Mosaic fungal individuals have the potential to evolve within a single generation.

Although cells of mushroom-producing fungi typically contain paired haploid nuclei (n + n), most Armillaria gallica vegetative cells are uninucleate. As vegetative nuclei are produced by fusions of paired haploid nuclei, they are thought to be diploid (2n). Here we report finding haploid vegetative nuclei in A. gallica at multiple sites in southeastern Massachusetts, USA. Sequencing multiple clones of a single-copy gene isolated from single hyphal filaments revealed nuclear heterogeneity both among and within hyphae. Cytoplasmic bridges connected hyphae in field-collected and cultured samples, and we propose nuclear migration through bridges maintains this nuclear heterogeneity. Growth studies demonstrate among- and within-hypha phenotypic variation for growth in response to gallic acid, a plant-produced antifungal compound. The existence of both genetic and phenotypic variation within vegetative hyphae suggests that fungal individuals have the potential to evolve within a single generation in response to environmental variation over time and space.

Time matching between grooming partners: Do methodological distinctions between short versus long-term reciprocation matter?

Primatologists have long focused on grooming exchanges to examine aspects of social relationships, co-operation, and social cognition. One particular interest is the extent to which reciprocating grooming partners time match, and the time frame over which they do so. Conclusions about time matching vary across species. Generally, researchers focus on the duration of pauses between grooming episodes that involve a switch in partner roles and choose a cut-off point to distinguish short from longer-term reciprocation. Problematically, researchers have made inconsistent choices about cut-offs. Such methodological variations are potentially concerning, as it is unclear whether inconsistent conclusions about short-term time matching are attributable to species/ecological differences, or are due in part to methodological inconsistency. We ask whether various criteria for separating short versus long-term reciprocation influence conclusions about short-term time matching using data from free-ranging rhesus ( Macaca mulatta) and captive-crested macaques ( Macaca nigra). We compare several commonly used cut-offs to ones generated by the currently preferred approach-survival analysis. Crested macaques displayed a mild degree of time matching regardless of the cutoff used. For rhesus macaques, whereas most cut-offs yielded similar degrees of time matching as the one derived from survival analysis, very short ones significantly underestimated both the degree of time matching and the influence of rank distance on time matching. Although researchers may have some flexibility in their choice of cut-offs, we suggest that they employ caution by using survival analysis when possible, and when not possible, by avoiding very short time windows.

Vocal and anatomical evidence for two-voiced sound production in the greater sage-grouse Centrocercus urophasianus.

Greater sage-grouse, Centrocercus urophasianus, have been a model system in studies of sexual selection and lek evolution. Mate choice in this species depends on acoustic displays during courtship, yet we know little about how males produce these sounds. Here we present evidence for previously undescribed two-voiced sound production in the sage-grouse. We detected this ;double whistle' (DW) using multi-channel audio recordings combined with video recordings of male behavior. Of 28 males examined, all males produced at least one DW during observation; variation in DW production did not correlate with observed male mating success. We examined recordings from six additional populations throughout the species' range and found evidence of DW in all six populations, suggesting that the DW is widespread. To examine the possible mechanism of DW production, we dissected two male and female sage-grouse; the syrinx in both sexes differed noticeably from that of the domestic fowl, and notably had two sound sources where the bronchi join the syrinx. Additionally, we found males possess a region of pliable rings at the base of the trachea, as well as a prominent syringeal muscle that is much reduced or absent in females. Experiments with a live phonating bird will be necessary to determine how the syrinx functions to produce the whistle, and whether the DW might be the result of biphonation of a single sound source. We conclude that undiscovered morphological and behavioral complexity may exist even within well-studied species, and that integrative research approaches may aid in the understanding of this type of complexity.

Haploid vegetative mycelia of Armillaria gallica show among-cell-line variation for growth and phenotypic plasticity.

Vegetative mycelial cells of Armillaria are expected to have diploid nuclei. Cells from a single mycelium therefore would not be expected to differ from one another for ecologically relevant quantitative traits. We isolated two sets of basidiome cell lines (from spores and stipe cells) and one set of vegetative cell lines (from an attached rhizomorph) from a single contiguous Armillaria gallica mycelium. We isolated a second set of vegetative cell lines from the soil 20 cm from the above basidiome-rhizomorph complex. In all four sets of cell lines in situ DAPI-DNA measurements showed cells are haploid and quantitative-trait analyses of cell lines grown at different water potentials revealed high levels of among-cell-line genetic variation for both growth and phenotypic plasticity. Haploidy and the existence of ecologically relevant genetic variation within vegetative individuals are unexpected and mean that a process similar to evolutionary adaptation could take place within the soma of a genetic individual. We believe this is a key to understanding how large A. gallica mycelia survive exposure to variation in ecological conditions during lives that potentially span several tree (host) generations.

Phenotypic plasticity and evolutionary potential in somatic cells of Armillaria gallica.

Somatic cells of Armillaria gallica fruit bodies have been shown to possess different genotypes for molecular-marker and mating-type loci. Here we report experiments on six quantitative traits and demonstrate that somatic cells of fruit bodies possess almost as much genetic variation for growth rate and phenotypic plasticity as do spores, the products of meiosis. Genetically distinct somatic cells therefore have the potential to grow at different rates relative to one another during primordial fruit body formation. This may confer an advantage on all cell lines within a fruit body, not just those that happen to grow better under a particular set of conditions. To our knowledge, genetic variation for fitness-related traits that make up a single genetic individual has not been reported before.