Технология укоренения удвоенных гаплоидов редиса европейского, полученных в культуре микроспор in vitro

Relevance. Doubled haploids (DH-plants) are excellent material for genetic research and breeding due to their complete homozygosity. The genus Raphanus from the Brassicaceae family is the toughest to produce doubled haploid plants through isolated microspore culture in vitro (IMC). The study of the causes of disturbed root formation and the development of elements of this stage of technology will significantly increase the effectiveness of the IMC technology for European radish.Methods. The study included three varieties from the collection of the Federal State Budgetary Scientific Institution Federal Scientific Vegetable Center (FSBSI FSVC): Teplichny Gribovsky, Rozovo-krasniy s belim konchikom and Rhodes. The experiments used a standard protocol for obtaining DH plants using IMC technology in a standard form and with a modification of the rooting stage. The solid MS medium (with agar 7g/L): MS without hormones, MS medium supplemented with IAA at concentrations of 0.5; 1 and 2 mg / L and liquid MSm medium supplemented with 0.1 mg / L kinetin were used for rooting of regenerated plants. All media were supplemented with 20 g/L sucrose. We used three types of techniques for transplanting plant explants onto a solid hormonefree MS medium: planting micro-shoots with their basal part immersed by 2-3 mm into the medium; planting in a well made in a nutrient medium using tweezers under sterile conditions; and landing on the surface of the medium without embedment.Results. In this work, we studied the features of the stage of rooting of regenerated European radish plants in vitro conditions. The transplant technique has been proven to be important for the successful establishment of radish micro-shoots. Plant explants must be planted strictly on the surface of a solid hormone-free nutrient medium MS, without embedment. The use of tubes with bridges made of filter paper and MSm liquid medium with the addition of 0.1 mg/L kinetin for the induction of root formation also showed high efficiency. For plants prone to the formation of root-like structures (RLS) with secondary tumors (ST), multiple dissection of abnormal formations with successive transplants is necessary. Modification at the rooting stage of micro-shoots growing has increased the percentage of successfully adapted DH plants in vivo conditions from 0-14% to 95-98%.Актуальность. Удвоенные гаплоиды (DH-растения), являются превосходным материалом для генетических исследований и селекции за счет полной гомозиготности. Род Rathanus в семействе Brassicaceae является самым не отзывчивым к технологии получения удвоенных гаплоидов с помощью культуры изолированных микроспор in vitro (IMC). Изучение причин нарушения корнеобразования у растений-регенерантов и отработка элементов этого этапа технологии, позволит значительно повысить эффективность IMC технологии для редиса европейского.Методы. В исследование были включены три сортообразца из коллекции ФГБНУ «Федеральный научный центр овощеводства» (ФГБНУ ФНЦО): Тепличный Грибовский, Розово-красный с белым кончиком и Родос. В ходе экспериментов использовали стандартный протокол получения DH-растений с помощью IMC технологии в стандартном виде и с модификацией этапа укоренения. Для укоренения использовали твердую (агар 7 г/л) среду MS: безгормональная и MS с добавлением ИУК (в концентрациях 0,5; 1 и 2 мг/л) и жидкая питательная среда MSm с 0,1 мг/л кинетина. Концентрация сахарозы во всех средах составляла 20 г/л. Использовали три вида техники пересадки растительных эксплантов на агаризованную безгормональную среду MS: посадка микропобегов с погружением их базальной части на 2-3 мм в среду; посадка в лунку, сделанную в питательной среде с помощью пинцета в стерильных условиях; и посадка на поверхность среды без заглубления.Результаты. В настоящем исследовании изучены особенности этапа укоренения растенийрегенерантов редиса европейского в условиях in vitro. Показано, что для успешного укоренения микропобегов редиса важна техника пересадки. Растительные экспланты необходимо высаживать строго на поверхность твердой безгормональной питательной среды MS без заглубления. Использование для индукции корнеобразования жидкой среды MSm с добавлением 0,1 мг/л кинетина в пробирках с мостиками из фильтровальной бумаги также показала высокую эффективность. Для растений, склонных к образованию корнеплодоподобных структур (КС) с вторичными опухолями (ВО), необходима многократная диссекция аномальных образований с последовательными пересадками. Модификация этапа укоренения микропобегов повысила процент успешной адаптации DH-растений в условиях in vivo с 0-14% до 9598%

External high-frequency control of combustion instability

The article presents the results of experimental studies of combustion instability in the pulse combustor. Propane-air mixture is burned in the chamber with the flame holder. It was experimentally found that feeding high-frequency sound vibrations into the combustion chamber causes the suppression of pulsating combustion. The oscillation frequency ranges in 870 to 1400 Hz. This corresponds to 9-12 resonance frequencies of oscillations in the combustor. The physical mechanism of the observed phenomenon consists in changing the conditions of formation and destruction of fuel jets in the vortex zone behind the flame holder

Factors affecting DH plants <i>in vitro</i> production from microspores of European radish

Over the recent years the market demand for scaling up the production of European radish (Raphanus sativus L.) varieties and hybrids for open and protected production, varying in ripeness group, root shape and color, has drastically increased. Therefore, the expansion of genetic diversity and acceleration of the selection process are important. Doubled haploid technology considerably curtails the time required for creation of homozygous constant parental cell lines when in vitro microspore culture is used as the most promising method. For the first time, we were able to realize the full production cycle of DH plants of European radish by in vitro microspore culture up to inclusion of the produced material into the selection process. We have selected: preferable flower bud size, heat shock parameters, induction and regeneration media. It was revealed that linear length on the flower buds with the best possible stage of microspore development is genotype-specific: the flower bud length 2.8-3.3 mm is optimal for accessions of Rhodes and 3.7-4.2 mm is optimal for accessions of Teplichny Gribovsky. Heat shock at 32 °C for 48 hours is the most suitable for most genotypes. For the first time Murashige and Skoog based culture medium has been used for embryogenesis induction, and a major dependence of embryogenesis induction on the genotype x medium interaction was found. At regeneration and tiller stage it is advisable to add 1 mg/mL of benzylaminopurine and 0.1 mg/L of gibberellic acid to the medium, and rotting of micro-sprouts is performed with the use of hormone-free medium. Analysis of the produced regenerant plants by chromosome count and cell nucleus flow cytometry showed that 69 % of plants have a diploid chromosome set, 9 % have a haploid chromosome set, and 22 % have mixoploids and aneu-ploids chromosome sets. The seed progeny from doubled haploids and mixoploids were obtained by self-pollination, where all R1 plants had a doubled set of chromosomes. This study launches the development of an efficient method of radish doubled haploid production to be used in the selection process

Features of vanadium geochemistry in oils from the oil and gas fields of Eastern region of Ukraine

The results of long-term studies of the geochemical features of vanadium in the oil from 36 deposits of the Eastern oil and gas region of Ukraine. Based on the results of correlation and regression analyses, the nature and forms of the relationship between the vanadium content in oil and nickel, zinc, chromium, manganese, cobalt, iron, mercury, aluminum and the total content of these metals were determined; average capacity of the productive horizon; oil density value; oil viscosity value; resin content; the density of formation water from the productive horizon; sulfur content in oil; actual depth of the productive horizon; actual temperature of the productive horizon; actual pressure of the productive horizon; oil boiling point; paraffin content; the value of mineralization for formation water from the productive horizon; content of asphaltenes. Correlation coefficients, pairwise linear regression equations as well as curves that illustrate relationships between these parameters are obtained. Based on the results of the cluster analysis, a dendrogram of the results for clustering by means of the weighted centroid method of the considered fields by the content of vanadium in the oils was elaborated. Based on the results of clustering by means of the weighted centroid method, the first natural classification of oil and gas fields of eastern region of Ukraine according to the content of vanadium in oils was developed. It has been proven that heteroatomic low-molecular sulfur-containing components of the petroleum system are the main concentrators of vanadium in the oil for the studied deposits. It has been shown that, based on the results of the cluster analysis, sample average values of vanadium concentrations that vary significantly between individual deposits or groups of deposits for established ranges could be interpreted in terms of qualitative assessment as follows: abnormally low; low; below average; average; above average; high; abnormally high. The implementation of this kind of approach makes it possible to visually compare and interpret in terms of geology various scale and various indicators of oil deposits obtained by means of experimental study

Mechanism of influence water vapor on combustion characteristics of propane-air mixture

The article discusses the results of an experimental study of the effect of water vapor at the flame temperature. Propane-butane mixture with air is burning on a modified Bunsen burner. Steam temperature was varied from 180 to 260 degrees. Combustion parameters changed by steam temperature and its proportion in the mixture with the fuel. The fuel-air mixture is burned in the excess air ratio of 0.1. It has been established that the injection of steam changes the characteristics of combustion fuel-air mixture and increase the combustion temperature. The concentration of CO in the combustion products is substantially reduced. Raising the temperature in the combustion zone is associated with increased enthalpy of the fuel by the added steam enthalpy. Reducing the concentration of CO is caused by decrease in the average temperature in the combustion zone by applying steam. Concentration of active hydrogen radicals and oxygen increases in the combustion zone. That has a positive effect on the process of combustion

Factors to affect inbred beet plants while developing material for linear selection

Considering its capacities, the generative system of Beta vulgaris L. is regarded as highly productive. While inbreeding, the reproductive potential of cross-pollinated beet plants with gametophytic self-incompatibility (SI) changes significantly and is determined by a joint effect of multiple factors including the level of inbred depression. In the present study, original data have been obtained revealing relationships between inbred beet seed productivity, its self-incompatibility and microgametophyte parameters, which is crucial for developing and maintaining constant fertile beet lines. It has been discovered that inbred depression increases the number of sterile microgametes and anomalous pollen grains, reduces pollen fertility and the length of pollen tubes. As a result, the seed yield in inbred beet progeny, including SI ones, reduces significantly just after the third inbreeding. At the same time, highly productive inbred beet is characterized by a lower rate of pollen tube growth in vitro. In inbred plants, there is no close relationship between pollen viability and seed productivity, because the elimination of germinated male gametes and degeneration of seed embryos may go over the entire period of fertilization starting its progamic phase. The SI plants have more degenerating embryos than self-fertile ones, but seed vessel outgrowth in the seeds with abortive embryos makes them morphologically similar to fertile seeds. For that reason, when assessing inbred beet plants based on their self-incompatibility/self-fertility, one should consider the qualitative characteristics of the seeds. Using the method of recurrent selection based on such factors as seed productivity, pollen tube length and field germination rate increase the output of plant forms with a potentially high self-compatibility in their progeny. To support such genotypes in the progeny, one has to, starting from the third inbreeding, perform sib crossing to reduce the negative effect of inbred depression and self-incompatibility

Методы кластеризации данных при распознавании эндоскопических снимков в задачах компьютерной медицинской диагностики

This paper presents the results of the analysis of existing methods for clustering data obtained during endoscopy of a larynx. A modification of the Viola-Jones method for image recognition using the flexible exit criterion is proposed. The Viola-Jones method explores all areas in the image and decides whether the recognized area belongs to the desired one by passing through a classified cascade. Endoscopic images have a large number of features, such as flare, noise, etc., which degrade the quality of recognition. To improve the quality of recognition, clustering with a flexible exit criterion was proposed, which satisfies the scalability criteria: changing the decision of the solution, instead of moving to another recognition area. It has been established that the proposed modification of the Viola-Jones method shows higher recognition results for endoscopic images.Представлены результаты анализа существующих методов кластеризации данных, получаемых при эндоскопии гортани. Предложена модификация метода Виолы-Джонса для распознавания изображений с использованием критерия гибкого выхода. Метод Виолы-Джонса исследует все области на изображении и принимает решение о принадлежности распознаваемой области к искомой путем прохождения через классифицированный каскад. Эндоскопические снимки имеют большое количество особенностей, таких как засветка, шумы и другие, которые ухудшают качество распознавания. Для улучшения качества распознавания предложена кластеризация с критерием гибкого выхода, которая удовлетворяет критериям масштабируемости – изменение решения вместо перехода к другой области распознавания. Установлено, что предложенная модификация метода Виолы-Джонса показывает высокие результаты распознавания для эндоскопических снимков

Recognition of medical imaging in the computer diagnostic problems

Рассмотрена модификация оригинального метода Собеля. Данная модификация учитывает особенности медицинских изображений, поэтому позволяет получать лучшие результаты при распознавании. The features of medical image recognition are considered

ЭМБРИОГЕНЕЗ В КУЛЬТУРЕ МИКРОСПОР БРОККОЛИ

The process of embryogenesis and technological experimental protocol has been studied and applied to produce doubled haploid plants from microspore cultured in vitro in broccoli B. oleracea L. convar. botrytis (L.) Alef. var. italica Plenck. It was shown that successful embryoid development occurred from microspore isolated from buds 4-5 mm long, containing microspores at late vacuolated stage and pollen grain at twocell stage. The optimal temperature of treatment was 32 Cᵒ within 2 days after culture was launched. The embryoids were produced from the following broccoli accessions: Arcadia F1, Everest, Green Valiant, Marathon F1, and Furio. The highest embryoid yield was obtained from accession Green Valiant, and consisted of 140 embryoids per Petri dish, whereas the lowest yield was in Furio, up to 3 embryoids per Petri dish. The first microspore division was observed in all accessions in 2-3 days of cultivation. Further development of embryoids went either directly into usual embryoid or into suspensor-like structures. The embryoids with suspensor developed more slowly than embryoids without one. We observed the embryoid formation not only at distal end towards microspore originated the suspensor-like structure but also the formation of chain of embryoids, and all variation of twin embryoid combinations. The study of process of embryogenesis in isolated microspores in vitro showed that this method can be used both to produce doubled haploid plants and study the developmental stages of zygotic embryos and suspensors.Изучен процесс эмбриогенеза и отработаны элементы технологии получения удвоенных гаплоидов брокколи B.  oleracea  L. convar. botrytis (L.) Alef. var. italica Plenck в культуре микроспор in vitro. Было выявлено, что успешное развитие эмбриоидов происходит из микроспор, изолированных из бутонов длиной 4 и 5 мм, где преимущественно содержатся микроспоры – на поздней вакуолизированной, пыльца – на ранней двухклеточной стадии развития. Оптимальным режимом температурной обработки является обработка 32°С в течение первых 2-х суток после введения в культуру. Эмбриоиды были получены из пяти образцов брокколи: Arcadia F1, Everest, Green Valiant, Marathon F1, Furio. Наибольший выход эмбриоидов был получен у образца Green Valiant, где он составил до 140 эмбриоидов на чашку Петри, а наименьший – у Furio (до 3 эмбриоидов/чашку Петри). Первые деления в культуре микроспор у всех образцов наблюдали уже на 2-3 сутки культивирования. Дальнейшее развитие эмбриоидов шло по двум направлениям – путем прямого развития или с образованием суспензороподобных структур. Эмбриоиды, содержащие суспензор, развивались медленнее, чем бессуспензорные. Мы наблюдали образование эмбриоида не только на дистальном (по отношению к микроспоре) конце суспензороподобной структуры, но и образование цепочки из эмбриоидов, а также все возможные близнецовые комбинации эмбриоидов. Было показано, что метод получения эмбриоидов в культуре микроспор in vitro может быть использован не только для получения удвоенных гаплоидных растений, но и служить моделью для фундаментальных исследований по изучению этапов развития зиготических эмбриоидов и суспензоров.

СОЗДАНИЕ СЕЛЕКЦИОННО ЦЕННЫХ БИОТИПОВ СВЁКЛЫ СТОЛОВОЙ С ИСПОЛЬЗОВАНИЕМ КУЛЬТУРЫ ШТЕКЛИНГОВ

The long period of time needed for production of homozygous ms-and mf-lines in biennial crop is a problem in development of heterotic hybrids in red beet. The use of steckling culture and protected cultivation technology are the  suitable methods  to  speed  up  the  development  of breeding lines. However, the relationship between exhibition of CMS trait in seed plants and horticulturally valuable traits of steckling is very important aim of the study to be considered. The study was performed in 2012-2014. The red  beet  inbreeding  lines (I2-I5), obtained  through steckling culture in low-volume technology under protected cultivation condition were used as a plant material for the study. The generations that were obtained were from cultivar population ‘Nezhnost’ and a hybrid population of foreign origin. As a result, it was shown that one of criterions of indirect selection of ms-form in inbreeding generation may be the fast-maturity of stecklings that determined their yield weight at harvesting. The average weight of steckling belonging to sterile plants in progenies of different inbreeding generations, not depending on the time of harvesting was higher than in fertile plants. Therefore, probability to select the valuable ms-forms among groups of fast-maturing  plants with  greater weight  of steckling was higher. The selection of stecklings from smaller size groups of middle maturing and late maturing inbreeding progenies is required to  develop valuable mf-lines with high ability for fertility maintaining.Проблемой при создании гетерозисных гибридов свеклы столовой как двулетней овощной культуры, является длительность получения гомозиготных ms- и mf-линий.   Для ускорения создания линейного материала  перспективно использование культуры штеклингов и защищенного грунта.  Однако  при  использовании  данного способа  важен характер взаимосвязи между проявлением признака ЦМС семенных растений и селекционно значимыми признаками штеклингов,  что и явилось целью данных исследований. Исследования  проводили  в  2012-2014  годах. Материал для исследований  – инбредные потомства  (I2-I5)  свёклы столовой, полученные  через  культуру штеклингов, с использованием малообъемной технологии  в  защищенном грунте. Потомства получены на основе сортопопуляции Нежность и гибридных популяций иностранного происхождения. В результате исследований установлено, что одним  из  критериев косвенного  отбора ms-форм  в инбредных  потомствах  может являться скороспелость штеклингов, которая определяет их массу к моменту уборки.  Средняя  масса  штеклингов  стерильных  растений в потомствах разных поколений инбридинга больше, чем у фертильных, не зависимо от срока уборки. Поэтому, вероятность отбора ценных ms-форм выше в группе скороспелых растений с большей массой штеклинга. Для создания перспективных  mf-линий  с высокой  закрепляющей способностью,  наоборот, целесообразно вести отбор штеклингов в более мелкой фракции среди среднеспелых и позднеспелых инбредных потомств