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УЛЬТРАСТРУКТУРНА ХАРАКТЕРИСТИКА ПРОВІДНИХ ПУЧКІВ ЛИСТКІВ ТРАНСГЕННИХ РОСЛИН ЦИКОРІЮ З ГЕНОМ ІНТЕРФЕРОНУ АЛЬФА 2b ЛЮДИНИ, ІНФІКОВАНИХ ВІРУСОМ ТЮТЮНОВОЇ МОЗАЇКИ

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А. А. Потрохов, Д. А. Климчук, С. Н. Щербаков, Е. П. Трохименко

Институт клеточной биологии и генетической инженерии Национальной академии наук Украины, Институт ботаники им. М.Г. Холодного Национальной академии наук Украины, Национальна медицинская академия последипломного образования им. П. Л. Шупика

УЛЬТРАСТРУКТУРНАЯ ХАРАКТЕРИСТИКА ПРОВОДЯЩИХ ПУЧКОВ ЛИСТЬЕВ ТРАНСГЕННЫХ РАСТЕНИЙ ЦИКОРИЯ С ГЕНОМ ИНТЕРФЕРОНА АЛЬФА 2B ЧЕЛОВЕКА, ИНФИЦИРОВАНЫХ ВИРУСОМ ТАБАЧНОЙ МОЗАЙКИ

Осуществлена оценка активности экстрактов из трансгенных растений цикория с геном интерферона α-2b человека. При тестировании экстрактов, полученных из трансгенных растений, в культуре клеток перевивных текстикул поросят (ПТП), которая была инфицированные вирусом везикулярного стоматита (ВВС), обнаружили интерфероноподобную активность от 942 до 1884 МЕ / г массы. Вместе с тем, растения не приобрели устойчивость к фитовирусной инфекций ВТМ. Инфицирования трансгенных растений фитовирусом приводило к развитию заболевания. Электронно-микроскопическими методами была исследована ультраструктура клеток трансгенных растений. Не было обнаружило существенных ультраструктурых различий между клетками трансформированных растений и дикого типа как до так и после инфицирования.

Ключевые слова: интерферон, трансгенные растения, фитовирус

 

A. A. Potrokhov, D. A. Klimchuk, S. N. Sherbakov, E. P. Trohimenko

Institute of Cell Biology and Genetic Engineering of the National Academy of Sciences of Ukraine, Institute of Botany of the National Academy of Sciences of Ukraine, National medical academy of post-graduate education P.L.Shupik Heals ministry of Ukraine

ULTRASTRUCTURAL CHARACTERISTIC OF CONDUCTING BEAMS IN TRANSGENE CHICORY PLANTS WITH THE HUMAN INTERFERON ALPHA 2B GENE, INFECTED BY TOBACCO MOSIC VIRUS

Viral diseases lead to a significant decline in crop yields in the world. Using genetic engineering methods it is possible to create resistant plants to viral infections. But, despite success of these methods, influence of viral infection on the cells ultrastructure in transformed plants remains unclear. For example, in normal, viruses are localized in mesophilic cells in the cytoplasm and in vacuole. To improve viral RNA transport into the plant organism, TMV change plasmodes, expanding them. However, the biotechnological plants undergo a double stress action, caused by contact with vectors in the process of transformation and with the virus infection. Possible negative effects of genetic transformation on plants and, in particular, on the cells ultrastructures are still unknown. Also the question of the cells ultrastructure in transgenic plants after viral infected has not yet been studied. The aim of this work was to investigate the cells ultrastructure of intact chicory plants with the human ifn-α2b gene, infected by TMV. The transformed plants of chicory were obtained by agrobacterial transformation. The PCR method confirmed the presence of the ifn-α2b and nptII genes. Interferon antiviral activities were obtained from the extracts from transgenic plants. Activity analysis was performed using a substrate-dependent culture of PTP cells infected by VSV. It was found that the extracts actually had interferon-like activity in the range from 942 to 1884 IU / g. Despite this activity transgenic plants were susceptible to tobacco mosaic virus. The conducted electron microscopic analysis of transgenic and control chicory plants did not show any significant differences in cells ultrastructure. Genetic transformation did not lead to changes in the ultrastructure in plant cells. The cells ultrastructure in transgenic plants, infected by TMV, did not significantly differ from the cells ultrastructure in control infected plants. In infected cells, the accumulation of virions, their localization and morphological changes in plants were the same. As a result, it has been shown that transgenic plants in their morphology and cell structure were similar with wild-type plants. Viral infection in such plants proceeds in the same way as in wild-type plants and does not lead to a significant difference. We ascertain the fact that cell ultrastructure between transgenic type and wild type plants was the same. This fact gives us reason to argue that genetic engineering does not lead to negative consequences on the cell ultrastructure. Also that the development of pathological reactions caused by virus in transgenic plants and wild-type plants was similar. Consequently, based on these parameters transgenic plants are not altered from wild-type plants and can be used in agriculture.

Keywords: interferon, transgenic plants, phytovirus

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