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SSR based characterization of peach (Prunus persica L.) and apricot (Prunus armeniaca L.) varieties cultivated in Hungary
17-24Views:263The SSR (Simple Sequence Repeat) markers allow the discrimination of the cultivars and determination its specific DNA fingerprints. The aim of this research was to evaluate fifteen apricot (Prunus armeniaca L.) and fifty-one peach (Prunus persica L.) genotypes cultivated in Hungary to obtain their DNA fingerprints in 6 SSR (Simple Sequence Repeats) loci by allele numbers and sizes.DNAs were extracted from leaves. PCR was carried out with CY-5 fluorescent labeled Prunus microsatellite markers and the products were separated on polyacrylamide gel with ALF (Automated Laser Flourometer)-Express II.According to our results, in the case of peach genotypes, all 6 SSRs were able to amplify alleles. UDP 96 005 was the most informative marker and UCDCH 17 was the least due to its monomorphic pattern. Regarding the apricot samples BPPCT 041 did not amplify any allele. In the case of P. armeniaca UDP 96 005 had the highest heterozygosity index as well and the highest number of alleles. The least informative marker was the UCDCH 17. Since the 6 SSR were not enough to discriminate the apricot and peach genotypes, it is suggested to use more SSR primers. -
Sequence stability at SSR, ISSR and mtDNA loci of common millet (Panicum miliaceum) from the middle ages
10-19Views:86Seed remains of medieval millet, recovered from a 15th century layer (King’s Palace, Budapest, Hungary), showed reddish yellow grain color after rehydrating on tissue culture medium that was close to grain color of modern cultivar Omszkoje. aDNA of medieval c. millet was extracted successfully, analyzed and compared to modern common millets by ISSR, SSR, CAPS and mtDNA. Analyses of fragments and sequences revealed
polymorphism at seven ISSR loci (22 alleles) and at the 5S-18S rDNA locus of mtDNA. CAPS analysis of the 5S-18S rDNA fragment revealed no SNPs in the restriction sites of six endonucleases TaqI, BsuRI, HinfI, MboI, AluI and RsaI. Sequence alignments of the restriction fragments RsaI also revealed
consensus sequence in the medieval sample compared to a modern variety. Morphological characterization of twenty common millet (Panicum miliaceum L., 2n=4×=36) cultivars and landraces revealed four distinct clusters which were apparently consistent with the grain colors of black, black and brown, red, yellow, and white. In the comparative AFLP, SSR and mtDNA analysis modern millet cv. ‘Topáz’ was used. AFLP analysis revealed that extensive DNA degradation had occurred in the 4th CENT. ancient millet resulting in only 2 (1.2%) AFLP fragments (98.8% degradation),
compared to the 15th CENT. medieval millet with 158 (40%) fragments (60% degradation) and modern millet cv. ‘Topáz’ with 264 fragments (100%). Eight AFLP fragments were sequenced after reamplification and cloning. Microsatellite (SSR) analysis at the nuclear gln4, sh1, rps28 and rps15 loci of the medieval DNA revealed one SNP (single nucleotide polymorphism) at the 29th position (A to G) of rps28 locus compared to modern millet.
Mitochondrial (mtDNA) fragment (MboI) amplified at the 5S-18S-rDNA locus in the medieval millet showed no molecular changes compared to modern millet. The results underline the significance of survived aDNA extraction and analysis of excavated seeds for comparative analysis and molecular reconstruction of ancient and extinct plant genotypes. An attempted phenotype reconstruction indicated that medieval common millet showed the closest morphological similarity to modern millet cultivar Omszkoje. -
Use of molecular marker methods in the classification of bamboo taxa: A review
51-59Views:154Bamboo plants are currently attractive to researchers because of their versatile uses. Understanding the bamboos’ genetic level is needed to develop new varieties. Taxonomic identification is the basis for plant development. Bamboos were identified as their taxonomical morphological characters which are dependent on environmental factors. Molecular Marker techniques can be used to perform accurate genotype identification, which can be used for genetic diversity analyses. The RFLP, RAPD, AFLP, SSR, ISSR, iPBS, SCARS, SCoT, SRAP marker systems have been shown to be able to efficiently determine the genetic diversity of bamboo species based on genotyping. This paper summarizes research that aims to analyze the genetic diversity of bamboo species on a molecular basis.
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Microsatellite Diversity of Androgenic Black Poplar (Populus nigra)
60-67Views:69Genetic variation of somatic clones (1 to 35) of black poplar (Populus nigra) developed from two anther-donor trees N-SL and N-309 was determined by five SSR primer pairs. Twenty SSR alleles were detected, the number of alleles per marker ranged from 1 to 6, with an average of 3.3 including WPMS-2 (5 alleles), WPMS-4 (6 alleles), WPMS-6 (2 alleles), WPMS-20 (6 alleles) and PTR-4 (1 allele) detected by ALF (automatic laser fluorometer). A
dendrogram produced by SPSS11 based on the presence versus absence of SSR alleles discriminated the groups of somatic clones of N-SL from somatic clones of N-309. The polymorphic markers of WPMS-2 (5 alleles), WPMS-4 (6 alleles) and WPMS-20 (6 alleles) revealed clonal variation in 1 clone (37) out of the 6 from the N-309 tree, and three subgroups out of the 29 somatic clones from the N-SL tree (17 and 24), (2 and 14) and (10 and 15). The remaining 23 of the 29 N-SL somatic clones with uniform genetic similarity suggests a good degree of genetic stability in black poplar. Nevertheless, the new SSR-clones may provide useful new genetic resources for poplar breeding.