Recombination and introgression of nuclear and chloroplast genomes between the peat mosses, Sphagnum capillifolium and Sphagnum quinquefarium

Haploid hybrid gametophytes are often present at low frequencies in sympatric populations of Sphagnum capillifolium and Sphagnum quinquefarium. We used intersimple sequence repeat (ISSR) markers and polymerase chain reaction-restriction fragment length polymorphism of the trnL(UAA) intron of the chloroplast genome to reveal the nuclear and chloroplast composition of mature hybrid gametophytes from natural populations and of gametophytes derived from spores of hybrid sporophytes collected in nature. Asymmetrical nuclear inheritance was found in the progeny of the hybrid sporophytes, indicating that only spores with a low level of recombination of parental genomes were viable. A similarly skewed nuclear composition was found among the naturally occurring hybrid gametophytes. All hybrid genomes contained a larger proportion of S. capillifolium ISSR markers, combined with only two to five S. quinquefarium markers together with a chloroplast haplotype derived from S. quinquefarium. In this way, a pattern resembling introgression is created within a single generation. Some individuals possessed nuclear genomes typical for S. capillifolium in combination with the chloroplast haplotype of S. quinquefarium, possibly indicating backcrossing. Our results indicate that hybridization between S. capillifolium and S. quinquefarium is relatively common, but the resistance of large parts of the genome against heterospecific genes maintains the genetic distinctness of the species. Further evolutionary and phylogenetic consequences of restricted interspecific gene exchange are discussed.     

Morphological and isozyme divergence in Korean Hepatica sensu stricto (Ranunculaceae)

 Morphological and electrophoretic data were studied to examine species delimitation, patterns of morphological and genetic variation in three Korean Hepatica including two endemics, H.␣maxima and H. insularis. Based on a phenogram using 15 morphological characters, taxa were distinct; it was consistent with the phenogram based on genetic distance. In the enzyme electrophoresis study, the genetic identities suggested that three taxa were genetically divergent enough to be recognized as different species, falling within the range expected␣for congeners. The genetic identity between H.␣asiatica and H. insularis was higher than the values between these two taxa and H.␣maxima, a restricted endemic of Ulleung Island. The least genetic variation was found in H. maxima and the greatest in widespread H. asiatica. These data are consistent with theoretical expectations that small populations are more likely to be genetically depauperate. Received November 13, 2001; accepted May 10, 2002 Published online: December 11, 2002     

Population structure and interspecific differentiation of the peat moss sister speciesSphagnum rubellum andS. capillifolium (Sphagnaceae) in northern Europe

Isozyme electrophoresis was used to study the morphologically similar sister speciesSphagnum rubellum andS. capillifolium from a sample of 1313 plants representing 37 populations from Scandinavia, Great Britain and S Germany. The mean pairwise genetic identities (I) among conspecific populations were 0.976 forS. rubellum and 0.969 forS. capillifolium, versus 0.627 between populations of the two species. Interspecific gene flow was indicated by the observation of occasional plants in sympatric populations with alleles otherwise unique to the other species. Populations of bisexualS. capillifolium were significantly more variable than populations of unisexualS. rubellum. Alpine populations ofS. rubellum andS. capillifolium were dominated by few genotypes, and differentiation among populations was pronounced, indicating a low level of sexual recombination. InS. rubellum, maximum variability was found in western areas with high annual precipitation. Distribution of alleles inS. rubellum indicated restricted gene flow between Great Britain and Scandinavia. Postglacial migration from separate refugia may explain large-scale variation inS. rubellum.     

Hybridization in staminate and pistillate Salix alba and S. fragilis (Salicaceae): morphology versus RAPDs

 The polyploid Salix alba–Salix fragilis hybrid complex is rather difficult to study when using only morphological characters. Most of the characters have a low diagnostic value for unambiguously identifying the hybrids, introgression patterns and population structures. Morphology and molecular variation determined with random amplified polymorphic DNAs (RAPDs) were investigated in a set of staminate and pistillate willows from Belgium. A thorough screening of possible RAPD markers was done to select homologous amplification products. The selected amplified products proved to be useful in a principal coordinate analysis for the identification of individuals from a morphological continuum comprising presumed pure species and introgressants. The RAPD based identity of the individuals or clones was checked against those based on morphological characters. A correspondence analysis indicated that all pubescence related characters were associated but separated from the size related characters. The RAPDs also revealed that the S. fragilis genotypes mainly consisted of staminate individuals whereas most of the pistillate trees belonged to the S. alba genotype cluster. It was suggested that both species have kept their gene pools well separated and that morphologically intermediate plants are not necessarily genetically intermediate. Received August 31, 1999 Accepted December 12, 2000     

The arctic-alpine polyploids Cerastium alpinum and C. nigrescens (Caryophyllaceae) in a sympatric situation: breakdown of species integrity?

 The high polyploids Cerastium alpinum (8x) and C. nigrescens (12x) were investigated in a mixed population in central Norway to evaluate whether hybridisation has resulted in continuous variation in morphology and genetic markers, a hypothesis previously proposed to account for the extensive taxonomic confusion in this species group. Isozyme, fertility, and morphological (37 characters) variation were examined among 347, 265, and 237 plants, respectively. A PCO analysis based on 23 quantitative morphological characters identified two main groups, corresponding to C. alpinum and C. nigrescens. The groups were also clearly separated in isozyme markers, several qualitative morphological characters, and chromosome numbers. Only 20 plants (8.4%) were more or less intermediate in the PCO analysis. These plants had a parental or hybrid isozyme multilocus phenotype and typically few well-developed anthers, low pollen stainability, and no seed set. Several plants within the C. nigrescens group also had reduced pollen stainability. These results indicate that later-generation hybrids and/or backcrosses towards C. nigrescens are formed. Thus, interspecific, interploidal gene flow probably occurs, but at rates that are insufficient to break down species integrity. Received July 5, 2000 Accepted August 2, 2001     

Molecular differentiation of two vicariant neotropical tree species, Plathymenia foliolosa and P. reticulata (Mimosoideae), inferred using RAPD markers

 Plathymenia foliolosa and P. reticulata are two tropical tree species economically important for their high quality wood. P. foliolosa occurs mainly in areas of Atlantic Forest (a tropical forest) whereas P. reticulata occurs mainly in the Cerrado region (a savanna vegetation), both Brazilian biomes classified as hotspots for conservation priorities. In the present study RAPD markers were used to differentiate P. foliolosa from the vicariant species P. reticulata. Using 10 random primers it was possible to obtain 84 markers, of which 22 (26.2%) were present in just one of the two species. Nine of the ten primers were successful in discriminating between the species through the detection of diagnostic markers or band frequency differences. One of the populations of P. foliolosa, located in a region where the two species co-occur, presented several markers that were characteristic of P. reticulata, suggesting the occurrence of gene flow between the two species. Received September 3, 2001; accepted August 19, 2002 Published online: November 22, 2002 Addresses of the authors: Daniela Rodrigues Lacerda (dlacerda@dedalus.lcc.ufmg.br), Maria Bernadete Lovato (lovatomb@icb.ufmg.br), Maria Dolores Porto Acedo (acedomd@icb.ufmg.br), Dept. Biologia Geral, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, CP: 486, Belo Horizonte, MG, 31270-901, Brasil. José Pires de Lemos Filho (lemos@icb.ufmg.br), Dept. Botanica, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, CP: 486, Belo Horizonte, MG, 31270-901, Brasil.     

Cladistic and phenetic analysis of relationships in Vicia subgenus Vicia (Fabaceae) by morphology and isozymes

 Variation of 80 multistate morphological characters and isozymes encoded by 13 loci among 23 vetch species of the type subgenus of the genus Vicia in comparison with V. dumetorum, V. pisiformis and V. sylvatica of the subgenus Cracca is described and analyzed with cladistic parsimony and phenetic neighbour-joining methods by using two different ways of coding. Morphological analyses showed the subgenus Vicia monophyletic and revealed subgroups in a general agreement with traditionally recognized sections, except showing V. faba nested within section Narbonensis and ambiguity in the position of V. lathyroides and V. bithynica. Parsimony analysis of orthozymes as presence/absence characters revealed in the subgenus two basic monophyletic clades: 1) V. faba and three species of the section Peregrinae, V. michauxii, V. aintabensis and V. peregrina, in one subclade linked with species of the Narbonensis and Hyperchusa sections together with V. pisiformis of subgenus Cracca in a second subclade; 2) species belonging to sections Vicia, Sepium, Pseudolathyrus and Lathyroides together with V. sylvatica of the subgenus Cracca. Neighbour-joining analysis of orthozymes revealed the same two basic groups, differing only in the relative position of some species in subclusters. Both isozyme analyses showed paraphyly of the subgenera Vicia and Cracca. Parsimony analysis of orthozymes as character states of isozymes yielded a largely unresolved strict consensus cladogram of 209 most parsimonious trees, and reweighting of characters failed to produce a stable tree. Phylogenetic congruence and discordance among morphological and isozyme analyses, coding ways, homoplasy and weighting of characters are discussed. Received November 20, 2001 Accepted January 31, 2002     

Genomic relationships within Boronia (Rutaceae) as revealed by karyotype analysis and RAPD molecular markers

 The genus Boronia Sm. section Boronia series Boronia contains species with n=7 (B. megastigma), n=7 or 8 (B. heterophylla), n=8 (B. molloyae) and n=9 (B. purdieana), representing ideal species with which to examine comparative chromosome morphology. Between species there were few chromosomes with similar morphology, indicating numerous genome re-organisations. Karyotypes between and within species of Boronia could be distinguished and inheritance of some chromosomes was observed. Species and hybrids with 2n = 14 or 15 had at least one large chromosome. Chromosome morphology indicated a closer relationship between B. heterophylla and B. molloyae and between B. purdieana and B. megastigma than between these two groups. Whole genomic DNA was extracted from 9 genotypes of Boronia. RAPD bands were analysed and pairwise distance matrices between genotypes were computed. Dendrograms were generated and analysed using unweighted pair-group method with arithmetic average cluster analysis. Dendograms supported cytological results, indicating B. heterophylla and B. molloyae are closely related and clearly distinct from B. megastigma and B. purdieana. The evolution of boronias is discussed. Received February 16, 2001; accepted March 21, 2002 Published online: October 14, 2002 Address of the authors: G. Yan, F. Shan, J. A. Plummer (e-mail: jplummer@cyllene.uwa.edu.au), Plant Sciences, Faculty of Natural and Agricultural Science, The University of Western Australia, 35 Stirling Hwy, Crawley, WA 6009, Australia.     

Genetic variation in Scandinavian Anthericum liliago (Anthericaceae): allopolyploidy, hybridization and immigration history

 We investigated patterns of isozyme variation and the hierarchic structure of genetic diversity in 25 Scandinavian populations of the lilioid herb, Anthericum liliago. Isozyme data suggest that tetraploid A. liliago has an allopolyploid origin and that A. ramosum may be one of its diploid progenitors. Two populations contained known or suspected hybrids between A. liliago and A. ramosum. Isozymes show that one population from S Sweden contains both triploid (hybrid) and tetraploid individuals whereas a putatively hybrid Danish population represents diploid A. liliago. There is an overall northward and eastward decline in allelic richness in the tetraploid populations, with the highest numbers of alleles in Denmark and SW Sweden. This pattern is consistent with a progressive loss of allelic variation during the species' postglacial colonization of Scandinavia. The between-population component of genetic diversity is 4% (compared with 12% in diploid A. ramosum), the between-region diversity component is 7% and 89% of the total diversity is stored within populations. Received March 13, 2002; accepted September 24, 2002 Published online: December 11, 2002     

Thlaspi s.str. (Brassicaceae) versus Thlaspi s.l.: morphological and anatomical characters in the light of ITS nrDNA sequence data

 Nuclear ribosomal (ITS) DNA sequence data from representatives of eleven out of 12 genera previously included in Thlaspi sensu lato were analyzed to elucidate relationships within the former genus Thlaspi sensu lato. Sequences from Thlaspi segregates Noccaea, Masmenia, Callothlaspi, Kotschyella, Microthlaspi, and Noccidium were added to the existing data sets, and only some Thlaspi sensu lato segregates formed a monophyletic group. The recently renamed genus Pseudosempervivum, formerly part of the genus Cochlearia, has been shown to be closely related to this group as well. Microthlaspi consists of three independent lineages of which Microthlaspi umbellatum is closely related to Neurotropis. The Thlaspi s.str. taxa including the type species T. arvense, are closely related to Peltaria and Alliaria and they represent a monophyletic group. Kotschyella and Noccidium, which were also integrated into Thlaspi s.l. are not closely related to any Thlaspi lineage. Therefore, we suggest major taxonomical changes which are not in agreement with concepts based on morphological data. Our ITS based phylogeny demonstrates that subtribe Thlaspidinae comprises some additional genera such as Teesdalia, Peltaria, and Alliaria, the latter two even previously classified in other tribes than tribe Lepidieae. Received August 15, 2000 Accepted January 30, 2001     

The problematic systematic position of Ceratostratiotes Gregor (Hydrocharitaceae ?) – morphological, anatomical and biochemical comparison with Stratiotes L.

 The enigmatic fossil taxon Ceratostratiotes sinjanus (Kerner) Bužek has been considered as either a Hydrocharitaceae seed (monocot) or a Ceratophyllaceae fruit (dicot). The co-occurrence of seeds of Stratiotes kaltennordheimensis (Zenker) Keilhack (Hydrocharitaceae) and Ceratostratiotes in Early Miocene sediments in Langau (Lower Austria) enabled comparisons of morphological-anatomical features and lignin compositions, independent of diagenetic biases. The biochemistry of the Ceratostratiotes seed coat wall is not monocotyledon-like and is dissimilar to that of the co-occurring Stratiotes testae. In contrast, the Ceratostratiotes seed coat anatomy resembles that of Hydrocharitaceae genera with the micropyle and raphe being very similar to those in seeds of Stratiotes. Although the horizontal arrangement of the Ceratostratiotes spines is different from the longitudinal arrangement of the surface ridges in Stratiotes kaltennordheimensis and the spines in Blyxa (Hydrocharitaceae), the spine structures are very similar. A mixture of similarities and differences characterizes the morphological-anatomical features. Thus the systematic affinity of Ceratostratiotes still remains enigmatic. Received April 18, 2002; accepted October 18, 2002 Published online: February 4, 2003     

A multivariate and cladistic study of Vicia L. ser. Vicia (Fabaceae) based on analysis of morphological characters

 Vicia ser. Vicia, which includes the Vicia sativa aggregate, was studied by observing morphological variation. The study is an attempt to resolve the taxonomic relationships within the Vicia sativa aggregate by novel investigations along with a synthesis of the results obtained by previous researchers. The study has included some accessions and characters used previously, but has drawn accessions from a wider geographical range than has been attempted previously. Cluster analysis on the basis of 53 morphological characters justified the delimitation of four species within the series and six subspecies within the Vicia sativa aggregate. Accessions from South Asia could easily be incorporated in the classification, but some of the accessions from North Africa showed morphological differences. The morphometric data were coded using the gap weighting method of Thiele (1993) and used in a phylogenetic analysis to study the relationships between the taxa. The phylogenetic analysis, using 33 variables, showed Vicia pyrenaica closely related to Vicia sativa subsp. amphicarpa. A dichotomous key and a multi-access key to aid the identification of the taxa in the series are presented. Received January 21, 2002; accepted November 8, 2002 Published online: March 20, 2003     

Systematic relationships within and between Peucedanum and Angelica (Umbelliferae–Peucedaneae) inferred from immunological studies of seed proteins

 Immunochemical investigation of the seed storage proteins of some large genera in the tribe Peucedaneae (Umbelliferae) has clearly demonstrated the heterogeneity of Angelica and Peucedanum and the comparative homogeneity of Ferula. Among the taxa included in Angelica, Ostericum appears to be the most distant from the type species A. sylvestris, and should be treated as an independent genus. On the contrary, Archangelica is serologically similar to Angelica s. str. A rather complicated picture has been revealed within Peucedanum s. l. Serotaxonomical studies show a relationship pattern largely corresponding to the infrageneric classification of Thellung (1926). The splitting of satellite genera from Peucedanum s. l. is only partially supported. Received June 25, 2002; accepted October 3, 2002     

Gymnogrammitis dareiformis is a polygrammoid fern (Polypodiaceae) – Resolving an apparent conflict between morphological and molecular data

 Maximum parsimony and maximum likelihood analyses of the combined data sets of two chloroplast genes, rbcL and rps4, demonstrate that nk;the monotypic genus Gymnogrammitis is part of the polygrammoid clade (Polypodiaceae + Grammitidaceae), and not the Davalliaceae as proposed in most studies. The genus forms a clade together with two Asiatic genera of the Polypodiaceae, Arthromeris and Selliguea. These last two genera have either simple or once-pinnate leaves, whereas Gymnogrammitis has highly divided (3- to 4-pinnate) blades. Two characters of this genus, the basic chromosome number of x=36 and the absence of indusia, support a relationship with the Polypodiaceae. Neither feature is found within Davalliaceae. Three morphological characters support the placement of Gymnogrammitis within the selligueoid lineage of Polypodiaceae: spores with a thick perine extending in microspines, sclerenchymatous strands in the rhizome, and non-clathrate rhizome scales. These results demonstrate that molecular and morphological data are phylogenetically congruent with the exception of blade dissection. Our study clearly shows the pitfalls of classifications based on single characters, and illustrates the importance of phylogenetic assessment of all taxonomic conclusions. Received November 22, 2001; accepted May 21, 2002 Published online: November 14, 2002 Addresses of the authors: Harald Schneider (e-mail: hschneid@duke.edu), Department of Biology, Duke University, Durham, North Carolina, 27708, USA; Current address: Albrecht-von-Haller Institut für Pflanzenwissenschaften der Universit?t G?ttingen, Abteilung Systematische Botanik, Untere Karspüle 2, D-37073 G?ttingen, Germany. Alan R. Smith, Ray Cranfill, University Herbarium, University of California, Berkeley, California 94720-2465, USA. Christopher H. Haufler, Terri Hildebrand, Department of Ecology and Evolutionary Biology, University of Kansas, Lawrence, Kansas 66045, USA. Tom A. Ranker, University Museum and Department of Environmental, Population, and Organismic Biology, University of Colorado, Boulder, Colorado 80309, USA.     

A genetic linkage map of Quercus robur L. (pedunculate oak) based on RAPD, SCAR, microsatellite, minisatellite, isozyme and 5S rDNA markers

 A genetic map of Pedunculate oak (Quercus robur) was constructed based on one 5S rDNA, 271 RAPD, ten SCAR, 18 microsatellite, one minisatellite, and six isozyme markers. A total of 94 individuals from a full-sib family was genotyped. Two maps, including 307 markers, were constructed according to the “two-way pseudo-testcross” mapping strategy. Testcross markers segregating in the 1 : 1 ratio were first used to establish separate maternal (893.2 cM, 12 linkage groups) and paternal (921.7 cM, 12 linkage groups) maps. Both maps provided 85–90% genome coverage. Homologies between the male and female linkage groups were then identified based on 74 intercross markers segregating in the 3 : 1, 1 : 2 : 1 and 1 : 1 : 1 : 1 ratios (RAPDs, SCARs, SSRs, 5S rDNA and isozymes) in the hybrid progeny. In each map, approximately 18% of the studied markers showed segregation distortion. More than 60% of the skewed markers were due to an excess of heterozygote genotypes. This map will be used for: (1) studying the molecular organisation of genomic regions involved in inter- and intraspecific differentiation in oaks and (2) identification of QTLs for adaptive traits. Received: 30 January 1998 / Accepted: 12 May 1998     

Contribution to the taxonomy and phylogeny of Sarcocapnos DC. (Fumariaceae)

 To solve problems concerning the status of the taxa described in the genus Sarcocapnos, we have conducted a study using morphological, pollen morphology (light microscopy), cytogenetic and molecular techniques. Focusing on the last technique, we have sequenced ITS-1 and ITS-2 of nuclear rDNA. The species differ basically according to 5 morphological traits (leaf shape, flower spur, corolla colour, corolla size, and crest of the stigmatic surface). The cytogenetic analyses indicated n=16 to be the standard chromosome number. The ITS analyses showed that the genus is monophyletic, defining two main well-supported clades, one containing S. saetabensis and S. enneaphylla, and one containing the rest of the species. In this second clade, S. speciosa, S. pulcherrima, and S. baetica subsp. ardalii are related, as are S. integrifolia, S. crassifolia subsp. crassifolia, and S. crassifolia subsp. atlantis; S. baetica subsp. baetica forms a trichotomy with the foregoing groups. S. speciosa is shown to be a species separate from S. crassifolia subsp. crassifolia, as in the case of S. baetica with respect to S. integrifolia. Palynologically, the parameters used enabled us to establish clear differences between the taxa, often corroborating the macromorphological and genetic data. The flower spur has been reduced several times in different groups of the genus, for which the classifications established on the basis of this trait are paraphyletic. Received July 16, 2002; accepted December 11, 2002 Published online: March 31, 2003     

The correlation between development of atypical bisexual flowers and phylogeny in the Aroideae (Araceae)

 In the intermediate zone of the inflorescence of genera of Aroideae one can find flowers with male and female characteristics. Until now, two types of developmental sequences of atypical bisexual flowers (ABFs) have been recognized: the Philodendron type and the Cercestis type. In the Philodendron type, bisexual flowers generally consist of functional carpels and staminodes inserted on the same whorl. In the Cercestis type, the gynoecium and stamens are inserted on two different whorls. These different ontogenetic patterns represent two different pathways in the evolution of unisexual flowers in this subfamily. A molecular phylogenetic analysis of 33 genera of Araceae, based on the chloroplast trnL intron and trnL–F intergenic spacer sequences was carried out. We use this phylogenetic analysis and those published by French et al. (1995) and Mayo et al. (1997) to examine the distribution of the two types of ABFs in selected genera. Our results suggest that the two developmental patterns of ABFs in Aroideae sensu Mayo et al. (1997) do not correspond to two separate evolutionary lineages but rather are more or less consistent within clades. Although this new molecular phylogeny does not include all aroid genera, it corroborates in general, at the subfamily level, the molecular analysis of French et al. (1995) based on chloroplast DNA restriction site data and the analysis of Mayo et al. (1997) based on morphological and anatomical data. Received March 15, 2001 Accepted October 11, 2001     

The nectary as the primary site of infection by Erwinia amylovora (Burr.) Winslow et al.: a mini review

 Erwinia amylovora is the causative agent of fire blight, a bacterial disease existing as an unsolved problem in most countries where pome fruits like apple (Malus domestica) and pear (Pyrus communis) or ornamental plants of Rosaceae are grown. The primary site of colonization is the open flower. As for the establishment of the disease, the importance of various organs within the flowers is considerably different. The usual place for developing a large epiphytic population is the stigma. The actual infection will be attained by the external washing (rain, heavy dew) of bacteria from the stigma to the hypanthium. The bacteria penetrate through the openings of the nectary, so, the nectarthodes are the main entrance sites for them. Nectar is an excellent medium for growth of fire blight bacteria. Most often, however, the incidence of disease is significantly less than the percentage of colonized flowers. Little is known about the interrelationships of free moisture, nectar sugar concentration, ovary water potential, fine-structural characteristics of nectary versus the disease incidence and severity. The aim of this paper is to review the ecology and infection biology of Erwinia amylovora on floral surfaces and in floral tissues. Received July 8, 2002; accepted October 17, 2002 Published online: June 2, 2003