Eukaryotic-like Ser/Thr Protein Kinases SpkC/F/K Are Involved in Phosphorylation of GroES in the Cyanobacterium Synechocystis

Serine/threonine protein kinases (STPKs) are the major participants in intracellular signal transduction in eukaryotes, such as yeasts, fungi, plants, and animals. Genome sequences indicate that these kinases are also present in prokaryotes, such as cyanobacteria. However, their roles in signal transduction in prokaryotes remain poorly understood. We have attempted to identify the roles of STPKs in response to heat stress in the prokaryotic cyanobacterium Synechocystis sp. PCC 6803, which has 12 genes for STPKs. Each gene was individually inactivated to generate a gene-knockout library of STPKs. We applied in vitro Ser/Thr protein phosphorylation and phosphoproteomics and identified the methionyl-tRNA synthetase, large subunit of RuBisCO, 6-phosphogluconate dehydrogenase, translation elongation factor Tu, heat-shock protein GrpE, and small chaperonin GroES as the putative targets for Ser/Thr phosphorylation. The expressed and purified GroES was used as an external substrate to screen the protein extracts of the individual mutants for their Ser/Thr kinase activities. The mutants that lack one of the three protein kinases, SpkC, SpkF, and SpkK, were unable to phosphorylate GroES in vitro, suggesting possible interactions between them towards their substrate. Complementation of the mutated SpkC, SpkF, and SpkK leads to the restoration of the ability of cells to phosphorylate the GroES. This suggests that these three STPKs are organized in a sequential order or a cascade and they work one after another to finally phosphorylate the GroES

Несезонное производство ягодной продукции малины красной в условиях отапливаемых зимних теплиц

Relevance. Currently, in many countries of the world, the production of non-season raspberry berry products has become widespread. Recently, interest in this technology has arisen in Russia, which has great prospects for the development of industrial gardening. In our opinion, it is promising to develop elements of technology for the non-seasonal production of red raspberries, propagated by the method of clonal micropropagation with a traditional and remontant type of fruiting in the conditions of winter heated greenhouses.Material and methods. The experiments were carried out in the laboratory of clonal micropropagation of garden plants in the fruit growing laboratory of RGAU-MSHA named after K.A. Timiryazev. The objects of research were varieties of red raspberries with a traditional (variety Volnitsa) and remontant (varieties Orangevoe Chudo and Bryanskoe Divo) type of fruiting. The experimental plants were propagated by the method of clonal micropropagation and grown before distillation in open and protected ground; plants propagated by root offspring served as control. Experimental plants were planted in open ground for growing in mid-May, in mid-October they were transplanted into 10 liter containers and transferred to protected ground conditions. Then put in the refrigerator compartment with a temperature of + 1 ... + 5°C. For distillation, the raspberry repairing plants were exposed in the winter heated greenhouse on January 20, while the shoots of replacing the aboveground system were normalized: without normalization, 3 shoots per plant, complete pruning of the aboveground system. Raspberries with a traditional type of fruiting were exposed in a winter heated greenhouse in three periods on January 20, February 10, March 2. Accounting for the passage of the phenological phases of development and yield was made for 3 months every 5 days.Results. In the conditions of winter heated greenhouses, efficiency has been shown and elements of technology for non-season production of raspberry berries remontant and berries with a traditional type of fruiting, propagated in vitro and grown before open field distillation are developed. It was revealed that it is necessary to normalize the shoots before distillation of raspberry remontant, and the optimal timing for the start of distillation for raspberries with a traditional type of fruiting has been established.Актуальность. В настоящее время во многих странах мира широкое распространение получило производство несезонной ягодной продукции малины. В последнее время интерес к данной технологии возник и в России, что имеет большие перспективы для развития промышленного садоводства. На наш взгляд, перспективно разрабатывать элементы технологии несезонного производства ягод малины красной, размноженной методом клонального микроразмножения с традиционным и ремонтантным типом плодоношения в условиях зимних отапливаемых теплиц.Материал и методика. Опыты проводили в лаборатории клонального микроразмножения садовых растений лаборатории плодоводства РГАУ-МСХА им. К.А. Тимирязева. Объектами исследований служили сорта малины красной с традиционным (сорт Вольница) и ремонтантным (сорта Оранжевое чудо и Брянское диво) типом плодоношения. Опытные растения были размножены методом клонального микроразмножения и выращены перед выгонкой в открытом и защищенном грунте, контролем служили растения, размноженные корневыми отпрысками. В открытый грунт растения были высажены в середине мая, в середине октября их пересадили в контейнеры объемом 10 л и перенесли в условия защищенного грунта. Затем выставили в холодильный отсек с температурой 1…5°C. Для выгонки растения малины ремонтантной выставляли в зимнюю отапливаемую теплицу 20 января, при этом производили нормировку побегов замещения надземной системы: без нормировки, 3 побега на растение, полная обрезка надземной системы. Малину с традиционным типом плодоношения выставляли в зимнюю отапливаемую теплицу в три срока 20 января, 10 февраля, 2 марта. Учеты прохождения фенологических фаз развития и поступления урожая производили в течение 3 месяцев через каждые 5 дней.Результаты. В условиях зимних отапливаемых теплиц показана эффективность и разработаны элементы технологии несезонного производства ягод малины ремонтантной и с традиционным типом плодоношения, размноженных in vitro и выращенных перед выгонкой в открытом грунте. Выявлено, что необходимо провести нормировку побегов перед выгонкой малины ремонтантной и установлены оптимальные сроки начала выгонки для малины с традиционным типом плодоношения

Development of Legal Regulation of Arctic Marine Oil and Gas Resources: Soft-law Approach

In recent years there has been seen rapid growth of interest in the Arctic marine oil and gas, which supposedly constitute 30-40 % of the world’s undiscovered oil and gas resources. At the same time, accidents, which could take place while developing of the resources, can not only cause irreparable injury to the Arctic ecosystem, but have global consequences. This article analyzes current international law regulating the development of the Arctic marine oil and gas resources and examines one of the directions of its development ‒ cooperation amount the Arctic States in international organizations and forums, creating “soft-law” rules. The article also considers to acts of international non-governmental organizations (unions, associations and etc.). In conclusion, the article analyzes the advantages and disadvantages of using of the “soft-law approach” to regulate the development of the Arctic marine oil and gas resources

Non-season production of raspberry of red berry products in conditions of heated winter greenhouses

Relevance. Currently, in many countries of the world, the production of non-season raspberry berry products has become widespread. Recently, interest in this technology has arisen in Russia, which has great prospects for the development of industrial gardening. In our opinion, it is promising to develop elements of technology for the non-seasonal production of red raspberries, propagated by the method of clonal micropropagation with a traditional and remontant type of fruiting in the conditions of winter heated greenhouses.Material and methods. The experiments were carried out in the laboratory of clonal micropropagation of garden plants in the fruit growing laboratory of RGAU-MSHA named after K.A. Timiryazev. The objects of research were varieties of red raspberries with a traditional (variety Volnitsa) and remontant (varieties Orangevoe Chudo and Bryanskoe Divo) type of fruiting. The experimental plants were propagated by the method of clonal micropropagation and grown before distillation in open and protected ground; plants propagated by root offspring served as control. Experimental plants were planted in open ground for growing in mid-May, in mid-October they were transplanted into 10 liter containers and transferred to protected ground conditions. Then put in the refrigerator compartment with a temperature of + 1 ... + 5°C. For distillation, the raspberry repairing plants were exposed in the winter heated greenhouse on January 20, while the shoots of replacing the aboveground system were normalized: without normalization, 3 shoots per plant, complete pruning of the aboveground system. Raspberries with a traditional type of fruiting were exposed in a winter heated greenhouse in three periods on January 20, February 10, March 2. Accounting for the passage of the phenological phases of development and yield was made for 3 months every 5 days.Results. In the conditions of winter heated greenhouses, efficiency has been shown and elements of technology for non-season production of raspberry berries remontant and berries with a traditional type of fruiting, propagated in vitro and grown before open field distillation are developed. It was revealed that it is necessary to normalize the shoots before distillation of raspberry remontant, and the optimal timing for the start of distillation for raspberries with a traditional type of fruiting has been established