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Sequence stability at SSR, ISSR and mtDNA loci of common millet (Panicum miliaceum) from the middle ages
Published November 15, 2007
10-19

Seed 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 b...y 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. 

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Selection of powdery mildew resistant and susceptible grapevine genotypes with molecular markers
Published November 15, 2007
100-104

Incorporation of competitive quality and resistance against the most important fungal diseases (powdery and downy mildew) in a cultivar is one of the most important aims of grapevine breeding. In the 20th century, the most advanced results in grapevine resistance breeding were achieved by French researchers. They used resistant cultivars in mor...e than 30% of their growing areas. In these varieties, North American wild Vitis
species were the resistance gene sources. The discovery of immunity-like resistance of Muscadinia rotundifolia opened new perspectives in resistance breeding. M. rotundifolia harbours a dominant powdery mildew gene, providing resistance in highquality cultivars after back-crosses with V. vinifera varieties. M. rotundifolia has been involved in the Hungarian grape breeding programs since 1996, thanks to a French-Hungarian variety exchange. In addition to traditional selection methods, application of MAS (Marker Assisted Selection) based on various types of
molecular markers, can provide additional tools for these efforts. Run1 locus, responsible for powdery mildew resistance, was identified in Muscadinia rotundifolia. Molecular markers closely linked to this locus are very significant in screening progenies deriving from M. rotundifolia and V. vinifera crosses, making possible the discrimination between resistant and susceptible genotypes at DNA level. In our analyses BC5 progeny of {(M. rotundifola×V. vinifera) BC4}×Cardinal (V. vinifera) tested for powdery symptoms were analysed with PCR-RFLP (GLP1- 12P1P3) and microsatellite markers (VMC4f3.1, VMC8g9). Our results proved the applicability of the linked markers and reliability of marker assisted selection.

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Isolation of promoters of tissue and ripening-specific strawberry genes by TAIL-PCR and bioinformatic characterization of the sequences
Published November 15, 2007
91-99

Isolation of ripening- and tissue-specific promoters has become a very important subject of the genetic regulation and plant physiology research in recent years. It could be possible to reveal the regulation of gene expression, and it may be a very useful approach in the biotechnology. In our work, we have isolated promoter regions of genes exh...ibiting ripening- and tissuespecific expression in our previous experiments, and the data were
characterized by bioinformatic methods. In the sequence of the ripening-specific Spatula and ACC-oxidase promoters (ACCoxidase is one of the key-enzymes of ethylene biosynthesis, directly related to the process of ripening); we could identify auxin- and ethylene-related cis-regulatory elements. This suggests that there is an interaction between ripening and ethylene-synthesis, in case of non-climacteric strawberry, too. We investigated the promoter regions of three green receptacle-specific genes (putative nitrilaselike protein, Ring transcription factor and an aquaporin protein)
and we could identify several regulatory elements, which refer to hormonal regulation. Additionally, we could find several cisacting elements which associated with stress-responsiveness and endospermium-specific expression. 

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Morphological diversity of current melons (Cucumis melo) compared to a medieval type
Published November 15, 2007
84-90

Morphological diversity of melon (Cucumis melo); phenotype reconstruction of a medieval sample. Morphological diversity among 47 melon (Cucumis melo) cultivars and landraces from Hungarian germplasm collection (ABI, Tápiószele) were analyzed with an ultimate aim to characterize morphologically cv. Hógolyó, which showed the closest genetic s...imilarity to a medieval melon recovered from the 15th century. Cultivars based on fruit morphology were grouped into the three main types of melon as reticulatus, cantalupensis and inodorus. Cluster analysis (by SPSS-11) based on 23 morphological (quantitative and qualitative) traits recorded revealed an extreme diversity among accessions, nevertheless cultivars were clustered into main melon clusters with only two exceptions of inodorus type cv. Zimovka J. and Afghanistan. Cultivars Sweet ananas and Ezüst ananász; and  two Hungarian landraces Kisteleki and Nagycserkeszi showed close similarity. Cultivars Hógolyó and Túrkeve of inodorus type
were also grouped in one cluster, which provide insight into the morphological reconstruction of the medieval melon recovered from the 15th century. These results also indicate that old Hungarian landraces could be re-introduced into breeding programs for broadening genetic base of melon.

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Application of microsatellite fingerprints for pedigree analysis of Hungaricum grapevine varieties
Published November 15, 2007
71-77

the Carpathian Basin were involved into our examination, which aimed at genotyping their accessions. DNA fingerprints of 101 varieties were determined with 6 microsatellite markers till 2005, resulting in successful discrimination of the accessions. Based on these results for pedigree determination, even more cultivars and primers were involved... into the analyses. For studying the origin of Csabagyöngye and for proving the parent-progeny relations of Irsai Olivér and Mátrai muskotály, 19 microsatellite markers were applied, while 11 were selected for tracing the origin of Királyleányka. Genetic distances between the varieties were estimated with cluster analysis and demonstrated by dendrogram, proving that the varieties can be discriminated from each other based on the microsatellite allele sizes. Pedigree of Irsai Olivér and Mátrai muskotály has been confirmed by microsatellite allele size results, searching for the parents of Csabagyöngye and Királyleányka is in progress, since the molecular-marker based pedigree does not correspond with the putative origin of these cultivars. Our results excluded progeny-parent relationships in the
Csabagyöngye-Bronnerstraube-Muscat ottonel (Ottonel muskotály) and the Királyleányka-Kövérszőlő combinations. 

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Agricultural Biotechnology MSc. course and agriculture in the 21st century
Published November 15, 2007
208-214

The establishment of an Agricultural Biotechnology MSc. course creates a new direction in Hungarian higher education. As an introduction, the article summarizes the main theoretical and practical possibilities and results of biotechnology which have necessitated launching the course. Subsequently, the preliminaries, aims and requirements of the... Masters course are introduced. The main data of the agricultural MSc course at Szent István University in Gödöllő (Hungary) are the following: 4 semesters, 120 credits, 1200 contact hours, 4 weeks training in a biotech laboratory, 6 fundamental subjects, 8 basic subjects in biotechnology. Plant and animal biotechnology, are the two options for specialization and both have 10 separate professional subjects. The teaching staff consists of 34 teachers (93% of them have PhDs or higher scientific degrees) from 10 Departments of 3 Faculties of Szent István University.

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Sequence heterogeneity of nSSR and cpDNA loci of Cucurbits (Citrullus sp.)
Published November 15, 2007
125-134

The evolution of water melon (Citrullus lanatus) microsatellites from the 15th century (Debrecen); 13th (Buda); and 18th century, (Pannonhalma) were analyzed. Microsatellite (nSSR, nuclear simple sequence repeat) and cpDNA profiles of the aDNA (ancient DNA) of seed remains were compared to modern water melon cultivars and landraces. Sixteen pri...mer pairs were applied. Sequence analysis at the (CT)26 and cpDNA trnV loci revealed a (CT)3 and Adenin deletions, respectively, form the current water melon cultivar compared to the medieval sample. Cila-1), a new LTR retrotansposon has been described. For morphological reconstruction, a dendrogram produced by SPSS11 based on the presence versus absence of 24 phenotypic characters were also analyzed.

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Microsatellite Diversity of Androgenic Black Poplar (Populus nigra)
Published November 15, 2007
60-67

Genetic 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 alle...les), 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. 

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SNP analysis of ITS1-5.8S-ITS2 rDNA loci in modern and ancient melons (Cucumis melo)
Published November 15, 2007
120-124

ITS (internal tanscribed spacer) profiles of the aDNA (ancient DNA) of seed remains extracted from an extinct sample recovered from the 15th century (Budapest, Hungary) were compared to 31 modern melon cultivars and landraces. An aseptic incubation followed by ITS analysis was used to exclude the exogenously and endogenously contaminated (Asper...gillus) medieval seeds and to detect SNPs in ITS1-5.8S-ITS2 region of rDNA (ribosomal DNA). SNPs were observed at the 94–95 bp (GC to either RC, RS or AG) of ITS1; and at 414 bp (A-to-T substitution), 470 bp (T to Y or C), 610 bp (A to R or G) of ITS2. The results facilitate the final aim of molecular and morphological reconstructions of ancient melon tpyes.

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