scholarly journals Biosynthesis of Polyene Antibiotics and Phytohormones by Streptomyces netropsis IMV Ac-5025 under the Action of Exogenous Isopentenyladenosine

2021 ◽  
Vol 83 (2) ◽  
pp. 32-41
Author(s):  
M.I. Loboda ◽  
◽  
L.O. Biliavska ◽  
G.O. Iutynska ◽  
◽  
...  

Streptomyces are active producers of a wide range of metabolites with multidirectional biological activity. Streptomyces netropsis IMV Ac-5025 synthesizes a polyene antibiotic complex in which two fractions were identified: heptaene candidine and a new tetraene fraction of unknown structure. The influence of secondary metabolites on the polyene antibiotics biosynthesis by soil streptomycetes is insufficiently explored. The aim of this work was to research the effect of exogenous isopentenyladenosine on the biosynthesis of polyene antibiotics and cytokinins by S. netropsis IMV Ac-5025. Methods. The strain was cultured in submerged cultivation condition in organic (soy) and synthetic (starch-ammonia) liquid nutrient media. The studies of biomass accumulation (gravimetric method), glucose consumption (glucose oxidase method), pH changes of culture media (ionometric method), biosynthesis of polyene antibiotics, and phytohormones (quantitative and qualitative thin layer chromatography spectrodensitometric method) were conducted. The results were analyzed in Statisticav.6.0 program. Results. It was found that polyene antibiotics are synthesized after the first day of cultivation, which indicates their role in the metabolism of streptomycetes. The biggest amount of the polyene antibiotics was accumulated in the stationary phase of producer growth (on the 7th day). It was found the decrease of polyene antibiotics and cytokinins accumulation in the producer’s biomass with the increase of exogenous cytokinin concentration from 25 ng/mL to 500 ng/mL. The bioproduction of the tetraene fraction was suppressed to a greater extent – up to 92% in the synthetic and up to 23% – in organic nutrient media. However, the amount of producer biomass increased under the action of the exogenous substance that confirming the positive effect of exogenous cytokinin on cell division of S. netropsis IMV Ac-5025. Exogenous isopentenyladenosine reduced the accumulation of endogenous cytokinins in streptomycetes biomass. Conclusions. The obtained results indicate an indirect metabolic relationship between the biosynthesis of polyene antibiotics and cytokinins in soil streptomycetes and provide a basis for the regulation of the biotechnological process for bioproduct formation with the appropriate quantitative composition of its components.

Growth regulators, phytohormones, both natural and artificial, are the main means to control plant ontogenesis. They are involved in regulating the processes of cell differentiation and cell divisions, the formation of tissues and organs, the changes in the rate of growth and development, the duration of the certain stages of ontogenesis. The main classes of phytohormones used in plant biotechnology, in particular, in the induction of haploid structures, are auxins and cytokinins. The mechanism of action of phytohormones on a cell is rather complicated and may have a different character. Understanding the characteristics of the action of phytohormones is complicated by the fact that the system of hormonal regulation of plant life is multicomponent. This is manifested in the fact that the same physiological process is most often influenced not by one, but by several phytohormones, covering a wide range of aspects of cell metabolism. In connection with the foregoing, the purpose of our work was to test a set of nutrient media with different basic composition and different proportions of phytohormones to determine the patterns of their influence on the processes of haploid structure induction in rape anther culture using accessions, developed at the Institute of Oilseed Crops NAAS. The material used was two accessions of winter rapeseed (No. 1 and No. 2) and one sample of spring rapeseed, provided by the Rapeseed Breeding laboratory of the Institute of Oilseed Crops. Incised inflorescences were kept against the background of low temperature of 6–8 ° C for several days, and then, under aseptic conditions, anthers with unripe pollen grains were isolated and planted on nutrient media differing in both basic mineral composition and content of phytohormones. MS (Murashige & Skoog 1962) and B5 (Gamborg et al 1968) media were used as basic media. Phytohormones were added to the basic media in various combinations – BA, 2,4-D, NAA at the concentrations of 0.1-0.6 mg/l. In each treatment up to 300 anthers were cultivated. Differences between treatments were evaluated using standard t-test. Studies have shown that in the anther culture of rapeseed on the tested nutrient media, morphogenic structures of different types (embryoids and callus) were originated. Synthetic auxin 2,4-D, regardless of the composition of the basic medium, caused the formation of structures of both types, though with a low frequency. Phytohormone BA of the cytokinin type had a similar effect. In this case, the frequency of structures was slightly higher, and the developed structures were represented mainly by embryoids. The joint action of cytokinin and auxin was the most favorable for the initiation of morphogenic structures. Such combination of phytohormones caused the formation of these structures with a frequency of 24.5-14.7% in the studied genotypes of winter rape. A similar effect of phytohormones on the induction and development of morphogenic structures was also observed in spring rape. In this case, a single basic MS medium was used. The experiment included treatments where phytohormones were absent (control), as well as various combinations of auxin and cytokinin. In the control treatment, the formation of new structures was not noted. In treatments with phytohormones, in addition to the medium with the combination of auxin and cytokinin, the medium in which only cytokinin was present was also rather effective. The treatment in which the action of auxin 2,4-D was combined with the action of another auxin, NAA, turned out to be practically ineffective. Thus, it was found that for the induction of morphogenic structures from microspores in rape anther culture of the tested genotypes, the combination of cytokinin with auxin, or the use of only single cytokinin BA without other phytohormones, had the most positive effect.


Toxins ◽  
2019 ◽  
Vol 11 (11) ◽  
pp. 645 ◽  
Author(s):  
Hamed K. Abbas ◽  
Nacer Bellaloui ◽  
Cesare Accinelli ◽  
James R. Smith ◽  
W. Thomas Shier

Charcoal rot disease, caused by the fungus Macrophomina phaseolina, results in major economic losses in soybean production in southern USA. M. phaseolina has been proposed to use the toxin (-)-botryodiplodin in its root infection mechanism to create a necrotic zone in root tissue through which fungal hyphae can readily enter the plant. The majority (51.4%) of M. phaseolina isolates from plants with charcoal rot disease produced a wide range of (-)-botryodiplodin concentrations in a culture medium (0.14–6.11 µg/mL), 37.8% produced traces below the limit of quantification (0.01 µg/mL), and 10.8% produced no detectable (-)-botryodiplodin. Some culture media with traces or no (-)-botryodiplodin were nevertheless strongly phytotoxic in soybean leaf disc cultures, consistent with the production of another unidentified toxin(s). Widely ranging (-)-botryodiplodin levels (traces to 3.14 µg/g) were also observed in the roots, but not in the aerial parts, of soybean plants naturally infected with charcoal rot disease. This is the first report of (-)-botryodiplodin in plant tissues naturally infected with charcoal rot disease. No phaseolinone was detected in M. phaseolina culture media or naturally infected soybean tissues. These results are consistent with (-)-botryodiplodin playing a role in the pathology of some, but not all, M. phaseolina isolates from soybeans with charcoal rot disease in southern USA.


2021 ◽  
pp. 92-95
Author(s):  
Александр Николаевич Остриков ◽  
Наталья Леонидовна Клейменова ◽  
Инэсса Николаевна Болгова ◽  
Максим Васильевич Копылов ◽  
Екатерина Юрьевна Желтоухова

Использование растительных масел в рационе человека необходимо для удовлетворения энергетической потребности организма и регулирования биологических процессов. В настоящей работе представлен качественный и количественный состав витаминов и токоферолов различных видов растительных масел, полученных холодным отжимом из выращенных в нашей стране расторопши, горчицы, подсолнечника, рыжика, рапса. Проведен сравнительный анализ литературных источников о наличии витаминов и токоферолов в различных растительных маслах. Количественный и качественный витаминный состав для одного и того же вида масла, по данным разных авторов, варьируется в достаточно широком диапазоне. В ходе исследования изучен витаминный состав пяти растительных масел. Результаты свидетельствуют, что наибольшей витаминной активностью обладают масла рыжиковое, расторопши и горчичное. Установлено наличие витамина А в маслах: рыжиковом (27,15±0,002 мкг%), расторопши (19,07±0,02 мкг%), горчичном (24,77±0,02 мкг%). Витамины В и В присутствуют в маслах горчичном, рыжиковом и расторопши; витамин В определен в горчичном и рыжиковом, В - в рыжиковом и расторопши. В маслах горчичном и расторопши присутствует витамин В. Наиболее богаты витамином Е масла рыжиковое (52,8±0,02 мг%), подсолнечное (48,3±0,02 мг%), расторопши (47,12±0,02 мг%). Витамин К и токоферолы определены во всех растительных маслах. Присутствие b-каротина обнаружено в рыжиковом (1,237±0,004 мг%), расторопши (0,812±0,002 мг%), подсолнечном (0,22±0,02 мг%) и горчичном (0,148±0,002 мг%) маслах. Анализируемые масла холодного отжима можно рассматривать как ценный источник при проектировании новых многоцелевых продуктов или побочных продуктов для промышленного, косметического и фармацевтического использования. The use of vegetable oils in the human diet is necessary to meet the energy needs of the body and regulate biological processes. This work presents the qualitative and quantitative composition of vitamins and tocopherols of various types of vegetable oils obtained by cold pressing from silybum marianum, mustard, sunflower, camelina, rapeseed grown in our country. A comparative analysis of the literature on the presence of vitamins and tocopherols in various vegetable oils has been carried out. The quantitative and qualitative vitamin composition for the same type of oil, according to different authors, varies in a wide range. During the study, the vitamin composition of five vegetable oils was studied. The results indicate that the following oils have the highest vitamin activity: camelina, silybum marianum and mustard. The presence of vitamin A was found in oils: camelina (27.15±0.002 μg%), silybum marianum (19.07±0.02 μg%), mustard (24.77±0.02 μg%). Vitamins B and B are present in mustard, camelina and silybum marianum oils; vitamin B is found in mustard and camelina; B in camelina and silybum marianum. Mustard and silybum marianum oils contain vitamin B. The richest in vitamin E are camelina (52.8±0.02 mg%), sunflower (48.3±0.02 mg%), silybum marianum (47.12±0.02 mg%). Vitamin K and tocopherols are found in all vegetable oils. The presence of b-carotene was found in camelina (1.237±0.004 mg%), silybum marianum (0.812±0.002 mg%), sunflower (0.22±0.02 mg%) and mustard (0.148±0.002 mg%) oils. The analyzed cold-pressed oils can be considered as a valuable resource when designing new multipurpose products or by-products for industrial, cosmetic and pharmaceutical applications.


1926 ◽  
Vol 22 (1) ◽  
pp. 19-22
Author(s):  
V. M. Aristovskiy ◽  
R. R. Gel'ttser

In recent years, great strides have been made on the cultivation of spirochetes, and if relatively simple and completely reliable methods of growing them in vitro on both solid and liquid nutrient media have been developed for a number of pathogenic and non-pathogenic spirochetes, then on the issue of With the cultivation of the pale spirochete, we are moving forward very slowly, and the successes achieved recently in this area must be recognized as very modest.


2019 ◽  
Vol 14 (1) ◽  
pp. 90-98
Author(s):  
Vladimir E Nikitchenko ◽  
Ekaterina O Rystsova ◽  
Anastasiya N Chernysheva

At all stages of the production of microbiological nutrient media (MNM), the manufacturer, and in particular, the microbiological laboratories that manufacture the media on site, face many operational risks. The presence of such risks, in almost every critical point of production and further operation of the MNM, is due to the presence of common basic requirements for all MNM, which must be taken into account and respected during their development and preparation; as well as the complexity and laboriousness of the very process of manufacturing high-quality differential-diagnostic and other nutrient media, requiring the availability of all the components necessary for preparing these media, equipment, sterile conditions and qualified personnel. In this regard, there is a need to search for effective methods to identify and prevent undesirable situations associated with the production and use of MNM. The aim of this work was to adapt the risk assessment methodology based on the expert method for analyzing the types and consequences of FMEA failures (Failure Mode Effect Analysis) set out in GOST R ISO 31010-2011 for the needs of microbiological laboratories, including those for veterinary and sanitary expertise, producing microbiological nutrient environments and using them. As part of this work, a comparative analysis of risk assessment methods was carried out in order to select the optimal one; adaptation of the QMS principle - risk-oriented thinking and the FMEA method for risk assessment in the implementation of MNM manufacturing processes in a microbiological laboratory (for example, solid agar media); risk assessment protocol forms were developed; calculations of a quantitative assessment of risk levels were carried out in order to determine the need for preventive actions and their implementation in order to minimize the negative consequences of risk in case of its implementation using the developed protocols. The results showed that this technique can be successfully implemented and used in the claimed area.


2021 ◽  
pp. 301-308
Author(s):  
Olesya Nikolayevna Mazko ◽  
Lyudmila Ivanovna Tikhomirova ◽  
Lyudmila Vladimirovna Shcherbakova ◽  
Natal'ya Grigor'yevna Bazarnova ◽  
Dmitriy Alekseyevich Karpitsky

The aim of this study was to evaluate the effect of 6-benzylaminopurine (BAP) separately and in interaction with auxins on the change in the qualitative and quantitative composition of flavonoids in the raw materials of regenerating plants Iris sibirica L. Cambridge grade in comparison with aeroponic and intact raw materials using the method of high-performance liquid chromatography. Raw materials of I. sibirica Cambridge variety obtained in vitro culture had a richer qualitative composition of flavonoids than intact plants. The dependence of the accumulation of flavonoids on the concentration of 6-benzylaminopurine in nutrient media was noted. The presence of 13 compounds was observed in extracts of 70% ethyl alcohol from regenerating plants grown at the lowest concentration of BAP (1.0 µM) within the experiment. In quantitative terms, the flavonoid apigenin was maximally determined on a medium with BAP 1 µM, and kaempferol - on media with BAP 5.0 µM, supplemented with auxins. For a medium with 7.5 µM BAP, the lowest variety of compounds was observed (9) and the lowest kaempferol content. Auxins influenced the synthesis of flavonoids. The amount of flavonoids in all variants of the experiment increased by an average of 13% in the presence of auxins. The stages of the technological process of obtaining raw materials I. sibirica Cambridge variety on the basis of clonal micropropagation and cultivation in aeroponics conditions allowed to obtain raw materials that do not contain heavy and toxic metals, are not infected with pathogens and pests. With 1 m2 of useful area of aeroponics for 1 year, it is possible to collect 5 times more raw materials than with field cultivation. According to the qualitative composition of phenolic compounds, aeroponic raw materials are identical to intact plants.


2021 ◽  
pp. 11-13
Author(s):  
Дарья Владиславовна Андриевская ◽  
Екатерина Владимировна Ульянова

В настоящее время известны основные причины формирования различных видов помутнений в спиртных напитках виноградного происхождения. В значительной степени изучены компоненты образующихся при этом осадков. Вместе с тем одна из часто возникающих проблем современного производства заключается в потере стабильности продукции при транспортировании и хранении, что влечет дополнительные расходы предприятий по возврату и технологическим обработкам забракованной продукции. Целью настоящей работы стало сравнительное исследование минерального состава обработанных коньяков и их стабильности к различным видам помутнений. Объекты исследований - образцы обработанных коньяков отечественного производства, которые были разделены в зависимости от возраста на четыре группы: 1 группа - 3-летние, 4-летние и 5-летние; 2 группа - выдержанные «КВ» и выдержанные высшего качества «КВВК»; 3 группа - старые «КС», 4 группа - очень старые «ОС». На 1-м этапе было проведено исследование физико-химических и органолептических показателей продукции. В результате установлено, что все образцы соответствовали требованиям ГОСТ 31732-2014. Тесты на склонность к различным видам помутнений показали стабильность образцов к обратимым коллоидным, белковым и кальциевым помутнениям. Установлено, что качественный состав минеральных веществ обработанных коньяков различного возраста идентичен. При этом массовая концентрация отдельных компонентов варьирует в широком диапазоне. Доля образцов обработанных коньяков, в которых были превышены уровни концентраций отдельных ионов, гарантирующих стабильность продукции, составила в среднем 15%. Сравнительный анализ количественного состава катионов и анионов в обработанных коньяках дает основание предположить, что образцы с высоким содержанием ионов Са2+, Na+ и SO42- имеют высокий риск потери стабильности в дальнейшем. Таким образом, результаты проведенных исследований показали необходимость совершенствования методики определения склонности коньяков к различным видам помутнений в части расширения контролируемых показателей. Currently, the main reasons for the formation of various types of turbidity in alcoholic beverages of grape origin are known. The components of the precipitation formed during this process have been studied to a large extent. At the same time, one of the frequently occurring problems of modern production is the loss of product stability during transportation and storage, which entails additional costs for enterprises to return and process rejected products. The purpose of this work was a comparative study of the mineral composition of processed cognacs and their stability to various types of turbidity. The objects of research were samples of processed cognacs of domestic production, which were divided depending on age into four groups: group 1 - three-year-old, four-year-old and five-year-old; group 2 - aged «KV» and aged of the highest quality «KVVK»; group 3 - old «KS», group 4 - very old «OS». At the first stage, a study of the physico-chemical and organoleptic parameters of the products was carried out. As a result, it was found that all samples met the requirements of GOST 31732-2014. Tests for the propensity to various types of turbidity showed the stability of the samples to reversible colloidal, protein and calcium turbidity. It is established that the qualitative composition of the minerals of processed cognacs of different ages is identical. At the same time, the mass concentration of individual components varies in a wide range. The share of samples of processed cognacs in which the concentration levels of individual ions guaranteeing the stability of products were exceeded averaged 15%. A comparative analysis of the quantitative composition of cations and anions in processed cognacs suggests that samples with a high content of Ca2+, Na+ and SO42- ions have a high risk of loss of stability in the future. Thus, the results of the conducted studies have shown the need to improve the methodology for determining the propensity of cognacs to various types of turbidity in terms of expanding the controlled indicators.


Author(s):  
Deep Chhavi Anand ◽  
Rishikesh Meena ◽  
Vidya Patni

Objective: The aim of the present study was to develop a callus induction protocol and comparative study of therapeutic phytochemicals present in in vivo leaf and in vitro callus extracts through Gas Chromatography-Mass Spectrometry analysis.Methods: Murashige and Skoog media was used as culture media for callus induction. In vitro callus induction protocol was developed by studying the effects of various plant growth regulators like auxin, 2, 4-D (2,4-dichlorophenoxyacetic acid), NAA (naphthalic acetic acid), alone and in combination with cytokinin BAP (benzyl aminopurine), on leaf and stem explants. The GC-MS analysis of Ampelocissus latifolia was carried out on Shimadzu QP-2010 plus with thermal desorption system TD 20 to study the phytochemical profile.Results: In vitro callus induction protocol was developed for the plant and callusing was done from leaf and stem explants of Ampelocissus latifolia. The best result for callus induction was obtained using leaf explant, and callus production were maximum in Murashige and Skoog medium fortified with BAP (0.5 mg/l) and NAA (1.0 mg/l). Major compounds identified in the GC-MS analysis were Campesterol, Stigmasterol, Beta-Sitosterol, Docosanol, Dodecanoic acid, etc., in in vitro extract and Beta Sitosterol, Tocopherol, Squalene, Bergamot oil, Margarinic acid, Hexadecanoic acid, etc., in in vivo extract. The different active phytochemicals identified have been found to possess a wide range of biological activities, thus this analysis forms a basis for the biological characterization and importance of the compounds identified for human benefits.Conclusion: This is the first report on callus induction in Ampelocissus latifolia. From the results obtained through the in vitro callus induction and its comparative GCMS analysis with in vivo extract, it is revealed that Ampelocissus latifolia contains various bioactive compounds that are of importance for phytopharmaceutical uses. The GCMS analysis revealed that the amount of Beta-sitosterol and 5-Hydroxymethylfurfural (HMF) was very high in in vitro extract as compared to in vivo extract.


2014 ◽  
Vol 50 (No. 2) ◽  
pp. 123-129 ◽  
Author(s):  
S. Bobkov

Pea (Pisum sativum L.) is a species for which there is no efficient method for the recovery of haploid plants yet. This research investigated the influence of various genotypes, nutrient media, and stress treatments on callus formation, embryogenesis and plant regeneration in anther cultures of pea. A wide range of pea genotypes and nutrient media was studied. Morphogenic calli were initiated on media supplemented with &alpha;-naphthaleneacetic acid (NAA), 6-benzyladenine (BA), and 2,4-dichlorophenoxyacetic acid (2,4-D) without application of stress treatments. Embryogenic calli and embryos were regenerated on media with low sucrose content in the presence of 2,4-D or indole-3-butyric acid (IBA) after cold stress (4&deg;C) of isolated buds, alone or in combination with in&nbsp;vitro treatment of isolated anthers at higher temperatures (35&ndash;38&deg;C). The efficiency of regeneration via shoot morphogenesis on different nutrient media and the peculiarities of regeneration from embryogenic calli were investigated. Green embryogenic calli initiated on 2,4-D were able to develop through shoot morphogenesis on a medium supplemented with BA and NAA. This process led to regeneration of hypertrophic embryos at various developmental stages. The origin of regenerated plants (i.e. from microspores or somatic anther cells) was estimated using marker alleles determining morphological traits. Almost all R<sub>0</sub> regenerants derived from morphogenic calli originated from anther somatic cells.


2004 ◽  
Vol 44 (1) ◽  
pp. 27 ◽  
Author(s):  
M. Bajji ◽  
P. Bertin ◽  
S. Lutts ◽  
J-M. Kinet

Somaclonal variation associated with in vitro selection has been used as a source of variability to improve drought resistance of 3 durum wheat (Triticum durum Desf.) cultivars (Selbera, Sebou, and Kyperounda). In a previous study, R0 plants with improved drought resistance-related characters were regenerated after selection on culture media containing polyethylene glycol (PEG). This improvement was transmitted to the R1 progeny. The present study analysed the behaviour of the selected tissue culture-derived lines in subsequent R2, R3 and R4�generations. Differences in electrolyte leakage, chlorophyll fluorescence (Fv/Fm), stomatal conductance and days to heading were found between the parental cultivars and most of their in vitro-derived lines. The changes may differ from one cultivar to another. Many promising somaclonal lines still presented improvement for at least 3 of the 4�parameters measured comparatively to initial cultivars. Somaclonal variation thus appears to induce a wide range of modifications among individual components of drought-resistance mechanisms. These improved traits could be valuable if shown to be inherited and to give enhanced agronomic performances in future field studies.


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