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Optimization of RNA isolation from stone fruits at different ripening stages
101-104.Views:212This study was conducted to select the most appropriate RNA isolation method that can be used successfully in case of stone fruits. The changing pattern of gene expression during the ripening process of stone fruits may elucidate the molecular background of several phenotypical or phytochemical alterations present among different genotypes. Our laboratory aims to study the expression of genes encoding for enzymes that catalyze crucial steps in the flavonoid biosynthesis pathway. RNA isolation from fruit mesocarp is a challanging task due to high levels of sugars and polyphenolics accumulating during fruit development. Therefore, at first, the optimal techniques eligible for RNA isolation from fruit tissues at different ripening stages must be selected. Our study compares three different RNA isolation protocols and describes their potential applicability according to different fruit species and ripening stages.
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Development of microsatellite markers for Rhodiola rosea
37-42.Views:243Rhodiola rosea L. is an important adaptogen medicinal plant. In this study two new microsatellite markers were developed. The assessment of the genetic diversity of R. rosea has recently started with molecular markers, but only a few species-specific microsatellite markers have been published so far. However the small number of markers allows only a limited insight into the genetic variability of the species therefore the aim of our work was to develop new microsatellite markers for R. rosea with a microsatellite enrichment library technique. Genomic DNA was cleaved with an endonuclease enzyme followed by adaptor ligation and PCR amplification. DNA fragments that contained microsatellites were first isolated using a biotin-streptavidin linkage based magnetic selection and then cloned into plasmids. Out of forty-three sequenced clones three contained microsatellites, in these cases primers were designed for the amplification of the microsatellite repeats. The newly developed primer pairs were tested on individuals from distant R. rosea populations and the variability of the amplified fragments was estimated by fragment-length analysis. The locus RhpB14a was found to be monomorphic while RhpB14b and RhpB13 were polymorphic. As a result of the present study, two novel variable microsatellite loci were identified in the genome of R. rosea.
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Analyses of Hungarian sour cherry germplasm with simple sequence repeat markers
27-31.Views:326Twenty-four sour cherry cultivars (genotypes), belonging to four cultivar groups were fingerprinted using microsatellite markers. All genotypes have been arisen from the Carpathian basin, which could be secondary gene centre of sour cherry, since its progenitor species, ground cherry and sweet cherry overlap here. Five SSR primer pairs, earlier used for fingerprinting Turkish sour cherry germplasm were tested. None of the five primer pairs showed any polymorphism within the cultivar groups. The primer pairs were able to distinguish between the cultivar groups. The Oblacsinszka and the Cigánymeggy cultivar groups were the most difficult to separate, while the Pándy cultivar group was the most distinguishable.
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Agrobacterium transformation of Rhodiola sp.: current status and limitations
37-42.Views:79The study of secondary metabolites has led to the discovery of new drugs for treating human diseases. However, consistent plant supply can be challenging, leading to the use of plant tissue culture techniques such as hairy root culture. Hairy roots have stable genetics, lateral branching, and can produce secondary metabolites, including alkaloids, flavonoids, and terpenoids. Research on hairy roots as a subject began in the late 19th century, and for the last four decades, hairy roots have been utilized for producing secondary metabolites and recombinant proteins. This article focuses on Rhodiola species - genus of perennial plants that belongs to the family Crassulaceae - and its potential as a source of secondary metabolites using hairy root culture techniques. Rhodiola sp. is widely distributed throughout the Arctic regions of the Northern Hemisphere, with several species having significant medicinal properties. The article discusses the possible use of hairy root cultures for the production of Rhodiola secondary metabolites, including salidroside and rosavins, which have demonstrated significant pharmacological activity in various studies. The use of elicitation and genetic engineering techniques to boost secondary metabolite production in Rhodiola hairy roots is also explored. Overall, the article highlights the potential of Rhodiola hairy root cultures as a valuable source of secondary metabolites with medicinal properties. However, despite some studies Rhodiola hairy root induction and culturing still remains highly unexplored.
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Technologies developed to avoid frost damages caused by late frost during bloom in the fruit growing regions of Siófok and Debrecen
99-105.Views:404The aim of the study was to fi nd out which of the methods used to avoid damages of late frosts would be the most effective for the fruit growing practice. Three technologies have been tested in the regions of Siófok and Debrecen-Pallag. The antifrost irrigation proved to be the most advantageous. For that purpose microjet sprayers are used, which are thrifty and do not need for that purpose large containers. With the aim to secure an even distribution of water, the sprayers are distributed on three levels: above and inside of the crown as well as on the level of trunks. On a large scale, a single microjet above the crown level would be suffi cient. By means of a detailed analysis served to set the optimum intervals between spraying phases: with each 15, 10, 5 and 3 minutes during half a minute. The synchronous presence of water and ice below the freezing point, the released freezing heat plus the water used much above the freezing point (9–10 °C) altogether maintains the temperature above around 0°C near the fl owers or growing fruitlets, meanwhile, the surrounding air cools down to –8 °C. The effectiveness of the generally used anti-frost would be increased substantially by the former application of cooling irrigation, which delays the blooming date. The Frostbuster represents a new technology developed for the same purpose of frost defence. It uses butane burning and produces high (80-90°C) temperature with a strong blast of air. On the protected area sensors have been distributed for measuring temperature and relative humidity. The means of the measurements proved a rise of temperature, which was suffi cient to save the fruit trees until the difference is less than –3–3.5 °C. Our analyses stated that paraffi n candles avert the frost until –4 °C. Its success depends largely on the intensity of air movements. A small wind would be enough to frustrate the effect. The results show clearly the utility of Frostbuster and paraffi n candle combined is approved.