vestibИзвестия Национальной академии наук Беларуси. Серия биологических наукProceedings of the National Academy of Sciences of Belarus, Biological Series1029-89402524-230XThe Republican Unitary Enterprise Publishing House "Belaruskaya Navuka"vestib-307Research ArticleСтатьиХАРАКТЕРИСТИКА acdS-ГЕНА БАКТЕРИЙ PSEUDOMONAS PUTIDA B-37 И СОЗДАНИЕ ГЕНЕТИЧЕСКОЙ КОНСТРУКЦИИ ДЛЯ ОПРЕДЕЛЕНИЯ ЕГО ТРАНЗИЕНТНОЙ ЭКСПРЕССИИ В РАСТИТЕЛЬНЫХ КЛЕТКАХ NICOTIANA BENTHAMIANACHARACTERISTICS OF BACTERIAL ACDS-GENE FROM THE STRAIN PSEUDOMONAS PUTIDA B-37 AND THE CREATION OF A GENETIC CONSTRUCT FOR DETERMINING ITS TRANSIENT EXPRESSION IN THE PLANT CELLS NICOTIANA BENTHAMIANAМельниковаА. А.MelnikavaA. A.

аспирант, мл. науч. сотрудник

Postgraduate student, Junior researcher

alesiamelnikava@gmail.comВолковаД. С.VolkavaD. S.

студент

Student

95volkovads@gmail.comХрамцоваЕ. А.KhramtsovaE. A.

канд. биол. наук, доцент

Ph. D. (Biol.), Assistant Professor

elenakhramtsova@inbox.ruБелорусский государственный университетBelarusian State University201705082017036168Copyright © Мельникова А.А., Волкова Д.С., Храмцова Е.А., 20172017Мельникова А.А., Волкова Д.С., Храмцова Е.А.Melnikava A.A., Volkava D.S., Khramtsova E.A.Данная работа распространяется под лицензией Creative Commons Attribution 4.0.This work is licensed under a Creative Commons Attribution 4.0 License.https://vestibio.belnauka.by/jour/article/view/307

Продуктивное ведение мирового сельского хозяйства затруднено в связи с влиянием на растения широкого спектра факторов биотического и абиотического происхождения. Данные стрессовые воздействия вызывают в растении продукцию фитогормона этилена, накопление которого приводит к ускорению процессов старения, пожелтению и опаданию листьев и плодов. В настоящее время одним из наиболее перспективных подходов к снижению концентрации стрессового этилена является использование фермента 1-аминоциклопропан-1-карбоксилат дезаминазы (АЦК-дезаминазы), который разлагает предшественник этилена, АЦК, до аммиака и α-кетобутирата. Ген acdS, кодирующий данный фермент, представлен в геномах ризобактерий, в частности видов рода Pseudomonas. В настоящей работе ген acdS выделен из бактерий Pseudomonas putida B-37 и клонирован в составе вектора pBI121. Полученная векторная конструкция использована для проведения агробактериальной трансформации листьев растений Nicotiana benthamiana для установления временной экспрессии. Полученные данные об экспрессии acdS-гена в растительных клетках позволяют сделать вывод о том, что создание трансгенных растений, несущих бактериальный ген АЦК-дезаминазы, возможно и перспективно для народного хозяйства. 

Ethylene is an essential plant hormone also known as a stress hormone because its synthesis is accelerated by induction of a variety of biotic and abiotic stress. The plant growth promoting bacteria containing the enzyme 1-aminocyclopropane-1- carboxylate deaminase (ACC-deaminase) enhances plant growth by decreasing ethylene level under stress conditions. The expression of ACC-deaminase (acdS) gene in transgenic plants is an alternative approach to overcome the ethylene-induced stress. Agrobacterium-mediated DNA transfer is currently the most facile and versatile method to deliver gene constructs into the nucleus for gene function analysis in diverse plant species. Transient gene expression via Agrobacteriummediated DNA transfer in different plant tissues offers a simple and fast method to analyze transgene functions. In present work, the acdS-gene was amplified by PCR and then cloned into pBI121 vector under the control of the cauliflower mosaic virus (CaMV) 35s promoter. Agrobacterium tumefaciens GV3101 strain harboring pBI121-acdS vector jointly with the helper strain 19К were used for Agrobacterium-mediated leaf infiltration in Nicotiana benthamiana to infect 3-weeks-old plants. Monitoring of transient expression efficiency at 3 days post-infection was conducted by plant RNA extraction and RT-PCR. RNA was extracted from Nicotiana’s infiltrated zones and an amount of 1 μg total RNA was used to synthesize first-strand cDNA and then RT-PCR.

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