RESUMEN
Dactylis glomerata L. is an important forage species in the Mediterranean region, and in other regions with a similar climate. Genetic material from 3 locations in north, central, and south Greece was studied, using morphological traits, SSR, and ISSR molecular markers. Morphological analysis revealed differences among the geographic locations studied for all morphological traits, except the number of reproductive tillers. Moreover, the highest phenotypic variation was observed on the accessions from south, while the lowest was observed on the accessions from the north. Although the results of the molecular marker analysis are indicative, a high level of genetic diversity at the species level was revealed by ISSRs (GST=0.291) and SSRs (FST=0.186). Analysis of molecular variance showed that a high level of genetic diversity existed for ISSRs and SSRs within populations (62 and 83%, respectively), rather than among populations (38 and 17%, respectively). Cluster analysis divided the 3 populations in 2 groups, with the population originating from the island of Crete forming 1 group, while the populations from north Greece (Taxiarchis) and central Greece (Pertouli) were clustered in a 2nd group. In general, the results indicate that SSRs are more informative compared to ISSRs about the genetic variation within a population, whereas the ISSRs were more informative about the genetic diversity among populations However, a similar trend in diversity (genotypic and phenotypic) was observed in the morphological traits and microsatellite-based (SSR/ISSR) markers at the locations studied.
Asunto(s)
Dactylis/genética , Variación Genética , Repeticiones de Microsatélite , Semillas/genética , Análisis de Varianza , Análisis por Conglomerados , Dactylis/clasificación , Genoma de Planta , Análisis de Componente PrincipalRESUMEN
Local breeds are important for the maintenance of genetic diversity and future food security. Nowadays, the worldwide distribution of pigs is dominated by a few breeds, tending towards a severe loss of pig biodiversity. Thus, it is critical to maintain distinct populations of pig breeds. The Greek black pig, a breed raised locally and known for the high quality of its meat for cured products, is the only traditional indigenous pig breed reared in Greece. We investigated the genetic diversity, based on microsatellite analysis, of the Greek black pig and evaluated its genetic uniqueness. One hundred and three pigs from 12 Greek farms were analyzed using 11 microsatellites. The total number of alleles amounted to 135, with a mean number of alleles per locus of 12.27, ranging between 10 and 16 alleles. The observed heterozygosity ranged from 0.363 to 0.825 per locus. The expected heterozygosity ranged from 0.471 to 0.707. The inbreeding coefficient ranged from -0.329 to 0.229. We conclude that the Greek black pig, despite its low population size, has a high degree of genetic variability, which will be useful for breeding programs aimed at maintaining long-term survival of this ancient breed.
Asunto(s)
Variación Genética , Repeticiones de Microsatélite/genética , Filogenia , Porcinos/genética , Alelos , Animales , Cruzamiento , Grecia , Heterocigoto , Endogamia , Sus scrofa/genéticaRESUMEN
The ability to discriminate all species is the ultimate target in barcoding. The Mediterranean basin is a center of origin for legumes and thus they have played a key role in feeding the Mediterranean population. It is also a region with important protected designation of origin and protected geographical indication legumes that provide income in rural areas. We evaluated the use of two chloroplast regions, trnL and rpoC1, and a nuclear internal transcriber region, ITS2, for their efficiency to barcode the main Mediterranean leguminous crops. Twenty-five legume species were studied. Plant material of pasture and legumes was obtained from the Greek GenBank and the Fodder Crops and Pastures Institute (National Agricultural Research Foundation). DNA was extracted with the Qiagen DNeasy plant mini-kit and PCR amplification was performed using the Kapa Taq DNA polymerase using primers amplifying the chloroplast trnL and rpoC1 regions or the nuclear region ITS2. PCR products were sequenced and the sequences were aligned using CLUSTAL W. Species identification based on the sequence similarity approach was performed using the GenBank database. In order to evaluate intraspecific and interspecific divergence in legumes we used Molecular Evolutionary Genetics Analysis 5 and for pairwise Kimura 2-parameter distance calculations for all 3 DNA regions (2 chloroplast regions, trnL and rpoC1, and the nuclear region ITS2). Four tree-based methods (neighbor joining and maximum parsimony, maximum likelihood, and Bayesian inference analyses) were used to exhibit the molecular identification results to represent differences as an uprooted dendrogram. Additionally, the sequence character-based method was used with DnaSP and the information from each site was treated as a character to distinguish the species from one another. The DNA regions trnL and ITS2 successfully (100%) discriminated the Mediterranean crop legume species used, while rpoC1 identified only 72% of them. Furthermore, the use of the trnL region enabled the discrimination of even very closely related species, like Phaseolus lunatus and P. coccineus or Vicia faba subsp major with V. faba subsp minor, which are so closely related that even in NCBI they were both referred as Phaseolus vulgaris and V. faba, respectively. We conclude that trnL and ITS2 are efficient DNA barcoding target regions in order to discriminate Mediterranean leguminous crops and provide a reliable and efficient tool for the scientific, agricultural and industrial community.
Asunto(s)
Núcleo Celular/genética , Productos Agrícolas/genética , Código de Barras del ADN Taxonómico/métodos , ADN de Cloroplastos/genética , ADN de Plantas/genética , Fabaceae/clasificación , Fabaceae/genética , Secuencia de Bases , Región Mediterránea , Datos de Secuencia Molecular , Filogenia , Especificidad de la EspecieRESUMEN
Quince is a deciduous tree known to the countries around the Mediterranean since antiquity. Nowadays, quince is used as an ornamental plant, and as a rootstock for pear trees, with its fruit being appreciated mainly for production of jam and sweets rather than for raw consumption. Quince leaves contain compounds with antioxidant, antimicrobial and anticancerous properties that have been the focus of recent research on pharmaceutical and medical uses as well as for food preservatives. An orchard has been established in Greece, composed of quince varieties (Cydonia oblonga, N = 49) collected from different sites of the country (mainly from home gardens), constituting a unique quince gene bank collection for southeast Europe. We made a phenotypic analysis using 26 morphological plus seven agronomical descriptors coupled with molecular techniques in order to examine the genetic diversity within the collection. Principal component analysis using the 33 descriptors identified 10 components explaining the existence of more than 70% of the total variation. Subsequent cluster analysis classified most of the previously identified productive varieties of the quince orchard in the same clade of a dendrogram. Molecular analysis generated by 13 inter-simple sequence repeat primers amplified 139 bands, including 109 polymorphic bands, indicating a level of polymorphism of 79%; mean gene diversity was calculated to be 0.309. Using stepwise multiple regression analysis, a number of markers significantly associated with fire blight susceptibility, yield, mean fruit weight, citric acid content, soluble solid content, and fruit drop were identified. Hence, data extracted by multiple regression analysis could be useful in marker-assisted breeding programs, especially when no previous genetic information is available.
Asunto(s)
Frutas/genética , Filogenia , Polimorfismo Genético/fisiología , Sitios de Carácter Cuantitativo , Rosaceae/genética , Marcadores Genéticos , GreciaRESUMEN
Cotton is the most important textile plant in the world and is one of the most important crops for the production of oilseed. Because of its worldwide economic importance, new cultivars are constantly being released in the world and consequently in the Greek market, as Greece is the largest producer in Europe. We used simple sequence repeat (SSR) markers for the identification and the phylogenetic analysis of the most widely cultivated cotton cultivars in Greece. Initially, we used 12 pairs of SSR molecular markers for the analysis of 29 cultivars of Gossypium hirsutum and an interspecific hybrid (G. hirsutum x G. barbadense). Of the 12 pairs of SSR primers, 11 amplified polymorphic products, while one pair did not amplify any product. Globally, 17 polymorphic marker loci were identified. Two to four different alleles were amplified at each genomic locus, with a mean of 2.53 alleles per locus. Among the 30 genotypes that we analyzed, the polymorphism information content ranged from 0 to 0.548, with a mean of 0.293. Three main groups were formed among the 30 genotypes when a phylogenetic analysis was performed using UPGMA. Computational analysis of each molecular marker separately showed an association of SSR markers with agronomic traits such as fiber quality. To our knowledge, this is the first in-depth molecular analysis of cotton cultivars grown in Greece using SSR markers. An analysis of association of SSR markers with fiber quality traits of 29 cotton cultivars is reported for the first time.