AFLP-based differentiation of tropical African Festuca species compared to the European Festuca complex

For the first time amplified fragment length polymorphism (AFLP) fingerprinting is applied to classify tropical African Festuca species. Five afro-alpine narrow- and two afro-montane broad-leaved species from Uganda and Ethiopia are compared to ten European grass species. A principal coordinate analysis (PCoA) accounts for 62.5% with its first three coordinates. The PCoA and the neighbor-joining (NJ) distinguish the five narrow-leaved African Festuca species from all other species. The broad-leaved African Festuca africana and Festuca simensis are linked to the broad-leaved European species through Festuca altissima and Festuca gigantea, respectively. The narrow- and broad-leaved European species are separated as expected in the NJ. One narrow-leaved African alpine species recently described appears merged (i.e. Festuca richardii with Festuca abyssinica). We provide chromosome numbers for all seven Ugandan species and compare taxonomy and AFLP classification. Our most striking result is that the narrow-leaved African Festuca species are unique and not clustering with the narrow-leaved European species.     

Species distinction in Irish populations of Quercus petraea and Q. robur: morphological versus molecular analyses

BACKGROUND AND AIMS: Populations of oak (Quercus petraea and Q. robur) were investigated using morphological and molecular (AFLP) analyses to assess species distinction. The study aimed to describe species distinction in Irish oak populations and to situate this in a European context. METHODS: Populations were sampled from across the range of the island of Ireland. Leaf morphological characters were analysed through clustering and ordination methods. Putative neutral molecular markers (AFLPs) were used to analyse the molecular variation. Cluster and ordination analyses were also performed on the AFLP markers in addition to calculations of genetic diversity and F-statisitcs. KEY RESULTS: A notable divergence was uncovered between the morphological and molecular analyses. The morphological analysis clearly differentiated individuals into their respective species, whereas the molecular analysis did not. Twenty species-specific AFLP markers were observed from 123 plants in 24 populations but none of these was species-diagnostic. Principal Coordinate Analysis of the AFLP data revealed a clustering, across the first two axes, of individuals according to population rather than according to species. High F(ST) values calculated from AFLP markers also indicated population differentiation (F(ST) = 0.271). Species differentiation accounted for only 13 % of the variation in diversity compared with population differentiation, which accounted for 27 %. CONCLUSIONS: The results show that neutral molecular variation is partitioned more strongly between populations than between species. Although this could indicate that the populations of Q. petraea and Q. robur studied may not be distinct species at a molecular level, it is proposed that the difficulty in distinguishing the species in Irish oak populations using AFLP markers is due to population differentiation masking species differences. This could result from non-random mating in small, fragmented woodland populations. Hybridization and introgression between the species could also have a significant role.     

Festuca inops and Festuca gracilior (Poaceae): are they two different species?

According to current systematics, Festuca inops and F. gracilior are two distinct species. However, they are hardly distinguishable from each other on the basis of their morphological characters. Festuca inops is considered a diploid species endemic to Italy, while F. gracilior has a discontinuous distribution area, apparently related to chromosomal levels: diploid populations in Italy and south-east France, tetraploid populations in north-east Spain. The diploid populations of both taxa from Italy and south-east France are investigated in the present study. Nearly 1000 exsiccata were examined and morphometric analysis was carried out on macro- and micromorphological features of 119 specimens (including type-specimens) and on 20 natural populations (including loci classici ). All these data showed that the two species should be referred to a single taxon, for which the rank of species seems to be appropriate. This result is supported by karyological, ecological and chorological data and was confirmed by the results of ISSR analysis. According to nomenclatural rules, the legitimate name for the species is Festuca inops De Not.  © 2006 The Linnean Society of London, Botanical Journal of the Linnean Society , 2006, 151 , 239–258.     

Hybridization in the genera Vulpia and Festuca (Poaceae): the characteristics of artificial hybrids

The chromosome numbers and morphological characteristics, including life–form, vegetative and floral measurements and pollen fertility, of 70 artificial interspecific Vulpia hybrid plants and 49 artificial intergeneric Festuca rubra agg. x Vulpia hybrid plants are presented, together with comparable data from 60 artificial Festuca rubra agg. x Vulpia fasciculata plants produced by R. Cotton. The inheritance of various characters is summarized. It is concluded that most characters of the tetraploid Vulpia fasciculata are dominant over those of diploid species of Vulpia , but that most characters of the hexaploid Festuca rubra agg. are dominant over those of diploid and tetraploid species of Vulpia. In diploid x diploid Vulpia crosses the pubescent ovary tip is dominant to the glabrous state, but the inheritance of lemma pubescence is more complicated. The keeled lemmas of V. fasciculata and V. pyramidata are dominant to the rounded ones of other Vulpia species, but recessive to the rounded ones of F rubra agg. Bearing in mind such generalizations as these, the parentage of wild hybrids should be capable of correct interpretation. The significance of the results in our understanding of the origin of V. fasciculata is discussed.     

Molecules and morphology in concert. II. The Festuca brachyphylla complex (Poaceae) in Svalbard

We used a combined molecular and morphological approach to unravel variation in the autogamous Festuca brachyphylla polyploid complex in the arctic archipelago of Svalbard. Forty populations were analyzed for random amplified polymorphic DNAs (RAPDs) and 46 morphological characters. Eighteen RAPD multilocus phenotypes were observed in the 86 plants analyzed, based on 30 polymorphic markers. Multivariate analyses of the RAPD data revealed four distinct groups of multilocus phenotypes; in contrast, the variation was more or less continuous in multivariate analyses of the morphological data. However, we identified several individual morphological characters that unambiguously discriminated among the four groups of RAPD multilocus phenotypes. Analysis of type material suggests that the four groups in Svalbard can be referred to Festuca baffinensis, F. brachyphylla, F. hyperborea, and F. edlundiae. This study shows that concerted analysis of molecules and morphology is a powerful tool in low-level taxonomy.     

On the identity of Festuca jubata Lowe (Poaceae) and the description of a new Festuca species in the Azores Islands

The majority of authors consider Festuca jubata Lowe as an endemic species common to Madeira and the Azores. Saint-Yves proposed that F. jubata was an Azorean endemic and described a geovicarious taxon in Madeira: F. filiformis C. Sm. ex Link in Buch ssp. mandonii St.-Yves. We undertook a complete bibliographical revision of the taxonomy, nomenclature, and chorology of F. jubata s.l. , and contrasted it with morphological and anatomical studies performed on samples from the Azores and Madeira. Azorean plants usually identified as F. jubata had a character combination distinct from that of those with a Madeiran provenance. Saint-Yves' proposal of two independent taxa was correct, but he erroneously considered F. jubata as an Azorean endemic because the name F. jubata was based on Madeiran plants. Consequently, F. jubata auct. pl. from the Azores belongs to a new species.  © 2008 The Linnean Society of London, Botanical Journal of the Linnean Society , 2008, 157 , 493–499.     

Molecular tagging of a senescence gene by introgression mapping of a stay-green mutation from Festuca pratensis

* Intergeneric hybrids between Lolium multiflorum and Festuca pratensis (Lm/Fp) and their derivatives exhibit a unique combination of genetic and cytogenetic characteristics: chromosomes undergo a high frequency of homoeologous recombination at meiosis; the chromosomes of the two species can easily be discriminated by genomic in situ hybridization (GISH); recombination occurs along the entire length of homoeologous bivalents; a high frequency of marker polymorphism is observed between the two species. * This combination of characters has been used to transfer and isolate a F. pratensis chromosome segment carrying a mutant 'stay-green' gene conferring a disrupted leaf senescence phenotype into L. multiflorum. * The genetic location within the introgressed F. pratensis segment of the senescence gene has been mapped using amplified fragment length polymorphisms (AFLPs), and F. pratensis-specific AFLP markers closely flanking the green gene have been cloned. * The use of these cloned sequences as markers for the stay-green locus in marker-assisted selection programmes has been tested. The potential application of Lm/Fp introgressions as a tool for the map-based cloning of introgressed Fp genes is discussed.     

Taxonomy, distribution and nomenclature of three confused broad-leaved Potamogeton species occurring in Africa and on surrounding islands

A taxonomic revision of broad-leaved Potamogeton species ascribed to the ' P. schweinfurthii–thunbergii complex' occurring in Africa and on surrounding islands is presented. Three species, P. nodosus , P. richardii and P. schweinfurthii , are recognized in the African mainland. The widespread species P. nodosus has been widely overlooked in sub-Saharan Africa. It is recorded here for the first time from eight countries of tropical and southern Africa and from six surrounding islands. The distribution of P. richardii is critically revised and the species is recorded for the first time from Cameroon, Swaziland and Madagascar. P. schweinfurthii is recorded for the first time from Algeria, Tunisia, Burkina Faso and Niger. The nomenclature of all three species is revised. Lectotypes are designated for six names. All original material of the name P. thunbergii Cham. et Schltdl. actually belongs to P. nodosus Poir. The correct name for the East and southern African species called ' P. thunbergii ' is P. richardii Solms. The lectotype of P. schweinfurthii designated by Dandy proved to be P. nodosus . A new type is therefore proposed for the species generally named P. schweinfurthii and the name itself is proposed for conservation. The morphology and stem anatomy of P. nodosus , P. richardii and P. schweinfurthii are described. In spite of some overlaps in the morphological variation in their vegetative characters, a detailed analysis of the variation patterns and instructions for reliable identification are given. The distributions of all three species are described, based solely on reliably identified specimens, many of which were also examined anatomically. Distribution maps are provided.  © 2005 The Linnean Society of London, Botanical Journal of the Linnean Society , 2005, 148 , 329–357.     

Physical and genetic mapping in the grasses Lolium perenne and Festuca pratensis

A single chromosome of the grass species Festuca pratensis has been introgressed into Lolium perenne to produce a diploid monosomic substitution line 2n = 2x = 14. In this line recombination occurs throughout the length of the F. pratensis/L. perenne bivalent. The F. pratensis chromosome and recombinants between it and its L. perenne homeologue can be visualized using genomic in situ hybridization (GISH). GISH junctions represent the physical locations of sites of recombination, enabling a range of recombinant chromosomes to be used for physical mapping of the introgressed F. pratensis chromosome. The physical map, in conjunction with a genetic map composed of 104 F. pratensis-specific amplified fragment length polymorphisms (AFLPs), demonstrated: (1) the first large-scale analysis of the physical distribution of AFLPs; (2) variation in the relationship between genetic and physical distance from one part of the F. pratensis chromosome to another (e.g., variation was observed between and within chromosome arms); (3) that nucleolar organizer regions (NORs) and centromeres greatly reduce recombination; (4) that coding sequences are present close to the centromere and NORs in areas of low recombination in plant species with large genomes; and (5) apparent complete synteny between the F. pratensis chromosome and rice chromosome 1.     

QTL analysis reveals different and independent modes of inheritance for diagnostic achene characters in Microseris (Asteraceae)

There are few reliable diagnostic morphological characters for species of the asteracean genus Microseris, and quantitative differences in the shapes of the achenes and the paleaceous pappus parts play a decisive role in species recognition. The genetic basis of species and strain differences in various characters has been studied previously, but little is known about quantitative characters of the achenes.

We performed a quantitative trait locus (QTL) analysis in the F2 of an interspecific cross between Microseris douglasii and M. bigelovii for achene length, achene diameter, achene shape, palea length, awn length, and achene pigmentation.

Independent inheritance of the main heritable achene characters – achene length, palea length and achene pigmentation – was revealed by the detection of distinct and specific QTLs for these characters. For palea length five QTLs with about equal phenotypic effects were mapped on four different linkage groups. Achene length and achene shape (achene length / achene diameter) were determined by two different genetic systems with one major gene and two modifiers. The detection of QTLs with a polarity of the effects opposite to that in the parental strains for achene length and achene shape reveals genetic variation for a potential increase in species differences. For the highly heritable trait, achene pigmentation, the bimodal F2 distribution suggested single-factor inheritance for absence versus presence of spots, with dominance for the spotted condition. However, only relatively weak QTL effects on that trait could be detected. Additional molecular markers (RAPDs, AFLPs) have to be tested for cosegregation with that major gene. The results are discussed in the context of different theories for the evolution of morphological characters.     

A demonstration of a 1:1 correspondence between chiasma frequency and recombination using a Lolium perenne/Festuca pratensis substitution

A single chromosome of the grass species Festuca pratensis has been introgressed into Lolium perenne to produce a diploid monosomic substitution line 2n = 2x = 14. The chromatin of F. pratensis and L. perenne can be distinguished by genomic in situ hybridization (GISH), and it is therefore possible to visualize the substituted F. pratensis chromosome in the L. perenne background and to study chiasma formation in a single marked bivalent. Recombination occurs freely in the F. pratensis/L. perenne bivalent, and chiasma frequency counts give a predicted map length for this bivalent of 76 cM. The substituted F. pratensis chromosome was also mapped with 104 EcoRI/Tru91 and HindIII/Tru91 amplified fragment length polymorphisms (AFLPs), generating a marker map of 81 cM. This map length is almost identical to the map length of 76 cM predicted from the chiasma frequency data. The work demonstrates a 1:1 correspondence between chiasma frequency and recombination and, in addition, the absence of chromatid interference across the Festuca and Lolium centromeres.     

A new taxonomic treatment of the Festuca ovina L. aggregate (Poaceae) in the British Isles

A new taxonomic treatment of the Festuca ovina L. aggregate (Poaceae) in the British Isles. A wide range of morphological, anatomical and cytological characters was collected and analysed by a variety of methods, some taxometric. As a result nine species are recognized in the British Isles, one (F. glauca Vill.) only as a garden plant and two (F. huonii Auquier and F. armoricana Kerguélen) only in the Channel Isles. Two other species sometimes claimed for the British Isles (F. guestfalica Boenn. ex Reichb. and F. indigesta Boiss.) are excluded for various reasons given. One species (F. ovina L.) is divided into three subspecies: the diploid subsp. ovina (commonest in the north); and the tetraploid subsp. hirtula (Hackel ex Travis) M. Wilkinson (the commonest taxon of the aggregate in the British Isles) and subsp. ophioliticola (Kerguélen) M. Wilkinson (scattered throughout the British Isles, but commonest on chalk and limestone and not restricted to serpentine soils as once thought).     

A survey of the genus Festuca L. (Poaceae) in Italy. Festuca gamisansii Kerguélen subsp. aethaliae,subsp. nova

ABSTRACT

Festuca gamisansii subsp. aethaliae, endemic to the Isle of Elba (Tuscan Archipelago) is described. The morphological, anatomical, karyological, ecological and chorological characteristics of the species are given. The main characteristics distinguishing F. gamisansii subsp. aethaliae from F. gamisansii subsp. gamisansii occurring in Corsica are also summarized.     

A new species of Festuca L.from Spain

A new species of Festuca (Festuca rivas-martinezii de la Fuente & Ortúñez) is described from the Sierra de Ayllón, Sierra de Guadarrama, Sierra de Gredos and associated chains (northcentral Spain: provinces of Avila, Guadalajara, Madrid and Segovia. Some morphological, anatomical and cytological characters of the taxon are presented.     

Low variability of internal transcribed spacer rDNA and trnL (UAA) intron sequences of several taxa in the Festuca ovina aggregate (POACEAE)

Identification and classification of numerous Festuca species is still a difficult problem due to the close morphological resemblance. The most difficult fine fescues to identify belong to the Festuca ovina aggregate, which is the largest group in the genus Festuca. Many taxons are considered to be separate species based on quantitative taxonomic characters, differences in ploidy level or the structure of sclerenchyma cells. In order to evaluate the taxonomic value of DNA-based markers, sequence analysis of the internal transcribed spacer (ITS1-5.8S-ITS2) region and the chloroplast trnL (UAA) intron was performed in the ten most problematic fine fescues belonging to the Festuca ovina aggregate. Intraspecific ITS variants were found in a single case while in other cases only intragenomic ITS polymorphisms were detected with 1-2 ambiguous positions. Among the sequences of the trnL (UAA) intron even intragenomic polymorphisms were not detected in any of the Festuca species studied. Thus, the results do not support the species status of these ten taxa.     

QTL analysis of leaf morphological characters in a Quercus robur full-sib family (Q. robur x Q. robur ssp. slavonica)

The distinction between white oak species (section Quercus sensu stricto) is largely based on leaf morphological characters. There is, however, considerable within-species variation and no single species-diagnostic character, possibly due to phenotypic plasticity and/or underlying genetic variation. The aim of the present study was to identify quantitative trait loci (QTL) underlying the high within-species variation for leaf morphological characters in an F(1) full-sib family derived from a cross between Q. robur and Q. robur ssp. slavonica. In accordance with an earlier QTL mapping study in an intraspecific Q. robur full-sib family, polygenic inheritance was detected for leaf morphological characters that are used to discriminate between the species Quercus robur and Q. petraea. QTLs were distributed over ten linkage groups, showed a moderate effect in terms of phenotypic variance explained (PVE) in the mapping pedigree (3.6-9.6%), but accounted for a considerable amount of the parental differences. Co-localisation of QTLs on the same linkage group in different genetic backgrounds was found for the number and percentage of intercalary veins (NV, PV) on linkage group 3 and for NV on linkage group 5, revealing a high congruence in the relative QTL positions. The generally low correspondence of the other QTLs in the different mapping pedigrees may be an effect of the genetic background and of the environment. In conclusion, leaf morphological characters were found to be under polygenic control, and a comparison to earlier published results led to the identification of two QTLs that were stable across different genetic backgrounds.     

New data on the Cricetidae from the Miocene “Terre Rosse” of Gargano (Apulia,Italy)

The Gargano “Terre Rosse” deposits are paleokarst fissure fillings found in the Mesozoic limestone of the Apricena-Poggio Imperiale area. They are an important source of information for evolutionary and paleobiogeographic studies. The Late Miocene-Early Pliocene assemblages found in the Terre Rosse attest the complex history of endemic faunal distribution in a paleoarchipelago. Based on the cricetid samples from six distinct fissures (F15, F21a, F21b, F21c, F1, F9, NBS) stored in the Department of Earth Sciences of the University of Florence, three species of endemic cricetids are described: the small-sized Hattomys beetsi, the middle-sized Hattomys nazarii, and the large-sized Hattomys gargantua. The specimens from fissure F15 are attributed to H. beetsi, those from F21a-b to the transitional form Hattomys beetsi-nazarii, those belonging to NBS are ascribed to H. nazarii, whereas those from F1 and F9 are classified as H. gargantua. F21c is considered contaminated with material from different fissures. In line with previously published results, the analysis confirms that the endemic cricetids underwent a remarkable increase in size through time. The morphological variations show a marked trend towards increasing enamel thickness, but also the tendency of the cusps to assume a carved-in aspect. The variations of the morphological characters confirm that the three species likely belong to the same lineage.     

AFLPMax: a user-friendly application for computing the optimal number of amplified fragment length polymorphism markers needed in phylogenetic reconstruction

Amplified fragment length polymorphisms (AFLPs) are widely used for phylogenetic inference especially in non-model species. Frequently, trees obtained with other nuclear or mitochondrial markers or with morphological information need additional resolution, increased branch support, or independent data sources (i.e. unlinked loci). In such cases, the use of AFLPs is a quick and cheap option. Computer simulation has shown that dominant AFLP markers lead to less accurate tree topologies than bi-allelic codominant markers such as SNPs, but this difference becomes negligible for shallow trees when using AFLP data sets that include a sufficiently large number of characters. Thus, determining how many AFLP characters are required to recover a given phylogeny is a key issue regarding the appropriateness of AFLPs for phylogenetic reconstruction. Here, we present a user-friendly, java-based graphical interface, AFLPMax, which executes an automatic pipeline of different programs providing the user with the optimal number of AFLP characters needed to recover a given phylogeny with high accuracy and support. Executables for Windows, linux and MacOS X operating systems, source code and user manual are available from: http://webs.uvigo.es/acraaj/AFLPMax.htm.     

Comparative study (AFLP and morphology) of three species of Prosopis of the Section Algarobia: P. juliflora, P. pallida, and P. limensis. Evidence for resolution of the “P. pallida–P. juliflora complex”

The problems of delimitation of species of Prosopis originate from the few morphological discontinuities which exist among some of them; some, however, originated as a result of wide distribution of germplasm without proper knowledge of the species, in particular, much material catalogued as P. juliflora, but being of other species, was distributed for reforestation projects worldwide. This work tests the morphological results obtained for P. pallida and P. limensis of the Peruvian–Ecuadorian coast and for P. juliflora of the Caribbean Basin of Colombia and Venezuela utilizing a study of AFLPs and a study of the morphology of plantlets developed in a conventional garden study. The phenogram obtained for the AFLPs demonstrates each of the three species to be a well differentiated cluster and the molecular variance between them is significantly greater than the variance within each species. Study of the plantlets also indicates statistically significant differences for four morphological characters between P. juliflora and the other two species (P. pallida and P. limensis). These results, in addition to the morphological differentiation evident between adult plants of P. pallida and P. limensis and the clear separation of these two species from P. juliflora, corroborate the genetic identity of the three taxa analyzed.