Michael R. Whitehead

A narrow group of monophyletic Tulasnella (Tulasnellaceae) symbiont lineages are associated with multiple species of Chiloglottis (Orchidaceae): Implications for orchid diversity.

UNLABELLED PREMISE OF THE STUDY The Orchidaceae is characterized by exceptional species diversity. Obligate orchid mycorrhizae are predicted to determine orchid distributions, and highly specific relationships between orchids and fungi may drive orchid diversification. In this study, mycorrhizal diversity was examined in the terrestrial, photosynthetic orchid genus Chiloglottis to test the hypothesis of mycorrhizal-mediated diversification in the genus Chiloglottis. This orchid genus secures pollination by sexual deception, an obligate and highly specific pollination strategy. Here we asked whether the obligate orchid-fungal interactions are also specific. • METHODS Two sequenced loci, the internal transcribed spacer region (ITS) and mitochondrial large subunit (mtLSU), were used to identify fungal isolates and assess fungal species diversity. Symbiotic germination of two species Chiloglottis aff. jeanesii and C. valida were used to assess germination potential of isolates and confirm mycorrhizal association. • KEY RESULTS Phylogenetic analyses revealed that six representative Chiloglottis species spanning a broad survey of the genus were all associated with a narrow group of monophyletic Tulasnella fungal lineages. • CONCLUSIONS The Chiloglottis-Tulasnella interaction appears to be the first known case of such a narrow symbiont association across a broadly surveyed orchid genus. It appears that the specific pollination system of Chiloglottis, rather than specific orchid-fungal interactions has been the key driving force in the diversification of the genus. These findings also indicate that plant groups with highly specific mycorrhizal partners can have a widespread distribution.

New perspectives on the evolution of plant mating systems

BACKGROUND The remarkable diversity of mating patterns and sexual systems in flowering plants has fascinated evolutionary biologists for more than a century. Enduring questions about this topic include why sexual polymorphisms have evolved independently in over 100 plant families, and why proportions of self- and cross-fertilization often vary dramatically within and among populations. Important new insights concerning the evolutionary dynamics of plant mating systems have built upon a strong foundation of theoretical models and innovative field and laboratory experiments. However, as the pace of advancement in this field has accelerated, it has become increasingly difficult for researchers to follow developments outside their primary area of research expertise. SCOPE In this Viewpoint paper we highlight three important themes that span and integrate different subdisciplines: the changes in morphology, phenology, and physiology that accompany the transition to selfing; the evolutionary consequences of pollen pool diversity in flowering plants; and the evolutionary dynamics of sexual polymorphisms. We also highlight recent developments in molecular techniques that will facilitate more efficient and cost-effective study of mating patterns in large natural populations, research on the dynamics of pollen transport, and investigations on the genetic basis of sexual polymorphisms. This Viewpoint also serves as the introduction to a Special Issue on the Evolution of Plant Mating Systems. The 15 papers in this special issue provide inspiring examples of recent discoveries, and glimpses of exciting developments yet to come.

Floral odour chemistry defines species boundaries and underpins strong reproductive isolation in sexually deceptive orchids

BACKGROUND AND AIMS The events leading to speciation are best investigated in systems where speciation is ongoing or incomplete, such as incipient species. By examining reproductive barriers among incipient sister taxa and their congeners we can gain valuable insights into the relative timing and importance of the various barriers involved in the speciation process. The aim of this study was to identify the reproductive barriers among sexually deceptive orchid taxa in the genus Chiloglottis. METHODS The study targeted four closely related taxa with varying degrees of geographic overlap. Chemical, morphological and genetic evidence was combined to explore the basis of reproductive isolation. Of primary interest was the degree of genetic differentiation among taxa at both nuclear and chloroplast DNA markers. To objectively test whether or not species boundaries are defined by the chemistry that controls pollinator specificity, genetic analysis was restricted to samples of known odour chemistry. KEY RESULTS Floral odour chemical analysis was performed for 600+ flowers. The three sympatric taxa were defined by their specific chiloglottones, the semiochemicals responsible for pollinator attraction, and were found to be fully cross-compatible. Multivariate morphometric analysis could not reliably distinguish among the four taxa. Although varying from very low to moderate, significant levels of genetic differentiation were detected among all pairwise combinations of taxa at both nuclear and chloroplast loci. However, the levels of genetic differentiation were lower than expected for mature species. Critically, a lack of chloroplast DNA haplotype sharing among the morphologically indistinguishable and most closely related taxon pair confirmed that chemistry alone can define taxon boundaries. CONCLUSIONS The results confirmed that pollinator isolation, mediated by specific pollinator attraction, underpins strong reproductive isolation in these taxa. A combination of large effective population sizes, initial neutral mutations in the genes controlling floral scent, and a pool of available pollinators likely drives diversity in this system.

POLLINATOR SPECIFICITY DRIVES STRONG PREPOLLINATION REPRODUCTIVE ISOLATION IN SYMPATRIC SEXUALLY DECEPTIVE ORCHIDS

Few studies have quantified the full range of pre‐ and postzygotic barriers that limit introgression between closely related plant species. Here, we assess the strength of four isolating mechanisms operating between two morphologically similar and very closely related sympatric orchid taxa, Chiloglottis valida and C. aff. jeanesii. Each taxon sexually attracts its specific wasp pollinator via distinct floral volatile chemistry. Behavioral experiments with flowers and synthetic versions of their floral volatiles confirmed that very strong pollinator isolation is mediated by floral odor chemistry. However, artificially placing flowers of the two taxa in contact proximity revealed the potential for rare interspecific pollination. Although we found hybrid vigor in F1 hybrids produced by hand‐crossing, genetic analysis at both nuclear and chloroplast loci showed significant and moderate‐to‐strong genetic differentiation between taxa. A Bayesian clustering method for the detection of introgression at nuclear loci failed to find any evidence for hybridization across 571 unique genotypes at one site of sympatry. Rather than inhibiting gene flow, postpollination barriers surveyed here show no contribution to overall reproductive isolation. This demonstrates the primacy of pollinators in maintaining species boundaries in these orchids, which display one of the strongest known examples of prepollination floral isolation.

Testing single-sample estimators of effective population size in genetically structured populations

The effective population size (Ne) is a key parameter in evolutionary and population genetics. Single-sample Ne estimation provides an alternative to traditional approaches requiring two or more samples. Single-sample methods assume that the study population has no genetic sub-structure, which is unlikely to be true in wild populations. Here we empirically investigated two single-sample estimators (onesamp and LdNe) in replicated and controlled genetically structured populations of Drosophila melanogaster. Using experimentally controlled population parameters, we calculated the Wright–Fisher expected Ne for the structured population (TotalNe) and demonstrated that the loss of heterozygosity did not significantly differ from Wright’s model. We found that disregarding the population substructure resulted in TotalNe estimates with a low coefficient of variation but these estimates were systematically lower than the expected values, whereas hierarchical estimates accounting for population structure were closer to the expected values but had a higher coefficient of variation. Analysis of simulated populations demonstrated that incomplete sampling, initial allelic diversity and balancing selection may have contributed to deviations from the Wright–Fisher model. Overall the approximate-Bayesian onesamp method performed better than LdNe (with appropriate priors). Both methods performed best when dispersal rates were high and the population structure was approaching panmixia.

An informational diversity framework, illustrated with sexually deceptive orchids in early stages of speciation

Reconstructing evolutionary history for emerging species complexes is notoriously difficult, with newly isolated taxa often morphologically cryptic and the signature of reproductive isolation often restricted to a few genes. Evidence from multiple loci and genomes is highly desirable, but multiple inputs require ‘common currency’ translation. Here we deploy a Shannon information framework, converting into diversity analogue, which provides a common currency analysis for maternally inherited haploid and bi‐parentally inherited diploid nuclear markers, and then extend that analysis to construction of minimum‐spanning networks for both genomes. The new approach is illustrated with a quartet of cryptic congeners from the sexually deceptive Australian orchid genus Chiloglottis, still in the early stages of speciation. Divergence is more rapid for haploid plastids than for nuclear markers, consistent with the effective population size differential (Nep < Nen), but divergence patterns are broadly correlated for the two genomes. There are nevertheless intriguing discrepancies between the emerging plastid and nuclear signals of early phylogenetic radiation of these taxa, and neither pattern is entirely consistent with the available information on the sexual cues used by the orchids to lure the pollinators enforcing reproductive isolation. We describe possible extensions of this methodology to multiple ploidy levels and other types of markers, which should increase the range of application to any taxonomic assemblage in the very early stages of reproductive isolation and speciation.

A Balancing Act: Integrating Evidence‐Based Knowledge and Cultural Relevance in a Program of Prevention Parenting Research with Latino/a Immigrants

Family therapists have a unique opportunity to contribute toward the reduction of widespread mental health disparities impacting diverse populations by developing applied lines of research focused on cultural adaptation. For example, although evidence-based prevention parent training (PT) interventions have been found to be efficacious with various Euro-American populations, there is a pressing need to understand which specific components of PT interventions are perceived by ethnic minority parents as having the highest impact on their parenting practices. Equally important is to examine the perceived cultural relevance of adapted PT interventions. This qualitative investigation had the primary objective of comparing and contrasting the perceived relevance of two culturally adapted versions of the efficacious parenting intervention known as Parent Management Training, the Oregon Model (PMTO). According to feasibility indicators provided by 112 Latino/a immigrant parents, as well as findings from a qualitative thematic analysis, the core parenting components across both adapted interventions were identified by the majority of research participants as relevant to their parenting practices. Participants exposed to the culturally enhanced intervention, which included culture-specific sessions, also reported high satisfaction with components exclusively focused on cultural issues that directly impact their parenting practices (e.g., immigration challenges, biculturalism). This investigation illustrates the relevant contributions that family therapy scholars can offer toward addressing mental health disparities, particularly as it refers to developing community-based prevention interventions that achieve a balance between evidence-based knowledge and cultural relevance.

Pollination by sexual deception promotes outcrossing and mate diversity in self-compatible clonal orchids

The majority of flowering plants rely on animals as pollen vectors. Thus, plant mating systems and pollen dispersal are strongly influenced by pollinator behaviour. In Australian sexually deceptive orchids pollinated by male thynnine wasps, outcrossing and extensive pollen flow is predicted due to floral deception, which minimizes multiple flower visitations within patches, and the movement of pollinators under mate‐search rather than foraging behaviours. This hypothesis was tested using microsatellite markers to reconstruct and infer paternity in two clonal, self‐compatible orchids. Offspring from naturally pollinated Chiloglottis valida and C. aff. jeanesii were acquired through symbiotic culture of seeds collected over three seasons. In both species, outcrossing was extensive (tm = 0.924–1.00) despite clone sizes up to 11 m wide. The median pollen flow distance based on paternity for both taxa combined was 14.5 m (n = 18, range 0–69 m), being larger than typically found by paternity analyses in other herbaceous plants. Unexpectedly for orchids, some capsules were sired by more than one father, with an average of 1.35 pollen donors per fruit. This is the first genetic confirmation of polyandry in orchid capsules. Further, we report a possible link between multiple paternity and increased seed fitness. Together, these results demonstrate that deceptive pollination by mate‐searching wasps enhances offspring fitness by promoting both outcrossing and within‐fruit paternal diversity.

Microdot technology for individual marking of small arthropods

1 Individual mark–release–recapture is an important method for gathering data on insect movement, although it is limited by the constraints of tagging small insects with individual information. 2 Microdots, originally developed for covert security applications, are small polymer discs (diameter 0.5 mm) bearing up to 26 characters of information and have the potential as an alternative to the larger bee tags. In the present study, we test microdots for the individual marking of a 9‐mm parasitoid wasp. 3 We individually marked 505 wasps. The recapture rate was 24% of individuals over 189 recapture events, for which 84% retained legible microdot labels. Movement was in the range 0–161 m with a mean displacement 21.2 ± 2.7 m. A captive survival experiment showed no difference in lifespan between marked and unmarked wasps. 4 The present study shows that microdots can provide an effective, durable, low‐cost method for individually tagging small insects. The technique offers new opportunities by greatly expanding the capability for individually marking small insects, shifting the minimum size below that of bee tags, which is the only other manufactured option for individualized miniature marking.

Pollination: The Price of Attraction

Nectar is the major currency bringing together plants and pollinators; yet the costs and benefits of nectar production remain poorly understood. A low nectar line developed in Petunia offers an innovative approach to this problem and may offer clues to why some plants cheat and secure pollination via deception.