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The morphological diversity in 25 populations of two native species of Brachypodium occurring in Poland, was studied. Principal component analysis and cluster analysis of morphometric data segregate specimens or populations of Brachypodium into two groups, corresponding to the two species. In most of morphological characters, B. pinnatum and B. sylvaticum are similar to each other. However, in terms of characteristics, such as the hairiness of culms and leaf-sheaths both can also vary much within a single population. In comparison with B. pinnatum, B. sylvaticum always has pubescent abaxial palea surfaces. Apparent differences were not detected between the two ecotypes of B. pinnatum named "Grassland" and "Woodland".
Brachypodium pinnatum belongs to native grasses which could dramatically decrease the biodiversity of calcareous grasslands, therefore, an examination of the mechanisms of its expansion is of prime importance for their conservation. We studied the genetic structure of 12 subpopulations of the expansive grass B. pinnatum in a heterogeneous landscape with AFLP markers, aiming at determination whether spatial isolation influences the genetic diversity and the population genetic structure of this species. A high level of overall (79%) and within population polymorphic loci (38–44%) were found and 220 different genotypes were distinguished among the 252 samples analysed. No significant population structure nor isolation-bydistance were found, despite their long-time fragmentation history. This confirms that isolation of calcareous grasslands in a landscape cannot prevent the expansion of B. pinnatum grasses because of the high generative dispersal ability. Subsequent quick and extensive clonal growth of this species enables the successful establishment. In most cases mowing or grazing is sufficient to keep this species at a low density within ancient grasslands and to prevent seed and pollen dispersal, but not for restoration of species-rich calcareous grasslands.
The genus Brachypodium has become the target of extensive cytomolecular studies since one of its representatives, B. distachyon, has been accepted as a model plant for temperate cereals and forage grasses. Recent preliminary studies suggested that intraspecific rDNA polymorphism can occur in at least two members of the genus, B. sylvaticum and B. pinnatum, so the aim of this study was to further analyse this phenomenon. FISH with 25S rDNA and 5S rDNA probes was performed on somatic metaphase chromosomes, supplemented by the silver staining technique which distinguishes transcriptionally active from inactive 18S-5.8S-25S rDNA loci. The number, size and chromosomal distribution of 5S rDNA loci were very constant: two loci were invariably observed in all studied diploid accessions of both species, while four 5S rDNA loci were present in the tetraploid B. pinnatum. In contrast to 5S rDNA loci, those of the 35S rDNA were more variable. Two or three loci were observed in the diploid B. pinnatum and four in tetraploid accessions. In chromosome complements of B. sylvaticum accessions from two to six 35S rDNA sites were detected. Regardless of total rDNA locus number, only two were transcriptionally active in diploid accessions of both species, while two or four were active in the tetraploid B. pinnatum. Additionally, the fluorescent CMA/DAPI banding method was used to identify the relation between rDNA sites and CMA+ bands. It was revealed that the number and chromosomal distribution of CMA+ bands are in congruence only with 35S rDNA loci which gave strong FISH signals.
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