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Sequence stability at SSR, ISSR and mtDNA loci of common millet (Panicum miliaceum) from the middle ages
10-19Views:108Seed 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. -
Investigation of genetic diversity in irradiated maize lines and its relation to hybrid performance
20-26Views:227Knowledge of genetic diversity among available parental lines is fundamental for successful hybrid maize breeding. The aims of this study were to estimate (1) genetic similarity (GS) and genetic distance (GD) (based on Jaccard index) in four maize inbreed lines; (2) to classify the lines according to their GD and GS; (3) to determine hybrid performance based on GD and heterosis for yield ability in 4x4 full diallel system. We used morphological description and AFLP (amplified fragment length polymorphisms) for estimation genetic polymorphism in four maize inbred lines. We estimated the applicability of genetic similarity in SC and reciproc hybrids for prediction of their performance.
Three primer combinations were used to obtain AFLP markers, producing 207 bands, 70 of whit were polimorphic. The dendogram based on genetic similarities (GS) and genetic distance (GD) and morphological description separated four inbred lines into well-defined groups. Morphological description just with AFLP analysis showed reliable results. In view of genetic distance, the UDL 1 line and their linear and reciprocal crosses showed significant heterosis effect, which was confirmed by heterosis calculation based on grain yield. -
Comparison of RAPD and AFLP Analysis in Some Maize (Zea mays L.) Lines and Hybrids
3-7Views:114The use of molecular markers to enhance plant breeding efforts is being widely studied. DNA-based fingerprinting technologies (RAPD and AFLP) have proven useful in genetic similarity studies. We estimated different maize (Zea mays L.) inbred lines and hybrids originated from mutant ones based on their genetic differences.
We carried out RAPD analysis with different primers and the 707 (CCCAACACCC) and 792 (CAACCCACAC) primers with 50% similarities provided quite good DNA fragments. By applying the DNA based-AFLP technique, we had very dense DNA fingerprinting. We differentiated 15-32 polymorphic bands, the highest number of bands were found in P-T/H-CA (32). AFLP seems to be the more efficient method of comparing genetic similarities/differences among different genotypes. -
Introduction of DNA-based Methods to Agriculture Through Molecular Taxonomic Examination of Poa Species
139-142Views:114biological methods, one among them is AFLP that is well applicable for taxonomic research. Bluegrass species, that are important components of meadow associations, thus their thorough knowledge is necessary in maintaining biodiversity, were examined with bringing this method to perfection.
Taxonomic relationship of the members of Poa pratensis aggregation is a controversial issue. Present study aimes to identify the members of this group, with a developed AFLP method through molecular taxonomic examination of Poa species in meadows nearby Debrecen, revealing their genetical distances. Species of the aggregation show a great genetic variability, but their genetic proximity approves the use of the term aggregation. Results established wider geographical investigation of three species of the aggregation. The distinctness of the species based on their morphological features was confirmed according to their genetical basis as well. The method overall turned out to be appropriate for the taxonomic research of bluegrass species like Poa pratensis and its aggregation. -
Molecular Biological Approach of Crocus sativus L. and its Allies
112-115Views:168The hay saffron (Crocus sativus L.) is a sterile triploid plant, known in human culture only, with no fertile seeds produced. The origin of saffron is still a mist, however it is assumed to be an autopoliploid mutant or a hybrid. The recent classification and most of the former taxonomic publications define C. sativus to be derived from C. cartwrightianus, a wild species. Because of the sterility of hay saffron it seemed to be reasonable to apply molecular biological methods to complete classical taxonomic studies in examining its relations. The DNA polymorphism based AFLP method has confirmed the close relationship between these species.
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Use of molecular marker methods in the classification of bamboo taxa: A review
51-59Views:254Bamboo 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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Application of AFLP-Method in Plant Sample Identification
207-213Views:82One possible method for the determination of DNA-polymorphism is the PCR-based AFLP (Amplified Fragment Length Polymorphism). This method had been succesfully introduced to the Department of Botany at University of Debrecen in 2000-2001 with the examination of hay saffron (Crocus sativus L.) and its allies. Hay saffron is grown as a spice for some thousand years producing the most expensive spice in the world. This plant is sterile, triploid reproduces only vegetatively with no fertile seeds. However its origin is unknown it exists only in cultivation and it is a mutated variety of another species or an artificial or natural hybrid. Usual methods for the systematic examination are restricted hence it seemed to be reasonable to apply molecular biological methods in its case. Results of this work include the introduction and many fold application of the method beside ensuring the consequences of science literature with determining the C. cartwrightianus to give the most similar genetical pattern to C. sativus.