The traditional Hungarian horse breed, Gidran has been close to the edge of extinction several times. Despite the multiple bottleneck effect, the breed has retained a part of its genetic variability, and performed prominently in carriage driving and show-jumping competitions. Maintaining of the Gidran breed is important in the point of view of...world heritage; because besides Hungary, smaller Gidran populations exist only in Bulgaria and Romania. Taking advantage of the special inheritance features of mtDNA, our study focused on two mtDNA regions of Gidran mares. Altogether, 251 hair samples from various Hungarian studs were examined. The analysis was successfully made in case of 251 samples of the cytochrome b and in case of 246 samples of D-loop regions. Because of the distinct mutation rates of the two mtDNA markers, the number of the haplotypes and the way of grouping samples into haplotypes was different. Our key finding was that most haplotypes may be compatible with mare families of the stud book; however incidental mistakes in stud book have occurred only in a few cases. Our results indicate the importance of the preservation and breeding those mare families, which are molecular genetically more diverse than the others, and are in the edge of extinction.
The genes being in the mitochondrial DNA primarily encode the enzymes of cellular respiration. Fungicides belonging to the family of quinol oxidase inhibitors (QoIs) play on important role in the protection against several plant diseases caused by fungi. These fungicides bind to the cytochrome bc1 complex so they block electron transport betwee...n cytochrome b and cytochrome c1. This way these fungicides inhibit the ATP synthesis consequently they inhibit the mitochondrial respiration. The QoI resistance has two mechanisms. One of them is the point mutation of the cytochrome b gene (CYTB), e.g. the substitution of a single glycine by alanine at position 143 results in high-resistance. The other is the cyanide-resistant alternative respiration sustained by the alternative oxidase.
In a cell there are several mitochondria. The phenomenon when the genomes of all mitochondria in the cell are identical is called homoplazmy. If in the cell there is wild and mutant mitochondrial DNA this is called heteroplasmy. Whether the mutation in the mitochondria causes fenotypical diversity or does not depend on the dose, i.e. it depends on the percentage of the changed mitochondrials. During our work we investigated Botrytis cinerea single spore isolates which have been collected in 2008-2009 on different host plants. Our goal was to decide whether heteroplasmy influences the level of resistance. We managed to detect the change of the level of heteroplasmy, so the change the level of the resistance due to the treatment with fungicide.
In our study PCR-temporal temperature gelelectrophoresis (TTGE) and MeltINGENY bioinformatic program were used to analyse the mutations in the genes of melanocortin-1 receptor (MC1R) and pituitary adenylate-cyclase activating polypeptide (PACAP) in cattle. Amplification of target DNA by PCR was performed with GC-clamp primers and non-GC-clamp p...rimers in simplex PCR reactions. The fragments were separated by denaturing polyacrylamide gelelectrophoresis (denaturing agents: high temperature, urea) after PCR reactions.MC1R homozygous individuals were used for the reaction.
We concluded that MeltINGENY program makes the decision and detection system easier, and more simple as the melting profile of target sequence is determined by the software. In case of MC1R gene, PCR-TTGE method is appropriate for SNP detection, however PACAP gene polymorphism can not be identified by the method, because PACAP mutations are not included in melting domains, therefore PCR-TTGE cannot detect them.