scholarly journals INFLUENCE OF CULTIVATION CONDITIONS AND CULTURE MEDIUM GORMONAL COMPOSITION ON THE MICROPROPAGATION OF THYMUS VULGARIS L. IN VITRO

Author(s):  
A.Sh. Tevfik ◽  
◽  
N.A. Yegorova ◽  
2020 ◽  
Vol 224 ◽  
pp. 04001
Author(s):  
A Sh Tevfik ◽  
N. A. Yegorova

Thymus vulgaris L. is one of the widely known spicy aromatic and medicinal plants. Thyme plant material is widely used in medicine, cooking and perfumery. To increase the efficiency of breeding and seed production, it is necessary to develop biotechnological techniques, in particular, clonal micropropagation. The aim of the research is to optimize the composition of culture media for the main stages of propagation in vitro and to select adaptation ex vitro conditions for the development of Thymus vulgaris. clonal micropropagation. The article presents the results of studies of explant morphometric parameters cultivated on 20 variants of culture media at firstsecond stages of micropropagation. It was found that the optimal culture medium at the introduction stage is MS medium with 1.0 mg/l Kin and 1.0 mg/l GA3, on which, on average, 2.2 microshoots per explant with a length of 1.9 cm were obtained. Both high vitrification rate of microshoots and formation of small shoots (0.6-0.9 cm) were observed on media supplemented with BAP or TDZ. The most effective culture medium at the proper propagation stage is MS with 1.0 mg/l Kin, on which 4.6 shoots per explant and the multiplication index 12.8 were obtained. It is advisable to root microshoots at the 3rd stage of micropropagation on MS culture medium supplemented with 1.0 mg/l IBA or 1.0 mg/l IAA. It has been shown that it is possible to obtain high plant survival rate (89.5%) during adaptation ex vitro, using a substrate consisting of peat and perlite (1:1).


2021 ◽  
Vol 937 (4) ◽  
pp. 042093
Author(s):  
O V Yakimova ◽  
N A Yegorova

Abstract Melissa officinalis L. is a perennial herbaceous essential and medicinal plant widely used in pharmacology, perfumery and cosmetics, as well as in alcoholic beverage and food industries. The low content of essential oil in lemon balm raw material determines the selection work aimed at creating high-oil cultivars. The use of clonal micropropagation method in vitro will increase the efficiency of this process and accelerate promising breeding samples multiplication. The aim of our research was to study the influence of cultivation conditions and cultivar on the M. officinalis in vitro rhizogenesis and ex vitro adaptation. It was found that the maximum frequency of shoot rooting (up to 93.3%) in cultivars ‘Citronella’ and ‘Sobornaya’ was on MS culture medium supplemented with 0.5 mg/l NAA. The number of roots was 10.1 and 13.6 pcs. per shoot, respectively. The highest rates of root formation for the cv. ‘Crimchanka’ was found on a culture medium supplemented with 1.0 mg/l IAA (8.7 roots per shoot). The mixture of peat, sand and perlite (2:1:2) as a substrate provided up to 93% of adapted ex vitro lemon balm microplants. The presented studies were used to develop a technique for clonal micropropagation of M. officinalis.


2020 ◽  
pp. 298-303
Author(s):  
Игорь Владимирович Гавриленко ◽  
Юлия Сергеевна Матяш ◽  
Анжела Владимировна Гавриленко ◽  
Дмитрий Александрович Шанин ◽  
Ирина Александровна Павлова ◽  
...  

Сорт винограда Кобер 5 ББ (Берландиери x Рипариа Кобер 5ББ) - один из основных подвоев, используемых в питомниководстве для получения привитых саженцев, поэтому в настоящее время крайне актуально создание маточников данного подвоя посадочным материалом категории «оригинальный». Это объясняет необходимость проведения исследований, связанных с оптимизацией условий культивирования сорта Кобер 5ББ, для повышения эффективности массового клонального микроразмножения с сохранением его генетической однородности и стабильности. Целью исследования являлась оптимизация и подбор питательных сред для клонального микроразмножения сорта-подвоя Кобер 5 ББ на этапе тиражирования (микрочеренкования). Материалом для исследований служили растения in vitro сорта подвоя Кобер 5ББ, свободные от основной патогенной инфекции (по результатам тестирования). Исследования проводили на средах: МS; WPM; DKW; PG (контроль). В качестве регуляторов роста использовали GA (гиббереллиновая кислота) в концентрациях: 0,2; 0,6; 1; 1,4 мг/л в сочетании с NAA (α-нафтилуксусная кислота) 0,05 мг/л. Показано, что растения на среде WPM, содержащей NAA-0,05 мг/л, по биометрическим показателям превосходили развившиеся на среде PG с аналогичным гормональным составом. Проведенные исследования по оптимизации среды культивирования для ускорения ростовых процессов позволили по результатам биометрических показателей выделить блок сред с основой WPM для размножения сорта-подвоя Кобер 5 ББ на этапе микрочеренкования. После проведения дополнительных исследований с расширенной выборкой среду WPM можно будет рекомендовать для клонального микроразмножения винограда на этапе микрочеренкования. The grape variety ‘Kober 5 BB’ (‘Berlandieri x Riparia Kober 5BB’) is one of the main rootstocks used in rootstock-growing farming to obtain grafted seedlings. Currently it is a hot issue to create nurseries for the rootstock grapevine with planting material of the "original" category. This explains the need for research related to optimization of cultivation conditions of the variety ‘Kober 5BB’ to increase the efficiency of mass clonal micropropagation while retaining its genetic homogeny and stability. The aim of the study was to optimize and select nutrient media for clonal micropropagation of the rootstock variety ‘Kober 5 BB’ at the stage of tiraging (micropropagation by cutting). The material of research was the in vitro plants of ‘Kober 5BB’ rootstock variety, free from basic pathogenic infection (according to the test results). The studies were carried out on media: MS; WPM; DKW; PG (control). Gibberellic acid (GA) was used as a growth regulator at concentrations: 0.2; 0.6; 1; 1.4 mg/l in combination with NAA (α-naphthyl acetic acid) 0.05 mg/l. Plants on WPM medium containing NAA-0.05 mg/l in biometric parameters were superior to those grown on PG medium with a similar hormonal composition. According to the results of biometric parameters the studies on optimization the culture medium for accelerating growth processes made it possible to isolate a group of media with a WPM base for propagation of the rootstock variety ‘Kober 5 BB’ at the stage of microcutting. After additional studies with expanded selection, the WPM medium can be recommended for clonal micropropagation of grapes at the stage of microcutting.


1991 ◽  
Vol 125 (3) ◽  
pp. 280-285 ◽  
Author(s):  
J. Alan Talbot ◽  
Ann Lambert ◽  
Robert Mitchell ◽  
Marek Grabinski ◽  
David C. Anderson ◽  
...  

Abstract We have investigated the role of Ca2+ in the control of FSH-induced estradiol secretion by Sertoli cells isolated from 8-10 days old rats. Exogenous Ca2+ (4-8 mmol/1) inhibited FSH-stimulated E2 secretion such that, with 8 mmol/l Ca2+ and FSH (8 IU/l) E2 secretion decreased from 2091±322 to 1480±84 pmol/l (p<0.002), whilst chelation of Ca2+ in the culture medium with EGTA (3 mmol/l) increased E2 secretion from 360±45 to 1242±133 pmol/l) in the absence of FSH. Further, EGTA (3 mmol/l) markedly potentiated FSH (8 IU/l), forskolin (1 μmol/l) and dibutyryl cAMP (1 mmol/l)-stimulated E2 secretion. Addition of the Ca2+ ionophores, ionomycin (2-5 μmol/l) and A23187 (2 μmol/l), inhibited FSH (8 IU/l)-stimulated E2 secretion by >80%. The effect of ionomycin was totally reversible, whereas that of A23187 was irreversible. Ionomycin (5 μmol/l) had no effect on EGTA-induced E2 secretion in the absence of FSH, but reduced EGTA-provoked E2 secretion by 59% in the presence of FSH (8 IU/l). Similarly, forskolin- and dibutyryl cAMP-provoked E2 production was inhibited 46-50% by ionomycin (5 μmol/l). We conclude that FSH-induced E2 secretion from immature rat Sertoli cells is modulated by intra- and extracellular Ca2+.


Reproduction ◽  
2000 ◽  
pp. 391-396 ◽  
Author(s):  
AH Duittoz ◽  
M Batailler

The aim of this study was to investigate the development of pulsatile GnRH secretion by GnRH neurones in primary cultures of olfactory placodes from ovine embryos. Culture medium was collected every 10 min for 8 h to detect pulsatile secretion. In the first experiment, pulsatile secretion was studied in two different sets of cultures after 17 and 24 days in vitro. In the second experiment, a set of cultures was tested after 10, 17 and 24 days in vitro to investigate the development of pulsatile GnRH secretion in each individual culture. This study demonstrated that (i) primary cultures of GnRH neurones from olfactory explants secreted GnRH in a pulsatile manner and that the frequency and mean interpulse duration were similar to those reported in castrated ewes, and (ii) pulsatile secretion was not present at the beginning of the culture but was observed between 17 and 24 days in vitro, indicating the maturation of individual neurones and the development of their synchronization.


Author(s):  
С.В. Калиш ◽  
С.В. Лямина ◽  
А.А. Раецкая ◽  
И.Ю. Малышев

Цель исследования. Репрограммирование М1 фенотипа макрофагов с ингибированными факторами транскрипции М2 фенотипа STAT3, STAТ6 и SMAD и оценка их влияния на развитие карциномы Эрлиха (КЭ) in vitro и in vivo. Методика. Рост опухоли иницировали in vitro путем добавления клеток КЭ в среду культивирования RPMI-1640 и in vivo путем внутрибрюшинной инъекции клеток КЭ мышам. Результаты. Установлено, что M1макрофаги и in vitro, и in vivo оказывают выраженный противоопухолевый эффект, который превосходит антиопухолевые эффекты М1, M1, M1 макрофагов и цисплатина. Заключение. М1 макрофаги с ингибированными STAT3, STAT6 и/или SMAD3 эффективно ограничивают рост опухоли. Полученные данные обосновывают разработку новой технологии противоопухолевой клеточной терапии. Objective. Reprogramming of M1 macrophage phenotype with inhibited M2 phenotype transcription factors, such as STAT3, STAT6 and SMAD and assess their impact on the development of Ehrlich carcinoma (EC) in vitro and in vivo . Methods. Tumor growth in vitro was initiated by addition of EC cells in RPMI-1640 culture medium and in vivo by intraperitoneal of EC cell injection into mice. Results. It was found that M1 macrophages have a pronounced anti-tumor effect in vitro , and in vivo , which was greater than anti-tumor effects of M1, M1, M1 macrophages and cisplatin. Conclusion. M1 macrophages with inhibited STAT3, STAT6 and/or SMAD3 effectively restrict tumor growth. The findings justify the development of new anti-tumor cell therapy technology.


Agrologia ◽  
2018 ◽  
Vol 1 (1) ◽  
Author(s):  
S. Tuhuteru ◽  
Meity L Hehanussa ◽  
Simon H.T Raharjo

Dendrobium anosmum is one of natural orchids in Indonesia. Optimization of medium composition for orchid propagation through in vitro culture is necessary to enhance propagule multiplication capabilities and quality. This study was aimed to study the influence of concentration of coconut water in culture medium on in vitro growth and development of D. anosmum orchid species and to determine the optimal coconut water concentration in culture media.  The experiment were arranged in a Completely Randomized Design with four treatments and eight replications. The treatments consisted of the addition of coconut water with concentrations: 0 ml•l -1 (control), 50 ml•l-1, 100 ml•l-1 and 150 ml•l-1. The results showed that addition of coconut water in culture medium gave different effect on shoot growth and multiplication of D. anosmum orchids.  Coconut water concentration of 100 ml•l-1 was the best concentration for growth and multiplication of D. anosmum orchids, based on both shoots and roots growth, plantlet height and wet weight.


Author(s):  
Bruna O. S. Câmara ◽  
Bruno M. Bertassoli ◽  
Natália M. Ocarino ◽  
Rogéria Serakides

The use of stem cells in cell therapies has shown promising results in the treatment of several diseases, including diabetes mellitus, in both humans and animals. Mesenchymal stem cells (MSCs) can be isolated from various locations, including bone marrow, adipose tissues, synovia, muscles, dental pulp, umbilical cords, and the placenta. In vitro, by manipulating the composition of the culture medium or transfection, MSCs can differentiate into several cell lineages, including insulin-producing cells (IPCs). Unlike osteogenic, chondrogenic, and adipogenic differentiation, for which the culture medium and time are similar between studies, studies involving the induction of MSC differentiation in IPCs differ greatly. This divergence is usually evident in relation to the differentiation technique used, the composition of the culture medium, the cultivation time, which can vary from a few hours to several months, and the number of steps to complete differentiation. However, although there is no “gold standard” differentiation medium composition, most prominent studies mention the use of nicotinamide, exedin-4, ß-mercaptoethanol, fibroblast growth factor b (FGFb), and glucose in the culture medium to promote the differentiation of MSCs into IPCs. Therefore, the purpose of this review is to investigate the stages of MSC differentiation into IPCs both in vivo and in vitro, as well as address differentiation techniques and molecular actions and mechanisms by which some substances, such as nicotinamide, exedin-4, ßmercaptoethanol, FGFb, and glucose, participate in the differentiation process.


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