vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJ18.347vavilov-1441Research ArticleКЛЕТОЧНАЯ И МОЛЕКУЛЯРНАЯ БИОЛОГИЯCELL AND MOLECULAR BIOLOGYДифференциальный протеомный анализ корней гороха на ранних этапах развития симбиоза с клубеньковыми бактериямиDifferential proteome analysis of pea roots at the early stages of symbiosis with nodule bacteriaЛеппяненИ. В.LeppyanenI. V.

Пушкин, Санкт-Петербург

Pushkin, St. Petersburg

КириенкоА. Н.KirienkoA. N.

Пушкин, Санкт-Петербург

Pushkin, St. Petersburg

ЛобовА. А.LobovA. A.

Санкт-Петербург

St. Petersburg

ДолгихЕ. А.DolgikhE. A.

Пушкин, Санкт-Петербург

Pushkin, St. Petersburg

dol2helen@yahoo.comВсероссийский научно-исследовательский институт сельскохозяйственной микробиологииРоссияAll-Russian Scientific Research Institute of Agricultural MicrobiologyRussian FederationРесурсный центр «Развитие молекулярных и клеточных технологий», научный парк, Санкт-Петербургский государственный университетРоссияResource Center “Development of Molecular and Cellular Technologies”, Science Park, St. Petersburg State UniversityRussian Federation201806042018222196204Copyright © Леппянен И.В., Кириенко А.Н., Лобов А.А., Долгих Е.А., 20182018Леппянен И.В., Кириенко А.Н., Лобов А.А., Долгих Е.А.Leppyanen I.V., Kirienko A.N., Lobov A.A., Dolgikh E.A.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/1441

В настоящей работе проведен анализ изменений в протеомном спектре корней гороха Pisum sativum L. при инокуляции клубеньковыми бактериями, направленный на выявление новых регуляторов развития симбиоза. Для изучения изменений в протеомном спектре корней гороха использован дифференциальный двумерный (2-D) электрофорез с флуоресцентными метками Cy2 и Cy5. Полученные изображения позволили выявить различия между контрольным вариантом (неинокулированные корни) и корнями, инокулированными ризобиями Rhizobium leguminosarum bv. viciae RCAM 1026 (24 ч после инокуляции). Выявлено и идентифицировано 20 белков, синтез которых усиливался в процессе инокуляции корней гороха клубеньковыми бактериями. Для идентификации белков использовали масс-спектрометрический анализ триптических пептидов на квадруполь-времяпролетном масс-спектрометре, совмещенном с высокоэффективным жидкостным хроматографом. Среди таких белков впервые найдены бета-субъединица G-белка и дисульфид изомераза/фосфолипаза С, функция которых может быть связана с сигнальной регуляцией симбиоза. Это показывает, что G-белки и фосфолипазы могут играть ключевую роль в развитии ранних стадий симбиоза у гороха. Дальнейшие эксперименты должны выявить, происходит ли взаимодействие бета-субъединицы G-белка с рецепторами к Nod-факторам и как это влияет на дальнейшую передачу сигнала. Среди других белков, изучение которых представляет интерес, обнаружены аннексин D8 и D1, протеинкиназа, взаимодействующая с кальцинерином B, актин-связывающий белок профилин, ГТФ-связывающий белок Ran1. Они могут быть вовлечены в регуляцию реакций с участием кальция, реорганизацию актинового цитоскелета и другие важные процессы у растений. Изучение роли таких белков-регуляторов в дальнейшем станет основой для понимания сложной системы сигнальной регуляции, которая активируется у растений гороха при взаимодействии с клубеньковыми бактериями.

In this paper, we have analyzed changes in the proteomic spectrum of pea Pisum sativum L. roots during inoculation with rhizobial bacteria with the aim of revealing new regulators of symbiosis development. To study the changes in the proteome spectrum of pea roots, a differential twodimensional (2-D) electrophoresis was performed using fluorescent labels Cy2 and Cy5. The images obtained made it possible to identify differences between the control variant (uninoculated roots) and the root variant after inoculation with Rhizobium leguminosarum bv. viciae RCAM 1026 (24 hours after treatment). 20 proteins were revealed and identified, the synthesis of which was enhanced during the inoculation of pea roots by nodule bacteria. To identify the proteins, a mass spectrometric analysis of tryptic peptides was performed on a quadrupole-time-of-flight mass spectrometer combined with a high-performance liquid chromatograph. Among such proteins, the beta-subunit of the G protein and the disulfide isomerase/phospholipase C were first found, whose function can be related to the signal regulation of symbiosis. This indicates that G-proteins and phospholipases can play a key role in the development of early stages of symbiosis in peas. Further experiments are expected to show whether the beta-subunit of the G protein interacts with the receptors to Nod factors, and how this affects the further signaling. Other proteins that might be interesting were annexin D8 and D1, protein kinase interacting with calcinerin B, actin-binding protein profilin, GTP-binding protein Ran1. They may be involved in the regulation of reactions with calcium, the reorganization of the actin cytoskeleton and other important processes in plants. The study of the role of such regulatory proteins will later become the basis for understanding the complex system of signal regulation, which is activated in pea plants by interaction with nodule bacteria.

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