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Detection of DNA mutations by PCR-TTGE method
21-25Views:510In 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 primers 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.
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Polymorphism of the bovine GH and LEP genes in a population of Slovak spotted bulls
19-23Views:278The aim of this study was the detection of polymorphism in the bovine growth hormone and leptin genes using the PCR-RFLP method. A
polymorphic site of the growth hormone gene (Alul loci) that results in amino acid change at position 127 of the protein chain (leucine, L to
valine, V) has been linked to differences in circulating metabolites, metabolic hormones and to milk yield. The polymorphism in bovine leptin
gene is situated in the intron between two exons, which results in an amino acid change at position 2059 of the protein chain (cytosine, C to
thymine, T). The polymorphisms were studied in a group of 58 bulls of the Slovak spotted breed. A strategy employing PCR was used to amplify 428 bp (GH gene) and 422 bp (LEP gene) products from blood samples. Digestion of PCR products with restriction enzymes AluI and Sau3AI revealed alleles: L and V; A and B for GH gene and LEP gene, respectively. The growth hormone gene is a candidate gene for body weight gain in cattle, since it plays a fundamental role in growth regulation. Leptin plays an important role in the regulation of feed intake, energy metabolism, growth and reproduction of cattle; therefore, animals with higher leptin gene expression will probably have lower daily weight gain than others with similar forage offer and nutritional condition and will also likely have longer calving intervals. -
Identification and biological examination of some inland Monilinia species
27-29Views:354The aim of this study was to identify and biologically analyse some Monilinia species from Hungary. 146 M. fructigena and 28 M. laxa species out 174 infectious fruit from all over the country were used for the study. For further study 10 isolates were used and apple fruit was inoculated with them according to Koch postulate. 1–2 mm ochre exogen stromas were observbed on infectious plant parts and growing signs on culture of all isolates were identical to M. fructigena. To affirm classical identification, isolates with molecular biological method were also prepared using PCR reaction. Control isolates of M. laxa, M. fructigena and M. fructicola were used. The size of PCR product showed that all isolates had a 415 bp band which was typical of M. fructigena. Results support the previous observation that M. fructigena and M. laxa species occure all over Hungary.
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Identification of cow’s and buffalo’s milk and dairy product using a DNA-based method
279-282Views:500Aim of our study was the optimization of a DNA method, that is appropriate for reliable, low cost identification of animal species in milk and dairy product (cheese) and to determine the ratio of species. Mitochondrial DNA was used in our work to analyse buffalo/cow milk mixtures contained different ratio of bovine milk such as 0.1%, 0.5%, 1%, 1.5%, 2%, 5%, 10%, 15% (v/v%). Buffalo cheese were produced using buffalo and cows milk (0%, 2%, 5%, 10%, 15% – v/v% cows milk in buffalo milk). In case of milk mixtures, using species specific primers, the PCR assay showed a 0.5 v/v% detection limit. Cattle, in the buffalo/cows milk 99.9/0.1 v/v% mixture, was not detectable. The identification of buffalo and cows DNA in cheese was successful. The intensity of eletroforetic PCR fragment indicated the increase of cow milk ratio in milk and cheese samples as well.
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Search for polymorphism in exon 5 of cattle pituitary adenylate cyclase activating polypeptide gene
17-20Views:619Pituitary adenylate cyclase-activating polypeptide is a neuropeptide expressed primarily in the hypothalamus and in other tissues, which divergent and extensive physiological functions are proved. Large number of SNPs in PACAP are published involved SNP that is associated with phenotypic trait of cattle as well. Hungarian Grey, Hungarian Simmental, Holstein, Charolais and Angus cattle breeds were involved in this study to search for polymorphism in exon 5 1–391 bp and G/A transition. Our results of PCR RFLP and PCR SSCP did not prove the occurrence neither G/A transition, nor any other SNP in cattle breeds involved.
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Preliminary Research Concerning the Association Between the Genotypes at the Kappa-Caseine Locus and Milk Production Traits in Cattle
45-48Views:179The genotypes for kappa-casein locus were established using PCR-RFLP. Genome DNA for PCR amplification was isolated from hair roots in cows (30 Schwitz individuals), and from frozen semen in bulls (42 bulls of different breeds).
A higher frequency of the B allele in Schwitz (pA = 0.45 and qB = 0.55 in cows and pA = 0.25 and qB = 0.75 in bulls), and a higher frequency of the A allele in bulls belonging to different breeds (pA = 0.61 and qB = 0.39 in Deutsch Simmental, pA = 0.84 and qB = 0.16 in Romanian Simmental; pA = 1.00 and qB = 0.00 in Holstein, and pA = 0.67 and qB = 0.33 in Pinzgauer) were estimated. The associations between the genotypes and alleles determined at the kappa-casein locus and milk production were tested. They emphasized significant associations only for milk and milk fat quantity, in advantage of individual carriers of the B allele in homozygous and heterozygous states. -
Qualitative detection of genetically modified organisms in plant samples
309-313Views:614We analysed the GMO content of corn samples by polymerase chain reaction following the appropriate optimization of the reaction. The analysis included two main steps: extraction of DNA from the sample, and detection of the GMO content by polymerase chain reaction. The polymerase chain reaction is an in vitro method to multiply chromosomatic or cloned DNA (cDNA) sequences through the enzymatic pathway. The reaction is sensitive enough to produce DNA in sufficient amount for the analysis from a single DNA. We identified the PCR products by agarose gel electrophoresis. When optimizing the reaction, the MgCl2 concentration, reaction time and temperature have to be taken into consideration. The temperature of the anellation has to be increased until the highest specificity and yield is reached. If the temperature of the anellation is too high, the primer is linked to non-specific sites as well; in the gel visualization, more lines can be seen at one sample. If the temperature of the anellation is too high, the primer is insufficiently linked or is not linked at all (too few lines in the gel visualization). After optimization, the GMO content in the unknown sample can be determined along with the appropriate positive and negative controls.
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Genetic polymorphism of candidate genes in pig meat production
37-40Views:205H-FABP, LEPR and MC5R genes were suggested as candidate genes for fat content in pig meat. The aim of this study was to detect genetic variation in the porcine H-FABP, LEPR and MC5R genes by PCR-RFLP method in a group of pigs. Genotyping of pigs was done by PCRRFLP methods. We identified three genotypes in the set of pigs, HH (0.504), Hh (0.412) and hh (0.084) for H-FABP (HinfI). Allele H showed higher frequency than allele h (0.710 vs. 0.290). Three genotypes were identified for the H-FABP (HaeIII) gene (DD - 0.194, Dd - 0.494, dd - 0.312). The allele D (0.441) showed slightly lower frequency than allele d (0.559). All three genotypes were identified for LEPR (HpaII) in the group of pigs (AA – 0.137, AB - 0.314, BB – 0.549). Higher frequency of LEPR gene was confirmed for allele B (0.706), as compared with allele A (0.294). We identified two genotypes for MC5R (BsaHI) in the group of pigs (AA - 0.348 and AG - 0.652), genotype GG was not found. As conforms with genotype structure, we recognize a higher frequency of allele A (0.674) as compared with allele G (0.326).
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The effect of β-glucan, carotenoids, oligosaccharides and anthocyanins on bacteria groups of excreta in broiler chickens
15-20Views:777This study was conducted to examine the effect of natural compounds, such as β-glucan, carotenoids, oligosaccharides, and anthocyanins in the diet on bacteria gropus of excreta in Ross 308 broiler chickens. Chickens were fed 5 diets: control (basal) diet, a diet supplemented by β-glucan at 0.05%, and diets supplemented by carotenoids, oligosaccharides, or anthocyanins at 0.5% of each compound. On experimental day 19, excreta were collected to determine the proportion of Lactobacillus, Bifidobacterium, Campylobacter, Clostridium, and Escherichia coli. Samples were collected aseptically and snap-frozen in liquid nitrogen. Bacterial DNA was isolated from samples, then polymerase chain reaction using primer pairs designed to the 16S rDNA of bacterial groups were applied to define the proportion of the mentioned bacteria. Another universal primer pair was used to amplify a region of 16S rDNA of all the examined bacteria. Proportion of each bacterial groups was determined relatively to the intensity of universal PCR product band by gel documenting system and ImageLab software. Based on the results, carotenoids and anthocyanins increased the proportion of Bifidobacterium, which might imply the beneficial effects of the mentioned compounds on the bacteria composition of excreta.
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DHAC-induced transgene overexpression in 35S-gshI GMO poplar (Populus × canescens)
78-83Views:127Relative gene expression levels of transgene gshI (γ-glutamylcysteine synthetase cloned from E. coli) were analyzed by qRT-PCR in two transgenic poplar (Populus × canescens) clones (11ggs and 6lgl) and wild type (WT). An extremely high expression level of transgene gshI was observed in the 6lgl clone (13.5-fold) compared to 11ggs (1.0) samples, which level was doubled (1.8-fold) in the DHAC (5.6-dihydro-5'-azacytidine hydrochloride) treated (at 10-4 M for 7 days) 6lgl samples but not in the 11ggs clone (0.4-fold). Contrary to this result, relative copy number of transgene gshI in the 6lgl clone was found to be less 60% less (1.0) then in the 11ggs samples (1.6). Relative expression levels of proper poplar gene gsh1-poplar showed significantly higher responsiveness to DHAC treatment than transgene gshI with the highest expression level in the untransformed (WT) poplar
clone (19.7-fold) compared to transformed 6lgl (8.7-fold) and 11ggs (2.5-fold) clones. For internal controls constitutively expressed housekeeping genes a-tubulin were applied. For data analysis, the 2−ΔΔCt method was used. DHAC applied in long-term cultures (27 days) at low concentrations (10-8 – 10-6 M) showed morphogenetic activity by initiating de novo root development of leaf discs. -
Selection of powdery mildew resistant and susceptible grapevine genotypes with molecular markers
100-104Views:323Incorporation 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. -
Effect of G2548A polymorphism in the leptin gene on the BMI level in human population
5-10Views:293The polymorphism in leptin (LEP 2548A) seems to influence obesity among others genes. The aim of this study is to investigate the effect of the G2548A polymorphism on body mass index. We included 79 people from Slovakia with some genetic relatedness and used barrels kit to isolate the genomic DNA from an adenoblast swab- from the salivary. PCR products were amplified by pursued polymorphisms and G2548A, we restriction-analyzed them and then we identified the specific fragments describing the presence of chosen SNP polymorphism by the agarose electrophoresis, to analyze SNP polymorphism by PCR-RFLP method.
The LEP gene had increased frequency of G allele (0.5506). The most common genotype occurring in the gene LEP was heterozygous genotype (AG) and the least frequent genotype in LEP was AA (0.1899). Taking the age into account the BMI is higher if the G allele occurs in the LEP gene. Moreover, if the G allele genotype was situated in dominant form, then the highest average BMI was present.
According to the results we can assume that the AA genotype (LEP) has a protective effect on the prevalence of obesity compared to the other genotypes.
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How does the S-locus determining self-incompatibility in stone fruits work in self-compatible peach?
93-100Views:334The majority of stone fruit species are self-incompatible, a feature that is determined by a specific recognition mechanism between the S-ribonuclease enzymes residing in the pistils and the F-box proteins expressed in the pollen tubes. Failure in the function of any component of this bipartite system resulted in self-compatibility (SC) in many cultivars of Prunus species. Peach (Prunus persica (L.) Batsch.) is the only species in the Prunoideae subfamily that is traditionally known to be self-compatible, but its molecular background is completely unknown. Isoelectric focusing and S-gene specific PCR revealed that SC is not due to functional inability of pistil ribonucleases. We hypothesize that SC may be a consequence of a kind of pollen-part mutation or the action of one or more currently unknown modifier gene(s). Only two S-alleles were identified in a set of peach genotypes of various origin and phenotypes in contrast to the 17–30 alleles described in self-incompatible fruit trees. Most important commercial cultivars carry the same S-allele and are in a homozygote state. This indicates the common origin of these cultivars and also the consequence of self-fertilization. According to the available information, this is the first report to elucidate the role of S-locus in the fertilization process of peach.
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Dieback of apricot plantations caused by 'Ca. Phytoplasma prunorum' in Borsod-Abaúj-Zemplén county (Northern-Hungary)
34-41Views:317Plant diseases caused by phytoplasmas have increasing importance in all over the world for fruit growers. Lately, phytoplasma diseases occur on many fruit varieties and responsible for serious losses both in quality and quantity of fruit production. In the long-run these diseases cause destruction of fruit trees. The apricot phytoplasma disease (Ca. Phytoplasma prunorum) was first reported in Europe in 1924 from France. In 1992 the disease has also been identified in Hungary. On the base of growers' signals serious damages of "Candidatus Phytoplasma prunorum" Seemüller and Schneider, 2004 (formerly: European stone fruit yellows phytoplasma) could be observed in different stone fruit plantations in the famous apricot-growing area nearby Gönc town, Northern-Hungary. Field examinations have been begun in 2009 in several stone fruit plantations in Borsod-Abaúj-Zemplén County mainly in Gönc region which is one of the most important apricot growing regions in Hungary, named “Gönc Apricot Growing Area”. Our goals were to diagnose the occurrence of Ca. Phytoplasma prunorum on stone fruits (especially on apricot) in the North-Hungarian growing areas by visual diagnostics and confirm data by laboratory PCR-based examinations. All the 28 collected samples were tested in laboratory trials and at 13 samples from apricot, peach, sour cherry and wild plum were confirmed the presence of phytoplasma (ESFY). On the base of observations it seems evident that the notable losses caused by "Ca. Phytoplasma prunorum" is a new plant health problem to manage for fruit growers, especially apricot producers in Hungary.
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Application of AFLP-Method in Plant Sample Identification
207-213Views:343One 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.
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Association analysis of TNNI1/XbaI polimorphism on carcass quality in hybrid pigs
59-62Views:554The contractile protein, which is encoded by troponin I 1 (TNNI1) gene, is located on the thin filaments of slow fibres in striated muscle. TNNI1 protein is a part of the troponin complex which plays an important role in regulation of muscle contraction by preventing actin-myosin interaction in absence of calcium. According to biological role, this gene can be potential marker for meat production related traits. The aim of this study is to define whether the previously reported gene polymorphism (EU743939:g.5174T>C) is connected with the slaughter traits measured in a standard slaughterhouse of the examined four-line European hybrid. The study included data from 404 gilts and barrows from 2 different samples. The polymorphism was detected using PCR-RFLP (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism) method with XbaI restriction enzyme. In this study the allele frequencies were found as follows: C: 0.84 and 0.808; T: 0.16 and 0.192. Based on result of the present study no significant impact of polymorphisms on production parameters was found.
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Comparative analysis of Staphylococcus aureus strains by molecular microbiology methods
34-39Views:546Staphylococcus aureus is a very important pathogen for dairy farms and milk processing plants. Subclinical mastitis is often caused by this species, and it can contaminate bulk tank milk when milking cows are suffering from mastitis. Additionally, thermostable enterotoxins (SE) produced by some types of this bacterium can cause food poisoning.
The aim of our research was to examine the number of S. aureus in bulk tank milk in two dairy farms and the enterotoxin-producing ability, genetic relation (pulsotype) and antibiotic resistance of S. aureus strains from different sources (bulk tank milk, udder quarter milk and environment).
The results show that the mean number of S. aureus of bulk tank milk of two farms significantly differed (P<0.05). Fourteen isolates were selected for further molecular genetic studies (five isolates were from bulk tank milk and nine isolates were from udder quarter milk). S. aureus was not recovered from the environmental samples. Three of the fourteen isolates (21.4%) tested by multiplex PCR were positive for SE genes. Two isolates carried one gene (seb) and one isolate carried two genes (seg and sei). The fourteen strains were classified into three pulsotypes and two subtypes at 86% similarity level. Isolates from bulk tank milk (n=5), were divided into 2 pulsotypes (A, C) and one subtype (C1). The isolates from udder quarter milk (n=9) belonged to three different pulsotypes (A, B, C) and two subtypes (A1, C1). The distribution of pulsotypes in the present study revealed genetic relationship between S. aureus isolated from udder quarter milk and bulk tank milk. This could be explained by the fact that in farms with a high number of infected cows, these cows could represent the main source of contamination. The results of the antibiotic resistance investigations show, that all strains were susceptible to methicillin, cefoxitin, lincomycin, tetracycline, erythromycin and sulfamethoxazole/trimethoprim. Thirteen out of fourteen strains were resistant to penicillin (A and C pulsotypes, A1 and C1 subtypes) and just one isolate was susceptible (B pulsotype) to all antibiotics tested. -
Identification of Hucul mare families by mtDNA markers
75-79Views:610Hundred animal species have disappeared during the last century. By this time, approximately one-third of domestic animals have been in the endangered category. Hucul horses are also in this category; furthermore saving the genetic diversity beside the race preservation is an important challenge as well. The number of mares and stallions is only one of the expressive elements of genetic diversity; together with their quality determine the genetic variability of this breed. Beyond that, if an exact breed can originates from more founders, it can be more renewed genetically. Stud book documents these data by registering the mare families and stallions’ genealogical lineage. Molecular genetics, especially mitochondrial DNA analysis can make the precise identification of mare families possible. As a result of these molecular based methods, protection of genetic diversity, as well as breed preservation became more reliable. After the primer designing, the optimal primer pair was chosen which targets a 1092 bp length DNA sequence in the cytochrome b region. After the successful PCR optimalisation, we determined 170 Hucul mares’ sequences. According to our results, the samples compose ten haplotypes, which are much less, than the registered number of mare families in the stud book. Further investigations are needed to reach more representative results, and drawn the further consequences.
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Influence of 17-alpha methyl testosterone on the production parameters of common carp (Cyprinus carpio L.) fry
37-43Views:579During our research we aimed at finding an answer as to what extent the different concentrations of 17-alpha methyl testosterone incorporated in the diet of common carp fries can influence the production parameters of the species, as well as how efficient their sexreversal can be with the use of this method. To this end, an aquarium experiment was conducted in the course of which four different hormone treatments were set and monitored. The fish feed was enriched with 17-alpha methyl testosterone in 50 ppm, 75 ppm, 100 ppm, 500 ppm dosages.
The obtained figures revealed that the hormone treatments had no influence on the production parameters and conservation of the common carp fries. Further on, our team is to determine the sex of the fish through the examination of gonads during autopsy when they reach the 500 g average weight.
Furthermore, a male specific test method which was supposed to be of great help in our attempt to select the sex-reversed specimens in the subsequent processes was also put to the trial. During the experiment the DNA-isolation of different sample types (muscle tissue, fin, mucus) of common carp with identified sex was successfully carried out. The extracted PCR product was examined with agarose gel. Our results indicated that the ccmf2 marker was applicable, however, the obtained figures were not reliable.
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Q-PCR analysis of the resistance of Hungarian Botrytis cinerea isolates toward azoxystrobin
41-44Views:283The 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 between 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. -
Recovery and confirmation of Haemonchus contortus from abomasal contents of roe deer (Capreolus capreolus) in Eastern-Hungary (Biharugra): A diagnostic case study
59-62Views:393Gastrointestinal parasites are ubiquitous. They occur both in wild and domesticated animals. Among such parasites of veterinary importance is the trichostrongyle worms, out of which the Haemonchus contortus species is regarded as the most pathogenic one in the small ruminant industry. The occurrence of this parasite in the sheep flock is now very well documented and an established fact in Europe, although the parasite was original of the warmer climatic region. Studies on the cross-transmission of H. contortus between the wild and domesticated animals are also on the rise although the question of the direction of transmission is still debated. This is an important area that needs to be addressed as it could potentially contribute indirectly to mitigating anthelmintic resistance. Hungary also has reported its share of the occurrence of the parasite, mainly in the sheep flock and a certain population of roe deer. The study presented here is the preliminary results of a diagnostic case study that confirms the presence of H. contortus in wild ruminant deer species that are close to the domesticated sheep population.
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Associations analysis of production traits with leptin gene T3469C polymorphisms in pig
39-43Views:518The aim of this study was to define the connection between the leptin (LEP) gene T3469C polymorphism and its potential association with production traits in improved hybrid pigs. The study included data from 397 gilts and barrows from 2 different sample. The polymorphism was identified by using the polymerase chain reaction-restriction fragment-length polymorphism (PCR–RFLP) method with HinfI restriction enzyme. Two alleles of LEP gene were identified: T (0.93) an C (0.07). The analysis of values of production traits, depending on LEP genotype did not reveal significant (P≤0.05) differences. In the examination the loin diameter (between the 2nd and 3rd ribs), the live weight at slaughter and the averege daily gain during fattening were higher at pigs with C allele than pigs with TT genotipe. Accordingt to our data the effect of C allele was favourable in this population, because these animals had bigger bodyweight without valuable change of lean meat percent.
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Isolation of promoters of tissue and ripening-specific strawberry genes by TAIL-PCR and bioinformatic characterization of the sequences
91-99Views:175Isolation 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 exhibiting 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. -
SSR based characterization of peach (Prunus persica L.) and apricot (Prunus armeniaca L.) varieties cultivated in Hungary
17-24Views:470The 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. -
Fruit melanotic ringspot (FMRS) – a disease of resistant Capsicum genotypes infected with Tomato spotted wilt virus (TSWV) on the fruits
64-69Views:167Etiology of pepper fruit melanotic ringspot (FMRS) disease (Salamon, 2009) was studied on fruit samples collected in forced pepper populations. It was noticed that in spite of heavy thrips (Frankliniella occidentalis) infestations and of TSWV epidemy detected in the forcing houses, FMRS occurred only in plants having healthy foliage. Symptomatological surveys strongly suggested that FMRS appeared exclusively in specific pepper genotypes. The size of melanotic ringspots has been observed to grow at room temperature during postripening of diseased fruits. A mechanically transmitted plant virus was isolated from symptomatic parts of 9 white pepper fruits affected by FMRS. On test plants each of the virus isolates caused systemic symptoms characteristic to TSWV. Using cDNA/PCR technique and TSWV N-gene specific primers a ca. 300 bp long DNA fragment has been amplified from total nucleic acid extracted from symptomatic tissues but never from asymptomatic parts of the fruits showing FMRS. Plant progenies grown from seeds of FMRS diseased fruits segregated in respect of resistance and/or susceptibility to TSWV infection. TSWV was also detected in and isolated from three fruits showed non-melanotic yellow rings (one of them was infected with a tobamovirus, too). Seedlings derived from these fruits proved to be susceptible to TSWV. Based on the above results we could conclude that the FMRS disease developed on fruits of “cecei” type white peppers that carry a TSWV resistance gene, most likely the Tsw gene in heterozygous form. These fruits were infected with thrips transmitted TSWV and FRMS appeared as a hypersentive reaction (HR) manifested in fruits.