Acta Fytotechnica et Zootechnica Online (Faculty of Agrobiology and Food Sciences, Slovak University of Agriculture in Nitra)
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Comparative analysis of different methods of Hedera helix DNA extraction and molecular evidence of the functionality in PCR
Get PDFReceived: 2016-06-25 | Accepted: 2016-07-29 | Available online: 2016-12-22http://dx.doi.org/10.15414/afz.2016.19.04.144-149The most suitable method of total DNA extraction still remains the crucial step for many plant species, although there are manydifferent protocols and commercial kits for DNA isolation. In this study, five different extraction protocols were analysed to find outthe most appropriate method for DNA extraction from Hedera helix L. This species has numerous medical and pharmaceutical usesand is also characterized by antioxidant effects on human body. In spite of its wide medical utilization, it belongs to those plantspecies, where the genomic information is very limited. Comparing of different protocols resulted in the yield of extracted DNAthat has ranged from 6.3 to 487 ng μl-1. The purity of extracted DNA has ranged from 1.4 up to 2.0 A260/A280. All the extractionmethods used in this study were evaluated not only in term of quantity and purity of DNA but also its functionality in the restrictionendonuclease digestion and polymerase chain reaction based downstream analysis was performed.Keywords: DNA extraction, CTAB, SDS,commercial kit, Hedera helix L.ReferencesABU-ROMMAN, S. (2011). Comparison of methods for isolating high quality DNA from sage(Salvia officinalis). Journal of Medicinal Plants Research, vol 5, no. 6, pp. 938-941BALANSARD, G. et al. (1980). Douvicidal and Antifungal Activities of α-Hederin Extracted fromHedera helix leaves. Planta Medica, vol. 39, p. 234.BARRA, M. et al. (2012). Simple and robust DNA extraction mehod for the large-scale analysis ofgenotypes containing high polyphenolic content, such as landraces of Solanum tuberosum and Zeamays. Plant breeding, genetic and genetic resources, vol. 39, no. 3, pp. 593-601.BEDIR, E. et al. (2000). Triterpene Saponins from the Fruits of Hedera helix. Phytochemistry, vol.53, pp. 905-909.CLARKE, M.M. et al., (2006). Prevalence of different horticultural taxa of ivy (Hedera spp.,Araliaceae) in invading populations. Biological Invasions, vol. 8, pp. 149–157.COTA-SÁNCHEZ, H. J. et al. (2006). Ready-to-Use DNA Extracted with a CTAB Method Adaptedfor Herbarium Specimens and Mucilaginous Plant Tissue. Plant Molecular Biology Reporter, vol. 24,no. 2, pp.161-167.CRESPIN, F. et al. (1995). Identification of 3O-beta-D-Glucopyranosyl Hederagenin from theLeaves of Hedera helix. Fitoterapia, vol. 66, pp. 47.DABO, S. M. et al. (1993). A method for the isolation of nuclear DNA from cotton (Gossypium)leaves. Analytical Biochemistry, vol. 210, pp. 34-38.DELLAPORTA, S. L. et al. (1983). A Plant DNA Minipreparation: Version II. Plant Mol. Biol. Rep.,vol. 1, pp. 19-21.DEMEKE, T. and JENKINS, R. (2010). Influence of DNA extraction methods, PCR inhibitors andquantification methods on Real time PCR assay of biotechnology-derived traits. Analytical andBioanalytical Chemistry, vol. 396, pp. 1977-1990.DOOSTY, B. et al. (2012). Comparative Analysis and Optimization of Different DNA ExtractionProtocols in Satureja khuzistanica. International Journal of Biology, vol. 4, no. 4,pp. 112-116.ELIAS, R. et al. (1991). Triterpenoid saponins from the leaves of Hedera helix. Journal of NaturalProducts, vol. 54, pp. 98-103.GREEN, A. F. et al. (2011). Phylogeny and Biogeography of Ivies ( Hedera spp., Araliaceae), aPolyploid Complex of Woody Vines. Systematic Botany, vol 36, no. 4, pp. 1114-1127.HUAQIANG, T. et al. (2013). Comparative Analysis of Six DNA Extraction Methods in Cowpea(Vigna unguiculata L.Walp). Journal of Agricultural Science, vol 5, pp. 82-90.IVERSEN, J. (1944). Viscum, Hedera and Ilex as climatic indicators. Geologiska FöreningersStockholm Förhandlinger, vol. 66, pp. 463–483.JOBES, D. V. et al. (1995) Plant DNA isolation: A Method to Efficiently Remove Polyphenolics,Polysaccharides, and RNA. Taxon, vol. 44, pp. 379-386.KALENDAR, R. et al. 2010. iPBS: a universal method for DNA Fingerprinting and retrotransposonisolation. Theoretical and Applied Genetics, vol. 121, pp. 1419–1430.KRAJEWSKA, A. (1981). Phytochemical Studies of Hedera helix Herbage, Flowers and Fruits.Herba Polonica, vol. 27, pp. 303-309.LADE, B.D. et al., (2014). Efficient genomic DNA extraction protocol from medicinal rich Passiflorafoetida containing high level f polysacharide and polyphenol. SpingerPlus, vol. 3, p. 457.LUTSENKO, Y. et al. (2010). Hedera helix as a medicinal plant. Herba Polonica, vol. 56, pp. 83-96.MAJESTERSAVORNIN, B. et al. (1991). Saponins of the ivy plant. Planta Medica, vol. 57, p. 260-262.
Influence of the Year Season and Lairage Length in the Slaughterhouse on the Quality of Pig Meat
Get PDFThe impact of the length of pig housing in the slaughterhouse and the season on the meat quality of pigs was analyzed in the experiment. There were altogether 104 commercial pig hybrids used in the experiment weighing from 100 to 130 kilograms. The acidity of meat was assessed 45 minutes after slaughter in the longest back muscle - pH1 MLD and the semimembranosus thigh muscle – pH1 MSM. We found the lower pH1 value in MLD muscle (6.07) compared to MSM muscle (6.22) (P<0.05). The lowest occurrence of PSE meat in MLD and MSM was found out in the pigs slaughtered two hours after arrival to the slaughterhouse. The length of housing lasting for eight hours and more has negatively affected the quality of meat, which resulted in decreasing the average values of pH1 in MLD (P<0.05) as well as in MSM (statistically insignificant).The lower average values of pH1 MLD (5.98) were recorded in pigs slaughtered in the summer season compared to the winter period (6.16) (P<0.05). During the summer season, there was a threefold higher occurrence of the PSE meat in MLD muscle than in winter. The season had no impact on the values of pH1 MSM. This work has been created with the support of KEGA 006SPU-4/2014 and VEGA 1/0818/16.Keywords: acidity, lairage, pork, PSE, seaso
Winter wheat noodle colour evaluation in sustainable farming systems
Get PDFThe aim of this work was to evaluate colour of noodles that were prepared from winter wheat grown in ecological and integrated arable farming systems after different forecrops with two levels of fertilization (fertilized and unfertilized) during the years 2009 and 2010. Winter wheat noodles were prepared from white flour and wholegrain flour and it’s colour was evaluated using the spectro-colorimeter. Colour of white flour noodles was significantly influenced by forecrop, crop nutrition (fertilized and unfertilized variants) and by meteorological conditions during experimental years. Farming systems had no statistical effect on white flour noodles colour values. The colour of wholegrain noodles was affected by farming system, forecrop, crop nutrition and also the year of growing. Whole grain noodles from ecological system were darker, with lower lightness, higher redness compared to noodles from integrated system. Fertilization increased redness, yellowness and decrease the lightness of white flour noodles, on the contrary, fertilization increase the lightness and decrease the redness of wholegrain noodles.Keywords: ecological, integrated arable system, wheat noodle colou
Socio-economic effects of the organic certification system
Get PDFThe function of the organic certification system is to enable systemic trust in organic agriculture and products. The growth of the organic sector over the past two decades has required therefore a change in the certification system replacing the first party, or peer review, system by a third party certification, in which a disinterested party provides the certification. Third party inspection is based on an institutionalized mistrust order, which induces several effects. Consumers trust in organic labelling schemes, when there is a substantial active and visible state involvement. But there are also other effects within both the certification system and the inspection bodies. Furthermore there are effects in interactions with producers or customers. The literature review shows that so far studies dealing both with processes in and effects of certification systems have been rare.Keywords: certification system, organic agriculture, confidence buildin
Preliminary analysis of utilization of genomic relationship in mating plan of Old Kladruber horse
Get PDFThe study analyzed 48 Old Kladruber horses genotyped by Illumina Equine SNP70 BeadChip for usefulness of genomic data in determining of mating plan. Totally 12 variants of data filtering and their impact on calculations in dependence of different parameters of GenCall Score, Minor Allele Frequency and assumed average values of loci of ancestors (l) was investigated. For possibility of comparison between genomic and commonly evaluated relationships, pedigree based relationship matrix was constructed and subsequently subtraction of pedigree from genomic matrix was performed. All matrices were thoroughly inspected and most suitable setting of parameters was chosen. Evaluation of genomic relationships can be successfully implemented in more precise method of mating plan design of Old Kladruber horses. Further genotyping and development of method for rescaling of differences between genomic and pedigree relationship matrices´ elements is advised for a purpose of better interpretation of results by breeders
Natural Fusarium occurrence qualitative evaluation on cereals under conventional and organic system in the Czech Republic
Get PDFCereals are the most growing crops under organic conditions. Cereals are supposed to be a source of mycotoxins produced by Fusarium species. Our research was led to describe the differences among natural Fusarium species occurrence in two localities and two growing systems – organic and conventional. The main differences were observed among years and cereal species. No statistically significant differences were determined between localities and farming systems.Keywords: Fusarium natural occurrence, organic farming, conventional farmin
Selected candidate genes affecting milk fatty acids
Get PDFCow´s milk consisted of relatively high quantity of saturated fatty acids (about 70%), which are considered less beneficial health. The content of more desirable fatty acids is about 25% (monounsaturated) and 5% (polyunsaturated). The aim of this research was to find ways how to increase the content of mono- and polyunsaturated fatty acids in milk fat. One of the ways how to alter the content of fatty acids is the monitoring of candidate genes and their polymorphism. Therefore, the manuscript deals with selected candidate genes, particularly DGAT1, SCD1 and FASN, which are associated with fatty acid biosynthesis. The results suggest that observing of this issue could be helpful for improving the quality of milk fat.
The canine melanophilin gene polymorphisms in Slovakian Rough-haired Pointer
Get PDFThe aim of this study was to determine the population genotypic structure based on polymorphisms located in exon 1 (c.-22G>A) and exon 7 (R199H) of canine gene encoding melanophilin (MLPH). The genomic DNA were obtained from in total 49 samples of Slovakian Rough-haired Pointer and analysed using PCR-RFLP methods. The prevalence of G allele has been found in case of both identified loci. The highest frequency has been observed for heterozygous animals and its sufficient proportion was confirmed also by the coefficient of heterozygosity (0.71 or 0.67) and the FIS index values (-0.45 or -0.47). The FST index showed expected high degree of genetic similarity between analysed populations resulted from their breeding purposes and common founders. The genotyping of such polymorphisms can be perspective mainly due to the fact that the MLPH gene was associated with the coat colour dilution and also alopecia not only in human but as well as in dog populations
P uptake, root properties and biomass of six cover-crops
Get PDFPhosphorus (P) is an essential and limited nutrient. To avoid shortages in the coming decades, P recycling and efficiency of P use will have to be enhanced. Cover crops may help to transform less available P forms into bioavailable P forms. The aim of this investigation was the comparison of P uptake with root traits of six cover crops. Root and shoot biomass was analysed for P content. Root hair length and root length was measured on the basis of 6400 dpi root scans using ImageJ and WinRhizo. In addition, the root hair cylinder was calculated. P uptake was found to be strongly correlated with the root hair cylinder and the total crop biomass.Keywords: cover crops, phosphorus uptake, root hair cylinder, root length, root hair
The effect of protein metabolism on weanlings blood parameters level
Get PDF DOI:10.15414/afz.2015.18.03.76-78 Received 14. July 2015 ǀ Accepted 28. August 2015 ǀ Available online 14. October 2015This study was conducted to determine the effects of low-protein diets supplemented with crystalline amino acids (AA) on biochemical parameters and performance in 10 crossbred piglets weaned at 28 days of age ( 2 groups of 5 each, 8.8±0.6 kg and 8.6±0.7 kg live weight). The treatments were a control diet containing 210.8 g.kg -1CP (crude protein) and low protein diet containing 186.4g.kg-1 supplemented with crystalline AA (lysine, threonine, methionine) to achieve an ideal AA pattern. Blood from all piglets was taken for determining biochemical parameters 5 weeks after weaning. The decrease in the diet CP content was manifested signification P< 0,01 decrease concentrations of blood urea (average concentrations 2.61 mmol.l-1 and 4.21 mmol.l-1), which means the increase of biological value in the feed mixture