Wild sunflower species are adapted to diverse habitats and possess considerable variability for biotic and abiotic traits, therefore wild germplasm can be a real source for improving the narrow genetic base of the cultivated sunflower crop by introgression of new genes which were previously identified in wild genotypes. In order to effectively exploit these genes in cultivated genotypes, researchers need more information regarding the quantity and distribution of the genetic variability available within the wild sunflower species, which can offer a multitudine of valuable traits for traditional or molecular breeding. This study was conducted to reveal the genetic diversity of wild and cultivated sunflower genotypes using ISSR markers. From a total of 19 ISSR markers, which were used to evaluate the genetic diversity, eight markers (UBC808, UBC823, UBC834, UBC836, UBC840, UBC845, UBC853, 17899A) clearly differentiated the wild sunflower genotypes from the wild sunflower and/or the cultivated sunflower. The primers amplified a total of 120 alleles ranging from 10 to 22 alleles per marker. A wide range of fragment length was detected among the accessions, from 140 to over 1500 bp. The neighbor-joining dendrogram, based on Rogers genetic distance, of the genotypes studied consisted of two main clusters of different sizes: five entries were grouped into cluster A and the remaining six entries were grouped into cluster B. It is interesting that genotypes of Helianthus anuus L., Helianthus maximiliani and Helianthus argophyllus were clustered together. Another aspect observed, refers to interspecific variability for Helianthus debilis genotypes. Regarding the PCA, the first two principal axes accounted for 26.8% (CP1) and 16.1% (CP2) of the total variation, respectively, together explaining 42.9% of the total variability. In the future, this molecular genetic information can be combined with morphological and biochemical data to improve the sunflower breeding program.
The hexaploid European plums (Prunus domestica L. and Prunus domestica subsp. insititia (L.) C.K.Schneid.) are main traditional fruit tree culture in Romania and represent an economically important fruit species with limited information on its genetic structure. Our objective was to fingerprint 33 cultivars using four Sequence-Related Amplified Polymorphism (SRAP) primer pairs to estimate the genetic relationships among local and international cultivars. The primer pairs amplified a total of 70 alleles ranging from 14 to 21 alleles per marker. A wide range of fragment length was detected among the accessions, from 65 to 2000 bp. The mean number of alleles per primer combination was 17.5, with the most alleles obtained with SRAP80 (21 alleles), range between 65-1800 bp and the fewest alleles (14) were obtained with SRAP95, range between 150-2000 bp. The neighbor-joining dendrogram, based on Rogers genetic distance, of the plum germplasm studied, consisted of two main clusters of different sizes: 23 entries were grouped into cluster 1 and the remaining 10 entries were grouped into cluster 2. It is interesting that cultivars representing plum species with 6x and 2x ploidy levels were clustered together. Another interesting aspect observed refers to plum genotypes clustered closely on the dendrogram according to their pedigree, such as 'Dani' (P14) - 'Tita' (P15), 'Roman' (P7) - 'Early Rivers' (P9), 'Zamfira' (P6) - 'Pescăruș' (P33). Accessions within the Romanian landrace/old autochthonous cultivars group were not clustered together. Regarding the PCA, the first two principal axes accounted for 12.91% (CP1) and 10.46% (CP2) of the total variation, respectively, together explaining 23.37% of the total variability. Our results showed that SRAP markers represent valuable tools for genetic diversity study on Prunus domestica. To our knowledge, this is the first study using SRAP markers for characterization of P. domestica germplasm. In the future, this molecular genetic information can be combined with phenological and biochemical data to identify genes, quantitative trait loci (QTL) and molecular markers that can be used to improve the plum crop breeding program.
Pea (Pisum sativum L.) is an important legume grown and consumed extensively worldwide. As a rich source of proteins, carbohydrates and vitamins, peas are important in human nutrition. In this study, we compared four different DNA isolation methods from four pea cultivars (F95-927, Specter, Windham and Nicoleta), with some modifications to the original extraction protocols: one SDS extraction buffer without PVP and one SDS extraction buffer with 1% PVP; one CTAB extraction buffer without PVP and one with 1% (1.5%) PVP. For all four extraction methods we used the same quantity of plant material (0.05 g) and an equal quantity of extraction buffer, so the comparison between methods to be very accurate. To establish which is the most efficient extraction method, after DNA isolation and purification, we submitted our samples to PCR analyzes with two markers: ISSR marker 17899A and SSR marker AA175. In my study, based on spectrophotometric measurements and PCR results, I concluded that both CTAB extraction methods were not suitable for DNA extraction from dry pea seeds because they did not show amplification products. The most appropriate DNA extraction method was SDS1 which provided a good quality DNA.
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