CHROMATOGRAPHIC EVIDENCE OF HYBRIDIZATION AND EVOLUTION IN PHLOX MACULATA

Representatives of P. maculata ssp. maculata and P. glaberrima ssp. interior and 36 putative hybrids were collected from a population in central Indiana. Chromatographic analysis revealed the presence of nine diagnostic components in P. maculata ssp. maculata and eight such components in P. glaberrima ssp. interior. Putative F₁ hybrids contained all or most of the parental marker components; markers of the non-recurrent parent were absent in the two classes of putative backcross progeny. The characters of P. maculata ssp. pyramidalis are a composite of those present in P. maculata ssp. maculata and P. glaberrima ssp. interior, and the former is virtually identical to certain hybrids from the population in question. The contention that P. maculata ssp. pyramidalis is a stabilized derivative of natural hybridization between these taxa is substantiated by the presence of all of the diagnostic constituents of the putative parents on its chromatograms. It is the author's opinion that the definitive data which chromatography affords will be invaluable in elucidating the complex pathways of evolution in Phlox.     

PHYSIOLOGICAL EVIDENCE OF HYBRIDIZATION AND RETICULATE EVOLUTION IN PHLOX MACULATA

A comparative physiological analysis was conducted on a hybrid swarm involving P. maculata subsp. maculata and P. glaberrima subsp. interior. The population contained an array of segregates and introgressants. Hybrids displayed a high degree of heterogeneity in their physiological attributes, some characters being intermediate to, similar to, or transgressing those of the parental taxa. For example, both net photosynthetic rates and net photosynthetic/dark respiratory ratios were enhanced in the hybrids, and the level of enhancement was associated with the level of morphological intermediacy. Conversely, no enhancement was noted in either respiration rates or foliar fresh weight/ovendry weight ratios of the hybrids. Because of physiological imbalance, about one-third of the hybrids were eliminated early in their development. Representatives of P. maculata subsp. pyramidalis, a putative stabilized derivative of hybridization between the above two parental species, were intermediate in most physiological features and closely resembled some natural hybrids. The data verify that the physiological features of subspecies pyramidalis can be assembled through recombination of the P. maculata and P. glaberrima genomes.     

RETICULATE EVOLUTION IN PHLOX AS SEEN THROUGH PROTEIN ELECTROPHORESIS

Seed protein profiles were used to document the genome constitutions of seven Phlox taxa believed to be stabilized derivatives of hybridization between P. pilosa and P. carolina, P. drummondii or P. amoena. These species have very distinctive seed protein profiles. Analyses of profiles from these and other phloxes not involved in the reticulum (P. maculata, P. glaberrima and their hybrid derivative P. maculata subsp. pyramidalis) demonstrated that profile components are inherited as co-dominant units, alteration of ploidal level does not alter the profile, profile components may undergo segregation and recombination, and profile components in hybrids do not re-combine in vivo or in vitro to produce new proteins. The putative derivatives of hybridization between P. pilosa and P. carolin, i.e., P. pilosa subsp. detonsa, P. pilosa subsp. sangamonensis, P. floridana and P. pulcherrima, display protein profiles which are nearly summations of the parental patterns as do the putative derivatives of P. pilosa and P. drummondii. i.e., P. villosissima and P. aspera. The putative derivative of P. pilosa and P. amoena, i.e., P. pilosa subsp. deamii, displays a completely additive profile. Genome constitutions judged from morphological, cytological, and chromatographical data are fully corroborated by the protein profiles.     

QUANTITATIVE VARIATION IN PHLOX: COMPARISON OF SELFING AND OUTCROSSING SPECIES

Variation of 20 quantitative characters was examined within and among 10 populations of the predominantly outcrossing Phlox drummondii and 4 populations of the predominantly selfing P. cuspidata grown in a greenhouse. Multivariate analysis of variance, considering all characters simultaneously, indicated that there were significant differences among populations in both species while analysis of individual characters demonstrated that there were significant population differences for 19 characters in P. drummondii and 13 characters in P. cuspidata. On average, 16% of the total phenotypic variation in P. drummondii occurred among populations compared to less than 4% of the total variation in P. cuspidata. In addition, P. drummondii exhibited significant differences among families within populations more frequently than P. cuspidata. Most observed variation in both species occurred within families where environmental and genetic sources of variation could not be partitioned. There was a trend for P. drummondii to have higher heritabilities than P. cuspidata for most characters even when assumptions about breeding systems were relaxed. Thus, the outbreeding species exhibited larger genetic differences among populations and among families within populations than the selfing species in the greenhouse environment. These data suggest that P. drummondii has the greater evolutionary potential of the two species and are consistent with the hypothesis that differences in population structure result from differences in the breeding systems of the two species.     

Evolutionary history of the Dactylorhiza maculata polyploid complex (Orchidaceae)

Taxonomic complexity may be associated with migration history and polyploidy. We used plastid and nuclear DNA markers to investigate the evolutionary history of the systematically challenging Dactylorhiza maculata polyploid complex. A total of 1833 individuals from 298 populations from throughout Europe were analysed. We found that gene flow was limited between the two major taxa, diploid ssp. fuchsii (including ssp. saccifera) and tetraploid ssp. maculata. A minimum of three autotetraploid lineages were discerned: (1) southern/western ssp. maculata; (2) northern/eastern ssp. maculata; and (3) Central European ssp. fuchsii. The two ssp. maculata lineages, which probably pre‐date the last glaciation, form a contact zone with high genetic diversity in central Scandinavia. Intermediate plastid haplotypes in the contact zone hint at recombination. Central Europe may have been a source area for the postglacial migration for the southern/western lineage of ssp. maculata, as well as for ssp. fuchsii. The northern/eastern lineage of ssp. maculata may have survived the LGM in central Russia west of the Urals. The tetraploid lineage of ssp. fuchsii is indistinguishable from diploid ssp. fuchsii, and is probably of postglacial origin. The Mediterranean region and the Caucasus have not contributed to the northward migration of either ssp. fuchsii or ssp. maculata. © 2010 The Linnean Society of London, Biological Journal of the Linnean Society, 2010, 101 , 503–525.     

INBREEDING DEPRESSION IN PARTIALLY SELF-FERTILIZING PHLOX

Inbreeding depression was measured in terms of the relative survivorship of self versus cross seed in 14 Phlox drummondii cultivars and 10 populations of P. cuspidata. The cultivars are predominantly outcrossing; P. cuspidata is predominantly self-fertilizing. The relative survivorship of self versus cross seed averages 0.84 in the cultivars and 0.99 in P. cuspidata. In wild P. drummondii, the average is 0.83. The mean number of lethal equivalents per zygote averages 0.69 in the cultivars and 0.05 in P. cuspidata, versus 0.79 in the wild P. drummondii. There is a significant negative correlation between the amount of automatic self-seed produced by a cultivar and the number of lethal equivalents it carries (r = ?0.60). Phlox cuspidata has a lower level of cross-seed abortion than P. drummondii, which probably is not due to a difference in the incidence of lethal genes in their populations.     

DYNAMICS OF A HYBRID ZONE IN PHLOX: AN EXPERIMENTAL DEMOGRAPHIC INVESTIGATION

The dynamics of a narrow hybrid zone between Phlox drummondii ssp. drummondii and ssp. mcallisteri was investigated using an experimental demographic approach. Three populations of each subspecies as well as those from the hybrid zone were involved. Experimental plots containing seeds from each of the nine populations were established in each of the eight sites. Germination, survivorship to reproduction, and fecundity were monitored at 4–6 wk intervals. Finite rates of increase were estimated from these data. There were significant site effects for each demographic variable and rate of increase as well as significant population effects for germination and finite rate of increase. Although there were population effects, no significant differences were found between plant types in sites of the subspecies or the hybrid zone. We reject the hypothesis that the narrow width of the hybrid zone is the result of hybrid disadvantage. We propose that the narrow width is a result of restricted gene flow and discuss the supporting evidence.     

RESPONSE TO SELECTION ON AUTOGAMY IN PHLOX

Two cycles of artificial selection were performed to increase autogamous fruiting in two wild populations of the self-incompatible Phlox drummondii, to decrease autogamous fruiting in two wild populations of the self-compatible Phlox cuspidata, and to both increase and decrease autogamous fruiting in a cultivar of P. drummondii which is pseudo-self-compatible. The breeding systems were determined to be genetically quite flexible, independent of inbreeding depression and other genetic phenomena which could hinder a breeding system shift. This is especially true for increasing autogamy. Self-pollen-pistil compatibility seems to be the single character affected by selection. Based on the continuous variation in both autogamy and self-compatibility, we suggest that the change has been due to genes which modify the self-incompatibility reaction rather than to the simple segregation of alleles at the S-locus.     

HYBRIDIZATION BETWEEN ANNUAL SPECIES OF PHLOX: POPULATION STRUCTURE

Phlox cuspidata (n = 7) and P. drummondii subsp. drummondii (n = 7) are closely related annuals which are indigenous to eastern and central Texas. The species typically occupy different ecological niches but may form contiguous or confluent populations in disturbed habitats and hybridize therein. On the basis of correlative interpretations of exomorphic, chromatographic and fertility information, hybridizing populations can be segregated into three distinct classes: (1) highly fertile plants with the morphological and phenolic attributes of P. drummondii; (2) highly fertile plants with the morphological and chemical attributes of P. cuspidata; (3) sterile plants with manifestly intermediate morphology and complementary chromatographic patterns. These data strongly suggest that hybridizing populations of P. drummondii and P. cuspidata are tritypic, being composed of “pure” or essentially “pure” parental species and a group of plants which has all of the attributes characteristic of an F₁ hybrid.     

POPULATION AND GENOTYPE NICHE WIDTH IN CLONAL PHLOX PANICULATA

We use a model population comprised of five genotypes of Phlox paniculata L. to investigate the contribution of individuals to the response breadth (niche) of the population on a light gradient and a moisture gradient. Analysis of within- and between-genotype components of population response width showed up to 20% of the response is due to between-genotype effects, depending upon the character considered. Since the only way a sedentary organism can deal with a variable environment is through plasticity of response, differences in levels of phenotypic plasticity between genotypes on the two resource gradients were also investigated. There was no correlation between level of phenotypic plasticity and success over a range of environments. Niche breadth calculated as Levins' (B) and Roughgarden's (w²) indicated that flowering, and hence sexual reproduction, was limited to a much narrower range of environments than was vegetative growth. We also found significant genotype × environment interactions on both gradients, indicating differences in genetic response to the environment.     

DYNAMICS OF SYNTHETIC PHLOX DRUMMONDII POPULATIONS AT THE SPECIES MARGIN

We compare the dynamics of artificial populations of the annual Phlox drummondii introduced along a transect extending from within to beyond the species margin southwest of San Antonio, Texas. The margin is defined by a shift from sandy, nutrient poor soils to clay, nutrient-rich soils. Fourteen sites were chosen along the 6.5-mi transect. In each site seeds were planted into 1.5-m-square cleared plots and uncleared 10-m transects. Transplants also were introduced into cleared plots. Survival, flower production and seed set were monitored over the course of the growing season. Individual fitnesses and the reproductive rates of artificial populations generally declined as the distance from the margin increased. The results indicate soil type per se is not limiting Phlox and that plants can successfully grow and reproduce, albeit at a reduced level, in sites beyond their natural margin.     

Habitat differentiation, hybridization and gene flow patterns in mixed populations of diploid and autotetraploid Dactylorhiza maculata s.l. (Orchidaceae)

Detailed ecological, morphological and molecular analyses were performed in mixed populations of diploid and autotetraploid Dactylorhiza maculata s.l. in Scandinavia. Comparisons were made with pure populations of either diploid ssp. fuchsii or tetraploid ssp. maculata. It was shown that mixed populations are the result of secondary contact between ssp. fuchsii and ssp. maculata. No patterns of recent and local autopolyploidization were found. Morphology and nuclear DNA markers (internal transcribed spacers of nuclear ribosomal DNA) showed that diploids and tetraploids from mixed populations have similar levels of differentiation to diploids and tetraploids from pure populations. Vegetation analyses, as well as analyses of environmental variables, revealed that diploid and tetraploid individuals in mixed populations are ecologically well differentiated on a microhabitat level. Diploids and tetraploids in pure populations have wider ecological amplitudes than they do in mixed populations. Triploid hybrids grew in intermediate microhabitats between diploids and tetraploids in the mixed populations. Plastid DNA markers indicated that both diploids and tetraploids may act as the maternal parent. Based on morphology and nuclear markers triploids are more similar to tetraploids than to diploids. There were indications of introgressive gene flow between ploidy levels. Plastid markers indicated that gene flow from diploid to tetraploid level is most common, but nuclear markers suggested that gene flow in opposite direction also may occur. Similar patterns of differentiation and gene flow appeared in localities that represented contrasting biogeographic regions. Disturbance and topography may explain why hybridization was slightly more common and the differentiation patterns somewhat less clear in the Scandinavian mountains than in the coastal lowland. An erratum to this article can be found at     

DIFFERENTIATION AMONG NINE POPULATIONS OF PHLOX. I. ELECTROPHORETIC AND QUANTITATIVE VARIATION

The organization of genetic variation in Phlox drummondii was investigated using both allozyme electrophoresis and quantitative genetics. Variation at five polymorphic enzyme loci was characterized in nine populations, and variation in 16 morphological and life-history characters was examined using an analysis of full- and half-sibs in seven populations. Significant levels of genetic variation were found at enzyme loci and for metric characters. Significant heritabilities were observed for 15 of the 16 characters examined. Genetic differences among populations were revealed both by Nei's genetic distance and by phenotypic differences, summarized by discriminant analysis. Partitioning variance in allozyme frequencies among hierarchical levels of genetic organization indicated that 94% of this variance lay within populations, 4% between populations within varieties, and 2% between varieties. Partitioning phenotypic variance for metric characters indicated that 73% lay within populations, 24% lay between populations within varieties, and 3% lay between varieties. Thus, both electrophoretic and metric characters indicated that despite extensive genetic differentiation among populations, most of the evolutionary potential of the species lies within populations.     

A biosystematic study of the Interiores species group of the genus Vernonia (Compositae)

The taxa of the Interiores species group are treated as two species, one of which is divided into two subspecies.Vernonia missurica, a widespread taxon, has large flower heads (32-58 flowers per head), obtuse involucral bract tips, and a characteristic flavonoid Chromatographic profile.Vernonia baldwinii ssp.baldwinii, occurring throughout the Ozarks and Ouachitas, andV. baldwinii ssp.interior of the Central Plains possess smaller flower heads (17-34 flowers per head), acute or acuminate involucral bracts, and a different Chromatographic profile.Vernonia baldwinii ssp.baldwinii is distinguished from ssp.interior by the recurved involucral bract tips and by the possession of more flowers per head. Morphological data reveal that an intergradation zone exists between the population systems of the two subspecies along the edges of the Ozarks and Ouachitas. Although much gene exchange occurs between these two taxa, they maintain a level of genetic integrity that permits their ready recognition throughout wide ranges. All three taxa have a gametic chromosome number ofn = 17 and hybridize extensively with sympatric species.     

Evolution,phylogeography and taxonomy of allopolyploid Dactylorhiza (Orchidaceae) and its implications for conservation

This review is based on recent molecular studies of Dactylorhiza (Orchidaceae). Most of the studies have focused on the allotetraploid members of the genus in general and on D. majalis ssp. lapponica in particular. It was concluded that most of the allotetraploid taxa have derived from hybridizations between the parental lineages D. maculata s.l. and D. incarnata s.l., with D. maculata s.l. serving as the seed parent. Evidence of multiple origins was found both among northern European allotetraploids as well as among Greek allotetraploids. Introgression from both parental lineages and hybridizations between independently derived polyploid lineages was also detected. The three morphologically similar taxa D. majalis ssp. traunsteineri, ssp. lapponica and ssp. russowii should be treated as one and most of the Greek allotetraploids should be regarded as regional variants of the southeastern European D. majalis ssp. cordigera. The Balkans and the Alps most probably served as refugia for the genus during the last glaciations and at least two waves of immigration reached Scandinavia. Finally, we suggest that the conservation of allotetraploid Dactylorhiza should emphasize important geographic areas and habitats and that the allopolyploids should have the same conservation status as the diploids.     

INTERSPECIFIC HYBRIDIZATION IN HAPLOPAPPUS AND ITS BEARING ON CHROMOSOME EVOLUTION IN THE BLEPHARODON SECTION

Jackson , R. C. (U. Kansas, Lawrence.) Interspecific hybridization in Haplopappus and its bearing on chromosome evolution in the Blepharodon section. Amer. Jour. Bot. 49 (2) : 119–132. Illus. 1962.—Cytological analyses of interspecific hybrids between H. gracilis (n = 2) and H. spinulosus ssp. australis (n = 8) indicate that ssp. australis is a segmental allotetraploid, derived from past hybridization between 2 taxa with chromosome numbers of n = 4. Analysis of hybrids between H. gracilis (n = 2) and H. ravenii (n = 4), a previously undescribed species, has shown that the chromosome segments of these 2 species are almost completely homologous. Differential contraction is suggested as the explanation for the disappearance in late pachytene of presumed non-homologous segments which were evident in some cells at early pachytene. The pairing relationship of gracilis and ravenii chromosomes at pachytene and later prophase I stages of meiosis indicates that gracilis has evolved from ravenii by an aneuploid reduction process similar to that described for Crepis. The close morphological relationship of the 2 species adds further support to this proposition. Data from the cytological analysis of both interspecific hybrids indicate that x = 4 is the basic chromosome number for the Blepharodon section of Haplopappus.     

Systematics and conservation genetics of Dactylorhiza majalis ssp. elatior (Orchidaceae) on Gotland

A tall allotetraploid member of the Dactylorhiza incarnata/maculata complex with unspotted leaves and large pinkish flowers from the island of Gotland in the Baltic was examined for molecular variation patterns at five nuclear microsatellite loci, nuclear ITS and in plastid haplotypes. The allotetraploid was well separated from allopatric allotetraploids of similar appearance, including the western European D. majalis ssp. integrata (syn. D. praetermissa) and forms of D. majalis ssp. lapponica from mainland Sweden. It also differed from other allotetraploids distributed in the Baltic Sea region, including D. majalis ssp. baltica and D. majalis ssp. lapponica. It is here recognized as D. majalis ssp. elatior (Fr.) Hedrén & H. A. Pedersen. Dactylorhiza osiliensis Pikner, described from Saaremaa (Estonia) is regarded as a synonym. The distribution covers Gotland, Saaremaa and possibly Hiiumaa. Dactylorhiza majalis ssp. elatior may have one or several recent origins within its present distribution area, and it contains no other molecular markers than those found in the parental D. incarnata var. incarnata and D. maculata ssp. fuchsii in the same area. It appears to have weak barriers towards secondary hybridization with its parental lineages. The situation is reminiscent to that of other young allotetraploids in the D. majalis s.l. complex, suggesting that introgression may be an underestimated process explaining the accumulation of genetic diversity in evolving allopolyploid plants.     

Systematics and phylogeography of the Dactylorhiza maculata complex (Orchidaceae) in Scandinavia: insights from cytological, morphological and molecular data

Flow cytometry, morphometry and molecular markers [plastid DNA and internal transcribed spacers (ITS) of nuclear ribosomal DNA] were used to determine taxonomic and phylogeographic patterns in Dactylorhiza maculata s.l. from Scandinavia. A total of 238 individuals from 27 populations from throughout all of Scandinavia, including the adjacent Kola Peninsula of Russia, were analyzed. Diploid D. maculata ssp. fuchsii and autotetraploid D. maculata ssp. maculata are morphologically differentiated. Fragment size variants from 10 plastid DNA loci (seven microsatellite loci and three loci with indel variation) were combined to give 43 haplotypes. Three major groups of haplotypes were found. Group I haplotypes were prevalent in the north and the northeast, whereas Group II haplotypes were prevalent in the south and the southwest. Group III was represented by only a single haplotype and appeared to be the result of introgression from D. incarnata s.l. Group I and Group II haplotypes did not correspond with cytologically and morphologically defined D. maculata ssp. fuchsii or D. maculata ssp. maculata. Past introgressive gene flow rather than recent hybridization is envisaged. Intermediate Group I haplotypes between Group II and the rest of Group I were detected in a zone of contact in central Sweden, which may suggest plastid DNA recombination. The six ITS alleles scored showed strong positive correlation with taxonomy. All data sets obtained for ssp. maculata were significantly correlated with geography. Three different autotetraploid lineages are hypothesized. One lineage may represent postglacial immigration from the south and the other two lineages may represent eastern immigration routes. Morphology and ITS data suggested that subarctic populations of ssp. maculata should be recognized as var. kolaënsis.     

ULTRASTRUCTURE OF VEGETATIVE AND DIVIDING CELLS OF THE UNICELLULAR RED ALGAE RHODELLA VIOLACEA AND RHODELLA MACULATA1

Rhodella violacea (Kornmann) Wehrmeyer and Rhodella maculata Evans were investigated for ultrastructural details of vegetative and dividing cells. Rhodella violacea has a nuclear projection into the pyrenoid similar to that found in R. maculata, although the nuclear projection in R. maculata traverses a starch-lined area before contacting the pyrenoid. Unlike most, red algae, the two Rhodella species lack a peripheral encircling thylakoid in the chloroplast and have dictyosomes associated solely with endoplasmic reticulum (ER) instead of with both mitochondria and ER. Both species also have a well-developed peripheral system of ER connected to the plasmalemma by tubules, a situation found only in red algal unicells, Cell division was studied primarily in R. violacea; a less thorough examination of R. maculata showed no essential differences. Both have small, double-ringed, nucleus-associated organ files (NAOs) surrounded by moderately electron-dense material, metaphase–anaphase polar gaps in the nuclear envelope, absence of perinuclear ER. and short interzonal spindles. This pattern of mitosis is similar in most respects to that reported in the unicell Flintiella. Following mitosis, microtubules extend from the region of each NAO to its associated nucleus and to the undivided pyrenoid. The NAOs appear to apply tension to the nuclei and the pyrenoid and may be the mechanism for ensuring equal partitioning of both organdies. Two different forms of pyrenoid-nucleus association occur during mitosis. Nuclear projections into the pyrenoid, prevalent during interphase and early stages of mitosis, recede at metaphase. Then, the pyrenoid extends protrusions into the nuclear polar areas, forming a cup that partially surrounds the nucleus. Cell division and vegetative characters confirm the close taxonomic affinity of these two species of Rhodella and support their separation from the genus Porphyridium.     

Cold tolerance of invasive freshwater snails,Pomacea canaliculata,P. maculata,and their hybrids helps explain their different distributions

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