Douglas H. Goldman

A phylogenetic analysis of the Orchidaceae: evidence from rbcL nucleotide sequences

Cladistic parsimony analyses of rbcL nucleotide sequence data from 171 taxa representing nearly all tribes and subtribes of Orchidaceae are presented here. These analyses divide the family into five primary monophyletic clades: apostasioid, cypripedioid, vanilloid, orchidoid, and epidendroid orchids, arranged in that order. These clades, with the exception of the vanilloids, essentially correspond to currently recognized subfamilies. A distinct subfamily, based upon tribe Vanilleae, is supported for Vanilla and its allies. The general tree topology is, for the most part, congruent with previously published hypotheses of intrafamilial relationships; however, there is no evidence supporting the previously recognized subfamilies Spiranthoideae, Neottioideae, or Vandoideae. Subfamily Spiranthoideae is embedded within a single clade containing members of Orchidoideae and sister to tribe Diurideae. Genera representing tribe Tropideae are placed within the epidendroid clade. Most traditional subtribal units are supported within each clade, but few tribes, as currently circumscribed, are monophyletic. Although powerful in assessing monophyly of clades within the family, in this case rbcL fails to provide strong support for the interrelationships of the subfamilies (i.e., along the spine of the tree). The cladograms presented here should serve as a standard to which future morphological and molecular studies can be compared.

A Phylogeny of the Monocots, as Inferred from rbcL and atpA Sequence Variation, and a Comparison of Methods for Calculating Jackknife and Bootstrap Values

Abstract A phylogenetic analysis of the monocots was conducted on the basis of nucleotide sequence variation in two genes (atpA, encoded in the mitochondrial genome, and rbcL, encoded in the plastid genome). The taxon sample of 218 angiosperm terminals included 177 monocots and 41 dicots. Among the major results of the analysis are the resolution of a clade comprising four magnoliid lineages (Canellales, Piperales, Magnoliales, and Laurales) as sister of the monocots, with the deepest branch within the monocots between a clade consisting of Araceae, Tofieldiaceae, Acorus, and Alismatales, and a clade that includes all other monocots. Nartheciaceae are placed as the sister of Pandanales, and Corsiaceae as the sister of Liliales. The Triuridaceae, represented by three genera, including Lacandonia, are resolved as monophyletic and placed in a range of positions, generally within Pandanales. Dasypogonaceae and Arecaceae diverge sequentially from a clade that includes all other commelinid taxa, and within the latter group Poales s. lat. are sister of a clade in which Zingiberales and Commelinales are sisters. Within Poales s. lat., Trithuria (Hydatellaceae) and Mayaca appear to be closely related to some or all elements of Xyridaceae. A comparison was conducted of jackknife and bootstrap values, as computed using strict-consensus (SC) and frequency-within-replicates (FWR) approaches. Jackknife values tend to be higher than bootstrap values, and for each of these methods support values obtained with the FWR approach tend to exceed those obtained with the SC approach.

AN EXPANDED PLASTID DNA PHYLOGENY OF ORCHIDACEAE AND ANALYSIS OF JACKKNIFE BRANCH SUPPORT STRATEGY

An expanded plastid DNA phylogeny for Orchidaceae was generated from sequences of rbcL and matK for representatives of all five subfamilies. The data were analyzed using equally weighted parsimony, and branch support was assessed with jackknifing. The analysis supports recognition of five subfamilies with the following relationships: (Apostasioideae (Vanilloideae (Cypripedioideae (Orchidoideae (Epidendroideae))))). Support for many tribal-level groups within Epidendroideae is evident, but relationships among these groups remain uncertain, probably due to a rapid radiation in the subfamily that resulted in short branches along the spine of the tree. A series of experiments examined jackknife parameters and strategies to determine a reasonable balance between computational effort and results. We found that support values plateau rapidly with increased search effort. Tree bisection-reconnection swapping in a single search replicate per jackknife replicate and saving only two trees resulted in values that were close to those obtained in the most extensive searches. Although this approach uses considerably more computational effort than less extensive (or no) swapping, the results were also distinctly better. The effect of saving a maximal number of trees in each jackknife replicate can also be pronounced and is important for representing support accurately.

An overview of the phylogenetic relationships within Epidendroideae inferred from multiple DNA regions and recircumscription of Epidendreae and Arethuseae (Orchidaceae).

Phylogenetic relationships within the epidendroid orchids with emphasis on tribes Epidendreae and Arethuseae were assessed with parsimony and model-based analyses of individual and combined DNA sequence data from ITS nuclear ribosomal DNA and plastid trnL intron, the trnL-F spacer, matK (gene and spacers), and rbcL regions. Despite the absence of boostrap support for some of the relationships, a well-resolved and supported consensus was found, for which most clades were present in more than one individual analysis. Most clades of this consensus attained high posterior probabilities with a Bayesian approach. Circumscription of Arethuseae and Epidendreae are different from most orchid systems based on morphology, but they correspond to a combination of patterns from several less comprehensive orchid phylogenetic analyses previously published. A new circumscription of Epidendreae includes only Neotropical subtribes (Bletiinae, Chysiinae, Laeliinae, Ponerinae, and Pleurothallidinae), whereas Arethuseae include Coelogyninae (all Old World) and Arethusinae (pantropical). Many previously included genera will need to be moved to other tribes. Taxa previously assigned to be Old World Epidendreae are related to different groups of Old World orchids, and this study can serve as a guide for sampling strategies in future studies to resolve troublesome epidendroid orchid clades.

Phylogenetics of Arethuseae (Orchidaceae) Based on Plastid matK and rbcL Sequences

Abstract Circumscriptions of Arethuseae have varied since the tribe was first described by John Lindley in 1840, containing over ninety genera among the different authors. The latest system of Arethuseae defined by Dressler, including around thirty genera, is the most commonly accepted today. The goals of this study are to assess whether Arethuseae sensu Dressler and component subtribes are monophyletic and evaluate the position(s) of Arethuseae within Orchidaceae. Sequences of two plastid genes, matK and rbcL, have been obtained for 24 representative genera of Arethuseae in Dresslers latest two taxonomic systems for the tribe, plus 46 other genera throughout Orchidaceae. Both separate and combined analyses of the matK and rbcL data indicate that the tribe may not be monophyletic, which is also true for most subtribes within Arethuseae. Furthermore, matK data suggest that this gene may be non-functional within Orchidaceae. Communicating Editor: Kathleen A. Kron

Molecular and cytological examination of Calopogon (Orchidaceae, Epidendroideae): circumscription, phylogeny, polyploidy, and possible hybrid speciation.

The orchid genus Calopogon R.Br. (Orchidaceae), native to eastern North America and the northern Caribbean, currently contains five species and up to three varieties. Using nuclear internal transcribed spacer (ITS) ribosomal DNA sequences, amplified fragment length polymorphisms (AFLPs), chloroplast DNA restriction fragments, and chromosome counts, we present a phylogenetic and taxonomic study of the genus. Calopogon multiflorus and C. pallidus are consistently sister species, but the relationships of C. barbatus, C. oklahomensis, and C. tuberosus are not as clear. In the ITS analysis C. oklahomensis is sister to C. barbatus, whereas it is sister to C. tuberosus in the plastid restriction fragment analysis. Furthermore, all species were found to have chromosome numbers of 2n = 38 and 40, with the exception of the putatively hybrid-derived C. oklahomensis with 2n = 114 and 120. The hexaploidy of the latter, plus the discrepancy in its position between the ITS and plastid restriction fragment trees, could suggest that it is of hybrid origin. However, the presence of unique morphological and molecular characters might indicate that it is either an ancient hybrid or not of hybrid derivation at all. Finally, using these molecular methods all taxa appear to generally be discrete groups, with the exception of C. tuberosus vars. latifolius and tuberosus, the former of which is best combined with the latter.

Are mitochondrial genes useful for the analysis of monocot relationships

A phylogenetic analysis of monocots and related dicots was conducted, using a four-gene matrix consisting of two genes from the plastid genome (matK and rbcL) and two from the mitochondrial genome (atpA/atp1 and cob). The taxon sample includes 101 monocots and 36 dicots, and all four genes were sampled for all 137 taxa. Jackknife support was assessed for clades resolved by the four-gene analysis, and compared to support for the same clades by each of the four three-gene subset matrices, in order to quantify the degree to which each gene contributed to or detracted from support for each clade. Instances of positively and negatively correlated support for clades by genes of the same and different genomes were observed. In particular, the placement of Acorus within a clade that also includes Tofieldiaceae, Araceae, and Alismatales s.s., as opposed to its frequent placement as sister of all other monocots, is supported by atpA and matK. The results indicate that genes from the mitochondrial genome provide a unique test of relationships that have been inferred with plastid-encoded genes.

Isozyme Variation and Species Delimitation among Diploid Populations of the Puccinellia nuttalliana Complex (Poaceae): Character Fixation and the Discovery of Phylogenetic Species

Phylogenetic species, the minimal elements appropriate as terminals in phylogenetic analysis, are population systems that are differentiated by unique (i.e., diagnostic) character combinations. Within the predominantly polyploid Puccinellia nuttalliana species complex, two diploid taxonomic species are recognized on the basis of morphology. Isozyme variation was analysed in 11 diploid populations identified as P. lemmonii and one identified as the rare P. parishii, and variation profiles were generated for each population, reflecting each allele as either absent, present and fixed, or present and not fixed. Population Aggregation Analysis (PAA) grouped these populations into two distinct isozyme species that correspond precisely in membership to the species recognized previously on the basis of morphology; thus, isozyme patterns are congruent with those of morphology. Mean genetic identities among populations of P. lemmonii are not unusually high for conspecific plant populations, nor are genetic identities between populations of the two species unusually low.

Morphometric circumscription of species and infraspecific taxa in Calopogon R. Br. (Orchidaceae)

Abstract.The orchid genus Calopogon contains five species, the widespread C. tuberosus containing three varieties. The members of this small genus have a history of circumscriptional uncertainty, suggesting the need for further taxonomic evaluation. To determine characters distinguishing different groups and to examine their circumscription, we performed morphometric analyses on two different subsets of Calopogon, one of all the currently recognized species of the genus and another of only C. tuberosus, based on 40 vegetative and floral variables measured from live plants. Principal components analyses (PCA) generally did not detect highly distinct groups, but canonical variate analyses (CVA) identified numerous characters that readily distinguish most of these groups, the least distinguishable of these being C. tuberosus var. latifolius. Furthermore, generalized patterns of similarity between taxonomic and geographic subdivisions within C. tuberosus were further examined on a plant-by-plant basis using squared Mahalanobis distances determined from the CVA of this species. Finally, using three different tests, the degree of morphological diversity was examined for each of the different groups of C. tuberosus, finding that the northern range of the species contained less diversity whereas the southern range generally contained more. We maintain the recognition of all Calopogon taxa except C. tuberosus var. latifolius, and do not recognize any new taxa based on our examinations of geographic partitions of the typical variety of C. tuberosus. However, morphology was observed to vary substantially over the geographic range of the typical variety and this species overall.