Marcelo Guerra

Patterns of heterochromatin distribution in plant chromosomes

The C-band distribution patterns of 105 angiosperm species were compared to identify general patterns or preferential sites for heterochromatin. The base-specific fluorochrome reaction of heterochromatin for 58 of these species and the role played by the average chromosome size in band distribution were also considered. The results showed that heterochromatin was preferentially located in similar chromosome regions, regardless of the distance from the centromere. This trend results in generalized bands, with heterochromatin distribution being identical in most chromosomes of a karyotype. Such bands very often displayed the same fluorochrome reaction, suggesting possible repeat transfer between non-homologous sites. Chromosome size may also play a role in heterochromatin location, since proximal bands were much more common in small-sized chromosomes.

Chromosome numbers in plant cytotaxonomy: concepts and implications.

Chromosome number is the karyotype feature most commonly used in cytotaxonomical analyses. The chromosome number can be a plesiomorphic characteristic of a large clade or a recurrent trait which arose independently in two or more clades. Some concepts regarding chromosome number variation, such as base number, aneuploidy, paleopolyploidy, and neopolyploidy have been used by different authors in quite different ways. Therefore, its use in cytotaxonomy and karyotype evolution deserves much attention. In this paper, these terms are reappraised and their meaning and implication for plant cytotaxonomy are discussed.

Extensive ribosomal DNA amplification during Andean common bean (Phaseolus vulgaris L.) evolution

The extent of 5S and 45S ribosomal DNA (rDNA) variation was investigated in wild and domesticated common beans (Phaseolus vulgaris) chosen to represent the known genetic diversity of the species. 5S and 45S rDNA probes were localized on mitotic chromosomes of 37 accessions by fluorescent in situ hybridization (FISH). The two 5S rDNA loci were largely conserved within the species, whereas a high variation in the number of 45S rDNA loci and changes in position of loci and number of repeats per locus were observed. Domesticated accessions from the Mesoamerican gene pool frequently had three 45S rDNA loci per haploid genome, and rarely four. Domesticated accessions from Andean gene pool, particularly from the race Peru, showed six, seven, eight or nine loci, but seven loci were found in all three races of this gene pool. Between three and eight loci were observed in accessions resulting from crosses between Andean and Mesoamerican genotypes. The presence of two to eight 45S rDNA loci in wild common beans from different geographic locations indicates that the 45S rDNA amplification observed in the Andean lineage took place before domestication. Our data suggest that ectopic recombination between terminal chromosomal regions might be the mechanism responsible for this variation.

Distribution of 45S rDNA sites in chromosomes of plants: Structural and evolutionary implications

Background45S rDNA sites are the most widely documented chromosomal regions in eukaryotes. The analysis of the distribution of these sites along the chromosome in several genera has suggested some bias in their distribution. In order to evaluate if these loci are in fact non-randomly distributed and what is the influence of some chromosomal and karyotypic features on the distribution of these sites, a database was built with the position and number of 45S rDNA sites obtained by FISH together with other karyotypic data from 846 plant species.ResultsIn angiosperms the most frequent numbers of sites per diploid karyotype were two and four, suggesting that in spite of the wide dispersion capacity of these sequences the number of rDNA sites tends to be restricted. The sites showed a preferential distribution on the short arms, mainly in the terminal regions. Curiously, these sites were frequently found on the short arms of acrocentric chromosomes where they usually occupy the whole arm. The trend to occupy the terminal region is especially evident in holokinetic chromosomes, where all of them were terminally located. In polyploids there is a trend towards reduction in the number of sites per monoploid complement. In gymnosperms, however, the distribution of rDNA sites varied strongly among the sampled families.ConclusionsThe location of 45S rDNA sites do not vary randomly, occurring preferentially on the short arm and in the terminal region of chromosomes in angiosperms. The meaning of this preferential location is not known, but some hypotheses are considered and the observed trends are discussed.

Cytogenetics of Rutaceae. V. High chromosomal variability in Citrus species revealed by CMA/DAPI staining

The CMA/DAPI chromosome banding pattern of six Citrus species was analysed. All of them showed a large amount of heterochromatin and heterozygosity. Most of the heterochromatin was CMA+/DAPI− and was located mainly in the terminal region of the long arms. Each individual was heterozygous for at least one chromosome pair and each species presented a different banding pattern. The greatest amount of information on karyotype variation can be obtained from the four largest pairs. Pairs I and III showed the highest amount of heterochromatin and heterozygosity whereas pair II was highly stable and almost completely lacked CMA+ heterochromatin. On the other hand, some features common to all species suggest a strong chromosome homoeology conservation. The CMA staining revealed some chromosome markers for each Citrus species that may be very useful in the identification of zygotic versus nucellar embryos, as well as for further evolutionary studies.

Karyotype diversity and the origin of grapefruit

Grapefruit is a group of citrus of recent origin, probably resulting from a cross between pummelo and sweet-orange. Aiming to investigate this putative origin and the genetic variability among grapefruit cultivars, the karyotype of six grapefruits, two pummelos, and one tangelo cultivar (grapefruit × tangerine) were analyzed using sequential CMA/DAPI double staining and FISH with rDNA probes. The karyotypes of grapefruit ‘Duncan’ and ‘Foster’ differ from those of ‘Flame’, ‘Henderson’, ‘Marsh’ and ‘Rio Red’. The former have two chromosomes with a single CMA+ band in both terminal regions (C type chromosome) and six chromosomes with only one CMA+ terminal band (D type), whereas the latter have three C and five D type chromosomes. All accessions investigated exhibited two chromosomes with 5S rDNA but a variable number of 45S rDNA. The two former grapefruits displayed four 45S rDNA sites, whereas the remaining grapefruit cultivars had five. The two pummelos showed identical karyotypes, homozygous for CMA+ bands and their four rDNA sites. From each pummelo chromosome pair one chromosome seems to be present in grapefruit karyotypes. The different grapefruit karyotypes might result from independent crosses between pummelos of different karyotypic constitution and sweet-oranges. The chromosome markers found in the tangelo ‘Orlando’ and the position of their two 45S rDNA confirm the grapefruit ‘Duncan’ and the tangerine ‘Dancy’ as their parents.

Cytogenetics of Angiosperms collected in the State of Pernambuco: V

Thirty three native and introduced species from 20 families of angiosperms collected in the State of Pernambuco were analysed. The karyotype description of the majority of the species was based on chromosome number and morphology, condensation pattern of prophase chromosomes as well as interphase nuclear structure. In five species (Cereus jamacaru, 2n=22; Clitoria fairchildiana, 2n=22; Eugenia luschnathiana, 2n=22; Licania tomentosa, 2n=22; and Spondias tuberosa, n=16) the chromosome number is reported here for the first time. In the case of Licania tomentosa, this is also the first report for the genus. Among the other 28 species, three (Cecropia cf. palmata, 2n=26; Crinum erubescens, 2n=70; and Schinus terebentifolius, 2n=28) showed chromosome numbers different from what has previously been reported.

High Dynamics of rDNA Cluster Location in Kissing Bug Holocentric Chromosomes (Triatominae, Heteroptera)

In this paper, we determine by fluorescent in situ hybridization the variability in the chromosomal location of 45S rDNA clusters in 38 species belonging to 7 genera of the Triatominae subfamily, using a triatomine-specific 18S rDNA probe. Our results show a striking variability at the inter- and intraspecific level, never reported so far in holocentric chromosomes, revealing the extraordinary genomic dynamics that occurred during the evolution in this group of insects. Our results also demonstrate that the chromosomal position of rDNA clusters is an important marker to disclose chromosomal differentiation in species karyotypically homogenous in their chromosome number.

The relationships among lemons, limes and citron: a chromosomal comparison

Lemons, limes and citron constitute a group of closely related Citrus species, whose species delimitations and taxonomic relationships are unclear. In order to identify karyotypic similarities and species relationships within this group, the CMA+/DAPI– banding pattern and the distribution of the 5S and 45S rDNA sites of 10 accessions of lime, lemon, and citron were investigated. The four cultivars of C. limon analyzed showed the same pattern of CMA+ bands and rDNA sites, suggesting that they originated from a single germplasm, later differentiated by distinct somatic mutations. The lemons C. jambhiri, C. limonia and C. volkameriana displayed karyotypes very similar to each other, but they differed from C. limon by the absence of a single chromosome with one band in each telomere. The limes, C. aurantifolia and C. limettioides, seemed less related to each other and exhibited different heteromorphic chromosome pairs. In C. aurantifolia, the presence of a chromosome type unknown in all other Citrus species cytologically known so far supports the assumption that this accession may be derived from a hybrid with a species from the subgenus Papeda or from another genus. Citrus medica was the only homozygous accession of this group and all of its chromosome types were clearly represented in limes and lemons, some of them forming heteromorphic pairs. The analysis of the distribution of rDNA sites allowed a further refinement of the comparison among accessions. The lemons and limes were heterozygous for all rDNA sites, whereas C. medica was entirely homozygous. These data support the hypothesis that C. medica is a true species while the other nine accessions are hybrids.

Cytotaxonomy: The end of childhood

Abstract Detailed comparative karyotype analyses have proved to be an important tool for taxonomy and for understanding chromosome evolution. The basic principle of cytotaxonomy is that closely related species share a more similar karyotype than less related ones. However, phylogenetic studies have demonstrated that some karyotype features are more often recurrent than previously thought, meaning that similarity is not all. Two other criticisms to the cytotaxonomical interpretation are intraspecific variation, traditionally neglected, and the interpretation of chromosomal alterations based only on classical structural changes, without any investigative support. Chromosome numbers and nuclear DNA content are among the most reliable and practical characters for cytotaxonomical analyses involving a large number of species, whereas chromosome mapping with molecular probes is more recommended for a restricted number of species. In both cases, a robust phylogenetic tree is necessary to understand the relationships between species and their karyotype evolution. Evolutionary inferences based only on putative karyological trends are no longer sustainable.