Richard P. Shefferson

An empirical test of partner choice mechanisms in a wild legume–rhizobium interaction

Mutualisms can be viewed as biological markets in which partners of different species exchange goods and services to their mutual benefit. Trade between partners with conflicting interests requires mechanisms to prevent exploitation. Partner choice theory proposes that individuals might foil exploiters by preferentially directing benefits to cooperative partners. Here, we test this theory in a wild legume–rhizobium symbiosis. Rhizobial bacteria inhabit legume root nodules and convert atmospheric dinitrogen (N2) to a plant available form in exchange for photosynthates. Biological market theory suits this interaction because individual plants exchange resources with multiple rhizobia. Several authors have argued that microbial cooperation could be maintained if plants preferentially allocated resources to nodules harbouring cooperative rhizobial strains. It is well known that crop legumes nodulate non-fixing rhizobia, but allocate few resources to those nodules. However, this hypothesis has not been tested in wild legumes which encounter partners exhibiting natural, continuous variation in symbiotic benefit. Our greenhouse experiment with a wild legume, Lupinus arboreus, showed that although plants frequently hosted less cooperative strains, the nodules occupied by these strains were smaller. Our survey of wild-grown plants showed that larger nodules house more Bradyrhizobia, indicating that plants may prevent the spread of exploitation by favouring better cooperators.

High specificity generally characterizes mycorrhizal association in rare lady's slipper orchids, genus Cypripedium

Ladys slipper orchids (Cypripedium spp.) are rare terrestrial plants that grow throughout the temperate Northern Hemisphere. Like all orchids, they require mycorrhizal fungi for germination and seedling nutrition. The nutritional relationships of adult Cypripedium mycorrhizae are unclear; however, Cypripedium distribution may be limited by mycorrhizal specificity, whether this specificity occurs only during the seedling stage or carries on into adulthood. We attempted to identify the primary mycorrhizal symbionts for 100 Cypripedium plants, and successfully did so with two Cypripedium calceolus, 10 Cypripedium californicum, six Cypripedium candidum, 16 Cypripedium fasciculatum, two Cypripedium guttatum, 12 Cypripedium montanum, and 11 Cypripedium parviflorum plants from a total of 44 populations in Europe and North America, yielding fungal nuclear large subunit and mitochondrial large subunit sequence and RFLP (restriction fragment length polymorphism) data for 59 plants. Because orchid mycorrhizal fungi are typically observed without fruiting structures, we assessed fungal identity through direct PCR (polymerase chain reaction) amplification of fungal genes from mycorrhizally colonized root tissue. Phylogenetic analysis revealed that the great majority of Cypripedium mycorrhizal fungi are members of narrow clades within the fungal family Tulasnellaceae. Rarely occurring root endophytes include members of the Sebacinaceae, Ceratobasidiaceae, and the ascomycetous genus, Phialophora. C. californicum was the only orchid species with apparently low specificity, as it associated with tulasnelloid, ceratobasidioid, and sebacinoid fungi in roughly equal proportion. Our results add support to the growing literature showing that high specificity is not limited to nonphotosynthetic plants, but also occurs in photosynthetic ones.

THE EVOLUTIONARY HISTORY OF MYCORRHIZAL SPECIFICITY AMONG LADY'S SLIPPER ORCHIDS

Abstract Although coevolution is acknowledged to occur in nature, coevolutionary patterns in symbioses not involving species-to-species relationships are poorly understood. Mycorrhizal plants are thought to be too generalist to coevolve with their symbiotic fungi; yet some plants, including some orchids, exhibit strikingly narrow mycorrhizal specificity. Here, we assess the evolutionary history of mycorrhizal specificity in the ladys slipper orchid genus, Cypripedium. We sampled 90 populations of 15 taxa across three continents, using DNA methods to identify fungal symbionts and quantify mycorrhizal specificity. We assessed phylogenetic relationships among sampled Cypripedium taxa, onto which we mapped mycorrhizal specificity. Cypripedium taxa associated almost exclusively with fungi within family Tulasnellaceae. Ancestral specificity appears to have been narrow, followed by a broadening after the divergence of C. debile. Specificity then narrowed, resulting in strikingly narrow specificity in most of the taxa in this study, with no taxon rewidening to the same extant as basal members of the genus. Sympatric taxa generally associated with different sets of fungi, and most clades of Cypripedium-mycorrhizal fungi were found throughout much of the northern hemisphere, suggesting that these evolutionary patterns in specificity are not the result of biogeographic lack of opportunity to associate with potential partners. Mycorrhizal specificity in genus Cypripedium appears to be an evolvable trait, and associations with particular fungi are phylogenetically conserved.

ESTIMATING DORMANCY AND SURVIVAL OF A RARE HERBACEOUS PERENNIAL USING MARK-RECAPTURE MODELS

Mark-recapture statistics have rarely been applied to plants, yet they can be useful for estimating plant demographic traits where individuals may be missed or unob- servable. We applied mark-recapture statistics based on an information theory approach to estimate annual probabilities of dormancy in a five-year study of a threatened, perennial plant, the small yellow ladys slipper orchid ( Cypripedium calceolusssp. parviflorum (Sal- isb.) Fernald). The aboveground states of 548 genets in eight patches were monitored over five years. Apparent survival probabilities (f), corrected for dormancy, were calculated. The best-fit model suggested that apparent survival was constant throughout the study for all patches, while dormancy varied additively with time among patches. The mean prob- ability of dormancy was 0.320 6 0.024, with a mean maximum overestimation of 0.067 as calculated using an estimate of the probability of detection. Dormancy typically lasted for no longer than two consecutive years, although dormancy as long as four years was observed. Dormancy displayed a strong covariate relationship with spring frost days, al- though effects of precipitation and mean spring temperature were almost equally strong. Mean apparent survival probabilities were high in each patch ( f5 0.878), but dormancy probabilities varied considerably among patches (d 5 0.188-0.672). Conventional re- sprouting probabilities underestimated apparent survival by a mean difference of 0.288 (range: 0.150-0.589). This novel application of mark-recapture statistics to plant demog- raphy allowed robust survival estimates that accounted for uncertainty due to an unob- servable, dormant life stage.

LIFE HISTORY TRADE‐OFFS IN A RARE ORCHID: THE COSTS OF FLOWERING, DORMANCY, AND SPROUTING

We tested for life history trade-offs among dormancy, sprouting, and flowering in a seven-year study of a threatened, perennial plant, the small yellow ladys slipper orchid (Cypripedium calceolus ssp. parviflorum (Salisb.) Fernald). The aboveground states of 629 genets were monitored over seven years in a wet meadow in northeastern Illinois, USA. With mark–recapture statistics, survival, resighting, and stage transitions were calculated among three stage classes of individuals: dormant, vegetative, and flowering. The best-fit and most parsimonious models suggested that (1) survival was constant among years, but varied by stage; (2) dormant individuals suffered significantly higher mortality and were more likely to become dormant in future years than sprouting or flowering individuals; (3) flowering individuals had significantly higher survival and were more likely to flower in the future than sprouting and dormant individuals; and (4) sprouting individuals had a significantly higher stage transition to d...

Mycorrhizal interactions of orchids colonizing Estonian mine tailings hills

Northeastern Estonia is home to extensive oil shale mines. Associated with these are desolate and environmentally damaging hills of ash and semicoke tailings. Interestingly, some of the first plants to colonize these hills are rare orchids. Here, we assess the identities of the mycorrhizal fungi associated with these orchids, in particular Epipactis atrorubens, Orchis militaris, and Dactylorhiza baltica, and compare them with mycorrhizal fungi from orchids from pristine habitat. Epipactis atrorubens associated with the widest breadth of fungi, including unnamed members of the basidiomycete family Tulasnellaceae and the potentially ectomycorrhizal ascomycetes Trichophaea woolhopeia and Geopora cooperi. Orchis militaris also associated with unnamed members of the Tulasnellaceae. Dactylorhiza baltica associated with Ceratobasidium albasitensis. In Epipactis and Orchis, the same fungi associated with plants in the pristine habitat as with those on ash hills. The tulasnelloid and ceratobasidioid fungi mycorrhizal with these orchids appear closely related to common orchid mycorrhizal fungi, while one of the ascomycetes mycorrhizal with E. atrorubens is closely related to a mycorrhizal fungus with E. microphylla. Our results suggest that these orchids and their fungi are not limited to pristine habitats and that environmentally polluted sites may present novel habitats that may be exploited for endangered plant conservation.

ADULT WHOLE-PLANT DORMANCY INDUCED BY STRESS IN LONG-LIVED ORCHIDS

Dormancy is a condition in which an herbaceous perennial does not sprout for one or more growing seasons. To test whether dormancy is an adaptive response to environmental stress, we defoliated and shaded individuals of two rare geophytic orchids, Cypripedium calceolus and Cephalanthera longifolia, in five Estonian populations early in the growing season in 2002 and 2003. We also censused plants at the same time, and conducted one more census in 2004. Mark–recapture models were used to estimate the probabilities of dormancy (d, the complement to resighting, p), and apparent survival (ϕ). Apparent survival varied little by treatment, with Cypripedium and Cephalanthera surviving at 0.986 ± 0.014 and 0.974 ± 0.021 (mean ± se), respectively. In contrast, treatment impacted dormancy dramatically. For both Cephalanthera and Cypripedium, defoliated (def.) plants were most dormant (0.320 ± 0.055 and 0.095 ± 0.036, respectively). However, while both control (cont.) and shaded (sh.) plants were roughly equally leas...

Evolution of host breadth in broad interactions: mycorrhizal specificity in East Asian and North American rattlesnake plantains (Goodyera spp.) and their fungal hosts

Host breadth is often assumed to have no evolutionary significance in broad interactions because of the lack of cophylogenetic patterns between interacting species. Nonetheless, the breadth and suite of hosts utilized by one species may have adaptive value, particularly if it underlies a common ecological niche among hosts. Here, we present a preliminary assessment of the evolution of mycorrhizal specificity in 12 closely related orchid species (genera Goodyera and Hetaeria) using DNA‐based methods. We mapped specificity onto a plant phylogeny that we estimated to infer the evolutionary history of the mycorrhiza from the plant perspective, and hypothesized that phylogeny would explain a significant portion of the variance in specificity of plants on their host fungi. Sampled plants overwhelmingly associated with genus Ceratobasidium, but also occasionally with some ascomycetes. Ancestral mycorrhizal specificity was narrow in the orchids, and broadened rarely as Goodyera speciated. Statistical tests of phylogenetic inertia suggested some support for specificity varying with increasing phylogenetic distance, though only when the phylogenetic distance between suites of fungi interacting with each plant taxon were taken into account. These patterns suggest a role for phylogenetic conservatism in maintaining suits of fungal hosts among plants. We stress the evolutionary importance of host breadth in these organisms, and suggest that even generalists are likely to be constrained evolutionarily to maintaining associations with their symbionts.

Impact of herbivory on flowering behaviour and life history trade-offs in a polycarpic herb: a 10-year experiment

Herbivores can have strong deleterious effects on plant growth, reproduction, and even survival. Because these effects might be strongly interrelated, the direct consumptive effects of herbivores and a variety of indirect effects are difficult to untangle. Reductions in growth, for example, may strongly impact the flowering behaviour of plant species in the current season, but at the same time incur costs to survival, growth and reproduction in the next growing season(s). To get better insights in the effects of herbivory on the flowering behaviour of the long-lived polycarpic grassland herb Primula veris L., flowering patterns were monitored over ten consecutive years under two treatments (grazing and control mowing regimes). We tested the hypothesis that the size at flowering was affected by the presence of herbivores, and whether this translated into costs to future reproduction and survival. Overall, grazed plants were significantly smaller than control plants, and the size at which plants flowered was also significantly smaller when herbivores were present. The transition probability of flowering and of surviving into the next year was significantly smaller for all plants in the current year if they had been grazed than if they had been mown, indicating that herbivory incurred costs to both flowering and survival. Grazed plants also needed longer to start flowering, had fewer flowers and flowered less frequently, causing a significantly lower proportion of flowering adults in the population. These results suggest that the observed regression in plant size due to herbivory does not allow plants to capture enough resources to guarantee regular flowering in the longer run.

Plants do not count… or do they? New perspectives on the universality of senescence

1. Senescence, the physiological decline that results in decreasing survival and/or reproduction with age, remains one of the most perplexing topics in biology. Most theories explaining the evolution of senescence (i.e. antagonistic pleiotropy, accumulation of mutations, disposable soma) were developed decades ago. Even though these theories have implicitly focused on unitary animals, they have also been used as the foundation from which the universality of senescence across the tree of life is assumed. 2. Surprisingly, little is known about the general patterns, causes and consequences of whole-individual senescence in the plant kingdom. There are important differences between plants and most animals, including modular architecture, the absence of early determination of cell lines between the soma and gametes, and cellular division that does not always shorten telomere length. These characteristics violate the basic assumptions of the classical theories of senescence and therefore call the generality of senescence theories into question. 3. This Special Feature contributes to the field of whole-individual plant senescence with five research articles addressing topics ranging from physiology to demographic modelling and comparative analyses. These articles critically examine the basic assumptions of senescence theories such as age-specific gene action, the evolution of senescence regardless of the organisms architecture and environmental filtering, and the role of abiotic agents on mortality trajectories. 4. Synthesis. Understanding the conditions under which senescence has evolved is of general importance across biology, ecology, evolution, conservation biology, medicine, gerontology, law and social sciences. The question ‘why is senescence universal or why is it not?’ naturally calls for an evolutionary perspective. Senescence is a puzzling phenomenon, and new insights will be gained by uniting methods, theories and observations from formal demography, animal demography and plant population ecology. Plants are more amenable than animals to experiments investigating senescence, and there is a wealth of published plant demographic data that enable interpretation of experimental results in the context of their full life cycles. It is time to make plants count in the field of senescence.