Transcription factors from the WOX family are well-known regulators of cell proliferation and differentiation in plants. Herein, we focused on several WOX genes from the intermediate clade and checked their impact on somatic embryogenesis using the model legume object Medicago truncatula. As a result, we show that MtWOX9-1 overexpression not only stimulates somatic embryogenesis in the embryogenic M. truncatula line, as it was shown previously, but can also induce somatic embryogenesis in the non-embryogenic line. Other intermediate clade WOX, including the close paralog of MtWOX9-1, as well as WOX11 homologs, did not have any significant impact on somatic embryogenesis in our in vitro cultivation system. Together, our results give new information about the diversity of the WOX family proteins and their specific functions. These data can be used for the search of new regeneration stimulators.

WOX family transcription factors are well-known regulators of plant development, controlling cell proliferation and differentiation in diverse organs and tissues. Several WOX genes have been shown to participate in regeneration processes which take place in plant cell cultures in vitro, but the effects of most of them on tissue culture development have not been discovered yet. In this study, we evaluated the effects of MtWOX2 gene overexpression on the embryogenic callus development and transcriptomic state in Medicago truncatula. According to our results, overexpression of MtWOX2 leads to an increase in callus weight. Furthermore, transcriptomic changes in MtWOX2 overexpressing calli are, to a large extent, opposite to the changes caused by overexpression of MtWOX9-1, a somatic embryogenesis stimulator. These results add new information about the mechanisms of interaction between different WOX genes and can be useful for the search of new regeneration regulators.

Genome modification of legumes, peas in particular, is accompanied by significant challenges. Establishing a reliable reporter system to identify tissue that expresses foreign DNA may help to optimize and develop transformation protocols for these species. The RUBY system, based on the synthesis of red betalain from tyrosine, offers a convenient solution for monitoring the efficiency of transgene introduction. To evaluate the effectiveness of RUBY application in pea tissue culture, we combined agrobacterial transformation with an in vitro cultivation system, inducing callus development. Transformed explants demonstrated RUBY pigmentation, but it disappeared during cultivation. We hypothesized that this issue is caused by tyrosine depletion. To check this suggestion, we tested whether tyrosine supplementation could maintain RUBY coloring. In the later stages, pigmentation still could not be preserved. However, our modified conditions increased the percent of colored shoot apex explants during the early cultivation stages. Thus, it is likely that some explants transformed with the RUBY cassette do not synthesize a sufficient amount of betalain due to the deficit of endogenous tyrosine. In this case, adding exogenous tyrosine would enhance betalain production and improve the detectability of tissues containing the RUBY cassette. These data can be used for the optimization of RUBY application conditions for peas and other species.

Мембранные процессы всегда являлись неотъемлемой частью биотехнологических процессов. Сегодня без них не обходится ни одно биотехнологическое производство: обратный осмос, ультрафильтрация, микрофильтрация, разделение газов, хроматография, первапорация и электродиализ – все эти процессы играют важную роль в тех или иных технологических решениях. Несмотря на то, что первые мембранные системы были заимствованы из технологий, которые разрабатывались для других промышленных применений, в течение последних 45 лет новые материалы и элементы создаются специально для нужд биотехнологической промышленности. Мембраны хорошо подходят для работы с биологическими молекулами: относительно низкие температуры и давления, отсутствие необходимости фазовых переходов и введения химических добавок минимизируют степень денатурации, дезактивации, и/или деградации биологических продуктов. В настоящее время мембранные технологии занимают лидирующие позиции в процессах стерилизующей фильтрации, осветления, культивирования клеточных культур, удаления вирусов, концентрирования и очистки белков. Все большее распространение получают процессы с применением мембранных биореакторов и мембранной хроматографии. В обзоре показаны возможности существующих разработок в мембранных технологиях, акцентируется внимание на используемых мембранных материалах и характеристиках мембранных систем, применяемых для коммерческого производства. Процессы мембранного разделения требуют более дорогостоящего оборудования по сравнению с традиционными процессами разделения, однако являются более энергоэффективными. По своей конструкции мембранные системы, как правило, компактны и имеют модульную архитектуру, благодаря которой появляется возможность использования одного и того же оборудования для решения различных задач. Имеются предпосылки того, что новые разработки в мембранных технологиях смогут удовлетворить растущие нужды биотехнологической отрасли в большей производительности и снижении стоимости производства. Для цитирования: Каталевский А.Д., Смирнов К.В., Смирнова Н.Н. Мембраны в биотехнологии: современное состояние и перспективы. Изв. вузов. Химия и хим. технология. 2025. Т. 68. Вып. 1. С. 6-22. DOI: 10.6060/ivkkt.20256801.7075.

BACKGROUND: Genetic modification of pea remains challenging, which can likely be attributed to its low regeneration efficiency. To obtain transgenic pea plants, shoot regeneration followed by rooting is usually applied. Another regeneration pathway, somatic embryogenesis, is not used for pea genome modification due to very low frequency of this process. If a gene stimulating somatic embryogenesis was identified, it could be used as a morphogenic regulator to enable regeneration of pea plants from genetically modified callus cells. The search for such genes relies on the development of a cultivation system, allowing production of a significant amount of callus tissue in which a potential morphogenic regulator is ectopically expressed. AIM: The aim of study was to evaluate the possibility of obtaining pea callus tissue expressing foreign DNA with or without usage of selective agents, specifically, kanamycin and hygromycin B. METHODS: In this study, we combined agrobacterial transformation protocol with a method of callus induction from pea shoot apex explants. To evaluate the effectiveness of this transformation system with different selective agents, we used two different reporters: RUBY and DsRed. RESULTS: Our results demonstrate that transformation of pea shoot apices using the developed system yields a significant percentage of calli containing tissue regions which express foreign DNA. Addition of the antibiotics as selective agents doesn’t increase frequency of calli expressing introduced DNA. CONCLUSION: Results obtained in this study suggest that searching for regeneration stimulators is feasible in Pisum sativum without usage of selective agents.

Currently, several species of the genus Gossypium are cultivated in agriculture to produce fiber. Cotton has been cultivated for a long time, however, many aspects of its cultivation and processing are still researched. Writing about the cultivation of cotton, it is worth mentioning the fundamental problems of its processing. For example, the amounts of pesticides used in the cultivation of cotton are greater than for any other crop. Chemicals sprayed on cotton fields are washed away from the fields and, reaching the fresh water sources, pollute them, causing significant damage to the environment. Fortunately, such challenges can be solved by switching to the cultivation of transgenic cotton. Transgenic cotton has already brought many important environmental, social and economic benefits, including reduce of the used pesticides, indirectly increasing of yield, minimizing environmental pollution, reducing the labor force involved and economic costs.Today, the main methods of obtaining transgenic cotton lines are still agrobacterial transformation and biolistics. In recent years, however, innovative methods of transformation have also been developed. For example, the introduction of genetic material mediated by a pollen tube for the cultivation of commercial transgenic cotton is actively used in China. Although in recent decades transgenic lines resistant to diseases, abiotic stresses and with improved fiber quality have been obtained, the dominant position in the market of transgenic cotton is still occupied by lines of plants resistant to insects and herbicides. All the above indicates an insufficient degree of integration between institutes that introduce new advanced developments and agricultural industry.In this review the results of research involving the cultivation and genetic modification of cotton were collected and summarized. The main methods of genetic transformation of cultivated representatives of the genus Gossypium, both actively used at present and still under development, were considered. The most remarkable transgenic lines were also described, among which both those that have already entered agricultural industry and those that have only recently been obtained. Thus, the reader will be able to get a general idea of the current achievements in the field of cotton genetic modification.