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Sampling for vegetative propagation: A phytosanitary status survey of grapevines collection by One Step RT-PCR method
2021
Ізейрай, М
Purpose. Grapevines (Vitis spp.) are affected by many viral diseases which cause serious pathological problems. GLRaV-3 is among the most widespread leafroll viruses, while Grapevine Fanleaf Virus (GFLV) is a destructive pathogen which reduces the lifespan of grapevine. Considering the impact and the spread of these diseases, our objective was to analyse the presence of these two viruses in several grapevine varieties in grapevine collection at ATTC Vlore. Data gathered from plant pathogens serve to better understand and prevent the spread of pathogens, as a mandatory rule for the quality control of certified plant material during vegetative propagation. Method. The presence of two common viruses were tested using virus specific primers; LC1/LC2 primer pair designed from the hHSP70 gene for detecting Grapevine Leafroll-associated Virus-3 (GLRaV3) and C3390/H2999 primer pair, designed from coat protein coding regions for detecting Grapevine Fanleaf Virus (GFLV), in six varieties; ‘Merlot’, ‘Kallmet’, ‘Shesh i zi’, ‘Shesh i bardhё’, ‘Debinё’, and ‘Pulёz’, provided through a randomised sampling procedure. One Step Reverse Transcription Polymerase Chain Reaction assay was used to detect the viral presence. Results showed a high (100%) prevalence of GLRaV3 virus in all of analysed samples, as the most frequent among the two pathogens. Analysis for of GFLV virus showed low infection rate, being present in only one sample. Conclusions. We herein show an efficient, fast and reproducible method for detecting grapevine viruses through one step RT-PCR. Our results suggest that sampling of the infected plant material should be avoided due to the presence of viral infections.
Afficher plus [+] Moins [-]Bioinformatic analysis of nucleotide sequences of the acetolactate synthase (als) gene of different members of the Poaceae family
2020
Кирієнко, А. В | Щербак, Н. Л | Симоненко, Ю. В
Purpose. Provide bioinformatic analysis and comparison of target regions of the acetolactate synthase (als) gene in several members of the Poaceae family and, on the basis of the obtained data, explore the possibility of creating a unified genetic construct for als gene editing using the CRISPR-Cas9 system. Methods. The als gene sequences of various members of the Poaceae family were obtained from the NCBI: Nucleotide database. For comparison, a fragment of the imi-2 gene of wheat of the soft line ‘TealIMI11A’ was used in two regions of the 367–390 and 1729–1749 nucleotide sequences. The Sequence Viewer 3.37.0 tool was used to assess the presence of nucleotide substitutions in the working sequence of the imi-2 gene. The dendrogram was built using the “Blast Tree” tool from the NCBI: Blast: Nucleotide resource. Results. A comparative analysis of the nucleotide sequences of seven different species was carried out: soft wheat (Triticum aestivum L.), common wild oat (Avena fatua L.), barley (Hordeum vulgare L.), Asian rice (Oryza sativa L.), maize (Zea mays L.), aleppo grass (Sorghum halepense Pers.) and Tausch’s goatgrass (Aegilops tauschii Coss.). The dendrogram is based on the gene sequence als, showed that all studied genotypes can be divided into two blocks: the first block included maize and aleppo grass, and the second block, a separate branch includes Asian rice and common wild oat, barley, soft wheat and Tausch’s goatgrass. 367–390 nucleotide sequences of soft wheat showed the highest 100% homology to Asian rice, Tausch’s goatgrass and common wild oat. The lowest homology was for maize and aleppo grass at 83.3%. Evaluation of the nucleotide sequence 1729–1749 showed no complete homology at the 100% level. It was the highest for barley and Tausch’s goatgrass – 95.2%, and the lowest for rice, maize and aleppo grass – 80.9% each. Conclusions. The analysis confirms a significant degree of homology of the als gene sequence for various species of the Poaceae family, which allows us to create a universal genetic vector. However, taking into account the high degree of sequence homology for species such as soft wheat, Tausch’s goatgrass, barley, Asian rice and common wild oat, it can be assumed that the corresponding genetic vector can be used with the greatest efficiency to alter the als gene of these genotypes.
Afficher plus [+] Moins [-]Study of some biochemical parameters and productivity of Silphium L. genotypes as perspective energetic crops
2020
Рахметов, Д. Б | Вергун, О. М | Рахметова, С. О | Шиманська, О. В | Фіщенко, В. В
Purpose. This study was aimed to determine some biochemical parameters and productivity of the gene fund of Silphium L. genus in the M. M. Gryshko National Botanical Garden of the NAS of Ukraine. Methods. Plant raw material investigated at the flowering stage (17 genotypes) and the end of vegetation (20 genotypes) of Silphium spp. 3 species and 4 cultivars studied for the content of nutrients at the flowering. Determination of dry matter, ash, calcium, nitrogenfree extractives conducted according to Hrytsaienko et al. (2003), phosphorus, protein – according to Pochinok (1976), sugars – according to Krishchenko (1983), cellulose – according to Zheng et al. (2018), lipids – according to Zamowski, Suzuki (2004). It was used productivity parameters: yield of above-ground mass, the yield of dry mass, energetic value, yield of energy. Data analyzed statistically. Results. Investigation of nutrients content showed that content of dry matter was in the range of 18.90–29.3%, protein – in the range of 8.88–23.56%, cellulose – 15.10–36.14%, ash – 8.13–12.19%, lipids – 1.83–3.97%; yield of above-ground mass – 45.0–139.0 t/ha, the yield of dry matter – 10.31–36.92 t/ha, energy value – 3933–4249 cal/g, and yield of energy – 43.81–149.27 Gcal/ha. A study of genotypes at the flowering and end of vegetation identified that the content of dry matter for all samples was in a range of 18.38–67.49%, sugars – 2.78–19.0%, ash – 3.93–11.20%, calcium – 1.68–5.99%, phosphorus – 0.14–1.21%, energy value – 3153.36– 3770.28 cal/g. Conclusions. Plant raw material of genotypes of Silphium L. spp. is a valuable source of nutrients. The content of ash, its components, energetic value, and parameters of productivity depending on genotype and stage of growth. The results allow recommending selected Silphium genotypes as perspective energetic crops in Ukraine as well as other countries.
Afficher plus [+] Moins [-]Bioinformatic analysis of maize gene encoding starch branching enzyme SBEIIb
2016
Сліщук, Г. І | Жернаков, Т. Ю | Волкова, Н. Е
Purpose. Investigation of maize ae1 gene polymorphism by bioinformatic methods. Methods. Global and local alignment of the nucleotide and amino acid sequences, in silico translation and transcription, translates modeling, primers design, phylogenetic analysis. Results. 255 nucleotide sequences of maize аe1 gene, 500 amino acid sequences of homology translates of maize ae1 gene (SBEIIb enzyme homologs) and 100 mRNA expressed from the maize ae1 gene were analyzed to establish phylogenetic relationships. Polymorphism of maize ae1 gene different regions was investigated by bioinformatic methods. Modeling of the maize enzyme SBEIIb was performed. Conclusions. According to the results of amino acid sequences of SBEIIb enzyme homologs alignment, it was found that ae1 gene orthologs are present only in monocots, paralogs – in monocots, dicots, and other taxa, including algae and animals. Based on the results of alignment of plants mRNA from which enzyme SBEIIb is translated, maize ae1 gene orthologs and the nearest paralogs encoding starch branching enzymes with chloroplast localization were defined; this suggests a possible origin of ae1 gene due to duplication of the gene encoding the 1,4-alpha-glucan-branching enzyme 2 with chloroplast or amyloplast localization. In the maize ae1 gene structure, regions were found that include polymorphic sites not defined previously. For the polymorphic sites design primers were developed that allowed to differentiate the maize lines. It was determined that the detection of polymorphism in theory can influence the enzyme function and, as a result, change the concentration of amylopectin in maize grain.
Afficher plus [+] Moins [-]Peculiarities of the genetically modified organisms’ detection in seed batches of rape (Brassica napus L.)
2014
Король, Л. В | Гончарова, С. О | Піскова, О. В | Костенко, А. В | Коровко, І. І | Кожемякіна, Л. М
This paper presents the results of molecular studies on testing and evaluation of multiplex test systems for the genetically modified organisms’ (GMOs) identification by PCR in real time. For the detection of genetic modifications in plants of the cabbage family (Brassicaceae) screening of major regulatory sequences is insufficient, as in genetic engineering manipulations 35S promoter – of DNA-containing cauliflower mosaic virus (CaMV) is used, which presence in the test material can lead to false positive results. In this work the feasibility of testing for the presence of viral CaMV’s DNA and analysis of screened gene sequences of interest for the usage during the analysis of rapeseed (Brassica napus L.) samples. It was established that the analysis of the presence/absence of a positive result for the CaMV 35S promoter is not giving complete answers to the absence of GMO in the sample. The appropriateness of GMO’s screening in rape seeds with NOS terminator and genes of interest.
Afficher plus [+] Moins [-]Adaptability of F1 sunflower hybrids, created according to an integrated system of line selection for economically valuable traits in various agroclimatic zones
2021
Бабич, В. О | Боровська, І. Ю | Шарипіна, Я. Ю | Парій, Я. Ф | Симоненко, Ю. В
Purpose. Determine the ecological plasticity and productivity of F1 sunflower hybrids created on the basis of maternal and parental lines, selected according to an accelerated selection system of lines resistant to herbicides (imidazoline and sulfonylurea groups) and broomrape (Orobanche cumana Wallr.). Methods. Statistical analysis of F1 sunflower hybrids was carried out using the methods of variation statistics, regression and analysis of variance using the Microsoft Office Excel 2016 application package. Molecular biological, biotechnological and classical selection methods were used for the accelerated system of line selection. Thus, for the purpose of targeted selection of sunflower sterility fixers, we used HRG01 molecular SCAR marker to identify the gene for the restoration of pollen fertility (Rf1). To accelerate the creation of parental lines resistant to tribenuron-methyl, we used a culture of immature embryos in vitro. Results. The results of testing of F1 sunflower hybrids at Kyiv, Chernihiv, Cherkasy (Uman and Shpolianskyi districts), Khmelnytskyi, Kharkiv, Kherson and Odesa regions. The hybrids were created on the basis of selected lines, chosen according to an accelerated selection system for herbicide-resistant lines (imidazoline (IMI-hybrids) and sulfonylurea (SU-hybrids) groups) and broomrape (Orobanche cumana Wall). The standards for comparison with hybrids were: for IMI hybrids – hybrids ‘NK Neoma’ (Syngenta) and ‘ES Genesis’ (Euralis), and for SU-hybrids – ‘SY Sumiko’ (Syngenta) and ‘P64LE25’ (Pioneer). As a result, it was found that among SU-hybrids, UA 2/106 had a 3.9% higher yield when compared to the standards (‘SY Sumiko’ and ‘P64LE25’). And for IMI-hybrids it was found that hybrids UA 1/67, UA 1/66, UA 1/84 have the same yield of 2.76 t/ha as the ‘NK Neoma’ standard. IMI hybrids UA 1/92, UA 1/102 have the same yield of 2.91 t/ha as ‘ES Genesis’. Conclusions. F1 hybrids were created on the basis of the original breeding material selected due to the accelerated system of sunflower lines selection. The hybrids were analyzed according to the yield indicator. The most productive among the tested SU-hybrids was UA 2/106 hybrid, among the IMI hybrids – UA 1/67, UA 1/66, UA 1/84, UA 1/92 and UA 1/102.
Afficher plus [+] Moins [-]Combining ability of tetraploid pollinator lines of sugar beet (Beta vulgaris L. ssp. vulgaris var. altissima Doell) by the elements of productivity
2013
Корнєєва, М. О | Чемерис, Л. М | Мацук, М. Б
Evaluation of the genetic component values considered to be the important part of the selection process on creating hybrids of sugar beet and The aim of the study is to determine tetraploid pollinators combining ability Bilotserkovskoy breeding on yield and sugar content and the genetic determination of the productivity elements and their phenotypic manifestations in first generation hybrids of sugar beet. The methods of test crossing on the type top cross of pollen sterile lines of Uuladivsky and Ivanovsky origin and stabilized ployidnistyu tetraploid pollinators of beet sugar Belotserkovskoy selection on yielding of sugar beet tetraploid pollinators of Bilotserkovskoy breeding have been applied. A key role belongs to non additive gene effects in gene structure of characteristics variability yielding has been determined.The part of additive gene action pollinators is predominant in genetic control of top cross hybrids sugar content. The 16 hybrid combinations are differentiated by their parental components combining ability. Pollinators of 1007 (the yield) and 1013 (for the sugar content) have been characterized by their valuable additive gene complexes. Perspective hybrid combinations for their further breeding study are revealed, genetic determination of productivity elements and sugar yield is defined Analysis of gene interactions in sugar beet hybrids confirms the theory of genetic balance MV Turbine
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