Search
Search Results
-
Application of microsatellite fingerprints for pedigree analysis of Hungaricum grapevine varieties
71-77Views:99the 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. -
Examination of microsatellite markers of Dorper sheep breed
57-61Views:176Number of not woolly and molty sheep exceeds 60 million throughout the world. Their numbers and their importance is growing, still they have appeared in the past two decades all over in North-America, Australia, New-Zealand and also in Europe. The South African Dorper has been a pioneer among them in Hungary. It was introduced in 2006 in the country. The Dorper sheep is the second largest breed in South Africa, which was developed from the crossing of Dorset Horn and the Blackhead Persian. The aim of the EU Member States in terms of this specific breed is increasing the small populations, improving the productive qualities, in addition to this avoiding inbreeding. However, finding appropriate breeding stock is difficult due to the small size of available populations and also to the suspected common of origin. With the help of various molecular genetic methods we could get a total view of the genetic background of these flocks. Nowadays the most commonly known and used genetic markers are microsatellites, because their applications give fast, accurate and easily reproducible results. There is no specific descriptive information on the genetic background of Dorper populations in the various EU countries , also regarding diversity between populations. Therefore in our work we want to optimize the conditions of applicability of 31 selected microsatellite reactions as a first step of mapping the entire genetic background of the different EU Dorper populations.
-
Genetic diversity study of Hungarian honey bee (Apis mellifera L.) colonies – Previous announcement
61-64Views:146In Hungary, the Apismelliferacarnica is the native breed which is the only recognised and breedable honey bee in the country. It is assumed that there are a number of non-native and hybrid honey bee breeds in Hungary. The microsatellite and mitochondrial DNA surveys applied here will be utilised to easily and accurately identify the various sprads, and open up new ways in the research of honey bees. The isolation of the genomial DNS from 5 to 7 day old larvae samples was successfully carried out. In the future the plan is to carry out the measure with microsatellite markers. As an initial step the optimal annealing temperature was identified. In the mitochondrial DNA survey the COI-COII mytochonrial regional primer due to its low anneling temperature cannot be used with any normal sequencing methods. By using these method the aim of this research is the measurement of genetic variance.
-
Examination for Susceptibility of CAE
33-36Views:95Caprine Arthritis Encephalitis (CAE) is a group of diseases in goats caused by a retrovirus, namely the Caprine Arthritis Encephalitis Virus. CAEV belongs to the lentivirus group within the retrovirus family. The CAEV is intimately associated with white blood cells, therfore any body secretions which contain white blood cells are a potential source of virus to other goats in the flock. Once goats are infected with CAEV, it remains infected for life and has to be eliminated.
Since not all goats infected with CAEV progress to disease, very important to test goats for infection using a serology test (ELISA, AGID, IDT). These serological tests demonstrate the presence of antibodies to CAEV in goat serum or detect the virus genome in the white blood cells. However, Swiss results (Dolf and Ruff, 1994) pointed out individual and species differencies in predisposition for this disease. This result was considered as a starting point to our examination. Microsatellite analysis was used in order to find whether there was any association between genotype and serological results, and to look for a marker associated with this disease. To date, altogether 135 goats have been examined. Unfortunately, a significant association between serological results and genotype was not found using the Chi2 test -
Selection of powdery mildew resistant and susceptible grapevine genotypes with molecular markers
100-104Views:108Incorporation 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 more 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. -
SSR based characterization of peach (Prunus persica L.) and apricot (Prunus armeniaca L.) varieties cultivated in Hungary
17-24Views:262The 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 heterogeneity of nSSR and cpDNA loci of Cucurbits (Citrullus sp.)
125-134Views:79The 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 primer 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.
-
Study of a Turkey Population for Gene Preservation
48-52Views:85Genetic variability is very important in small populations. We examined an indigenous bronze turkey population which is bred for gene conservation in order to see if the current mating system maintains genetic variability. The present generation was surveyed using microsatellite markers and a computer model was used to simulate changes in the population over 100 generations.
The data was analysed using the concept of entrophy from information theory instead of genetic variance so that we could more accurately measure genetic variability.
The results indicate that the breeding method currently in use, rotational line mating, is acceptable with respect to preserving genetic variability, but new selection methods may provide additional protection against the loss of alleles. -
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. -
Microsatellite Diversity of Androgenic Black Poplar (Populus nigra)
60-67Views:67Genetic 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. -
Application of molecular genetic methods for species Equus caballus – Literature review
21-28Views:151In this study our aim was to provide a comprihensive overview of the most commonly used methods in molecular genetic studies related to Equus caballus. Thus we are dealing with the D-loop region of mitochondrial DNA, with microsatellites and also with single nucleotid polimorphism as SNP. The advantages and drawbacks of each method were also explored.