ПНЕВМАТИЧЕСКИЙ СЕПАРАТОР И РЕЗУЛЬТАТЫ СОРТИРОВКИ СЕМЯН СВЕКЛЫ СТОЛОВОЙ С ЕГО ПРИМЕНЕНИЕМ

The results of the air separator tests on the beet seeds are given.  Приведена конструкция пневматического сепаратора для сортировки и очистки семян овощных культур и приведены результаты испытаний пневмосепаратора на семенах свеклы столовой.

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.

РАЗРАБОТКА ЭЛЕМЕНТОВ ТЕХНОЛОГИИ ПОЛУЧЕНИЯ ПОСАДОЧНОГО МАТЕРИАЛА САЛАТА (LACTUCA SATIVA L.) НА БЕЗВИРУСНОЙ ОСНОВЕ С ИСПОЛЬЗОВАНИЕМ МЕТОДОВ БИОТЕХНОЛОГИИ

The article presents the results of research on the production in vitro of regenerated plants from the seeds of cultivars of lettuce (Lactuca sativa L.) Emerald, Bouquet, Chameleon (FSBSI Federal Scientific Vegetable Center), susceptible of aspermia tomato (Tomato aspermy cucumovirus) – AsTV. Seeds of strongly susceptible to AsTV varieties of salad Chameleon and Bouquet were subjected to thermotherapy at different temperatures (37°C, 38°C, 40°C) for a different time interval (1, 3, 5, 7, 10 days) in dry form and when moistened. Marked varietal specificity during germination of seeds after thermotherapy. Thus, the greatest number of seedlings in the emerald variety was obtained after 5 days of thermotherapy (10.0±0), while the Bouquet variety had the best results after 3 days of thermotherapy (9.3±1.2) with moisture. After thermotherapy of dry seeds by 40°C plant material of cultivar Emerald was planted on solid and liquid culture media. The conditions of step sterilization of lettuce seeds for introduction into the culture in vitro were chosen: washing in 96% ethanol, then in 50% aqueous solution of "Whiteness" with the addition of Twin-20, after in sterile distilled water. The nutrient medium for germination of lettuce seeds was used: Gamborg B5 (2% sucrose, 7.0 g/l agar), and the liquid nutrient medium was of that composition. The obtained seedlings were cutted and transferred to medium MS (2% sucrose, 0.1 mg/l ha and 1 mg/l BAP, 3.0 g/l phytogel). The formed shoots for rooting were transferred to the MS medium (2% sucrose, 3.0 g/l phytogel). In the future, lettuce plants will be adapted in vivo and tested for the presence of tomato aspermia virus in the planting material.В статье представлены результаты исследований по получению в культуре in vitro растений-регенерантов из семян сортов салата (Lactuca sativa L.) Изумрудный, Букет, Хамелеон (селекции ФГБНУ ФНЦО), восприимчивых к вирусу аспермии томата (Tomato aspermy cucumovirus) – AsTV. Семена сильновосприимчивых к AsTV сортов салата Хамелеон и Букет были подвергнуты термотерапии при разных температурных режимах (37°С, 38°С, 40°С) в течение различного временного интервала (1, 3, 5, 7, 10 суток) в сухом виде и при увлажнении. Отмечена сортовая специфичность при прорастании семян после термотерапии. Так, наибольшее количество проростков у сорта Изумрудный получено после 5 суток термотерапии (10,0±0), тогда как у сорта Букет лучшие показатели были после 3 суток термотерапии (9,3±1,2) при увлажнении. После термотерапии сухих семян при 40°С сорта Изумрудный растительный материал был высажен на твердые и жидкие питательные среды. Подобраны условия ступенчатой стерилизации семян салата для введения в культуру in vitro: промывание в 96% этаноле, затем в 50% водном растворе «Белизны» с добавлением Твина-20, после в стерильной дистиллированной воде. Использована питательная среда для проращивания семян салата: Gamborg В5 (2% сахароза, 7,0 г/л агара), жидкая питательная среда была того состава. Полученные проростки черенковали и переносили на среду МС (2% сахароза, 0,1 мг/л ГК и 1 мг/л БАП, 3,0 г/л фитогеля). Образовавшиеся побеги для укоренения были перенесены на среду МС (2% сахароза, 3,0 г/л фитогеля). В дальнейшем будет проведена адаптация растений салата в условиях in vivo и тестирование на наличие вируса аспермии томата в посадочном материале

PNEUMATIC SEPARATOR AND SORT RESULTS TABLE BEET SEED WITH ITS USE

The results of the air separator tests on the beet seeds are given

EMBRYOGENESIS IN CULTURE OF ISOLATED MICROSPORE OF BROCCOLI

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

DEVELOPMENT OF ELEMENTS OF TECHNOLOGY FOR PLANTING MATERIAL OF LETTUCE (<i>LACTUCA SATIVA L.</i>) ON VIRUS-FREE BASIS USING METHODS OF BIOTECHNOLOGY

The article presents the results of research on the production in vitro of regenerated plants from the seeds of cultivars of lettuce (Lactuca sativa L.) Emerald, Bouquet, Chameleon (FSBSI Federal Scientific Vegetable Center), susceptible of aspermia tomato (Tomato aspermy cucumovirus) – AsTV. Seeds of strongly susceptible to AsTV varieties of salad Chameleon and Bouquet were subjected to thermotherapy at different temperatures (37°C, 38°C, 40°C) for a different time interval (1, 3, 5, 7, 10 days) in dry form and when moistened. Marked varietal specificity during germination of seeds after thermotherapy. Thus, the greatest number of seedlings in the emerald variety was obtained after 5 days of thermotherapy (10.0±0), while the Bouquet variety had the best results after 3 days of thermotherapy (9.3±1.2) with moisture. After thermotherapy of dry seeds by 40°C plant material of cultivar Emerald was planted on solid and liquid culture media. The conditions of step sterilization of lettuce seeds for introduction into the culture in vitro were chosen: washing in 96% ethanol, then in 50% aqueous solution of "Whiteness" with the addition of Twin-20, after in sterile distilled water. The nutrient medium for germination of lettuce seeds was used: Gamborg B5 (2% sucrose, 7.0 g/l agar), and the liquid nutrient medium was of that composition. The obtained seedlings were cutted and transferred to medium MS (2% sucrose, 0.1 mg/l ha and 1 mg/l BAP, 3.0 g/l phytogel). The formed shoots for rooting were transferred to the MS medium (2% sucrose, 3.0 g/l phytogel). In the future, lettuce plants will be adapted in vivo and tested for the presence of tomato aspermia virus in the planting material

Embryogenesis induction of carrot (Daucus carota L.) in isolated microspore culture

Haploid technologies are used to create homozygous lines for accelerated breeding. We aimed to optimize the technology for using the isolated microspore culture in vitro to obtain doubled haploids of the carrot (Daucus carota L.). We studied two carrot varieties with different responsiveness to embryogenesis, Altajskaya lakomka and Breeding line 17. Carrot microspores were isolated from buds and cultivated in liquid nutrient media supplemented with an antibiotic and activated carbon in vitro. They were exposed to different thermal treatments. The experiment showed the benefits of combining cold pre-treatment of buds (5°C for 1 day) with heat shock of isolated microspores in vitro (32°C for 2 days). The induction of embryogenesis on the NLN-13 medium was twice as high as on the MSm-13 medium. The use of 1% activated carbon in 0.5% agarose increased the yield of embryoids by more than 1.5 times. 100 mg/L of ampicillin was found to be the most efficient concentration. After 30 days of cultivation under optimized conditions, the yield was 161.3 and 44.0 embryoids per Petri dish for the cultivar Altajskaya lakomka and Breeding line 17, respectively. The induction of carrot embryogenesis is determined by the type and duration of thermal stress, the composition of the nutrient medium, the use of activated carbon as a sorbent, the addition of β-lactam antibiotics, and the type of explant exposed to thermal treatment. Our technology enabled us to obtain homozygous doubled haploid lines of carrots during a year, and these lines were included in the breeding process to create F1 hybrids