Analysis of the small chromosomal Prionium serratum (Cyperid) demonstrates the importance of reliable methods to differentiate between mono- and holocentricity.

For a long time, the Cyperid clade (Thurniceae-Juncaceae-Cyperaceae) was considered a group of species possessing holocentromeres exclusively. The basal phylogenetic position of Prionium serratum (Thunb.) Drège (Thurniceae) within Cyperids makes this species an important specimen to understand the centromere evolution within this clade. In contrast to the expectation, the chromosomal distribution of the centromere-specific histone H3 (CENH3), alpha-tubulin and different centromere-associated post-translational histone modifications (H3S10ph, H3S28ph and H2AT120ph) demonstrate a monocentromeric organisation of P. serratum chromosomes. Analysis of the high-copy repeat composition resulted in the identification of two centromere-localised satellite repeats. Hence, monocentricity was the ancestral condition for the Juncaceae-Cyperaceae-Thurniaceae Cyperid clade, and holocentricity in this clade has independently arisen at least twice after differentiation of the three families, once in Juncaceae and the other one in Cyperaceae. In this context, methods suitable for the identification of holocentromeres are discussed.

Comparative analyses in Lotus: the cytogenetic map of Lotus uliginosus Schkuhr.

A comparative cytogenetic map was built for Lotusuliginosus (2n = 12), expanding previous analyses that revealed intra- and interspecific chromosomal rearrangements in the model legume L. japonicus, L. filicaulis, and L. burttii. This species is positioned in a sister clade of the previously-mapped species and is proposed as one of the progenitors of L. corniculatus, the main forage crop of the genus. The cytogenetic map allowed the location of 12 genomic regions to be compared between these species. A high macrosynteny was revealed, but it was interrupted by a translocation involving chromosomes 3 and 5, a new rearrangement for the genus. Also, a transposition on chromosome 2 was found in L. japonicus 'Miyakojima'. Furthermore, changes in the number, size, and position of rDNA sites were observed, as well as an intraspecific size heteromorphism of the 5S rDNA site on L. uliginosus chromosome 6. The karyotype differences observed are proportional to the phylogenetic distance among these species.

Contrasting rDNA evolution in lima bean (Phaseolus lunatus L.) and common bean (P. vulgaris L., Fabaceae).

Phaseolus vulgaris has two 5S rDNA sites in chromosomes 6 and 10 and from two up to nine 45S rDNA sites depending on the accession. The presence of three 45S rDNA sites, in chromosomes 6, 9 and 10, is considered the ancestral state for the species. For P. lunatus, only one 5S and one 45S rDNA sites in distinct chromosomes were known. In order to investigate the homeologies among these rDNA-bearing chromosomes and the stability of the rDNA sites in P. lunatus, rDNA and P. vulgaris chromosome-specific probes were hybridized in situ to P. lunatus. The chromosomes bearing the 5S and the 45S rDNA of P. lunatus are homeologous to chromosomes 10 and 6 of P. vulgaris, respectively. In contrast to the common bean, no variation in the number of rDNA loci was detected, except for a duplication of the 5S rDNA in the same chromosome in a small group of cultivars. These results suggest that the 5S rDNA site in chromosome 10 and the 45S rDNA site in chromosome 6 represent the ancestral loci in the genus. The 5S rDNA site in chromosome 10 of P. vulgaris is located in the long arm, while in P. lunatus it is present in the short arm, suggesting the occurrence of a transposition or a pericentric inversion after separation of both lineages.

The anthracnose resistance locus Co-4 of common bean is located on chromosome 3 and contains putative disease resistance-related genes.

The broadest based resistance to anthracnose of common bean ( Phaseolus vulgaris L.) is conferred by the Co-4 locus. We sequenced a bacterial artificial chromosome clone harboring part of the Co-4 locus of the bean genotype Sprite and assembled a single contig of 106.5 kb for functional annotation. This region contained five copies of the COK-4 gene that encodes for a serine threonine kinase protein previously mapped to the Co-4 locus and 19 novel genes with no similarity to any previously identified genes of common bean. Several putative genes of the Co-4 locus seemed to be expressed as they matched perfectly with bean expressed sequence tags. The expression of the COK-4 genes was assessed by reverse transcription (RT)-PCR, and a single 850-bp cDNA fragment was sequenced and compared with the genomic sequences of the COK-4 homologs. Although the COK-4 cDNA was isolated from a different bean cultivar, it showed high similarity (95%) to the exons of genes BA17 and BA21, suggesting that they were expressed. In a phylogenetic tree including all currently available Pto-like sequences from Phaseolus species, the COK-4 homologs formed a single cluster with the Pto gene, whereas two sequences from P. coccineus and all sequences of P. vulgaris formed two closely related clusters. The Co-4 locus was physically mapped to the short arm of bean chromosome 3, which corresponds to linkage group B8. This study represents a first step in gaining an understanding of the genomic organization of an anthracnose resistance locus of common bean and provides molecular data for comparative analysis with other plant species.