Molecular phylogeny and systematics of the pseudoxyrhophiine snake genus Liopholidophis (Reptilia, Colubridae): evolution of its exceptional sexual dimorphism and descriptions of new taxa

The pseudoxyrhophiine snake genus Liopholidophis Mocquard, 1904 is endemic to Madagascar and consists of two distinct species groups. We performed molecular phylogenetic analyses using nucleotide sequences of c . 2200 bp from two mitochondrial genes (16S rRNA, cyt b ) and one nuclear gene (c-mos) to test the monophyly of the genus Liopholidophis and to investigate the relationships of the known species and an unidentified Liopholidophis population. Our phylogeny strongly supports the polyphyly of the genus Liopholidophis , confirms the monophyly of both species groups, and reveals that the unidentified population belongs to a new species. We therefore transfer the species of the former L. stumpffi group ( L. epistibes , L. infrasignatus , L. lateralis , L. martae , and L. stumpffi ) to a new genus Bibilava gen. n. and describe Liopholidophis dimorphus sp. n. from the rainforests of Montagne d'Ambre National Park in northern Madagascar. Finally, we compare our phylogeny with previous hypotheses and discuss the evolution of the extreme sexual dimorphism in tail length of Liopholidophis s. str.     

Revision of the genus Cryptomonas (Cryptophyceae): a combination of molecular phylogeny and morphology provides insights into a long-hidden dimorphism

Seventy-three strains of cryptophytes assigned to the genera Cryptomonas, Campylomonas or Chilomonas were studied by light microscopy, spectrophotometry and whole-mount electron microscopy. Twelve groups of strains were distinguished by light and whole mount electron microscopy using a combination of characters, mainly cell size, type of periplast and presence/absence and number of pyrenoids. However, characters previously used to distinguish Cryptomonas from Campylomonas (e.g. the type of periplast: polygonal periplast plates vs. a continuous periplast sheet) were found to occur together in dimorphic strains, indicating that periplast types relate to different life-history stages of a single taxon. To evaluate the taxonomic significance of the type of periplast and other characters previously used to distinguish genera and species, representatives of each strain group were subjected to molecular phylogenetic analyses using two nuclear ribosomal DNA regions (ITS2, partial LSU rDNA) and a nucleomorph ribosomal gene (SSU rDNA). The results of the phylogenetic study provide molecular evidence for a life history-dependent dimorphism in the genus Cryptomonas: the genus Campylomonas represents the alternate morph of Cryptomonas. Campylomonas and Chilomonas are reduced to synonyms of Cryptomonas, the genus Cryptomonas is revised and typified, two new species are described and six species are emended.     

Genome comparison provides molecular insights into the phylogeny of the reassigned new genus Lysinibacillus

Background

Lysinibacillus sphaericus (formerly named Bacillus sphaericus) is incapable of polysaccharide utilization and some isolates produce active insecticidal proteins against mosquito larvae. Its taxonomic status was changed to the genus Lysinibacillus in 2007 with some other organisms previously regarded as members of Bacillus. However, this classification is mainly based on physiology and phenotype and there is limited genomic information to support it.

Results

In this study, four genomes of L. sphaericus were sequenced and compared with those of 24 representative strains belonging to Lysinibacillus and Bacillus. The results show that Lysinibacillus strains are phylogenetically related based on the genome sequences and composition of core genes. Comparison of gene function indicates the major difference between Lysinibacillus and the two Bacillus species is related to metabolism and cell wall/membrane biogenesis. Although L. sphaericus mosquitocidal isolates are highly conserved, other Lysinibacillus strains display a large heterogeneity. It was observed that mosquitocidal toxin genes in L. sphaericus were in close proximity to genome islands (GIs) and mobile genetic elements (MGEs). Furthermore, different copies and varying genomic location of the GIs containing binA/binB was observed amongst the different isolates. In addition, a plasmid highly similar to pBsph, but lacking the GI containing binA/binB, was found in L. sphaericus SSII-1.

Conclusions

Our results confirm the taxonomy of the new genus Lysinibacillus at the genome level and suggest a new species for mosquito-toxic L. sphaericus. Based on our findings, we hypothesize that (1) Lysinibacillus strains evolved from a common ancestor and the mosquitocidal L. sphaericus toxin genes were acquired by horizontal gene transfer (HGT), and (2) capture and loss of plasmids occurs in the population, which plays an important role in the transmission of binA/binB.

Electronic supplementary material

The online version of this article (doi:10.1186/s12864-015-1359-x) contains supplementary material, which is available to authorized users.Keyword: Lysinibacillus, Bacillus, Lysinibacillus sphaericus, Genome, Phylogeny     

New insights into and limitations of the molecular phylogeny in the taxon-rich land snail genus Montenegrina (Mollusca: Gastropoda: Clausiliidae)

Rock-dwelling gastropods are usually patchily distributed in limestone habitats, presumably have low active and passive dispersal ability and often represent narrow-ranged endemic taxa. Their current taxonomy is predominantly shell morphology based, and it remains unknown whether the morphologically differentiated and geographically separated populations represent phylogenetic clades. In this study, we analysed the hyperdiverse, terrestrial door snail genus Montenegrina. Based on the current taxonomy defined by shell morphology, it contains 29 species and 106 subspecies distributed in the Balkan region. The constructed phylogenetic tree using three mitochondrial markers was used to test whether it agrees with the current taxonomy. In this comprehensive tree, about half of the species and subspecies are monophyletic. Some of the paraphylies could be reasonably resolved by taxonomic changes; that is, some subspecies should be reassigned or raised to species level. Other incongruencies probably arose due to introgression even between distant clades. The histone genes turned out to be unsuitable for elucidating the phylogeny of Montenegrina. In the species-delimitation tests, considerably more molecular operational taxonomic units were delimited than the number of presently described species. The present data indicate that (a) shell morphology-based taxonomy and taxon recognition can be problematic in such a large and morphologically highly variable genus; (b) the potential error due to incomplete sampling presents a problem in a genus as variable as Montenegrina; (c) multi-locus analyses should be conducted to arrive at a better basis for species delimitation; and (d) integrative approaches including genetic as well as morphological/anatomical data from a comprehensive geographic sample are necessary.     

New insights into PKC family affairs: three novel phosphorylation sites in PKCepsilon and at least one is regulated by PKCalpha

The question of how Bax is activated during apoptosis to perform its role in permeabilization of mitochondrial membranes is intriguing for investigators in the wide field of cell death research. In their paper published in the Biochemical Journal in 2006, Capano and Crompton presented their discovery that simulated ischaemia causes rapid activation of AMPK (AMP-activated protein kinase) which phosphorylates and activates p38 MAPK (mitogen-activated protein kinase) leading to Bax activation and translocation to mitochondria in isolated cardiac myocytes. This was the first report on the molecular mechanism of Bax activation and migration during ischaemia-induced apoptosis in cardiomyocytes.     

New insights into the molecular phylogeny and taxonomy of mormyrids (Osteoglossiformes,Actinopterygii) in northern East Africa

Based on morphological data and analysis of mitochondrial cytochrome b gene and nuclear (S7 intron 1) DNA sequences, the phylogenetic relationships of all Pollimyrus species known from the Omo‐Turkana enclosed basin and Nile system below the Murchison Falls were solved. A mormyrid “Pollimyrus” petherici is distantly related to all other studied Pollimyrus species and clusters together with Cyphomyrus species forming with the later a monophyletic group. Moreover, the West African (but not the Congo River) populations of Cyphomyrus psittacus, the type species of the genus, seem to be conspecific to C. petherici. That is, the range of the genus Cyphomyrus is extended toward the Nile and Omo‐Turkana basins. This genus belongs to the large clade widely distributed in sub‐Saharian Africa and characterized by the presence of a chin appendage. Significance of this character for mormyrid phylogeny is discussed. Two distinct lineages of Pollimyrus occurring sympatrically in the White Nile tributaries and previously reported as the light and dark forms of Pollimyrus isidori together with five other congeneric species studied form a monophyletic group. The light form apparently represents P. isidori distributed in the Nile system downstream of the Murchison Falls and West Africa; the dark‐colored form (designated as Pollimyrus “D”) represents a distinct phylogenetic lineage inhabiting both the Omo‐Turkana and the White Nile basin. Morphological and ecological data suggest that this form may be conspecific to East African Pollimyrus nigricans or most probably represents a new species.     

Molecular phylogeny and phylogeography of genus Pseudois (Bovidae,Cetartiodactyla): New insights into the contrasting phylogeographic structure

Blue sheep, Pseudois nayaur, is endemic to the Tibetan Plateau and the surrounding mountains, which are the highest‐elevation areas in the world. Classical morphological taxonomy suggests that there are two subspecies in genus Pseudois (Bovidae, Artiodactyla), namely Pseudois nayaur nayaur and Pseudois nayaur szechuanensis. However, the validity and geographic characteristics of these subspecies have never been carefully discussed and analyzed. This may be partially because previous studies have mainly focused on the vague taxonomic status of Pseudois schaeferi (dwarf blue sheep). Thus, there is an urgent need to investigate the evolutionary relationship and taxonomy system of this genus. This study enriches a previous dataset by providing a large number of new samples, based on a total of 225 samples covering almost the entire distribution of blue sheep. Molecular data from cytochrome b and the mitochondrial control region sequences were used to reconstruct the phylogeny of this species. The phylogenetic inferences show that vicariance plays an important role in diversification within this genus. In terms of molecular dating results and biogeographic analyses, the striking biogeographic pattern coincides significantly with major geophysical events. Although the results raise doubt about the present recognized distribution range of blue sheep, they have corroborated the validity of the identified subspecies in genus Pseudois. Meanwhile, these results demonstrate that the two geographically distinct populations, the Helan Mountains and Pamir Plateau populations, have been significantly differentiated from the identified subspecies, a finding that challenges the conventional taxonomy of blue sheep.     

A new species of electric knifefish,genus Compsaraia (Gymnotiformes: Apteronotidae) from the Amazon River,with extreme sexual dimorphism in snout and jaw length

Abstract

A new species of the neotropical electric fish genus Compsaraia is described from the western Amazon of Peru and Brazil. Compsaraia samueli is distinguishable from all other apteronotids by sexual dimorphism in which mature males exhibit extreme elongation and slenderness of the snout and jaws. Compsaraia samueli is readily distinguishable from its only congener, C. compsa, by more caudal‐fin rays (17–18 vs. 13–16), a shorter caudal peduncle (mean length 9% vs. 34% body length to end of anal fin), a less tapering body shape in lateral profile (mean ratio of body depth at origins of anal fin and dorsal organ 93% vs. 75%), and a smaller maximum adult body size (230 vs. 305 mm). The genus Compsaraia is readily separated from other apteronotids by a pale antorbital stripe and a pale L‐shaped patch over the supra‐temporal canal. The phylogenetic position of C. samueliis estimated by inclusion in a previously published data matrixof osteological and other morphological characters. Comparisons of the cranial bones in apteronotids shows the derived morphology of C. samueli to be a composite of three developmentally and phylogenetically discrete characters: (1) positive allometric growth before sexual maturity in both sexes of the pre‐orbital region of the neurocranium, (2) positive allometric growth of the (oral) jaws, and (3) secondary sexual dimorphism of snout and jaw morphology. The genus Compsaraia represents one of at least three phylogenetically independent cases of snout elongation and one of at least four cases of jaw elongation within the Apteronotidae. Compsaraia samueli also represents one of at least four cases of secondary sexual dimorphism in snout and jaw length within the Apteronotidae. The phylogenetic distribution of snout and jaw characters within the Apteronotidae suggests the influence of both sexual and trophic functional influences on the evolution of head morphology.     

New insights into the systematics of Malagasy mongoose‐like carnivorans (Carnivora,Eupleridae, Galidiinae) based on mitochondrial and nuclear DNA sequences

The Malagasy carnivorans (Eupleridae) comprise seven genera and up to ten species, depending on the authority, and, within the past decades, two new taxa have been described. The family is divided into two subfamilies, the Galidiinae, mongoose‐like animals, and the Euplerinae, with diverse body forms. To verify the taxonomic status of Galidiinae species, including recently described taxa, as well as some recognized subspecies, we studied intrageneric genetic variation and structure, using both mitochondrial and nuclear markers. Our results suggest the recognition of four species in the Galidiinae, rendering each genus monospecific. We propose to recognize three subspecies in Galidia elegans (G. e. dambrensis, G. e. elegans, and G. e. occidentalis), two subspecies in Mungotictis decemlineata (M. d. decemlineata and M. d. lineata), and two subspecies in Galidictis fasciata (G. f. fasciata and G. f. grandidieri, the latter was recently described as a distinct species). Our results indicate also that Salanoia durrelli should be treated as a junior synonym of Salanoia concolor. Low levels of intraspecific divergence revealed some geographical structure for the Galidiinae taxa, suggesting that environmental barriers have isolated certain populations in recent geological time. All taxa, whether at the species or subspecies level, need urgent conservation attention, particularly those with limited geographical distributions, as all are threatened by forest habitat degradation.     

New insights into the phylogeny and biogeography of subfamily Orontioideae (Araceae)

Proto‐Araceae, the earliest diverged lineage within the family Araceae, includes two subfamilies, Gymnostachydoideae (one species) and Orontioideae (eight species). Based on an extensive sampling (a total of 198 accessions) of six chloroplast non‐coding regions (5799 aligned sites), we assessed phylogenetic relationships among the genera and species within subfamily Orontioideae and estimated the timing of intercontinental disjunct events in the Northern Hemisphere. Overall phylogenetic relationships among the genera were consistent with results from previous studies, but several new important findings were discovered, primarily within Symplocarpus Salisb. ex W. P. C. Barton. First, two major lineages within Symplocarpus were identified: one lineage included S. foetidus (L.) Salisb. ex W. Barton, S. nabekuraensis Otsuka & K. Inoue, and S. renifolius Schott ex Tzvelev (Japan), whereas the other included S. nipponicus Makino, S. egorovii N. S. Pavlova & V. A. Nechaev, and S. renifolius (Korea). Symplocarpus renifolius in Japan was tetraploid and closely related to the tetraploid S. foetidus in eastern North America. Populations of S. renifolius in Korea were confirmed to be diploid (2n = 30) and shared the most recent common ancestor with the other diploid species, S. nipponicus. Second, two recently described species, S. nabekuraensis and S. egorovii, were deeply embedded within S. renifolius in Japan and Korea, respectively, and their distinct taxonomic status requires further assessment. Finally, two intercontinental disjunction events in the subfamily, one in Lysichiton Schott between eastern Asia and western North America and the other in Symplocarpus between eastern Asia and eastern North America, were estimated to be between 4.5 and 1.4 million years ago (Pliocene and Pleistocene) and between 1.9 and 0.5 million years ago (Pleistocene), respectively.     

Towards a phylogeny and definition of species at the molecular level within the genus Mycobacterium

16S rRNA sequences from Mycobacterium tuberculosis, M. avium, M. gastri, M. kansasii, M. marinum, M. chelonae, M. smegmatis, M. terrae, M. gordonae, M. scrofulaceum, M. szulgai, M. intracellulare, M. nonchromogenicum, M. xenopi, M. malmoense, M. simiae, M. flavescens, M. fortuitum, and M. paratuberculosis were determined and compared. The sequence data were used to infer a phylogenetic tree, which provided the basis for a systematic phylogenetic analysis of the genus Mycobacterium. The groups of slow- and fast-growing mycobacteria could be differentiated as distinct entities. We found that M. simiae occupies phylogenetically an intermediate position between these two groups. The phylogenetic relatedness within the slow-growing species did not reflect the Runyon classification of photochromogenic, scotchromogenic, and nonchromogenic mycobacteria. In general, the phylogenetic units identified by using rRNA sequences confirmed the validity of phenotypically defined species; an exception was M. gastri, which was indistinguishable from M. kansasii when this kind of analysis was used.     

Distribution and molecular phylogeny of the dinoflagellate genus Ostreopsis in Thailand

The toxic benthic dinoflagellate genus Ostreopsis has been connected to the production of palytoxin and its analogs in many tropical and temperate areas. Although the type species, O. siamensis, was originally described from the Gulf of Thailand in 1901, little is known about the species composition and distribution of the genus Ostreopsis in Thailand. In this study, a total of 64 Ostreopsis strains isolated from the Andaman Sea as well as the Gulf of Thailand were investigated by analyzing the nucleotide sequences of the LSU rDNA D1/D2, D8/D10 and ITS-5.8S rDNA regions. Phylogenetic analyses (BI and ML) resulted in some of the strains being assigned to previously described clades, O. cf. ovata and Ostreopsis sp. 6, and revealed the existence of a novel clade named Ostreopsis sp. 7, which exhibited large genetic distances from the other clades. Among O. cf. ovata, several strains from Thailand were formed into a new subclade, the Thailand subclade, whereas a few strains belonged to the South China Sea subclade. Morphometric characteristics such as the cell sizes of the two O. cf. ovata subclades and those of Ostreopsis sp. 7 were not significantly different from each other (p > 0.05). Their characteristics were similar but slightly different from those of O. ovata and were significantly different from those of Ostreopsis sp. 6 (p < 0.05). Toxicities of Ostreopsis from Thailand were evaluated using mouse bioassay. Strains of Ostreopsis sp. 6 and Ostreopsis sp. 7 tested were highly toxic, while the two subclades of O. cf. ovata strains seemed to be nontoxic. This study suggests that toxic Ostreopsis sp. 7 is distributed in the Andaman Sea, whereas the two subclades of O. cf. ovata and toxic Ostreopsis sp. 6 are distributed in the Gulf of Thailand.     

Pholcid spider molecular systematics revisited,with new insights into the biogeography and the evolution of the group

We analysed seven genetic markers sampled from 165 pholcids and 34 outgroups in order to test and improve the recently revised classification of the family. Our results are based on the largest and most comprehensive set of molecular data so far to study pholcid relationships. The data were analysed using parsimony, maximum‐likelihood and Bayesian methods for phylogenetic reconstruction. We show that in several previously problematic cases molecular and morphological data are converging towards a single hypothesis. This is also the first study that explicitly addresses the age of pholcid diversification and intends to shed light on the factors that have shaped species diversity and distributions. Results from relaxed uncorrelated lognormal clock analyses suggest that the family is much older than revealed by the fossil record alone. The first pholcids appeared and diversified in the early Mesozoic about 207 Ma ago (185–228 Ma) before the breakup of the supercontinent Pangea. Vicariance events coupled with niche conservatism seem to have played an important role in setting distributional patterns of pholcids. Finally, our data provide further support for multiple convergent shifts in microhabitat preferences in several pholcid lineages. Our findings suggest that both adaptive and non‐adaptive speciation may have played an important role in the diversification of pholcid lineages.     

Molecular phylogeny and systematics of Neotropical toucanets in the genus Aulacorhynchus (Aves,Ramphastidae)

Bonaccorso, E., Guayasamin, J. M., Peterson, A. T. & Navarro‐Sigüenza, A. G. (2011). Molecular phylogeny and systematics of Neotropical toucanets in the genus Aulacorhynchus (Aves, Ramphastidae). —Zoologica Scripta, 40, 336–349. We studied the phylogenetic relationships in the genus Aulacorhynchus, an assemblage of Neotropical toucanets distributed from Mexico south to Bolivia. Based on mitochondrial and nuclear DNA characters, we obtained a robust hypothesis of relationships for all recognized species, including good representation of distinct geographic populations. Our results support the monophyly of the genus Aulacorhynchus, but contradict previous taxonomic arrangements. The genus is made up of three major clades: the Aulacorhynchus prasinus complex, Aulacorhynchus huallagae + Aulacorhynchus coeruleicinctis, and Aulacorhynchus haematopygus + Aulacorhynchus sulcatus + Aulacorhynchus derbianus. Andean populations of A. derbianus are more closely related to A. sulcatus than to Pantepuian populations of A. derbianus, rendering A. derbianus paraphyletic. Based on the molecular phylogeny, and information on geographic distributions and morphological and behavioural characters, we review the specific status of these taxa and propose a new taxonomic arrangement within Aulacorhynchus.     

Diet and sexual dimorphism in the very catholic lizard genus,Leiocephalus of the Bahamas

Summary Over 5,000 prey items from specimens of Bahamian Leiocephalus lizards were measured and identified taxonomically. The diet in general consists mainly of arthropods, but much plant matter is also eaten, including flowers and buds as well as fruit. Lizards comprise about 2% of the diet by volume. Individuals inhabiting relatively small islands are more likely to have eaten plant matter than those from relatively large islands. Within the most widespread species (carinatus), sexual dimorphism in size is greater, the smaller the number of sympatric species in its structural habitat. Prey-size differences between differently sized Leiocephalus are greater, the greater the dimorphism. However, even the most dimorphic sexes take rather similar prey sizes. For all Bahamian species combined, the inverse correlation of sexual dimorphism with sympatric species is not as strong as an inverse correlation with latitude. We suggest that sexual selection on female size to increase the clutch size that can be carried may have affected sexual dimorphism in the genus.     

Structure and molecular phylogeny of sasA genes in cyanobacteria: insights into evolution of the prokaryotic circadian system

Cyanobacteria are the simplest organisms known to have a circadian system. In addition to the three well-studied kai genes, kaiA, kaiB, and kaiC, an important element of this system is a two-component sensory transduction histidine kinase sasA. Using publicly available data of complete prokaryotic genomes, we performed structural and phylogenetic analyses of the sasA genes. Results show that this gene has a triple-domain structure, and the domains are under different selective constraints. The sasA gene originated in cyanobacteria probably through the fusion of the ancestral kaiB gene with a double-domain, two-component sensory transduction histidine kinase. The results of the phylogenetic analyses suggest that sasA emerged before the kaiA gene, about 3,000-2,500 MYA, and has evolved in parallel with the evolution of the kaiBC cluster. The observed concordant patterns of the sasA and kaiBC evolution suggest that these genes might compose an ancient KaiBC-SasA-based circadian system, without the kaiA gene, and that such a system still exists in some unicellular cyanobacteria.     

Light at the end of the tunnel: insights into the molecular systematics of East African puddle frogs (Anura: Phrynobatrachidae)

To investigate intra- and interspecific variability of ‘Little Brown Frogs’ (LBFs) from East Africa, we analysed molecular data from 23 species of Phrynobatrachus using 16S rRNA, a mitochondrial gene considered a universal DNA barcoding marker in amphibians. Conspecific populations, excluding P. natalensis, exhibited low (0–0.5%) sequence divergences, while conspecific populations of P. natalensis exhibited a wide range of differentiation (0.2–6.4%) with a distinct bimodal distribution, affirming that this may be a complex of cryptic species. Sister species yielded a range of differentiation from 1.5–8.3% with median of 2.3%. Results revealed that at least three non-sister populations of miniaturized puddle frogs are currently identified as P. mababiensis, and a new species from Ethiopia was identified. This first step into revising East African Phrynobatrachus systematics illustrates the necessity of rigorous taxonomic study, especially for groups with confusing or highly incomplete taxonomy.     

New insights into the systematics of the enigmatic Polynesian sandpipers Aechmorhynchus parvirostris and Prosobonia leucoptera

With only a single extant representative, endemic to the Tuamotu Archipelago, the Polynesian sandpipers (Aechmorhynchus and Prosobonia) may have had a larger distribution in Eastern Polynesia in the past, with four endemic taxa. Although these aberrant sandpipers' membership to the Scolocapidae has been well supported, finding their closest living taxa has proved difficult and the phylogenetic relationships of these taxa have remained unresolved. We present the first molecular analysis of the Polynesian sandpipers, including sampling of the only known specimen of the extinct Prosobonia leucoptera, collected in 1773. Based on mitochondrial and nuclear gene sequence data, the phylogenetic analyses demonstrate that the Polynesian sandpipers were sister taxa and belonged to the clade that included the other sandpipers (Calidris and allies) and turnstones (Arenaria), although without a close relative among extant genera. Divergence time estimates suggested that the lineage leading to Prosobonia diverged from the other extant sandpipers during the Oligocene and that either the Line Islands or the Tuamotu Archipelago were probably the first archipelagos colonized by the Prosobonia lineage. On the basis of these results, we suggest that Aechmorhynchus parvirostris and Prosobonia leucoptera be regarded as related species within the same genus, and thus that the senior name Prosobonia be used for both taxa.     

New insights into molecular structure and function of ecto-nucleotidases in the nervous system

Nucleotides can be released as signaling substances in the nervous system from both neural and glial cells. Their function is terminated by ecto-nucleotidases and sequential extracellular metabolism to the nucleoside. Recently considerable progress has been made in unraveling the molecular structure of an ecto-ATPase and an ecto-ATP diphosphohydrolase, two closely related ecto-enzymes. Molecular structure, tissue distribution and functional properties of the ecto-nucleotidases are discussed with particular emphasis on the nervous system. © 1998 Elsevier Science Ltd. All rights reserved.     

Molecular phylogeny of the neotropical genus Christensonella (Orchidaceae, Maxillariinae): species delimitation and insights into chromosome evolution

Background and Aims

Species'' boundaries applied within Christensonella have varied due to the continuous pattern of variation and mosaic distribution of diagnostic characters. The main goals of this study were to revise the species'' delimitation and propose a more stable classification for this genus. In order to achieve these aims phylogenetic relationships were inferred using DNA sequence data and cytological diversity within Christensonella was examined based on chromosome counts and heterochromatin patterns. The results presented describe sets of diagnostic morphological characters that can be used for species'' identification.

Methods

Phylogenetic studies were based on sequence data of nuclear and plastid regions, analysed using maximum parsimony and maximum likelihood criteria. Cytogenetic observations of mitotic cells were conducted using CMA and DAPI fluorochromes.

Key Results

Six of 21 currently accepted species were recovered. The results also support recognition of the ‘C. pumila’ clade as a single species. Molecular phylogenetic relationships within the ‘C. acicularis–C. madida’ and ‘C. ferdinandiana–C. neowiedii’ species'' complexes were not resolved and require further study. Deeper relationships were incongruent between plastid and nuclear trees, but with no strong bootstrap support for either, except for the position of C. vernicosa. Cytogenetic data indicated chromosome numbers of 2n = 36, 38 and 76, and with substantial variation in the presence and location of CMA/DAPI heterochromatin bands.

Conclusions

The recognition of ten species of Christensonella is proposed according to the molecular and cytogenetic patterns observed. In addition, diagnostic morphological characters are presented for each recognized species. Banding patterns and chromosome counts suggest the occurrence of centric fusion/fission events, especially for C. ferdinandiana. The results suggest that 2n = 36 karyotypes evolved from 2n = 38 through descendent dysploidy. Patterns of heterochromatin distribution and other karyotypic data proved to be a valuable source of information to understand evolutionary patterns within Maxillariinae orchids.Key words: Chromosome number, Christensonella, Cymbidieae, cytotaxonomy, fluorochrome staining, Maxillaria, Maxillariinae, molecular phylogenetics, species delimitation