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Modulation of growth and chemical element accumulation in Fragaria × ananassa plants in vivo under the effect of silicon chelates 在硅螯合物的作用下调节 Fragaria × ananassa 植物的体内生长和化学元素积累
Pub Date : 2023-12-26 DOI: 10.21285/2227-2925-2023-13-4-494-505
E. Ambros, E. S. Krupovich, Yu. P. Kolmogorov, E. Trofimova, I. S. Gusev, B. Goldenberg
Due to the protective role played by silicon in plants against unfavorable environmental conditions, siliconcontaining preparations are of considerable interest as biostimulants. In this work, a mechanical composite of rice husk and green tea containing soluble silica chelate complexes was used as the source of silicon. The study aims to examine the effect of silicon chelates on the growth and physiological parameters and the chemical composition of Fragaria × ananassa plants (Solnechnaya Polyanka variety) under greenhouse conditions. The plants were watered using water without a mechanical composite (control) or an aqueous solution containing 0.3 g/L of mechanical composite twice per period. Sampling was carried out one week after the last treatment. In order to determine the concentration of chemical elements (Si, P, S, Cl, K, Ca, Ti, Mn, Fe, Ni, Cu, Zn, Br, Rb, Sr, and Mo) in the roots and shoots of garden strawberry, it was proposed to use synchrotron X-ray fluorescence analysis. The use of the mechanical composite was shown to increase the amount of chlorophyll a, chlorophylls a and b, and carotenoids; decrease the amount of hydrogen peroxide; and increase the activity of the main antioxidant enzymes (superoxide dismutase, catalase, and peroxidase). It was determined that under the effect of the preparation, silicon accumulates primarily in shoots, affecting the accumulation of micro- and macroelements in the shoots and roots of plants. The obtained results substantiate the use of silicon-containing “green chemistry” as a means of controlling the growth and development of garden strawberry plants under in vivo conditions.
由于硅对植物在不利环境条件下的保护作用,含硅制剂作为生物刺激剂备受关注。在这项研究中,使用了一种含有可溶性硅螯合物的稻壳和绿茶机械复合材料作为硅源。研究旨在考察硅螯合物对温室条件下 Fragaria × ananassa 植物(Solnechnaya Polyanka 品种)的生长和生理参数以及化学成分的影响。使用不含机械复合剂(对照)的水或含 0.3 克/升机械复合剂的水溶液浇灌植物,每期两次。取样在最后一次处理一周后进行。为了确定花园草莓根和芽中化学元素(Si、P、S、Cl、K、Ca、Ti、Mn、Fe、Ni、Cu、Zn、Br、Rb、Sr 和 Mo)的浓度,建议使用同步辐射 X 射线荧光分析法。结果表明,使用机械复合材料可以增加叶绿素 a、叶绿素 a 和叶绿素 b 以及类胡萝卜素的含量;降低过氧化氢的含量;提高主要抗氧化酶(超氧化物歧化酶、过氧化氢酶和过氧化物酶)的活性。据测定,在制剂的作用下,硅主要在芽中积累,影响植物芽和根中微量和大量元素的积累。所获得的结果证明,可以利用含硅的 "绿色化学 "作为一种手段,在体内条件下控制花园草莓植物的生长和发育。
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引用次数: 0
Effect of the overexpression of external alternative NADH dehydrogenase gene in Arabidopsis on the resistance of transformed tobacco plants to negative temperatures 拟南芥外部替代 NADH 脱氢酶基因的过表达对转化烟草植物抗负温度能力的影响
Pub Date : 2023-12-26 DOI: 10.21285/2227-2925-2023-13-4-516-522
G. B. Borovskii, E. L. Gorbyleva, A. Katyshev, N. A. Korotaeva
The study aims to confirm the functional activity and localization of the At_NDB2 transgenic protein of Arabidopsis in tobacco cells and to evaluate the effect produced by the permanent expression of external alternative NADH dehydrogenase on the resistance of a heat-loving plant to low temperatures. Proteins and mitochondria were isolated from the leaves of tobacco plants grown at 25 °С (day/night) at the 7-leaf stage. At_NDB2 protein localization in mitochondria was determined via electrophoresis and immunoblotting. The functional activity of At_NDB2 was confirmed through the polarography of isolated mitochondria and the specific mitochondrial complex I inhibitor (rotenone). It was also found that the cyanide-resistant respiration rate and the activity of an alternative oxidase enzyme were significantly higher in transgenic plants than in wild-type plants. In order to determine the resistance to low temperatures, the parent and transgenic tobacco plants were grown in soil until the 2–3 and 6–7 leaf stages, after which they were kept at 3 to -3 °С for a day in the dark and left to regrow at 25 °С for seven days. Although it was previously shown that oxidative stress is reduced in transgenic plants at low temperatures as compared to wild-type plants, it was established that the tolerance of transgenic and nontransformed plants does not differ. Thus, alternative NADH dehydrogenase activity was found to reduce oxidative stress and increase alternative oxidase activity, without enhancing the resistance of Nicotiana tabacum to negative temperatures.
本研究旨在确认拟南芥 At_NDB2 转基因蛋白在烟草细胞中的功能活性和定位,并评估永久表达外部替代 NADH 脱氢酶对喜热植物抗低温能力的影响。研究人员从 25 ℃(昼夜温差)条件下生长的烟草植株 7 叶期的叶片中分离出蛋白质和线粒体。通过电泳和免疫印迹测定了 At_NDB2 蛋白在线粒体中的定位。通过对分离的线粒体和特异性线粒体复合体 I 抑制剂(鱼藤酮)进行极谱分析,证实了 At_NDB2 的功能活性。研究还发现,转基因植株的耐氰呼吸速率和替代氧化酶的活性明显高于野生型植株。为了测定烟草对低温的抗性,亲本和转基因烟草植株在土壤中生长到 2-3 叶期和 6-7 叶期,然后在 3 至 -3 ℃ 的黑暗环境中保持一天,并在 25 ℃ 的环境中再生长七天。尽管之前有研究表明,与野生型植物相比,转基因植物在低温下的氧化应激会减少,但已证实转基因植物和非转基因植物的耐受性并无不同。因此,研究发现替代 NADH 脱氢酶活性可降低氧化应激,提高替代氧化酶活性,但不会增强烟草对负温的抗性。
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引用次数: 0
Effect of growth medium composition on the viability and genetic stability of Chrysanthemum × morifolium Ramat. explants under in vitro cold storage conditions 生长培养基成分对离体冷藏条件下菊花 × morifolium Ramat.外植体活力和遗传稳定性的影响
Pub Date : 2023-12-26 DOI: 10.21285/2227-2925-2023-13-4-483-493
N. N. Ivanova, V. A. Tsiupka, N. V. Korzina
Cultivation of plants at low positive temperatures is promising for the in vitro preservation of valuable Chrysanthemum × morifolium Ramat. varieties. The study aims to examine the effect of chlorocholine chloride concentrations in a specialized medium on the viability and the physiological and morpho-anatomical parameters of chrysanthemum plants (Excel, Sheer Purple, and William Seward varieties), as well as their genetic stability after 12 months of cold storage under in vitro gene bank conditions. In vitro microshoot tips (without leaves) were placed on a 1/4 Murashige and Skoog medium supplemented with 60.0 g/L sucrose and 0.2–0.8 g/L chlorocholine chloride. The explants were stored in cooling chambers at a temperature of 4 °C. After 12 months of preservation, a reduction was observed in growth kinetics with increasing concentration of chlorocholine chloride in the medium. The viability of explants decreased from 96 to 91% at higher concentrations of chlorocholine chloride. The effective photochemical quantum yield of photosystem II decreased more than twice under cold storage conditions as compared to the control. Several histological features were identified in the leaves of in vitro explants cold-stored in the gene bank: decreased in leaf thickness and mesophyll density in leaf blades, degradation of palisade tissue, and a high degree of transpiration regulation. The SSR and ISSR analyses were used to show that the microshoots of the examined chrysanthemum varieties were identical to the original plants following in vitro cold storage. The conducted studies revealed that the presence of 0.6–0.8 g/L chlorocholine chloride in the medium reduces the growth kinetics of explants while maintaining their high viability and genetic stability.
在低温条件下栽培植物对于离体保存珍贵的菊花品种很有前景。本研究的目的是考察在离体基因库条件下,氯化氯胆碱浓度对菊花植株(Excel、Sheer Purple 和 William Seward 品种)的存活率、生理和形态解剖参数的影响,以及低温贮藏 12 个月后的遗传稳定性。将离体显微芽尖(不含叶片)置于添加了 60.0 克/升蔗糖和 0.2-0.8 克/升氯化胆碱的 1/4 Murashige 和 Skoog 培养基上。外植体储存在温度为 4 °C 的冷却室中。保存 12 个月后,观察到随着培养基中氯代胆碱浓度的增加,生长动力学有所下降。氯化胆碱浓度越高,外植体的存活率从 96% 降至 91%。在冷藏条件下,光系统 II 的有效光化学量子产率比对照组下降了两倍多。在基因库中冷藏的离体外植体的叶片上发现了几个组织学特征:叶片厚度和叶片中叶密度下降、栅栏组织退化和高度蒸腾调节。SSR 和 ISSR 分析表明,受检菊花品种的小枝在离体冷藏后与原植株完全相同。研究表明,在培养基中加入 0.6-0.8 克/升氯化胆碱可降低外植体的生长动力学,同时保持其较高的存活率和遗传稳定性。
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引用次数: 0
New smart thermosensitive biomaterials on the basis of collagen modified by ethylene glycol vinyl glycidyl ether for 4D bioprinting 基于乙二醇乙烯基缩水甘油醚修饰的胶原蛋白的新型智能热敏生物材料,可用于 4D 生物打印
Pub Date : 2023-12-26 DOI: 10.21285/2227-2925-2023-13-4-469-475
I. A. Farion, A. S. Buinov, A. N. Nikishina, V. F. Burdukovskii
In this study, collagen reactive under UV radiation was obtained for the first time via the functionalization of the side amino groups of lysine moieties with unsaturated ethylene glycol vinyl glycidyl ether (Vinylox) containing, similarly to glycidyl methacrylate, an epoxy group. The presence of this group having a moderate reactivity provides selective grafting of the side amino groups of collagen in a neutral or weakly alkaline medium and excludes the formation of by-products in the Michael reaction and hydrolysis characteristic of the analog—glycidyl methacrylate. Photometry data indicate that with modification in a weakly basic aqueous solution at room temperature and a significant excess of Vinylox, a grafting degree of 23.4% can be achieved. For the first time, film-forming composites capable of photocuring were obtained by adding polyethylene glycol diacrylate and poly(N-isopropylacrylamide) to a weakly acidic aqueous solution of collagen. The presence of vinyl oxide groups provided satisfactory mechanical characteristics of films as the result of UV-initiated collagen cross-linking, while the presence of poly(N-isopropylacrylamide) ensured hydrophilic-hydrophobic smart sensitivity. The films have a well-developed fibrillar structure, and the size of voids enables the free movement of nutrients and other compounds. According to MTT assay data, the films release no cytotoxic components and maintain the metabolic activity of stem cells, providing sufficient density of stem cells on their surface. All of the factors mentioned above determine the prospects of using the films both as an artificial extracellular matrix scaffold and as smart thermosensitive scaffolds used to grow stem cells on their surfaces for subsequent laser transfer bioprinting.
在这项研究中,通过将赖氨酸分子的侧氨基与不饱和乙二醇乙烯基缩水甘油醚(Vinylox)进行官能化,首次获得了在紫外线辐射下具有活性的胶原蛋白,Vinylox 与甲基丙烯酸缩水甘油酯类似,含有一个环氧基团。该基团具有中等反应活性,可在中性或弱碱性介质中选择性地接枝胶原蛋白的侧氨基,并避免在迈克尔反应中形成副产物以及类似的甲基丙烯酸缩水甘油酯所特有的水解作用。光度测量数据表明,在室温下的弱碱性水溶液中进行改性,并加入大量过量的 Vinylox,接枝度可达 23.4%。在胶原蛋白的弱酸性水溶液中加入聚乙二醇二丙烯酸酯和聚(N-异丙基丙烯酰胺),首次获得了能够光固化的成膜复合材料。氧化乙烯基的存在使薄膜具有令人满意的机械特性,这是紫外线引发胶原交联的结果,而聚(N-异丙基丙烯酰胺)的存在则确保了亲水-疏水智能敏感性。薄膜具有发达的纤维状结构,空隙的大小使营养物质和其他化合物能够自由流动。根据 MTT 检测数据,薄膜不会释放细胞毒性成分,并能保持干细胞的新陈代谢活性,在其表面提供足够密度的干细胞。上述所有因素决定了薄膜既可用作人工细胞外基质支架,也可用作智能热敏支架,用于在其表面培育干细胞,然后进行激光转移生物打印。
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引用次数: 0
Effect of protatranes on the biosynthesis of extracellular enzymes of Candida ethanolica BKM Y-2300 T 原烷对乙醇念珠菌 BKM Y-2300 T 细胞外酶生物合成的影响
Pub Date : 2023-12-26 DOI: 10.21285/2227-2925-2023-13-4-506-515
A. S. Kiryukhina, T. S. Lozovaya, S. N. Adamovich
The study aims to explore the possibility of using synthetic biologically active compounds (protatranes) to increase the lipolytic and proteolytic activity of Candida ethanolica. Protatrane 1 (tris(2-hydroxyethyl) ammonium-4chlorophenyl-sulfanyl acetate) and protatrane 2 (tris(2-hydroxyethyl) ammonium-4-chlorophenyl-sulfonyl acetate) were added to the growth medium at trace concentrations of 1×10−6–1×10−8wt% separately and together. It was established that with the introduction to the growth medium, protatranes had a multidirectional effect on the biosynthesis of enzymes by Candida ethanolica yeast. This effect was dependent on the concentrations of these compounds, as well as on their separate or combined introduction. All the studied concentrations were found to be effective for lipase synthesis, as they improved enzyme synthesis by 1.7–8.6 times. The combined use of protatranes increased enzyme synthesis by 3.4–11.7 times. For protease formation, the combined introduction of the studied protatranes at a concentration of 10−6wt% was found to be the most effective, with enzyme synthesis reaching 184.8±7.02 U/mL in the culture broth. When co-introduced, the studied protatranes tris(2-hydroxyethyl) ammonium-4-chlorophenylsulfanyl acetate and tris(2-hydroxyethyl) ammonium-4-chlorophenyl-sulfonyl acetate can be used to increase the production efficiency of extracellular lipases and proteases by Candida ethanolica. The specific effect of protatranes was revealed to be the uneven dynamics of extracellular enzyme accumulation, manifested in the presence of sharp peaks during yeast cultivation. The reasons for this unevenness require further research.
本研究旨在探索利用合成生物活性化合物(原鹤草)提高乙醇念珠菌脂肪分解和蛋白分解活性的可能性。在生长培养基中分别和同时加入微量浓度为 1×10-6-1×10-8wt% 的原烷 1(三(2-羟乙基)铵-4-氯苯基硫酰乙酸酯)和原烷 2(三(2-羟乙基)铵-4-氯苯基硫酰乙酸酯)。结果表明,在生长培养基中加入原烷后,原烷对乙醇念珠菌酶的生物合成具有多向影响。这种影响取决于这些化合物的浓度,以及它们的单独或联合引入。研究发现,所有浓度的化合物都能有效促进脂肪酶的合成,因为它们能将酶的合成提高 1.7-8.6 倍。联合使用原烷可使酶的合成增加 3.4-11.7 倍。在蛋白酶的形成方面,以 10-6wt% 的浓度联合引入所研究的原膜是最有效的,在培养液中酶的合成量达到 184.8±7.02 U/mL。所研究的三(2-羟乙基)铵-4-氯苯基磺酰乙酸酯和三(2-羟乙基)铵-4-氯苯基磺酰乙酸酯原药共同加入后,可用于提高乙醇念珠菌胞外脂肪酶和蛋白酶的生产效率。研究发现,protatranes 的特殊效果是细胞外酶积累的动态不平衡,表现为酵母培养过程中出现尖峰。造成这种不均衡的原因需要进一步研究。
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引用次数: 0
Synthesis and antinociceptive activity of 2,2’-(1,4-phenylene)bis[3-aryl-2-azaspiro[3.5]nonan-1-ones] 2,2'-(1,4-亚苯基)双[3-芳基-2-氮杂螺[3.5]壬-1-酮]的合成和抗镇痛活性
Pub Date : 2023-12-26 DOI: 10.21285/2227-2925-2023-13-4-476-482
R. R. Makhmudov, E. A. Nikiforova, D. P. Zverev, L. A. Balyukina, A. Р. Skachkov
Over the years, azetidin-2-ones, or β-lactams, have received a lot of attention from scientists as potential drug candidates due to their diverse biological activity. Spiro-β-lactams also exhibit biological activity; therefore, it is of interest to synthesize and study the properties of new compounds belonging to this class. The study aims to examine the antinociceptive activity of several synthesized bis(spirolactams), specifically 2,2’-(1,4-phenylene)bis[3-aryl-2azaspiro[3.5]nonan-1-ones]. These bis(spiroazetidine-2-ones) were obtained in the interaction of a twofold excess of the Reformatsky reagent, derived from methyl 1-bromocyclohexane carboxylate and zinc, with N,N-(1,4-phenyle- ne)bis(1-arylmethanimines) by means of boiling them in a 10:1 mixture of toluene and hexamethylphosphorictriamide for four hours. Bis(spiro-β-lactams) on the basis of diimines derived from p-phenylenediamine, 2-methoxybenzaldehyde, p-tolualdehyde, and 3-bromobenzoic aldehyde were synthesized for the first time. The composition and structure of the previously undescribed products were established using IR, [1]H, and 13C NMR spectroscopy and elemental analysis. The antinociceptive activity of the obtained compounds was studied on outbred white mice of both sexes via the hot plate test with an intraperitoneal injection. The effect was estimated two hours after administration. Several synthesized compounds were found to exhibit antinociceptive activity at or above the level of the comparator product – metamizole sodium. Nerve endings can be considered the target of the antinociceptive activity of examined substances since under the effect of these substances, no signs of central action are observed in the behavior of animals. Thus, the conducted studies showed the promise of further search for biologically active substances among the compounds of this series.
多年来,氮杂环丁烷-2-酮或β-内酰胺类化合物因其多种多样的生物活性,作为潜在的候选药物受到了科学家们的广泛关注。螺-β-内酰胺类化合物也具有生物活性,因此合成和研究这一类新化合物的特性是非常有意义的。本研究旨在考察几种合成的双(螺内酰胺),特别是 2,2'-(1,4-亚苯基)双[3-芳基-2-氮杂螺[3.5]壬-1-酮]的抗痛觉活性。这些双(螺氮杂环丁烷-2-酮)是在 10:1 的甲苯和六甲基磷酰三酰胺混合物中沸腾四小时后,由 1-溴环己烷羧酸甲酯和锌衍生的两倍过量的 Reformatsky 试剂与 N,N-(1,4-苯基-氖)双(1-芳基甲亚胺)作用而得到的。以对苯二胺、2-甲氧基苯甲醛、对甲苯甲醛和 3-溴苯甲酸醛衍生的二亚胺为基础,首次合成了双(螺-β-内酰胺)。利用红外光谱、[1]H 光谱、13C NMR 光谱和元素分析确定了这些之前未曾描述过的产物的组成和结构。通过腹腔注射热板试验,研究了所获化合物对近亲繁殖的雌雄白鼠的抗痛觉活性。药效在给药两小时后进行评估。研究发现,几种合成化合物的抗痛觉活性达到或超过了对比产品--美他咪唑钠的水平。由于在这些物质的作用下,动物的行为没有出现中枢作用的迹象,因此可以认为神经末梢是所研究物质抗痛觉活性的靶点。因此,所进行的研究表明,有望在该系列化合物中进一步寻找生物活性物质。
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引用次数: 0
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Proceedings of Universities. Applied Chemistry and Biotechnology
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