Исследование регенерационной способности и микроклональное размножение Petunia hybrida in vitro

Scientific relevance. The garden petunia, Petunia hybrida, is a popular and wide spread ornamental crop from the family Solanaceae. It is a promising model plant for molecular and genetic research. In vitro micropropagation plays an important role in the distribution of the garden petunia because the survivability and quality of seed material decreases significantly in every subsequent generation. Besides, micropropagation reduces the cost of production substantially. Considering that very few researchers addressed this question in the Russian Federation, this direction of research is still worthy of attention.Materials and methods. The experiments were conducted by the Laboratory of Breeding and Genetic Research on Field Crops at FSBSI “Federal Scientific Center of Agricultural Biotechnology of the Far East named after A.K. Chaiki”. Seeds of Petunia hybrida (double-flowered) were used as primary explants. Liquid bleacher ACE diluted with distilled water in the proportion 1:9 was used as a sterilizing agent (the working solution contained 0.50% NaOCl). The total time of exposure was 15 minutes. The primary explants were subcultured onto a hormone-free Murashige and Skoog basal medium containing 20 g/L sucrose and 6 g/L agar. Isolated in vitro objects were cultured in test tubes with cotton-gauze plugs at an illuminance of 4000 lx, a temperature of 22–25 °C, and a 16h photoperiod in a culture room. The duration of one passage was 60 days. Micropropagation was carried out using 7- 10 mm cuttings with one or two nodes. The pot culture of the regenerants was established under controlled conditions in a light room (photoperiod was 16 hours, temperature was 23°С).Results. The optimal method for introducing Petunia hybrida into cell culture is the use of seeds treated with the solution of bleacher ACE that was diluted with distilled water in the proportion 1:9. The optimal time of exposure is 15 minutes. Petunia hybrida demonstrated a high regeneration rate on the hormone-free MS medium – it had a fast growth and development rate, and good rhizogenesis; the reproductive rate was 8.77. For the micropropagation of the garden petunia, it is advisable to use cuttings of test tube plants, which should be placed onto a hormone-free MS medium. The test tube plants of Petunia hybrida acclimatized successfully on a soil substrate. This shows the high plasticity of the culture. Актуальность. Петуния гибридная – популярная, широко культивируемая во всем мире декоративная культура семейства пасленовые и перспективное модельное растение для молекулярно-генетических исследований. Важную роль в тиражировании гибридной петунии имеет микроклональное размножение in vitro, поскольку жизнеспособность и качество посевного материала резко ухудшается в последующих поколениях. Немаловажный аспект при этом – существенное снижение экономических затрат. Учитывая, что в РФ исследований по данному вопросу крайне мало, данное направление работ является весьма актуальным.Материалы и методы. Исследования проводили в лаборатории селекционно-генетических исследований полевых культур ФГБНУ «ФНЦ агробиотехнологий Дальнего Востока им. А.К. Чайки». В качестве первичных эксплантов использовали семена Petunia hybrida (махровая форма). Стерилизующий агент – бытовой отбеливатель АСЕ, разбавленный дистиллированной водой в соотношении 1 : 9 (0,50% содержание NaOCl в рабочем растворе). Время экспликации – 15 мин. Первичные экспланты пассировали на безгормональную питательную среду с минеральной основой по Мурасиге-Скуга, содержащую 20 г/л сахарозы и 6 г/л агара. Изолированные in vitro объекты культивировались в пробирках с ватно-марлевыми пробками при освещенности 4 тыс. лк, температуре 22…25°С, фотопериоде 16 ч в условиях культуральной комнаты. Продолжительность одного пассажа – 60 суток. Микроклональное размножение проводили черенками длиной 7-10 мм с узлом. Перевод регенерантов в горшечную культуру осуществлён в контролируемые условия в световой комнате: фотопериод 16 ч – день, температура 23°С.Результаты. Для введения в культуру in vitro петунии гибридной оптимальным способом является использование семян, обработанных раствором бытового отбеливателя АСЕ, разбавленным дистиллированной водой в соотношении 1 : 9 при времени экспликации 15 мин. Petunia hybrida продемонстрировала высокие темпы регенерации на безгормональной питательной среде МС, выражающиеся в достаточной скорости роста и развития растений, хорошем ризогенезе, а также коэффициенте размножения 8,77. Для микроклонального размножения петунии целесообразно использовать черенки пробирочных растений, которые помещают на безгормональную среду МС. Пробирочные растения Petunia hybrida обладают высокой приживаемостью в почвенном субстрате, что свидетельствует о высокой пластичности культуры.

Фузариоз гречихи посевной в Приморском крае

Relevance. Common buckwheat is one of the most important cereal and melliferous crops being in demand both in Russia and overseas. Despite the fact that buckwheat has lower sus- ceptibility to infectious diseases in comparisons with other grain and cereal crops, research on its pathogens is a topical issue considering a high disease rate for this crop in Primorsky krai. Materials and methods. The study on pathogenic composition was conducted in selective crop rotations of FSBSI “FSC of Agricultural Biotechnology of the Far East named after A.K. Chaiki” at the territory of Ussuriysky district in Primorsky krai. Indeterminate (Izumrud, Pri 7, Bashkirskaya krasnostebel’naya) and determinate (Dikul) buckwheat varieties were used for the study. Fungi were isolated from rhizoplane using water washing technique, from soil and rhizosphere via the dilution method, from leaves and root systems by accumulation in a moist chamber with subsequent transfer of culture onto selective medium. Fungal isolates were cul- tivated on toxigenic medium (Myro) to determine their phytotoxicity. Phytopathogenic activity of living cultures was evaluated on potato sucrose agar according to the modified method of Chelkowski and Manka . All experiments were conducted in accordance with established methods.Results. Culture filtrates of F. avenaceum and F. graminearum isolates, obtained from different anatomical parts of common buckwheat, were characterized by low toxicity, and culture filtrates of F. oxysporum isolate were highly toxic. Phytotoxicity of pathogens manifested itself not only in decrease in laboratory germination ability of buckwheat seeds, but also in inhibition of buckwheat sprout development. It proved that metabolites exerted a prolonged effect on sensitive plants. F.avenaceum, F. oxysporum and F. graminearum have pronounced phy- topathogenic and aggressive properties in relation to buckwheat and test-plants in the laboratory conditions. The sum total of their studied phytotoxic properties is convincing enough to consider them potentially hazardous to buckwheat for wilt disease development.Актуальность. Гречиха посевная – важнейшая крупяная и медоносная культура, востребованная в России и за рубежом. Несмотря на меньшую степень поражения инфекционными болезнями по сравнению с другими зерновыми и крупяными культурами, исследование патогенов гречихи считается весьма актуальным, учитывая довольно высокую степень поражаемости культуры в Приморском крае.Материалы и методы. Исследования патогенного состава проводили в селекционных севооборотах ФГБНУ «ФНЦ агробиотехнологий Дальнего Востока им. А.К. Чайки» на территории Уссурийского района Приморского края. Использовался набор сортов гречихи индетерминантного (Изумруд, При 7, Башкирская красностебельная) и детерминантного (Дикуль) типа развития. Выделение грибов из ризопланы осуществляли методом водных смывов; выделение из почвы и ризосферы – методом разведений; выделение из листьев и корневой системы – методом накопления во влажной камере с последующим переносом культуры на питательную среду. Для определения фитотоксичности изоляты выращивали на токсигенной среде Myro, фитопатогенность живых культур – на КСА по модифицированной методике Челковского и Манки. Все опыты проводились по общепринятым методикам.Результаты. Культуральные фильтраты выделенных из различных анатомических частей гречихи посевной изолятов F. avenaceum и F. graminearum оказались слаботоксичными, а изолята F. oxysporum – высокотоксичными. Фитотоксическое действие патогенов проявилось не только в ограничении лабораторной всхожести семян гречихи, но и в ингибировании развития её проростков, что свидетельствует о пролонгированном действии метаболитов на чувствительные растения. F.avenaceum, F. oxysporum и F. graminearum обладают выраженными фитопатогенными и агрессивными свойствами в отношении гречихи и тест-растения в лабораторных условиях. По совокупности изученных фитотоксических свойств их следует считать потенциально опасными для возникновения сосудистого увядания гречихи

Influence of the toxic effect of zinc and mineral starvation on the growth and development of buckwheat plantlets <i>in vitro</i>

Background. Common buckwheat is a cereal crop with high potential for genetic improvement in terms of developing breeding material resistant to abiotic stressors. To date, there have been no reports on in vitro production of buckwheat plantlets resistant to high doses of zinc and a lack of macronutrients.   Materials and methods. Aseptic single-node cuttings from the obtained regenerated plants of common buckwheat cultivars ‘Dikul’ and ‘Izumrud’ were cultivated in vitro on nutrient media with the addition of the selection factor ZnSO4 × 7 H2O in a concentration of 808–1313 mg/L. Survived plants were microcloned on nutrient media without macrosalts for mineral starvation modeling. Morphological traits and general nonspecific adaptation reactions of the plantlets were evaluated for the following characteristics: plant height, the number of internodes, the number of leaves, leaf blade length, the presence of roots, and leaf color.   Results. According to the results of the 33-day cultivation of test-tube microcuttings on media with zinc toxicity, 33–91 % of lines resistant to ionic stress were selected in different variants. The secondary testing of the plantlets under conditions of mineral starvation in vitro turned out to be the strongest inhibitory factor for buckwheat. At the same time, high resistance to stress was observed in cv. ‘Dikul’. Cultivation of the obtained buckwheat lines on the MS nutrient medium for two passages showed a sufficiently high level of regeneration in the studied genotypes. The test-tube buckwheat plantlets obtained under selective conditions are promising material for further breeding as well as for studying the possibility of their use as phytoremediators

Sepharadim/conversos and premodern Global Hispanism

Sepharadim participated in the Hispanic vernacular culture of the Iberian Peninsula. Even in the time of al-Andalus many spoke Hispano-Romance, and even their Hebrew literature belies a deep familiarity with and love of their native Hispano-Romance languages. However, since the early sixteenth century the vast majority of Sepharadim have never lived in the Hispanic world. Sepharadim lived not in Spanish colonies defined by Spanish conquest, but in a network of Mediterranean Jewish communities defined by diasporic values and institutions. By contrast, the conversos, those Sepharadim who converted to Catholicism, whether in Spain or later in Portugal, Italy, or the New World, lived mostly in Spanish Imperial lands, were officially Catholic, and spoke normative Castilian. Their connections, both real and imagined, with Sephardic cultural practice put them at risk of social marginalization, incarceration, even death. Some were devout Catholics whose heritage and family history doomed them to these outcomes. Not surprisingly, many Spanish and Portugese conversos sought refuge in lands outside of Spanish control where they might live openly as Jews. This exodus (1600s) from the lands formerly known as Sefarad led to a parallel Sephardic community of what conversos who re-embraced Judaism in Amsterdam and Italy by a generation of conversos trained in Spanish universities. The Sephardic/Converso cultural complex exceeds the boundaries of Spanish imperial geography, confuses Spanish, Portuguese, Catholic, and Jewish subjectivities, and defies traditional categories practiced in Hispanic studies, and are a unique example of the Global Hispanophone

Fusarium blight of common buckwheat in Primorsky krai

Relevance. Common buckwheat is one of the most important cereal and melliferous crops being in demand both in Russia and overseas. Despite the fact that buckwheat has lower sus- ceptibility to infectious diseases in comparisons with other grain and cereal crops, research on its pathogens is a topical issue considering a high disease rate for this crop in Primorsky krai. Materials and methods. The study on pathogenic composition was conducted in selective crop rotations of FSBSI “FSC of Agricultural Biotechnology of the Far East named after A.K. Chaiki” at the territory of Ussuriysky district in Primorsky krai. Indeterminate (Izumrud, Pri 7, Bashkirskaya krasnostebel’naya) and determinate (Dikul) buckwheat varieties were used for the study. Fungi were isolated from rhizoplane using water washing technique, from soil and rhizosphere via the dilution method, from leaves and root systems by accumulation in a moist chamber with subsequent transfer of culture onto selective medium. Fungal isolates were cul- tivated on toxigenic medium (Myro) to determine their phytotoxicity. Phytopathogenic activity of living cultures was evaluated on potato sucrose agar according to the modified method of Chelkowski and Manka . All experiments were conducted in accordance with established methods.Results. Culture filtrates of F. avenaceum and F. graminearum isolates, obtained from different anatomical parts of common buckwheat, were characterized by low toxicity, and culture filtrates of F. oxysporum isolate were highly toxic. Phytotoxicity of pathogens manifested itself not only in decrease in laboratory germination ability of buckwheat seeds, but also in inhibition of buckwheat sprout development. It proved that metabolites exerted a prolonged effect on sensitive plants. F.avenaceum, F. oxysporum and F. graminearum have pronounced phy- topathogenic and aggressive properties in relation to buckwheat and test-plants in the laboratory conditions. The sum total of their studied phytotoxic properties is convincing enough to consider them potentially hazardous to buckwheat for wilt disease development

Regenerative ability and micropropagation of <I>Petunia hybrid</I> in vitro

Scientific relevance. The garden petunia, Petunia hybrida, is a popular and wide spread ornamental crop from the family Solanaceae. It is a promising model plant for molecular and genetic research. In vitro micropropagation plays an important role in the distribution of the garden petunia because the survivability and quality of seed material decreases significantly in every subsequent generation. Besides, micropropagation reduces the cost of production substantially. Considering that very few researchers addressed this question in the Russian Federation, this direction of research is still worthy of attention.Materials and methods. The experiments were conducted by the Laboratory of Breeding and Genetic Research on Field Crops at FSBSI “Federal Scientific Center of Agricultural Biotechnology of the Far East named after A.K. Chaiki”. Seeds of Petunia hybrida (double-flowered) were used as primary explants. Liquid bleacher ACE diluted with distilled water in the proportion 1:9 was used as a sterilizing agent (the working solution contained 0.50% NaOCl). The total time of exposure was 15 minutes. The primary explants were subcultured onto a hormone-free Murashige and Skoog basal medium containing 20 g/L sucrose and 6 g/L agar. Isolated in vitro objects were cultured in test tubes with cotton-gauze plugs at an illuminance of 4000 lx, a temperature of 22–25 °C, and a 16h photoperiod in a culture room. The duration of one passage was 60 days. Micropropagation was carried out using 7- 10 mm cuttings with one or two nodes. The pot culture of the regenerants was established under controlled conditions in a light room (photoperiod was 16 hours, temperature was 23°С).Results. The optimal method for introducing Petunia hybrida into cell culture is the use of seeds treated with the solution of bleacher ACE that was diluted with distilled water in the proportion 1:9. The optimal time of exposure is 15 minutes. Petunia hybrida demonstrated a high regeneration rate on the hormone-free MS medium – it had a fast growth and development rate, and good rhizogenesis; the reproductive rate was 8.77. For the micropropagation of the garden petunia, it is advisable to use cuttings of test tube plants, which should be placed onto a hormone-free MS medium. The test tube plants of Petunia hybrida acclimatized successfully on a soil substrate. This shows the high plasticity of the culture

Using plant extracts for the micropropagation of buckwheat

Background. Various plant hormones are used (cytokinins, auxins) to increase the regeneration efficiency and the net reproduction rate of buckwheat in vitro. However, the growth and development rates of plantlets have been noted to be low under these conditions. For this reason, search for the plant extracts that are able to stimulate the regenerative ability of plants is a promising direction of biotechnological research.Materials and methods. Aseptic single-node cuttings of common buckwheat plantlets (varieties Dikul and Izumrud) were grown on MS nutrient media with plant extracts from Fagopyrum esculentum and Reynoutria japonica (0.1, 0.5, and 1%) for 21 days. The following morphobiological paramaters of the plantlets were evaluated: plant height, the number of internodes, the number of leaves, leaf length, and the number and length of roots.Results. Dealcoholized aqueous solutions of the extracts from F. esculentum and R. japonica in the studied concentrations (0.1-1%) significantly stimulated the growth and development of the buckwheat plantlets increasing their net reproduction rate (4.00-6.00) and rhizogenesis. The media with the plant extracts in concentrations of 0.1-0.5% were observed to produce the strongest positive effect. As the result, the morphobiological characteristics of the plantlets and the success rate of the micropropagation were the highest

Establishing the <i>in vitro</i> culture of and micropropagating edible honeysuckle

Edible honeysuckle is a popular fruit crop. Its therapeutic and health-promoting effects are attributed to a high content of bioactive compounds in the fruits. Unlike the traditional plant multiplication methods, the in vitro propagation allows scientists to obtain high-quality planting material of honeysuckle in a great quantity and within a short time. The research was carried out at the Laboratory of Breeding and Genetic Research on Field Crops of the Federal Scientific Center of Agricultural Biotechnology of the Far East named after A.K. Chaiki. Honeysuckle variety Podarok amurchanam created by the Far Eastern State Agrarian University was used as the research object. The research materials were sterilized according to the methodology of N.I. Vavilov All-Russian Institute of Plant Genetic Resources with some modifications. Several products were used as chemical agents for sterilization in the following sequence: a 5% solution of surfactants, fungicide Fundazol, EC (1 g/l), the bleaching agent ACE freshly diluted with distilled water in the proportion 1:9 (0.50% of NaOCl in the working solution), and 70% ethanol. The primary explants were cultured on an MS containing 20 g/l sucrose and 6 g/l agar (hereafter – MS) and supplemented with 6-benzylaminopurine (BA) at a concentration of 0.5 mg/l. The pH of the medium was adjusted to 5.7-5.8 using 1N КОН. The explants (microcuttings with one-two internodes) were subcultured on an MS supplemented with BA (0.5 mg/l). The morphometric parameters of the plants were measured on the 35th day of cultivation. The sterilization of the explants with Fundazol (1 g/l) and the ACE diluted with distilled water in the proportion 1:9 allowed us to obtain a high number of viable microclones (50%). The elimination of leaves from the honeysuckle microcuttings drastically decreased the survival rate and led to the death of the microclones in most cases (the mortality rate was 98.7 %). Subculturing the microcuttings on the MS supplemented with BA at a concentration of 0.5 mg/l facilitated the normal growth and development of the regenerated honeysuckle plants (the average reproduction rate was 4.65)