RESUMO
We report the first comprehensive multilocus molecular characterization of cultivated Ilex paraguariensis plants belonging to a breeding program. Using nuclear and homologous chloroplastidic molecular markers, we have genotyped 158 plants from four plantation sites. Analyses of the nuclear data (187 variable dominant loci) allowed detecting high diversity (0.569), the occurrence of four distinct genetic clusters, and a low but significant differentiation among sites. Additionally, 20 chloroplastidic alleles were identified applying five microsatellite polymorphic markers, and a high chloroplastidic diversity was recognized (0.505); two haplogroups were distinguished amongst the 63 haplotypes detected. Our results from both nuclear and plastidic markers indicate that most genetic variation reside within plantations sites (≥ 95%), and that these plantations were established on highly variable materials (either as seeds or plantlets) derived from, at least, 63 maternal lineages. Moreover, our study suggests that the genetic structure within each plantation site was most likely shaped by past admixture favored by farmers´ practices during the establishment of each plantation. Also, subsequent constraints in gene flow and/or a low level of shared polymorphism among plantations could have contributed to current structure.
Assuntos
Cruzamento , DNA de Plantas/genética , Genes de Plantas/genética , Ilex paraguariensis/genética , Repetições de Microssatélites/genética , Alelos , GenótipoRESUMO
The Northwestern Argentina (NWA) highland region is one of the southernmost areas of native maize cultivation. We studied variations of different cytological parameters, such as DNA contents, presence/absence of B chromosomes (Bs), and number and sequence composition of heterochromatic knobs in ten accessions of four maize landraces growing along a broad altitudinal cline in NWA. The aim of this work was to assess variations in cytological parameters and their relationship with the crop altitude of cultivation, in an adaptive context. The A-DNA content of the A chromosome complements showed 40% of difference between the lowest (4.5 pg) and the highest (6.3 pg) 2C value. This variation could be attributed to differences in number and size of heterochromatic knobs. Fluorescent in situ hybridization studies revealed the sequence composition of each knob, with a higher proportion of knobs composed of 180-bp repeats rather than TR-1 repeats, in all accessions. We also found numerical polymorphisms and the highest frequency of Bs reported in maize to this date. These results lead us to propose that the frequencies and doses of Bs are influenced by the landrace genotypical make-up. The Bs might be maintained in higher frequencies in those accessions having lower heterochromatin content, so as to preserve an optimal nucleotype. Furthermore, selective forces acting along the altitudinal gradient might be modulating the cytological parameters studied, as suggested by the significant correlations found among them.
Assuntos
Altitude , Cromossomos de Plantas/genética , DNA de Plantas/genética , Variação Genética , Zea mays/genética , Argentina , Meio Ambiente , Genótipo , Heterocromatina/química , Hibridização in Situ FluorescenteRESUMO
The "yerba mate" tree, Ilex paraguariensis St. Hil., is a crop native to subtropical South America, marketed for the elaboration of the highly popular "mate" beverage. The Uruguayan germplasm occupies the southernmost area of the species distribution range and carries adaptations to environments that considerably differ from the current production area. We characterized the genetic variability of the germplasm from this unexplored area by jointly analyzing individuals from the diversification center (ABP, Argentina, Brazil and Paraguay) with 19 nuclear and 11 plastidic microsatellite markers. For the Uruguayan germplasm, we registered 55 alleles (18 % private), and 80 genotypes (44 % exclusive), whereas 63 alleles (28.6 % private) and 81 genotypes (42 % exclusive) were recorded for individuals from ABP. Only two plastidic haplotypes were detected. Distance-based and multilocus genotype analyses showed that individuals from ABP intermingle and that the Uruguayan germplasm is differentiated in three gene-pools. Significant positive correlations between genetic and geographic distances were detected. Our results concur in that ABP individuals harbor greater genetic variation than those from the tail of the distribution, as to the number of alleles (1.15-fold), He (1.19-fold), Rs (1.39-fold), and the between-group genetic distances (1.16-fold). Also the shape of the genetic landscape interpolation analysis suggests that the genetic variation decays southward towards the Uruguayan territory. We showed that Uruguayan germplasm hosts a combination of nuclear alleles not present in the central region, constituting a valuable breeding resource. Future conservation efforts should concentrate in collecting numerous individuals of "yerba mate" per site to gather the existent variation.
Assuntos
Pool Gênico , Variação Genética , Ilex paraguariensis/genética , Alelos , Núcleo Celular/genética , DNA de Cloroplastos/genética , DNA de Plantas/genética , Genética Populacional , Genótipo , Haplótipos , Repetições de Microssatélites , Análise de Sequência de DNA , UruguaiRESUMO
The "yerba mate" tree, Ilex paraguariensis, is a functionally dioecious crop species with economic relevance in several South American countries. We report a genomic screening accomplished through representational difference analysis (RDA) in male and female I. paraguariensis trees. The aim of the present paper was to investigate the occurrence of sex-related genomic differences in order to develop an early gender detection molecular method that could help reducing energy inputs during the "yerba mate" processing and that could be suitable for breeding programs. An intra-experiment redundancy was detected via SSCP analysis and sequence characterization. Taking together both reciprocal RDA assays, fragments isolated can be discriminated into three main categories. The first category of fragments shows spurious affinities with available deposited sequences and could be considered as specific to I. paraguariensis. The second category comprises sequences identified as organellar or ribosomal plant DNA. Sequences grouped in the third category involve clones akin to conserved domains of retrotransposons (RNaseH, integrases and/or chromodomains) from at least two distinct lineages of Ty3/Gypsy retrotransposons and one from Ty1/Copia retroelements, which in addition are associated to sex determination regions of the Solanaceae, Caricaceae and Salicaceae. A contig sequence was assembled that codes for an integrase core domain and a chromodomain. The phylogenetic analysis of the so-called IPRE (for I. paraguariensis retroelement) integrase domain indicates that it belongs to the Del lineage of the Chromoviridae. This is the first report of mobile elements isolated and detected from the "yerba mate" tree. Although RDA derived fragments, so far tested, have been retrieved from both sexes with similar sequences, association to sex related regions cannot be completely discarded. Implications of present results are further discussed.