Searching for regulators that interact with BELL1 transcription factor and control the legume-rhizobial symbiosis development

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The development of nitrogen-fixing nodule, which is formed during legume-rhizobial symbiosis, requires the involvement of cell cycle regulators, phytohormones and homeodomain-containing transcription factors as well as other organogenesis processes. Along with homedomain-containing transcription factors from KNOX family, which participation in the control of nodule development has been studied recently, the role of transcription factors from BELL family in this process remains under-explored. At the same time, transcriptomic data for legumes shows an increase in the expression levels of genes encoding proteins of this family in the plant roots inoculated by Rhizobium. In this work we performed screening of сDNA library, which was generated from inoculated pea roots, using BELL1 protein in yeast two-hybrid system. As a result, we detected two interacting proteins, which are highly interesting for future examination. In the first case we discovered interaction between BELL1 and LysM-containing receptor-like kinase LYK9. Another identified regulator interacting with BELL1 became the glycine-rich protein A3, which may be involved in the regulation of defense reactions in plants and their resistance to phytopathogens. Transcriptomic analysis for pea roots has revealed high expression level of gene, which encodes this protein in the nodules, that may demonstrate its important role in symbiosis regulation.

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About the authors

Alexandra Vyacheslavovna Dolgikh

All-Russia Research Institute for Agricultural Microbiology; Saint Petersburg State University

ORCID iD: 0000-0003-1845-9701
Scopus Author ID: 5719038282
ResearcherId: ABC-2930-2020

Russian Federation, 3 Podbelsky chausse, Pushkin, Saint Petersburg, 196608; Saint Petersburg


Elena A. Dolgikh

All-Russia Research Institute for Agricultural Microbiology

Author for correspondence.
ORCID iD: 0000-0002-5375-0943
SPIN-code: 4453-2060
Scopus Author ID: 6603496335
ResearcherId: G-6363-2017

Russian Federation, 3 Podbelsky chausse, Pushkin, Saint Petersburg, 196608

Dr. Sci. (Biol)


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Supplementary files

Supplementary Files Action
Fig. 1. Analysis of the interaction between the pea transcription factor BELL1 and proteins identified during screening libraries using yeast dihybrid system. The interaction was evaluated on selective SC medium without leucine, tryptophan and uracil (SC-LTU). Yeast growth on a selective medium shows the interaction of the studied proteins. As control, several pairs of vectors were used [pEXP32 / Krev1 and pEXP22 / RalGDS - wild type (wt), pEXP22 / RalGDS-m1 (m1 - mutant 1) and pEXP22 / RalGDS-m2 (m2 - mutant 2)] proposed by the manufacturer (Thermo Fisher Scientific) for study strong, weak interaction and undetectable interaction, respectively

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Fig. 2. Phylogenetic tree illustrating the relationship between the protein identified during the screening of the library Psat5g112080 and LysM-RPK model bean Medicago truncatula

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Fig. 3. Phylogenetic tree illustrating the relationship between the Psat4g107720 (URGI) protein detected during the library screening and glycine-proline-rich proteins of the A3 family of M. truncatula and peas

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Fig. 4. Graphical illustration of data on the level of expression of genes encoding glycine-proline-rich proteins of the A3 family in pea nodules (cv. Cameor) at different stages development based on transcriptome analysis (RNA-seq) using the P. sativum v1 genome as a reference [17]. A - laying of nodules, B - the period of the beginning of flowering, C - 10 days after flowering

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