vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJ16.179vavilov-698Research ArticleАктуальные технологии генетики и селекции. ОБЗОРCurrent technologies in genetics and breeding. REVIEWТрансгенные растения как генетические модели для изучения функций генов растенийTransgenic plants as genetic models for studying functions of plant genesКочетовА. В.KochetovA. V.

Новосибирск

Novosibirsk

ak@bionet.nsc.ruШумныйВ. К.ShumnyV. K.

Новосибирск

Novosibirsk

Федеральное государственное бюджетное научное учреждение «Федеральный исследовательский центр Институт цитологии и генетики Сибирского отделения Российской академии наук»; Федеральное государственное автономное образовательное учреждение высшего образования «Новосибирский национальный исследовательский государственный университет»РоссияInstitute of Cytology and Genetics SB RAS; Novosibirsk State UniversityRussian Federation201625092016204476481Copyright © Кочетов А.В., Шумный В.К., 20162016Кочетов А.В., Шумный В.К.Kochetov A.V., Shumny V.K.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/698

В настоящее время трансгенные растения широко используются как для изучения функций отдельных генов, так и для реконструк­ции сетей взаимодействующих генов, контролирующих формиро­ вание морфологических, биохимических и физиологических признаков в процессе развития и при воздействии внешних факторов различной природы. В статье обсуждается классический инструментарий генной инженерии, позволяющий получать линии растений с измененными параметрами экспрессии гена-мишени. Описана структура генетических конструкций (экспрессия белоккодирующего трансгена, антисмысловых и дцРНК-супрессоров). В качестве примера рассмотрены трансгенные растения, характе­ризующиеся увеличенным или сниженным уровнем экспрессии генов S-подобных рибонуклеаз, а также сниженным уровнем экспрессии гена пролиндегидрогеназы, отвечающего за катабо­лизм пролина. Функции S-подобных РНКаз связывали главным образом с ремобилизацией фосфата из стареющих и отмирающих частей растений, однако паттерн экспрессии некоторых генов из этой группы предполагал возможность их участия в защите от патогенов (индукция при повреждении тканей в районе повреж­ дения (локально) и в удаленных органах (системно)). Кроме того, некоторые белки семейства PR-4 (pathogenesis-related proteins 4) обладают рибонуклеазной и противогрибковой активностью. Исследование трансгенных линий растений табака показало, что повышенная РНКазная активность в апопласте приводит к повышению устойчивости к вирусу табачной мозаики, что говорит о новой функции S-подобных рибонуклеаз, связанной с участием в системе неспецифической защиты от вирусов. Другой набор линий трансгенных растений содержал антисмысловой супрессор гена пролиндегидрогеназы (ПДГ) на основе сегмента гена араби­ допсиса. Использование этой генетической конструкции для полу­ чения трансгенных растений других видов (табака, кукурузы, под­ солнечника) приводило к умеренному снижению активности ПДГ и повышению содержания пролина в норме в 1,5–3 раза. Оказа­ лось, что при этом также повышается устойчивость растений к различным видам абиотических стрессов (засуха, засоление, холод, соли тяжелых металлов), что может быть связано с защит­ным действием пролина на ранних этапах неблагоприятных воз­действий, предотвращающим повреждение белков клеточного аппарата экспрессии свободными радикалами.

Transgenic plants are widely used for the investigation of functions of particular genes as well as for reconstruction of complex gene networks controlling plant morphology, biochemistry, and physiology during different development stages and in response to various external stimuli. Gene engineering instruments for the design of transgenic plants with either elevated or suppressed expression of target genes are discussed. Genetic constructs for protein synthesis or antisense RNA/self-complementary double-stranded RNA transcription are described. Transgenic plants with elevated or decreased levels of expression of S-like ribonucleases and decreased expression of the proline dehydrogenase gene are considered as examples. It was believed that S-like RNase functions concern mainly phosphate remobilization from senescent organs. However, expression patterns of some genes coding for S-like RNases were similar to some pathogen-responsive genes (both local and systemic induction after wounding or pathogen inoculation). In addition, some pathogenesis-related proteins (PR-4 family) possess RNase activity and can inhibit growth of pathogenic fungi. Investigation of transgenic plants revealed that high ribonuclease activity in apoplast correlated with increased resistance against tobacco mosaic virus. Thus, S-like RNases may have a new function as a part of the plant basal antiviral defense mechanism. Another set of transgenic plants bears an antisense suppressor of the proline dehydrogenase gene (PDH) constructed with an Arabidopsis target gene segment. Tobacco, maize and sunflower plants with this heterologous suppressor were characterized with a moderate decrease in PDH activity and a mild (1.5–3-fold) increase in the proline content under normal conditions. It was also found that these plants were more tolerant to various abiotic stresses (drought, NaCl, cold, toxic heavy metals), which may result from the protective proline effect early in exposure to stress, preventing the cellular gene expression machinery from damage by stress-generated free radicals.

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The authors declare that there are no conflicts of interest present.