Abstract
Background. Legumes establish symbioses with nitrogen-fixing bacteria from the Rhizobium group. In exchange for nutrients, bacteria provide fixed nitrogen needed to support plant growth. At the moment, information about the involvement of alternative splicing (AS) in the establishment and maintenance this symbiotic relationships is almost absent, but, as it is a powerful mechanism for the regulation of proteome diversity of the cell, it therefore may participate in cellular response to microsymbionts.
 Materials and methods. Alternative splicing was analyzed using the assembly of supertranscripts and alignment of the reads from nodules and root tips to this reference. Target genes expression levels was estimated in tips of non-inoculated roots, and in nodules (2, 4, and 6 weeks post inoculation) with use of RT-qPCR.
 Results.In this study, the analysis of AS events in the nodules and root tips of the pea was carried out. The presence of isoforms of four pea genes (PsSIP1, PsIGN, PsWRKY40, PsPR-10) was confirmed and their expression level was estimated.
 Conclusion. Pea nodules were shown to be more enriched with AS events compared to root tips. Among the functional groups of genes that demonstrate AS events, one of the most enriched functional groups is the pathogens stress response. Intron retention probably leads to degradation of the transcript via NMD-system or to change of the protein function, that modulates the activity of genes in nodules.
Highlights
CC Горох посевной (Pisum sativum L.) способен вступать в симбиотические отношения с азотфиксирующими бактериями из группы ризобий (Rhizobia)
Information about the involvement of alternative splicing (AS) in the establishment and maintenance this symbiotic relationships is almost absent, but, as it is a powerful mechanism for the regulation of proteome diversity of the cell, it may participate in cellular response to microsymbionts
Target genes expression levels was estimated in tips of non-inoculated roots, and in nodules (2, 4, and 6 weeks post inoculation) with use of RT-qPCR
Summary
В рамках работы был проведен анализ событий АС в клубеньках и кончиках корней гороха посевного. ANALYSIS OF ALTERNATIVE SPLICING EVENTS IN THE ROOT TIPS AND NODULES OF PISUM SATIVUM L. For citation: Zorin EA, Kulaeva OA, Afonin AM, et al Analysis of alternative splicing events in the root tips and nodules of Pisum sativum L. В данной работе описаны события АС в клубеньках и кончиках корней гороха посевного с применением метода сборки «супертранскриптов» из имеющихся транскриптомных данных клубеньков и кончиков корней с последующим анализом паттернов картирования прочтений на собранные «супертранскрипты». С помощью методов ПЦР с обратной транскрипцией и количественной ПЦР в реальном времени подтверждено наличие АС и количественно оценена представленность мРНК различных изоформ в кончиках корней и клубеньках гороха на трех временных точках после инокуляции для четырех генов: PsSIP1, PsIGN1, PsWRKY40, PsPR‐10.
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