AGRIS - 

Data provenance by Country

2022

Egorova, N.A. | Stavtzeva, I.V.

The most important task of plant breeding is the creation of highly productive cultivars resistant to abiotic stress factors of the environment, in particular, to drought. To solve many breeding issues, biotechnological methods are being actively used. One of these methods is in vitro cell selection, which allows targeted selection of genotypes with desired traits. The purpose of the work was to study the features of the effect of osmotic stress on the development of isolated embryos of clary sage cultivars and samples to develop a cell technology for creating forms resistant to this stress factor in vitro. Cultivars and samples of sage (Salvia sclarea L.) differing in field drought resistance were used in the studies. Mature zygotic embryos were used as explants, which were cultivated on Murashige and Skooga (MS) culture media supplemented with osmotic agents (NaCl, mannitol, sorbitol, sucrose) at various concentrations. In the control, embryos were cultured on MS medium. It was shown that the cultivation of embryos of 3 cultivars (C-785, Ai-Todor and Taigan) on media with 0.9% NaCl, 4.0–5.0% mannitol or sorbitol and 7% sucrose made it possible to differentiate cultivars by resistance to osmotic stress. In the following experiment, the development of isolated embryos of 10 cultivars and samples of sage (with coefficients of drought resistance from 22.5 to 73.5%) was analyzed on a medium supplemented with a sublethal concentration of mannitol (4.5%). Herewith, a decrease in comparison with the control from 1.4 to 14.5 times was revealed for all the studied parameters, such as the frequency of embryos germination and seedlings formation, the length of the shoot and root. It was established that the maximum correlation coefficients (0.76–0.79) were between the field drought resistance of genotypes and the frequency of seedling development on the medium with mannitol. The conducted studies showed that the developed selective system for obtaining or evaluating in vitro forms of sage resistant to abiotic stress is rather promising. | Важнейшей задачей селекции растений является создание высокопродуктивных сортов, устойчивых к абиотическим стрессовым факторам среды, в частности к засухе. Методами клеточной селекции in vitro можно целенаправленно отобрать генотипы с заданными признаками. Цель работы – изучение особенностей действия осмотического стресса на развитие изолированных зародышей сортов и образцов шалфея мускатного для разработки клеточной технологии создания устойчивых к этому стрессовому фактору форм in vitro. В исследованиях использовали сорта и образцы шалфея (Salvia sclarea L.), различающиеся по полевой засухоустойчивости. Эксплантами служили зрелые зиготические зародыши, которые культивировали на питательных средах Мурасиге и Скуга (МС), дополненных осмотиками (NaCl, маннит, сорбит, сахароза) в различных концентрациях. В контроле зародыши культивировали на среде МС. Показано, что культивирование зародышей 3 сортов (С-785, Ай-Тодор и Тайган) на средах с введением 0,9% NaCl, 4,0–5,0% маннита или сорбита и 7,0% сахарозы позволило дифференцировать сорта по устойчивости к осмотическому стрессу. В следующем опыте проведен анализ развития изолированных зародышей 10 сортов и образцов шалфея (с коэффициентами засухоустойчивости от 22,5 до 73,5%) на среде, дополненной сублетальной концентрацией маннита (4,5%). При этом выявлено снижение по сравнению с контролем от 1,4 до 14,5 раз всех изученных параметров – частоты прорастания зародышей и образования проростков, длины побега и корня. Максимальные коэффициенты корреляции (0,76–0,79) установлены между полевой засухоустойчивостью генотипов и частотой развития проростков на среде с добавлением маннита. Проведенные исследования показали перспективность использования разработанной селективной системы для получения или оценки in vitro устойчивых к абиотическому стрессу форм шалфея.

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Central Scientific Agricultural Library - Russian Federation

2022

Singina, G.N. | Shedova, E.N. | Lopukhov, A.V.

Biotechnology of in vitro embryo production (IVP) and their transfer to recipient animals can increase the rate of genetic progress in the breeding of industrial cattle breeds and ensure the conservation of small breeds. Efficiency of IVP technology can be increased by optimizing the in vitro embryo culture conditions (IVC). There was studied of supplementation of fetal bovine serum (FBS) in BO-IVC medium from 5th day of bovine IVP embryos cultivation on their development and viability in vitro. The post mortem isolated bovine oocytes (n=250) were cultured in the maturation medium, in vitro fertilized with the frozen-thawed sperm and transferred to the culture medium for embryo development. On day 7 after oocytes fertilization the number of embryos developed to the blastocyst stage (blastocyst rate) and their quality (the rate of blastocyst of Grade 1) were evaluated, and at the end of cultivation (day 10), the number of blastocysts that reached the hatching stage (viability check) was determined. For developing embryos, there was used either commercial BO-IVC medium alone (System 1) or the same medium added with 5% FBS starting from the fifth day of culturing (System 2). In systems 1 and 2, the yield of blastocysts on the 7th day of culturing was the same and amounted to 34.5±2.8 and 30.4± 1.0%, and their number belonging to category 1 was 57.7±3.5 and 65.6± 5.3%, respectively. At the same time, the presence of FCS increased the portion of blastocysts that hatched on day 10 of culturing from 42.7±4.7 to 70.9±3.2% (p less than 0.01). It is concluded, that the BO-IVC medium is highly effective for obtaining IVP embryos in cattle. Also, it is obvious that the addition of FBS to this medium from the 5th day of culturing does not affect the development of embryos to the blastocyst stage, but increases their viability in vitro conditions. | Биотехнология, направленная на получение эмбрионов in vitro (in vitro embryo production, IVP) и их трансплантацию животным-реципиентам, позволяет увеличить темпы генетического прогресса в селекции индустриальных пород крупного рогатого скота и обеспечить сохранение малочисленных пород. Повысить эффективность данной технологии можно за счет моделирования условий культивирования эмбрионов in vitro (in vitro embryo culture, IVC). Было изучено влияние введения фетальной бычьей сыворотки (ФБС) в среду BO-IVC с 5-го дня культивирования IVP-эмбрионов на их развитие и жизнеспособность в условиях in vitro. Выделенные post mortem ооциты коров (n=250) инкубировали в среде созревания, оплодотворяли in vitro замороженно-оттаянной спермой и переносили в среду для развития эмбрионов. На 7-е сутки после оплодотворения ооцитов проводили подсчет числа эмбрионов, развившихся до стадии бластоцисты, и оценку их качества, а в конце культивирования (10-й день) определяли количество бластоцист, достигших стадии вылупления (тест на жизнеспособность). Для эмбрионального развития использовали либо только коммерческую среду BO-IVC (система 1), либо эту же среду, но с добавлением в нее с 5-го дня культивирования 5% ФБС (система 2). В системах 1 и 2 выход бластоцист на 7-е сут культивирования был одинаковым и составлял 34,5±2,8 и 30,4±1,0%, а их количество, относящееся к 1 категории, - 57,7±3,5 и 65,6±5,3% соответственно. В то же время, присутствие ФБС повышало долю вылупившихся бластоцист на 10-й день культивирования с 42,7±4,7 до 70,9±3,2% (P меньше 0,01). Сделан вывод о высокой эффективности среды BO-IVC для получения IVP-эмбрионов у крупного рогатого скота. Также очевидно, что добавление ФБС в данную среду c 5-го дня культивирования не влияет на развитие эмбрионов до стадии бластоцисты, но повышает их жизнеспособность в условиях in vitro.

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Central Scientific Agricultural Library - Russian Federation

2014

Kruglova, N.N., Russian Academy of Sciences. Ufa Scientific Center (Russian Federation). Institute of Biology

Revealed were the stages of wheat embryo autonomy of 10 hybrid combinations of spring soft wheat for further developing biotechnology of producing drought-resistant samples. The stage of autonomy was revealed according to a capability of embryos isolated at the last stages of embryogenesis of giving normal sprouts under in vitro condition on a hormone-free medium and forming from such sprouts full value fertile plants under ex vitro conditions. It is shown that the formed embryo corresponds to the autonomy stages (20 days after pollination, the length 2.1-2.2 mm) characterized by the presence of all organs that are typical for grain variety embryos. As a selective medium simulates deficit of water, it is possible to give an express evaluation of each newly wheat variety being created (originally – hybrid combination) as to the resistance to the stress factor "drought". Acceleration in this case is achieved due to that the hybrid combination is diagnosed for drought-resistance at the earliest stage of ontogenesis – embryo in the step of autonomy but not by laboratory evaluation of mellow grain or field evaluation of a plant as it is conventional in the routine selection practice. | Выявляли стадии автономности зародыша 10-ти гибридных комбинаций яровой мягкой пшеницы для дальнейшей разработки биотехнологии получения засухоустойчивых образцов. Стадию автономности выявляли по способности зародышей, изолированных на последовательных стадиях эмбриогенеза, завершить эмбриогенез, дать нормальные проростки в условиях in vitro на безгормональной среде и сформировать из таких проростков полноценные фертильные растения в условиях ex vitro. Показано, что стадии автономности соответствует сформированный зародыш (20 суток после опыления, длина 2,1–2,2 мм), характеризующийся наличием всех типичных для зародышей злаков органов. Т.к. селективная питательная среда имитирует дефицит влаги, возможно дать экспресс-оценку каждого вновь создаваемого сорта (первоначально – гибридной комбинации) пшеницы по устойчивости к стресс-фактору "засуха". Ускорение в данном случае достигается за счет того, что гибридная комбинация диагностируется на засухоустойчивость на самой ранней стадии онтогенеза – зародыше в стадии автономности, а не путем лабораторной оценки зрелого зерна или полевой оценки растения, как это принято в рутинной селекционной практике.

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Central Scientific Agricultural Library - Russian Federation

2013

Plaksina, T.V. | Kul'khanova, D.S. | Mat'kova, E.Yu., The M.A. Lisavenko Research and Development Inst. of Siberia Horticulture, Barnaul

Hybrid bank of cherry at Altai was extended by new original genotypes, including hybrids of complex genetic origin and ploidy. These forms often give nongerminating seeds at crossings, following to lower breeding efficiency. The use of in vitro embryoculture allows not only preservation but also propagation of valuable hybrid material. The aim of our work was to obtain experimental data on the influence of growth regulators on development of isolated cherry embryos in vitro at the stages of introduction and micropropagation for preservation of original hybrid genotypes. At the stage of introduction of isolated embryo low concentrations of growth regulators was optimal, 2.5µM 6-benzylaminopurine (BAP) in combination with 0.1 µM indole-3-butyric acid (IBA) and 1.0 µM gibberellic acid (GA3). In Murashige-Skoog (MS) nutritional medium with such ratio of growth regulators not only the growth of embryos was stimulated, but also additional shoots (2.8 ± 1.2 pcs/embryo) formed. Depending on crossing combination the ratio of growing embryo from al embryos introduced in culture in vitro ranged from 12.5 to 63.6%. At the micropropagation stage the application of combined MS media, BAP 2.5 µM + IBA 0.1 µM + GA3 1.0 µM made it possible to raise propagation coefficient of regenerated plants of the studied crossing combinations on the average 1.5 times. Shoots with height 1.2-2.0 cm were transferred to rooting media. The composition of the rooting medium was 1/2 MS without myoinositol, with IBA 5.0 µM and lnaphthleneacetic acid (NAA) 0.5 µM. All rooted plants within 3 weeks were transferred for acclimatization to vegetation room. Adapted plants (not less than ten plants for every cross combination) got through estimation under field condition. | Гибридный фонд вишни на Алтае пополнен новыми оригинальными генотипами, в том числе сложного генетического происхождения и плоидности. Эти формы при скрещиваниях часто дают невсхожие семена, что снижает эффективность селекции. Применение метода эмбриокультуры позволяет не только сохранить, но и размножить ценный гибридный материал. Исследовали влияние регуляторов роста на развитие изолированных зародышей вишни оригинальных гибридных генотипов в условиях in vitro на этапах введения и микроразмножения. На этапе введения в культуру in vitro оптимальны невысокие концентрации регуляторов роста – 6-бензиламинопурин (БАП) 2,5 мкМ в сочетании с индолилмасляной кислотой (ИМК) 0,1 мкМ и гиббереловой кислотой (ГК) 1,0 мкМ. На питательной среде Мурасиге - Скуга (МС) с таким соотношением регуляторов происходила не только стимуляция роста самого зародыша, но и формирование дополнительных побегов (2,8±1,2 шт./раст.). В зависимости от комбинации скрещивания доля тронувшихся в рост зародышей от числа введенных в культуру in vitro составила от 12,5 до 63,6%. На этапе микроразмножения применение комбинированных питательных сред МС (БАП 2,5 мкМ + ИМК 0,1 мкМ + ГК 1,0 мкМ) позволяет повысить коэффициент размножения регенерантов большинства гибридов в изученных комбинациях скрещивания в среднем в 1,5 раза. Микропобеги, достигшие 1,5-2 см высоты, укореняли на питательной среде МС, разбавленной вдвое по минеральному составу и сахарозе, без мезоинозита с добавлением 5,0 мкМ ИМК и 0,5 мкМ НУК. Через 3 недели растения с корнями высаживали в субстрат и адаптировали к условиям ex vitro в вегетационной комнате. Адаптированные растения (не менее 10 шт. каждой комбинации скрещивания) передавали для оценки в полевых условиях.

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Central Scientific Agricultural Library - Russian Federation

2013

Simonov, V.S. | Vysotskij, V.A., All-Russia Selection and Technological Inst. of Horticulture and Plant Breeding, Moscow (Russian Federation) | Kulemekov, S.N., Science and Practice Center Fitogenetika

The descriptions of two new winter hardy varieties Velichavaya and Tuliza obtained by means of interspecific hybridization and embryo culture are given. The hybrid plants were obtained from embryos isolated on earliest stages of development (22-30 days after pollination). Among hybrid seedlings obtained from crossing of myrobalan plum Kubanskaya Kometa with plum variety Narach several hybrid seedlings were selected for following trials. Two of these seedlings named later as Velichavaya and Tuliza were characterized with the high fruit quality. The evaluation of winter hardiness of these forms by means of artificial frizzing detected their high winter hardiness in comparison with control plum varieties. The molecular-genetic analysis proved hybrid nature of these new varieties and cytological investigations showed that these forms were tetraploids (4x=32). Thereby the possibility for effective using ob biotechnological techniques for breeding projects for stone fruits has been demonstrated for widening genetic base and new varieties creating met modern requirements of agricultural production. | Описаны 2 новых зимостойких сорта сливы – Величавая и Тулица, полученных методами межвидовой гибридизации и культуры изолированных зародышей. Гибридные растения были получены из зародышей, изолированных на ранних стадиях развития - 22-30 дн. после опыления. Среди гибридных сеянцев, полученных в результате гибридизации алычи сорта Кубанская комета с сортом сливы Нарач, было отобрано несколько гибридных сеянцев для дальнейшего испытания. Два из них, получившие в дальнейшем названия Величавая и Тулица, характеризовались высоким качеством плодов. Оценка зимостойкости этих форм с помощью искусственного промораживания выявила их повышенную по сравнению с контрольными сортами сливы зимостойкость. Молекулярно-генетический анализ подтвердил гибридную природу этих новых сортов, а цитологические исследования показали, что они являются тетраплоидами (4х=32.) Таким образом, продемонстрирована возможность эффективного использования биотехнологических приемов в селекционных программах по косточковым культурам для расширения генетической базы и создания новых сортов, отвечающих современным требованиям производства.

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Central Scientific Agricultural Library - Russian Federation

2011

Kruglova, N.N. | Sel'dimirova, O.A., Russian Academy of Sciences. Ufa Scientific Center (Russian Federation). Institute of Biology

Central Scientific Agricultural Library - Russian Federation

2010

Sitnikova, O.I. | Polyakov, A.V., All-Russia Research and Development Inst. of Vegetable Growing, Moscow Region (Russian Federation) | Sharafova, O.F.

To overcome the pro- and postgametic incompatibility of squash species, we have worked out an advanced complex method consisting of the combination of crafting, pollination of buds 1 day before flowering and the technique of embryo culture. As maternal and paternal forms, cultivars of C. maxima L. (K 236, K 4253) and C. moschata Duch. (K 4612, K 2657) were used. The use of crafting resulted, at best, in formation of 8,6—13,3%, pistils decapitation, 2,7—2,9%, buds pollination 1 day before flowering, 2,3—6,3% of fruits. The seeds of Fi hybrids were characterized by the lowered germination (from 8% to 27%) in comparison with the parental forms (from 47,2% to 96,6%). The use of sodium hypochlorite for sterilization of embryos and seeds allowed to obtain of 89,0% and 92,0% sterile morphogenic explants, respectively. The optimum age of embryos (30 days) for introduction in in vitro culture was establishled, allowing to save their vitality at the level 10,5—25,0%. Media (Sh-2 and MS) for effective cultivation of the embryos isolated from not well developed seeds were determined. The use of these media resulted in growth and development of 10,5—14,2% embryos of the C. maxima x C. moschata hybrid. The in vitro cultivation of immature, not well developed seeds and embryos resulting from the crossing permitted to produce 89 plant regenerants. The interspecific nature of the C. maxima x C. moschata hybrids was confirmed by morphological, molecular-genetic, immunological and biochemical analyses. The forms of squash, obtained from the hybrid, combined morphological characteristics of parental cultivars and high resistance to peronosporosis and bacteriosis, caused by Pseudopero-nospora cubensis and Pseudomonas syrinae, respectively

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Central Scientific Agricultural Library - Russian Federation

2009

Akhmetova, A.Sh. (al_sham@mail.ru) | Bajburina, R.K. | Mironova, l.N., Russian Academy of Sciences. Ufa Science Centre (Russian Federation). Botanical Garden-Inst.

Embryos from seed of tulip hybrid forms were tested. The hybrids were a result of 5 breeding combinations of 9 varieties. The total number of the inoculated embryos was 140. The seeds were sterilized by 70% ethanol solution during 30 sec and by 0.2% diacid solution during 240 sec. Chloramin B could be used instead of diacid during 120 sec. The optimal period of embryo isolation was the 53-56 days after pollination when it was possible to pull out embryos of 1.5-3.0 milli m. At that case 91% of embryos started regenerating. The most intensive shoot proliferation was observed on the mediums containing 6-BAP (3.00 milli gram/l) and naphthylacetic acid (NAA) (0.15 mg/l). After 7 months an average number of shoots per an explant was 5.4 while shoot length was 0.043 m. The shoots elongated up to 0.050-0.055 m after 10 months. Cytokines were excluded from the medium and NAA was replaced by indolebytic acid (0.5 mg/l) for stimulating microbulb forming. Cultivating at dark at 4 deg C during 12 weeks inducted bulb-forming shoots. Then the regenerates were placed at light and were exposed at 26 deg C till the complete forming of microbulbs (30-120 days). The maximal output of microbulbs (39.9%) was at the hybrid combination Alladin x Blue heron; at the other combinations it was from 10.9 to 16.1%. The output of microbulbs was 6-8 units per an explant. The conclusion was made that the culture of isolated embryos allowed to keep and to propagate tulip hybrid forms effectively.

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Central Scientific Agricultural Library - Russian Federation

2009

Tretyakova, I.N. | Izhboldina, M.V., Russian Academy of Sciences. Siberian Branch, Krasnoyarsk (Russian Federation). The V.N. Sukachiev Inst. of Forest Science

An embryogenic callus (EC) further giving somatic germs was obtained by cultivating immature zygotic germs of Siberian cedar (Pinus sibirica) growing in natural forest crop of the West Sayan Mountains and on clonal plantations. The formation of somatic germ comprises the initiation of embryogenic callus, proliferation of embryonic suspension mass, obtaining and ripening somatic germs. The basic media MS and ½ SM added with mesoinozite and phytohormones: 2,4-D and 6-BAP, saccharose, as well as agar or Gerleete to obtain EC from megagametophytes and zygotic germs. For proliferation e embryonic mass the concentration of 6-BAP, 2,4-D and saccharose reduced 2-4 times. The cultivation was carried out in darkness. Hormone-free basic media with activated carbon were used to pass somatic germ to ripening. The cultivation was carried out for 1 week at 16-hour photoperiod and temperature 24 degrees C. mesoinozite, L-glumatine, 2,4-D and ABK, saccharose, agar were added to the media to ripen somatic germs. Cytological investigation showed that at the stage of initiation there is active lengthening of hypocotyl cells. Then lengthened cells are asymmetrically divided and form a little embryonic cell (future somatic germ) and lengthened embryonic tube fulfilling a gaustorial function. The process of initiation of EC was observed 58% of explants of the genotypes of Siberian cedar studied. Active formation of EC was observed only in 3 of 8 genotypes tested, and the formation of somatic germs was observed after 2-3 months of cultivation

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Central Scientific Agricultural Library - Russian Federation

2007

Tyukavin G.B.

Methods of germ culture, androgenesis and in vitro gynogenesis, as well as genetic transformation are applied in carrot selection. Some techniques were developed to sterilize explants. In initiating germ culture the treatment with 96% ethanol and sterilization with 0.1 or 0.5% water mercury bichloride solution were used. In obtaining anther culture and seed-buds flower buds were sterilized with chloramine B. In regenerating carrots from zygotic germs by androgenesis and gynogenesis there was used a Murasige-Skuga medium modified and added with 0.2 mg/l 2.4 D at the stage of induction of callus or embryodides and 0.1 mg/l kinetin on the stage of regeneration as such. For cultivation of zygotic germs the same medium is optimum added with 2 g/l of casein hydrolysate having yeast extract, 0.5 mg/l kinetin, 0.1 mg/l IAA and 0.01 mg/l gibberelic acid. The induction of androgenesis requires dark cultivation period. Explantation of anthers is best to be done at the stage of vacuolated microspores. The induction of gynogenesis under explantation of germs at the stage of ripen germ sac requires cultivation of the germ in the darkness for 14 days, and seed-buds – in the light. The cultivation of regenerants at temperature 2-4 degrees C slows down their growth and development allowing to maintain the obtained material for a long time. The genetic transformation was performed by combined cultivation of zygotic germs with agrobacteria on filtrated paper for 2-3 days. The culture selection was carried out on a medium comprising 200 mg/l kanamicyne, regeneration and acceleration – on a medium with 100 mg/l of kanamicyne. Transgenic plants carrying gene thaumatin 2 were produced. The expression of this gene is confirmed in leaves and root crops. Seedlings from seeds obtained by self-pollination of transgenic plants on the infectious background in vitro were resistant to fungi g. Cladosporium and Alernaria.

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Central Scientific Agricultural Library - Russian Federation