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.

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. | Цель. Провести биоинформатический анализ и сравнить целевые участки гена ацетолактат синтазы (als) у нескольких представителей семейства Злаковых и на основе полученных данных исследовать возможность создания унифицированной генетической конструкции для направленного изменения гена als с помощью системы CRISPR-Cas9.Методы. Сиквенсы гена als различных представителей семейства Злаковых были получены из базы данных NCBI: Nucleotide. Для сравнения был использован фрагмент гена imi-2 пшеницы мягкой линии ‘TealIMI11A’ в двух участках сиквенса 367–390 и 1729–1749 нуклеотидов. Для оценки наличия нуклеотидных замен в рабочих сиквенсах гена imi-2 использовали инструмент ‘Sequence Viewer 3.37.0’. Дендрограмму строили с использованием инструмента “Blast Tree” с ресурса NCBI: Blast: Nucleotide. Результаты. Был проведен сравнительный анализ нуклео­тидных последовательностей семи различных видов: пшеницы мягкой (Triticum aestivum L.), овсюга обыкновенного (Avena fatua L.), ячменя обыкновенного (Hordeum vulgare L.), риса посевного (Oryza sativa L.), кукурузы (Zea mays L.), сорго алепского (Sorghum halepense Pers.) и эгилопса Тауша (Aegilops tauschii Coss.). На основе сравнительного анализа сиквенса участков гена іmi-2 для 7-ми генотипов построили филогенетическое дерево, которое показало, что исследованные виды можно разделить на два блока. В первый блок вошли кукуруза и сорго алеппское, а ко второму блоку – рис посевной, овсюг обыкновенный, ячмень обыкновенный, пшеница мягкая и эгилопс Тауша. Определение степени гомологии между последовательностью 367–390 нуклеотида пшеницы мягкой и другими видами показала, что абсолютной была гомология с соответствующими последовательностями риса посевного, эгилопса Тауша и овсюга обыкновенного (100%). Наименьшим нуклеотидное родство оказалось для кукурузы и сорго алеппского – по 83,3%. На участке 1729–1749 нук­леотидов гена іmi-2 никакой из 6 сиквенсов не показал 100% гомологии с последовательностью пшеницы мягкой. Самой высокой она была для ячменя обыкновенного и эгилопса Тауша – 95,2%, а наименьшей для риса посевного, кукурузы и сорго алеппского – по 80,9%.Выводы. Проведенный анализ подтверждает значительную степень гомологии последовательности гена als для различных видов семейства Злаковых. Это позволяет допустить возможность создания универсальной генетической конструкции, с помощью которой можно осуществлять редактирование генома и получения растений, устойчивых к гербициду разных представителей этой семьи. Учитывая высокую степень гомологии последовательностей для таких видов как пшеница мягкая, эгилопс Тауша, ячмень обыкновенный, рис посевной и овсюг обыкновенный, можно предположить, что с наибольшей эффективностью соответствующая генетическая конструкция может быть использована для редактирования гена als именно этих генотипов. | Провести біоінформатичний аналіз та порівняти цільові ділянки гена ацетолактат синтази (als) у декількох представників родини Злакових і на основі отриманих даних дослідити можливість створення уніфікованої генетичної конструкції для направленого редагування гена als за допомогою системи CRISPR-Cas9. Методи. Сиквенси гена als різних представників родини Злакових було отримано з бази даних NCBI: Nucleotide. Для порівняння було використано фрагмент гена іmi-2 пшениці м’якої лінії ‘TealIMI11A’ у двох ділянках сиквенсу: 367–390 та 1729–1749 нук­леотидів. Для оцінювання наявності нуклеотидних замін в робочих сиквенсах гена іmi-2 використовували інструмент “SequenceViewer 3.37.0”. Дендрограму будували з використанням інструменту ‘BlastTree’ з ресурсу NCBI: Blast: Nucleotide. Результати. Було проаналізовано нуклеотидні послідовності сімох різних видів: пшениці м’якої (TriticumaestivumL.), вівсюга звичайного (AvenafatuaL.), ячменю звичайного (HordeumvulgareL.), рису посівного (OryzasativaL.), кукурудзи (ZeamaysL.), сорго алепського (SorghumhalepensePers.) та егілопса Тауша (AegilopstauschiiCoss.). На основі порівняльного аналізу сиквенсу ділянок гена іmi-2 для 7-ми генотипів було побудовано філогенетичне дерево, яке показало, що досліджені види можна поділити на два блоки. До одного з них увійшли кукурудза та сорго алепське, а до другого – рис посівний, вівсюг звичайний, ячмінь звичайний, пшениця м’яка та егілопс Тауша. Визначення ступеня гомології між послідовністю 367–390 нуклеотиду пшениці м’якої та іншими видами показало, що абсолютною була гомологія з відповідними послідовностями рису посівного, егілопса Тауша та вісюга звичайного (100%). Найменшою нуклеотидна спорідненість виявилась для кукурудзи та сорго алепського – 83,3%. У ділянці 1729–1749 нуклеотидів гена іmi-2 жоден з 6 сиквенсів не показав 100% гомології з послідовністю пшениці м’якої. Найвищою вона була до ячменю звичайного та егілопсу Тауша – 95,2%, а найменшою для рису посівного, кукурудзи та сорго алепського – по 80,9%. Висновки. Проведений аналіз підтверджує значний ступінь гомології послідовності гена als для різних видів родини Злакових. Це дозволяє зробити припущення про можливість створення універсальної генетичної конструкції, за допомогою якої можна редагувати геном та отримувати рослини, стійкі до гербіцидів різних представників цієї родини. Зважаючи на вищий ступінь гомології послідовностей для таких видів як пшениця м’яка, егілопс Тауша, ячмінь звичайний, рис посівний та вівсюг звичайний, можна припустити, що ефективнішою відповідна генетична конструкція буде для редагування гена als саме цих генотипів.

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.

Purpose. To investigate rice blast resistance genes polymorphism by using bioinformatic methods. Methods. Global and local nucleotide alignment, phylogenetic analysis, HyPhy test. Results. For Pib gene, numerous single nucleotide substitutions and deletions of 1–3 bp were established. The phylo­geny of this gene has been studied and homologues have been found both in various rice species and in other cereals. These sequences can encode proteins that «recognize» the phytopathogens effectors, and can also be associated with resistance to phytopathogens. The Pi4 gene is characterized by single nucleotide substitutions, insertions and deletions; the number of non-synonymous substitutions exceeds the number of sy­nonymous ones. The Pi54 gene variability is significantly lower than that of the Pi4 and Pib genes. The predominant types of polymorphism were single nucleotide substitutions and small-sized indels. It was found that non-synonymous substitutions in Pi54, Pi4 and Pib genes were in close proximity, sometimes forming clusters, while some coding regions were either superconservative or contained predominantly synonymous substitutions. On philodendrograms, cultivated rice samples were clustered with samples of ancestral wild-growing species. Conclusions. Evolution of the rice blast resistance genes Pi4, Pib and Pi54 is characterized by diversification selection. Considering that tense coevolution and significant rate of adaptation and creation of new pathogen races are typical for a plant and a parasite, these genes are subjected to intensive selection aimed at increasing diversity for obtaining the resistance to new races of the pathogen.

It’s been shown the ways user impotent nucleotide plant of soybean, agriculture growing in the world last years. Revealed main point of formation nationality plant varieties resource of soybean.

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.

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.