented and disturbed forest by frugivorous birds

Most endangered plant species in a fragmented forest behave as a unique source population, with a high dependence on frugivorous birds for recruitment and persistence. In this study, we combined field data of dispersal behavior of birds and GIS information of patch attributes to estimate how frugivorous birds could affect the effective dispersal pattern of Chinese yew (Taxus chinensis) in a fragmented and disturbed forest. Nine bird species were observed to visit T. chinensis trees, with Urocissa erythrorhyncha, Zoothera dauma and Picus canus being the most common dispersers. After foraging, six disperser species exhibited different perching patterns. Three specialist species, P. canus, Turdus hortulorum, and Z. dauma stayed in the source patch, while three generalist species, U. erythrorhyncha, Hypsipetes mcclellandii, and H. castanonotus, could perch in bamboo patches and varied in movement ability due to body size. As a consequence of perching, dispersers significantly contributed to the seed bank, but indirectly affected seedling recruitment. Moreover, the recruitment of T. chinensis was also affected by patch attributes in a fragmented forest (distances to source patch, patch type, size). Our results highlighted the ability of unique source population regeneration of T. chinensis in a fragmented forest, with high dependence on both frugivorous birds and patch attributes, which should be considered in future planning for forest management and conservation.     

Fruiting phenology as a “triggering attribute” of invasion process: Do invasive species take advantage of seed dispersal service provided by native birds?

Mechanisms underlying biological invasion of highly disturbed ecosystems are well known, yet mechanisms responsible for biological invasion of undisturbed or weakly disturbed ecosystems are less understood. The triggering attribute (TA) approach, proposed as a mechanism that explains plant invasion success in undisturbed or weakly disturbed systems, considers that the spread of alien species depends on specific vegetative or regenerative traits in invasive species, discontinuously distributed in comparison to the resident community. In mountain Chaco woodland, fruiting phenology of ornithocorous invasive plants has been proposed as a TA, because it would allow invasive species to benefit from seed dispersal service, which is unused by native plants during a specific period of the year (winter). Under the seed dispersal ecology framework, we evaluated if fruiting phenology (fructification largely uncoupled with native species) of the fleshy-fruited invasive Pyracantha angustifolia affects bird fruit consumption, and allows the invasive to take advantage of the unused seed dispersal service during winter. If uncoupled fructification phenology represents a TA, seed disperser, seed predator, and pulp consumer diversity, abundance, and fruit consumption on P. angustifolia (which fructifies in winter), will be higher than on its exotic congeneric P. coccinea during summer, when fructification overlaps with native Celtis ehrenbergiana and many other native species. We found that: (1) disperser bird abundance and fruit consumption did not differ between P. angustifolia and P. coccinea; (2) the most diverse frugivorous assemblage was observed on C. ehrenbergiana, yet it had the lowest proportion of seed dispersers and the highest fruit consumption by seed predators and, (3) we also observed higher proportion of seed predators on P. angustifolia (uncoupled fructification scenario) than on P. coccinea (coupled fructification scenario). Our results suggest that invasive uncoupled fructification phenology does not represent a true TA which facilitates plant invasion processes in undisturbed or weakly disturbed ecosystem.     

Is induction ability of seed germination of <Emphasis Type="Italic">Phelipanche ramosa</Emphasis> phylogenetically structured among hosts? A case study on Fabaceae species

Phelipanche ramosa is a major root-holoparasitic damaging weed characterized by a broad host range, including numerous Fabaceae species. In France, the agricultural threat posed by P. ramosa has increased over two decades due to the appearance of a genetically differentiated pathovar presenting a clear host specificity for oilseed rape. The new pathovar has led to a massive expansion of P. ramosa in oilseed rape fields. The germination rate of P. ramosa seeds is currently known to vary among P. ramosa pathovars and host species. However, only a few studies have investigated whether phylogenetic relatedness among potential host species is a predictor of the ability of these species to induce the seed germination of parasitic weeds by testing for phylogenetic signal. We focused on a set of 12 Fabaceae species and we assessed the rate of induction of seed germination by these species for two pathovars based on in vitro co-cultivation experiments. All Fabaceae species tested induced the germination of P. ramosa seeds. The germination rate of P. ramosa seeds varied between Fabaceae species and tribes studied, while pathovars appeared non-influential. Considering oilseed rape as a reference species, we also highlighted a significant phylogenetic signal. Phylogenetically related species therefore showed more similar rates of induction of seed germination than species drawn at random from a phylogenetic tree. In in vitro conditions, only Lotus corniculatus induced a significantly higher germination rate than oilseed rape, and could potentially be used as a catch crop after confirmation of these results under field conditions.     

Seed Dispersal by Primates and Implications for the Conservation of a Biodiversity Hotspot,the Atlantic Forest of South America

Primates play a fundamental role as seed dispersers, particularly in tropical rainforests. Because defaunation and fragmentation are leading several primate species to local extinction, it is fundamental to understand the role of primates as effective seed dispersers. Here we present a systematic review of studies of seed dispersal by primates in a biodiversity hotspot, the Atlantic Forest of South America, to 1) highlight gaps in our knowledge, 2) determine species richness and proportion of seed species dispersed, and 3) test the relationship between primate body size and the size of dispersed species. Our review found 79 studies of the diet of six ecospecies (Callithrix, Leontopithecus, Callicebus, Sapajus, Alouatta, Brachyteles) but only 20 of these report information on seed dispersal, and none of these are on Callithrix or Callicebus. We found a strong bias in the distribution of species and regions, with most of the studies concentrated in southeastern Brazil. All ecospecies dispersed a large proportion of the seed species they handled (72.1–93.6%). Brachyteles dispersed the highest diversity of plants (N = 73), followed by Sapajus (N = 66), Leontopithecus (N = 49), and Alouatta (N = 26). Although we found no significant relationship between primate body size and the size of seeds dispersed, Brachyteles disperse a higher diversity of large-seeded species than smaller-bodied primates. These results suggest that the local extinction of large primate species may lead to dramatic changes in the plant community, as many large-seeded plants are inaccessible to smaller arboreal frugivores. We propose guidelines for future research on primate seed dispersal to enable the evaluation of seed dispersal effectiveness and to improve our understanding of the fundamental role of primates in this key ecosystem process.     

Temporal variations in seed dispersal patterns of a bird-dispersed tree, <Emphasis Type="Italic">Swida controversa</Emphasis> (Cornaceae), in a temperate forest

The seed dispersal patterns of bird-dispersed trees often show substantial seasonal and annual variation due to temporal changes in frugivorous bird and bird-dispersed fruit distributions. Elucidating such variation and how it affects plant regeneration is important for understanding the evolution and seed dispersal maintenance strategies of these plants. In this study, we investigated the seed dispersal quantity and distance of a bird-dispersed plant, Swida controversa, for 2 years and detected large seasonal variations in dispersal pattern. Early in the fruiting season, short seed dispersal distance and large amounts of fruit consumption by birds (seed dispersal quantity) were observed. In contrast, late in the fruiting season, a long seed dispersal distance and small seed dispersal quantity were observed. This relationship between seed dispersal distance and quantity may help to maintain constant seed dispersal effectiveness during the long S. controversa fruiting season. Annual variation was also detected for both seed dispersal quantity and distance. More effective seed dispersal was achieved in the masting year, because both seed dispersal quantity and distance were greater than that in the non-masting year. These seed dispersal dynamics may contribute to the evolution and maintenance of S. controversa masting behavior. Thus, we identified substantial temporal variation on both seasonal and annual scales in the seed dispersal pattern of a bird-dispersed plant. The temporal variation in seed dispersal pattern revealed in this study probably plays a substantial role in the life history and population dynamics of S. controversa.     

Two New Species and New Occurrences of <Emphasis Type="Italic">Syneches</Emphasis> Walker for Brazilian Biome of Caatinga (Diptera: Hybotidae: Hybotinae)

Syneches from Brazilian biome of Caatinga were studied, two new species are described, Syneches atratus sp. nov. and Syneches limeirai sp. nov., and three species, Syneches annulipes Bezzi, 1909, Syneches moraballi Smith, 1963, and Syneches rafaeli Ale-Rocha & Vieira, 2008, are recorded for the biome. An identification key for the species of Syneches from Caatinga biome is provided.     

Phylogeographic structure of <Emphasis Type="Italic">Spondias tuberosa</Emphasis> Arruda Câmara (Anacardiaceae): seasonally dry tropical forest as a large and continuous refuge

Spondias tuberosa occurs in the Caatinga domain (seasonally dry tropical forest biome) of north-eastern Brazil, a large biome with ecogeographic regions that may have modelled the population structure of the species. Here we studied the phylogeographic pattern of S. tuberosa using sequences of the accD-psaI plastid region and six SSR markers in individuals distributed across 20 localities. The results for accD-psaI demonstrated nine haplotypes: some of which were exclusive to Caatinga ecoregions, whereas others were found in all localities. Spatial analysis of molecular variance revealed two groups (F_ct?=?0.34, P?<?0.0039) with 33.91% variation between them. The SSR analyses displayed 2–5 alleles at each locus, some of which were unique to certain localities. As in the accD-psaI region, the population structure obtained using SSR markers fell into two groups: (1) a large group containing the majority of the geographic region of Caatinga and (2) a small group near the Atlantic forest. We demonstrate the population structure of S. tuberosa, identifying the Caatinga as large, continuous refuge and the region near the interface between the Caatinga and the Atlantic forest as second refuge.     

Role of indehiscent pericarp in formation of soil seed bank in five cold desert Brassicaceae species

The dispersal and germination unit of some Brassicaceae species is the fruit, and we hypothesized that it could affect germination phenology and promote formation of a soil seed bank. We determined the effects of the indehiscent pericarp on germination and longevity of buried seeds of five Brassicaceae species native to cold deserts of central Asia. Germination phenology (seedling emergence) was monitored for intact dispersal units and isolated seeds of Chorispora sibirica, Goldbachia laevigata, Spirorrhynchus sabulosus, Tauscheria lasiocarpa (annuals), and Sterigmostemum fuhaiense (perennial) at natural temperatures in watered and non-watered (natural precipitation) soil. Intact dispersal units and isolated seeds were buried under natural conditions and exhumed at regular intervals for 35 months to monitor germination, viability and moisture content of isolated seeds, seeds in dispersal units, and seeds removed from dispersal units after burial. Isolated seeds of Goldbachia, Spirorrhynchus, and Tauscheria germinated only the first autumn and those of Chorispora and Sterigmostemum the first autumn and first spring, with higher germination percentages in all species in watered than in non-watered soil. A high percentage of seeds in buried dispersal units of Chorispora, Goldbachia, and Sterigmostemum was viable after 35 months, and seeds exhibited a 6-month dormancy cycle, being non-dormant only in autumn and spring. Seeds in buried dispersal units of Spirorrhynchus and Tauscheria germinated when exhumed in the first spring, but all non-germinated seeds were dead after 1 year. Thus, the presence of the pericarp allows Chorispora, Goldbachia, and Sterigmostemum to form a persistent seed bank but not Spirorrhynchus and Tauscheria.     

Seed dispersal distances by ant partners reflect preferential recruitment patterns in two ant-dispersed sedges

An important feature of seed dispersal mutualism is the differentiation of dispersal-related seed traits (dispersal syndrome), which potentially contribute to partitioning of both seed dispersers and regeneration sites among sympatric plants. Yet, the selective factors underlying the diversity in dispersal syndromes are largely unknown. The differential requirements for seed dispersal distances are often proposed as a main factor in plant adaptations to disperser animals. Focusing on two sympatric ant-dispersed sedges Carex lanceolata and Carex tristachya (Cyperaceae), we tested the association of the adaptation to different dispersers with requirements for seed dispersal distances. We found that C. lanceolata was more frequently dispersed by the large ant Formica japonica (which had relatively long dispersal distances compared with other smaller ants) than by C. tristachya, and this was caused by the higher seed attractiveness of C. lanceolata to F. japonica. Pot experiments manipulating adult-to-seedling distances showed that isolation from conspecific adults only benefited C. lanceolata seedlings, and C. tristachya seedlings were not affected. These results support the importance of differential requirements for seed dispersal distances as a factor underlying the diversity in dispersal syndromes among animal-dispersed plants.     

Effects of <Emphasis Type="Italic">Prangos ferulacea</Emphasis> aqueous and hydroalcoholic extracts obtained from different organs on the regeneration of <Emphasis Type="Italic">Trifolium resupinatum</Emphasis>

Prangos ferulacea is one of the widely used, nutritional and popular fodders in livestock industry. This species is also considered as an important option in rangeland restoration and management. In this study, the comparative phytotoxic activity of aqueous and hydroalcoholic extracts obtained from different organs (flower, shoot and leaf) of P. ferulacea on proline content, seed germination and seedling growth of Trifolium resupinatum has been investigated. According to the results, the hydroalcoholic extract of P. ferulaceae flower possesses the highest total phenolic and flavonoid content and the uppermost phytotoxic effect on T. resupinatum. The extracts significantly decreased seed germination and seedling growth of T. resupinatum and increased the proline content. Our findings indicate that hydroalcoholic extract induced a stronger oxidative stress in T. resupinatum. Finally, based on the results, aqueous allelochemicals that originated from P. ferulacea played a significant role in the successful propagation and development of T. resupinatum in rehabilitated pastures. According to our results, the phytotoxicity effect of the hydroalcoholic extract was significantly higher than that of the aqueous extract. Since in nature, the allelopathic interaction between plants is closer to the aqueous method, primary evaluations of rangeland restoration using this method is suggested.     

Ants Associated with <Emphasis Type="Italic">Turnera subulata</Emphasis> (Turneraceae): Elaiosome Attraction,Seed Dispersion and Germination

Symbiosis between plants and ants include examples in which the plant provides shelter and/or food for ants that, in turn, act in the defense or in the dispersion of seeds from the host plant. Although traditionally referred as mutualistic, the results of these interactions may vary with the ecological context in which patterns are involved. A range of species have facultative association with Turnera subulata (Turneraceae). Here, using behavioral bioassays, we investigated the effects of the most frequent ant species associated with T. subulata (Brachymyrmex sp.1, Camponotus blandus (Smith), Dorymyrmex sp.1, Crematogaster obscurata Emery, and Solenopsis invicta Buren) in the dispersion of plant host seeds and in the number of seedlings around the associated ant nests. We also evaluated the effects of these ant species in the germination of T. subulata seeds, in the consumption of elaiosome, and in the attractiveness to elaiosome odor. Our results showed that the ant species associated with T. subulata presented variation in the attraction by the odor and in the rate of consumption of the elaiosomes. However, none of the ant species studied contributed significantly to the increase of seed germination and seedling growth. Our results suggest that the consumption of the elaiosome by ant species is not a determinant factor to the success of germination of T. subulata. However, such species could contribute indirectly to seed germination by carrying seeds to sites more fertile to germination. In general, our results help to elucidate the results of ecological interactions involving ants and plants.     

Landscape change alters reproductive phenology and sex expression in <Emphasis Type="Italic">Attalea</Emphasis> palms (Arecaceae) of southwestern Amazonia

As deforestation and land-use/land-cover change advance in tropical forest regions, an understanding of how plants adjust phenology and reproductive dynamics to altered landscapes can provide insights into plasticity, productivity, and population persistence. We compared the reproductive phenology, sex expression, and flower and fruit production of two monoecious Amazonian palms, Attalea phalerata and Attalea speciosa, in old-growth forest and as remnant trees growing in actively grazed pastures. Using 2 years of phenology data collected from natural populations near Vila Extrema, Rondônia, and eastern Acre, Brazil, we compared flowering and fruiting in the two habitats and tested for effects of palm height, crown size, and light availability on inflorescence and sex expression. Forest conversion to pasture stimulated greater overall flowering and fruiting in individual Attalea palms. As a population, remnant pasture palms continuously bore flowers and fruits year-round, while forest palms flowered seasonally in isolated peaks with consecutive months of inactivity. Crown size and greater light exposure affected flowering and fruiting dynamics in A. phalerata and A. speciosa, respectively, and increased light availability shifted A. speciosa sex expression towards greater female investment, primarily through regulation of sex determination and bud abortion. Removal of tropical forest does not always lead to the downfall of remnant tree populations, and under favorable conditions, such as abandonment of cropland and pasture, higher levels of reproduction can facilitate recovery of future generations. Tree species with flexible sex expression may be particularly resilient in the face of land-use and land-cover change.     

The effect of the dominant grass <Emphasis Type="Italic">Festuca rupicola</Emphasis> on the establishment of rare forbs in semi-dry grasslands

Establishment is the most sensitive process in the life-cycle of plant species, and each stage – from germination to survival and growth – can be affected by environmental factors and plant traits. We hypothesized that the dominant tussock grass Festuca rupicola negatively affects forb establishment in semi-dry grasslands where it has recently expanded into. Moreover, we expected that seedling recruitment is affected by grass density and seed size, with larger seeded forbs being more successful in areas of higher grass density. In a garden experiment, we sowed seven forb individuals of differing seed size (smallest to largest: Veronica spicata < Dianthus carthusianorum < Plantago media < Biscutella laevigata < Filipendula vulgaris < Scabiosa ochroleuca < Sanguisorba minor) into pots with zero, one, two and three grass individuals, and assessed germination, survival and growth over one year. As expected, increasing grass density negatively affected germination, survival and growth of forbs; however, contrary to our expectation, seed size did not influence any of parameters measured. The response of each individual species varied from zero or weak to strong with respect to grass density. Festuca rupicola therefore acts as a strong competitor against the establishment of forbs, irrespective of their seed size, and its spread lowers species diversity in semi-dry grasslands.     

Effect of cold stratification on seed germination in <Emphasis Type="Italic">Solidago</Emphasis> × <Emphasis Type="Italic">niederederi</Emphasis> (<Emphasis Type="Italic">Asteraceae</Emphasis>) and its parental species

In this study, we investigated the influence of cold stratification on seed germination in S. × niederederi, a hybrid between the North American S. canadensis and the European S. virgaurea, using fruit samples collected in 2016 in Poland. We aimed to test the hypothesis that the low temperature exposure decreases the final percentage and speed of seed germination in the hybrid and its parental species. For each species, sets of 100 achenes in three replications were mixed with dry sand and stored in Petri dishes in darkness for 12 weeks, at ?18 °C and?+?4 °C, and?+?25 °C. The seeds were incubated for 21 d at room temperature (+25 °C), under the 12 h photoperiod (630 lx). We showed a lack of significant differences in: (i) the final percentage of germinated seeds of studied species stored at the same conditions, (ii) the final percentage of germinated seeds between the applied stratification conditions in the hybrid and its parental species, and (iii) the mean values of Timson’s index, mean germination time, and coefficient of velocity of germination between the stratification conditions in each species. The statistically significant inter-specific differences in the mean germination time parameter after the +25 °C treatment suggest that the seeds of S. × niederederi are able to germinate faster than the seeds of its parental species. However, to improve our knowledge of naturalization and invasion abilities of S. × niederederi by sexual reproduction, the seed germination and seedling survival of the hybrid should be tested in the field.     

Variation in mycorrhizal performance in the epiphytic orchid <Emphasis Type="Italic">Tolumnia variegata in vitro</Emphasis>: the potential for natural selection

Symbiotic seed germination is a critical stage in orchid life histories. Natural selection may act to favor plants that efficiently use mycorrhizal fungi. However, the necessary conditions for natural selection – variation, heritability, and differences in fitness – have not been demonstrated for either orchid or fungus. With the epiphytic orchid Tolumnia variegata as a model system, we ask the following questions: (1) Do seeds from different individuals in a population differ in germination and seedling development in the presence of the same fungi? (2) Do different mycorrhizal fungi (Ceratobasidium spp.) differ in ability to stimulate seed germination and growth in T. variegata? And (3) are the Ceratobasidium isolates that best induce seed germination and seedling development more closely related to each other than to isolates that are less effective? We performed symbiotic seed germination experiments in vitro. The experiments were done using mycorrhizal fungi isolated from T. variegata; relationships among the fungi were inferred from nuclear ribosomal ITS sequences. We found significant variation for both symbiotic germination and seedling growth among biparental seed crops obtained from a population of T. variegata plants. Differences among Ceratobasidium fungi in seed germination were significant. The fungi that induced highest seed germination and seedling development belonged to two of four clades of Ceratobasidium. The two experiments show that there is potential for natural selection to act on orchid–fungus relationships. Given that orchids vary in performance, and that mycorrhizal fungi are not geographically distributed homogeneously, mycorrhizae may affect population size, distribution and evolution of orchids.