Satellite-DNA evolutionary patterns under a complex evolutionary scenario: the case of Acrolophus subgroup (Centaurea L., Compositae) from the western Mediterranean

Within the genus Centaurea (subtribe Centaureinae, tribe Cardueae, Compositae) hybridizations and reticulate-evolution phenomena have widely been recognized. This is especially true in the taxa included in the subgroup Acrolophus from the western Mediterranean area, in which recurrent hybridizations of parapatric ("microallopatric") lineages within the geographical range of a primary radiation have been suggested. The subgroup Acrolophus includes taxa from three sections (i.e. Acrolophus, Phalolepis and Willkommia), and, together with other subgroups, forms the named Jacea group (one of the three main groups into which Centaurea is divided). In this paper, we have studied the influence that the complex evolutionary scenario described for the Acrolophus subgroup from the western Mediterranean exerts on the evolutionary pattern of a satellite-DNA family, the HinfI family, which exists within the genomes of these taxa. To this end, we have analyzed the evolution of this satellite-DNA family in taxa from different taxonomic comparative levels: i) seven subspecies of the C. boissieri complex (one of which with two varieties) of the sect. Willkommia; ii) species of the sections Willkommia (10 species, 19 taxa), Acrolophus (two species), and Phalolepis (two species), all in the Acrolophus subgroup; iii) one external species to the Jacea group, C. granatensis from the group Acrocentron; iv) and species from other related genera from the Centaureinae subtribe (Phonus and Carthamus, both belonging to the Carthamus group). The influence of the suggested model for the origin and diversification of the Acrolophus subgroup is evidenced by the existence of three different HinfI satellite-DNA subfamilies coexisting in some genomes, and by the analysis that we have made by comparing site-by-site the transition stages in the process of concerted evolution between the sequences of the each subfamily. From this analysis, we can deduce that the HinfI repeated subfamilies evolved in a gradual manner, and that the different stages of concerted evolution fit quite well with the combined nuclear-chloroplast-DNA-deduced divergences and phylogeny of the subtribe Centaureinae. The HinfI satellite-DNA from the Carthamus species group (genera Carthamus and Phonus) and from the Acrocentron group (Centaurea granatensis) shows a high intraspecific conservation of the repeats, suggesting that the mechanisms producing concerted evolution have been efficient in these taxa. In addition, the comparison of individual nucleotide positions between related species shows a paucity in the spreading of variants in each subfamily with satellite-DNA divergence, an indication of a constant rate of homogenization of the repeated cluster. On the contrary, this trend is absent in the comparisons of the HinfI sequences from taxa of the subgroup Acrolophus. In this subgroup, we have found in this repetitive family similar representative average sequences for each taxon analyzed, polymorphic sites in each taxon being scant, most of them autapomorphic, representing early stages of genetic differentiation between taxa in the process of concerted evolution. The absence of concerted evolution was visualized by similar levels of intraspecific variation and interspecific divergence and by the lack of fixed species-diagnostic nucleotide sites. These facts might reflect the reticulate mode of evolution of Acrolophus.     

Karyology, generic delineation and dysploidy in the genera Carduncellus, Carthamus and Phonus (Asteraceae)

Within the Mediterranean complex Carduncellus Carthamus Phonus , taxonomic classification has proven problematical. A recent molecular analysis has clarified the generic delineation, while a revision of the abundant karyological data has suggested that correlation of taxonomic classification and karyology would be of great interest. In this paper, a confirmation of chromosome numbers in the complex is offered, with 44 populations of 30 taxa studied. Six of the counts are new. A short analysis of the correlation between base chromosome numbers and generic classification follows. The main conclusion is that descending dysploidy is the main mechanism of karyological evolution in the complex, as has been the case in other genera of the subtribe.     

Palynological Features as a Systematic Marker in Artemisia L. and Related Genera (Asteraceae, Anthemideae) - II: Implications for Subtribe Artemisiinae Delimitation

Abstract: Using optical and scanning electron microscopy, we completed a palynological study of the subtribe Artemisiinae (Asteraceae, Anthemideae), which we started in a previous paper. This subtribe contains different genera with a systematic position that, in many cases, has been controversial. There is a group of genera closely related to Artemisia, and another one more related to Chrysanthemum sensu lato. We confirm the existence of two pollen morphological patterns - concerning exine ornamentation - in the tribe Anthemideae and in the subtribe Artemisiinae as currently considered: one with long spines ( Anthemis type) and the other with short spinules ( Artemisia type). This feature is a good taxonomic marker, well correlated with other morphological and with molecular characters. This enables a new delimitation of the subtribe Artemisiinae, which is characterized by pollen grain ornamentation constituted by short spinules (microechinate pollen), and should be restricted to Artemisia and the closely related genera that share this trait.     

Generic Delimitation and Phylogeny of the Subtribe Centaureinae (Asteraceae): A Combined Nuclear and Chloroplast DNA Analysis

Taxonomic complexity has hindered the partitioning of the artificial genus Centaurea, even though it has long been recognized as a polyphyletic assemblage. On the basis of morphology, pollen type, karyology and DNA sequence analysis, previous workers have defined five informal groups in the genus (Acrocentron,Centaureasensu stricto , Cyanus, Jacea and Psephellus). However, the precise delimitation of these groups and their relationships remain largely unknown. Moreover, although some informal groups have been established among the rest of the subtribe (Amberboa, Carthamus and Stemmacantha), many genera cannot be classified in any group. Newer molecular approaches are essential for resolving these problems. Sequences of the internal transcribed spacers (ITS) of nuclear ribosomal DNA and the chloroplast gene mat K were analysed for a comprehensive sample of the whole subtribe, with the aim of clarifying the delimitation and the phylogeny of the groups of the Centaureinae. Results largely confirm the suggested informal entities as natural groups, with some interesting changes of placement of some genera, especially in the Acrocentron and the Stemmacantha groups. Our results confirm that the sections of Centaurea with Dealbata pollen type should be classified as a different genus, Psephellus. In addition to morphology and suggested pollen type evolution, the ITS and mat K phylogenies are also supported by karyological evidence. Our results confirm that the natural delimitation of Centaurea that minimizes nomenclatural changes is possible only if a new type of the genus is designated.     

Evolution of pollen morphology in Loranthaceae

Earlier studies indicate a strong correlation of pollen morphology and ultrastructure with taxonomy in Loranthaceae. Using high-resolution light microscopy and scanning electron microscopy imaging of the same pollen grains, we document pollen types of 35 genera including 15 studied for the first time. Using a molecular phylogenetic framework based on currently available sequence data with good genus-coverage, we reconstruct trends in the evolution of Loranthaceae pollen and pinpoint traits of high diagnostic value, partly confirming earlier intuitive hypotheses based on morphological observations. We find that pollen morphology in Loranthaceae is strongly linked to phylogenetic relationships. Some pollen types are diagnostic for discrete genera or evolutionary lineages, opening the avenue to recruit dispersed fossil pollen as age constraints for dated phylogenies and as independent data for testing biogeographic scenarios; so far based exclusively on modern-day data. Correspondences and discrepancies between palynological and molecular data and current taxonomic/systematic concepts are identified and suggestions made for future palynological and molecular investigations of Loranthaceae.     

Pollen Morphology of Eight Genera of the Subtribe Mutisiinae Less. sensu Bremer (Compositae) from Asia

The pollen morphology of 28 species and one variety representing eight genera of the subtribe Mutisiinae Less. sensu Bremer (Compositae) was investigated by scanning electron microscopy (SEM) and light microscopy. The pollen grains are usually radically symmetrical, isopolar, and tricolporate. The shape varies from spheroidal to subprolate in equatorial view and is three-lobed circular in polar view. Based on exine sculpturing observed under SEM, the pollen grains can be divided into three types: (i) type Ⅰ, with macrogranulate sculpture; (ii) type Ⅱ, with a finely granulate sculpture; and (iii) type Ⅲ, with spiny sculpture.The palynological data showed that the Macroclinidium Maxim. was remarkably distinguished from those of the other seven genera. The variation of pollen characteristics within the Mutisiinae in Asia is little at species level, but it proves some useful information for studying the relationships among genera and it has an important significance in further understanding the evolutionary history of Mutisiinae.     

Pollenmorphologie und Chromosomenzahlen der Poaceen-Subtribus Deschampsiinae Holub

Pollen morphology and ultrastructure of twenty-seven European species of the subtribe Deschampsiinae Holub were investigated by light, phase contrast and scanning electron microscopy. Genera are characterized by pollen size, development of the anuli and pollen surface. Comparison of the data with the known chromosome numbers and some karyotypes confirms the important influence of karyology on the size and fine structure of pollen.     

Pollen Morphology of Eight Genera of the Subtribe Mutisiinae Less. sensu Bremer （Compositae） from Asia

The pollen morphology of 28 species and one variety representing eight genera of the subtribe Mutisiinae Less. sensu Bremer （Compositae） was investigated by scanning electron microscopy （SEM） and light microscopy. The pollen grains are usually radically symmetrical, isopolar, and tricolporate. The shape varies from spheroidal to subprolate in equatorial view and is three-lobed circular in polar view. Based on exine sculpturing observed under SEM, the pollen grains can be divided into three types： （i） type Ⅰ, with macrogranulate sculpture; （ii） type Ⅱ, with a finely granulate sculpture; and （iii） type Ⅲ with spiny sculpture. The palynological data showed that the Macroclinidium Maxim. was remarkably distinguished from those of the other seven genera. The variation of pollen characteristics within the Mutisiinae in Asia is little at species level, but it proves some useful information for studying the relationships among genera and it has an important significance in further understanding the evolutionary history of Mutisiinae.     

POLLEN MORPHOLOGY AND DETAILED STRUCTURE OF FAMILY COMPOSITAE,TRIBE CICHORIEAE. I. SUBTRIBE STEPHANOMERIINAE

A survey of pollen morphology in 20 species representing the 11 genera of the North American subtribe Stephanomeriinae by light, scanning electron, and transmission electron microscopy revealed 10 of the 11 genera to have echinate, tricolporate pollen grains, Lygodesmia being the only genus with echinolophate pollen. Sectioned exines of most of the species examined are similar, being composed of ektexine and endexine. The ektexine surface is composed of spines which typically have globose perforate bases. A cavus occurs as a separation between the basis (foot layer) and the columellae in all of the genera examined except Chaetadelpha. Pollen of the two species of Glyptopleura were found to be strikingly different in exomorphology. Pollen of the putatively self-fertile G. marginata has much shorter spines than the closely related G. setulosa. Atrichoseris, Anisocoma, Calycoseris, Glyptopleura, Pinaropappus, Prenanthella, and most species of Malacothrix have pollen which lack paraporal ridges. The remaining genera, Chaetadelpha, Lygodesmia, Rafinesquia, and Stephanomeria have well-developed ridges of fused spine bases around the apertures. Pollen characters, particularly those of the aperture region, have been found to be systematically useful in the subtribe, therefore acetolyzed material gives more useful information than untreated pollen.     

POLLEN MORPHOLOGY OF HAPLOPAPPUS AND RELATED GENERA (COMPOSITAE—ASTEREAE)

Pollen grains of Haplopappus and related genera in the subtribe Solidaginae from North and South America were examined by light and scanning electron microscopy. The grains are consistently tricolporate and echinate. Some genera can be distinguished by pollen size, spine length, and number of spine rows between colpi. Based on these characters, the divergence of Benitoa from other members of the subtribe, as indicated by its morphology and secondary chemistry, is supported. Additionally, the recently suggested absence of a close relationship between Pyrrocoma and Oonopsis is indicated by their contrasting pollen types. This study demonstrates the potential of pollen studies in distinguishing some taxonomic groups in the Astereae.     

Pollen morphology of the Gardenieae-Gardeniinae (Rubiaceae)

Pollen grains of 107 species of 61 genera of the Gardeniinae were investigated by scanning electron and light microscopy. The subtribe is europalynous and grains occur either as monads, tetrads or massulae. Grains are porate, colporate or rarely pororate. The majority of the genera has 3-zonoaperturate pollen grains. However, deviating numbers of apertures occur in Byrsophyllum (4–8) and Pseudomantalania (4–5). Pantoporate pollen occurs in Randia . The average equatorial size (E) of monads varies between 13 μm in Anomanthodiu to 59 μm in Posoqueria . The tetrads vary from 37 μm in Casasia to 61 μm in Calochone and Euclinia . The exine is usually reticulate, but also foveolate, rugulate, perforate and psilate exine occurs in the subtribe. The thickness of the exine is only 0.3 μm in tetrad grains of Randia armata and in the pantoporate monad grains of R. ruiziana , whereas the thickest exine occurs in Posoqueria (2.8 pm) and Euclinia (2.9 μm). Tectal excrescenses are generally absent. However, three genera with pollen in tetrads, namely Euclinia, Gardenia and Oligoco-don , show this feature. Contrary to the subtribe most genera are fairly stenopalynous, but in the neotropical genera Alibertia and Randia several different types of pollen are encountered.     

Comparative pollen morphology and ultrastructure of Mentheae subtribe Nepetinae (Lamiaceae)

This study provides new pollen data of 52 representative species belonging to all 12 genera in the currently classification of the subtribe Nepetinae, and considers the possible presence of orbicules for the first time. Pollen morphology and ultrastructure were investigated with light, scanning electron and transmission electron microscopy. Nepetinae pollen is small to large (P = 16–65 µm, E = 17–53 µm), oblate to prolate (P/E = 0.7–1.6) in shape and mostly hexacolpate (sometimes octocolpate). The exine stratification in all taxa studied is similar and characterized by unbranched columellae and a continuous, granular endexine. Sexine ornamentation in the Nepetinae is bireticulate, microreticulate or perforate. In perforate and microreticulate pattern a tendency towards a bireticulum could be recognized due to trace of secondary tectal connections. The bireticulate pattern is most common with variations of primary muri and secondary reticulum. In Hymenocrater and Schizonepeta the observed variation of sexine ornamentation is particularly valuable at the generic level. Pollen data support Lophanthus and Nepeta as very closely allied and Lallemantia is clearly distinct from Dracocephalum. The formerly suggested infrageneric relationships within Dracocephalum and Nepeta are only partly corroborated by palynological characters. Orbicules are absent in the Nepetinae.     

Differential spreading of HinfI satellite DNA variants during radiation in Centaureinae

Background and Aims

Subtribe Centaureinae appears to be an excellent model group in which to analyse satellite DNA and assess the influence that the biology and/or the evolution of different lineages have had on the evolution of this class of repetitive DNA. Phylogenetic analyses of Centaureinae support two main phases of radiation, leading to two major groups of genera of different ages. Furthermore, different modes of evolution are observed in different lineages, reflected by morphology and DNA sequences.

Methods

The sequences of 502 repeat units of the HinfI satellite DNA family from 38 species belonging to ten genera of Centaureinae were isolated and compared. A phylogenetic reconstruction was carried out by maximum likelihood and Bayesian inference.

Key Results

Up to eight different HinfI subfamilies were found, based on the presence of a set of diagnostic positions given by a specific mutation shared by all the sequences of one group. Subfamilies V–VIII were mostly found in older genera (first phase of radiation in the subtribe, late Oligocene–Miocene), although some copies of these types of repeats were also found in some species of the derived genera. Subfamilies I–IV spread mostly in species of the derived clade (second phase of radiation, Pliocene to Pleistocene), although repeats of these subfamilies exist in older species. Phylogenetic trees did not group the repeats by taxonomic affinity, but sequences were grouped by subfamily provenance. Concerted evolution was observed in HinfI subfamilies spread in older genera, whereas no genetic differentiation was found between species, and several subfamilies even coexist within the same species, in recently radiated groups or in groups with a history of recurrent hybridization of lineages.

Conclusions

The results suggest that the eight HinfI subfamilies were present in the common ancestor of Centaureinae and that each spread differentially in different genera during the two main phases of radiation following the library model of satellite DNA evolution. Additionally, differential speciation pathways gave rise to differential patterns of sequence evolution in different lineages. Thus, the evolutionary history of each group of Centaureinae is reflected in HinfI satellite DNA evolution. The data reinforce the value of satellite DNA sequences as markers of evolutionary processes.     

Primary identifications and palynological observations of Perilla in China

Unresolved controversies concerning classification of the monotypic genus Perilla L. have hindered the complete understanding and subsequent sustainable use of these vital food, oil and medicinal plants to their full potential. We attempted to use scanning electron microscopy to obtain palynological evidence from Perilla plants of 21 samples from seven provinces in China as a potential extra attribute in classification. The findings showed that pollen grains from plants of 11 samples were oblate, while those of the other 10 were suboblate in shape, and there were no any type of prolate pollen grains being observed. Pollen grains of all the samples had diverse exine ornamentations. Based on whether having continuous tecta on the ornamentations, all of the pollen grains derived from the 21 samples were classified into two categories, fourteen of them with irregular reticulates, seven with continuous tecta with no perforations. None of the samples were bireticulates. The ornamentation pattern and size of pollen grains jointly provided evidence that it is appropriate to classify the genus Perilla into five varieties of one species. Furthermore, by comparison, it is concluded that shapes and exine ornamentations of Perilla are unique among those of the seven genera already investigated in the subfamily Lamioideae. Using these pollen features, Perilla could be easily distinguished from two other subtribes (Menthinae Briq. and Thyminae Briq.) in the same tribe, supporting the view that Perilla and other four related genera were divided into one subtribe (Perillinae).     

Pollen studies in the genus Echinops L. and Xeranthemum group (Asteraceae)

The phylogeny of the genus Echinops and the Xeranthemum complex were the subject of a previous molecular study of the tribe Cardueae. An electronic microscope survey of pollen morphology and ultrastructure is presented for all the genera belonging to these groups, allowing four pollen types to be identified. The exine pattern indicates that there are two pollen types in the genus Echinops (including Acantholepis ), although no differences have been found between sections in Echinops . First, the annual species of Echinops and Acantholepis orientalis show a pollen type that differs from that of perennial species of the former genus and, second, the genera Chardinia and Xeranthemum exhibit a pollen type distinct from that Amphoricarpos and Siebera . The internal structure of the pollen wall is the same for all the genera. Only slight differences were observed in the tectum, but in all cases these were of the anthemoid type. Statistically significant correlations between pollen size and the life cycle were found.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 154 , 549–557.     

Pollen morphology and systematics of Burseraceae

The Burseraceae are a medium‐sized family in which 18 genera are currently recognised. They are the subject of a long‐term project to describe the pollen morphology from light, scanning electron and transmission electron microscopy. The pollen morphology of tribe Protieae has been published, as well as an account of the pollen of the African taxa in the family. Pollen data for the other two tribes, Bursereae and Canarieae, are more or less complete. The pollen of all the genera have been examined, with the exception of the recently described Pseudodacryodes Pierlot for which, currently, there is no pollen material available. This paper summarises the results.

There is considerable variation in exine and aperture features between, and occasionally within, the genera and 14 major pollen types are defined, including two previously undescribed types: ‘Canarium oleiferum’ and ‘Canarium gracile’. The distribution of pollen characteristics throughout the family is compared with previously published tribal and subtribal groupings, as well as with current ideas of generic relationships from molecular analyses. Comparisons show notable congruence of pollen data with molecular data. To some extent pollen morphology is different for each of the subtribes. Nevertheless, there are some notable exceptions, for example, the pollen of Garuga and Boswellia are remarkably similar, although Garuga has been included, somewhat tenuously, in tribe Protieae, and Boswellia is included in tribe Bursereae, subtribe Boswelliinae. In a recent molecular tree Garuga and Boswellia appear to be closely related, and this supports the conclusion, based on several macromorphological characters as well as pollen, that Garuga should be transferred to tribe Bursereae.