Ecological geneticsEcological geneticsЭкологическая генетика1811-09322411-9202Eco-Vector69579110.17816/ecogen695791QNPESWGenetically modified organism. The history, achievements, social and environmental risksГМО: История, достижения, социальные и экологические рискиResearch ArticleEvaluation of selective agent requirements for pea callus culture expressing foreign DNAОценка необходимости использования селективных агентов для каллусной культуры гороха, экспрессирующей чужеродную ДНКhttps://orcid.org/0000-0002-9037-46849475-2980SimonovaVeronika Yu.СимоноваВероника Юрьевна
Russian Federation
nikasimonova14@gmail.comhttps://orcid.org/0000-0002-5045-26414484-7595PotsenkovskaiaElina A.ПоценковскаяЭлина Андреевна
Russian Federation
potsenkovskaya.ea@talantiuspeh.ruhttps://orcid.org/0009-0006-9753-20044763-4324VaninaAlexandra A.ВанинаАлександра Алексеевна
Russian Federation
alexandraspb15@gmail.comhttps://orcid.org/0000-0002-1425-70131478-4870KiselevaAnna S.КиселеваАнна Сергеевна
Russian Federation
anykisely@gmail.comhttps://orcid.org/0000-0002-9458-01949877-5352MatveenkoAndrew G.МатвеенкоАндрей Георгиевич
Russian Federation

Cand. Sci. (Biology)

канд. биол. наук

a.matveenko@spbu.ruhttps://orcid.org/0009-0006-7828-41058002-0762PavlovaDaria B.ПавловаДарья Борисовна
Russian Federation
db_pavlova@mail.ruhttps://orcid.org/0000-0002-2875-37982440-2405SmirnovKirill V.СмирновКирилл Вадимович
Russian Federation
kirill.vad.smirnov@gmail.comhttps://orcid.org/0000-0002-2565-11554033-2739EfremovaElena P.ЕфремоваЕлена Павловна
Russian Federation
elena.efremova@spbu.ruhttps://orcid.org/0009-0002-8358-81398700-4413BrynchikovaAnna V.БрынчиковаАнна Викторовна
Russian Federation
annbv19@gmail.comhttps://orcid.org/0000-0002-0549-14576489-5206TvorogovaVarvara E.ТвороговаВарвара Евгеньевна
Russian Federation

Cand. Sci. (Biology)

канд. биол. наук

krubaza@mail.ruSirius University of Science and TechnologyНаучно-технологический университет «Сириус»Saint Petersburg State UniversityСанкт-Петербургский государственный университетAll-Russia Research Institute for Agricultural MicrobiologyВсероссийский научно-исследовательский институт сельскохозяйственной микробиологии031220250305202624129420411202529112025Copyright ©; 2026, Eco-VectorCopyright ©; 2026, Эко-Вектор2026Eco-VectorЭко-Векторhttps://eco-vector.com/for_authors.php#07https://journals.eco-vector.com/ecolgenet/article/view/695791

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.

Обоснование. Генетическая модификация гороха остаётся сложной задачей, что, вероятно, связано с низкой эффективностью регенерации. Для получения трансгенных растений гороха обычно применяют регенерацию побегов с последующим укоренением. Другой путь регенерации, соматический эмбриогенез, не используется для трансформации гороха из-за крайне низкой частоты этого процесса. Идентификация генов, стимулирующих соматический эмбриогенез, позволила бы использовать их в качестве морфогенетических регуляторов, обеспечивающих регенерацию растений гороха из генетически модифицированных клеток каллуса. Для поиска таких генов необходима разработка системы культивирования, позволяющей получать значительное количество каллусной ткани, в которой эктопически экспрессируется потенциальный морфогенетический регулятор.

Цель исследования. Оценить возможность получения каллусной ткани гороха, экспрессирующей чужеродную ДНК, с использованием или без использования селективных агентов, включая канамицин и гигромицин.

Методы. В данном исследовании объединён протокол агробактериальной трансформации с методом индукции каллусообразования из эксплантатов верхушек побегов гороха. Для оценки эффективности такой системы трансформации с применением разных селективных агентов мы использовали два разных репортера: RUBY и DsRed.

Результаты. Полученные результаты показывают, что трансформация верхушек побегов гороха, согласно разработанной системе, позволяет получать значительный процент каллусов с клетками, экспрессирующими чужеродную ДНК. Добавление селективных агентов не увеличивает частоту каллусов с экспрессией внедрённой ДНК.

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