TY - JOUR

T1 - The Role of Heterotrimeric G-Protein Beta Subunits During Nodulation in Medicago truncatula Gaertn and Pisum sativum L.

AU - Bovin, Andrey D.

AU - Pavlova, Olga A.

AU - Dolgikh, Aleksandra V.

AU - Leppyanen, Irina V.

AU - Dolgikh, Elena A.

N1 - Publisher Copyright: Copyright © 2022 Bovin, Pavlova, Dolgikh, Leppyanen and Dolgikh.

PY - 2022/1/12

Y1 - 2022/1/12

N2 - Heterotrimeric G-proteins regulate plant growth and development as master regulators of signaling pathways. In legumes with indeterminate nodules (e.g., Medicago truncatula and Pisum sativum), the role of heterotrimeric G-proteins in symbiosis development has not been investigated extensively. Here, the involvement of heterotrimeric G-proteins in M. truncatula and P. sativum nodulation was evaluated. A genome-based search for G-protein subunit-coding genes revealed that M. truncatula and P. sativum harbored only one gene each for encoding the canonical heterotrimeric G-protein beta subunits, MtG beta 1 and PsG beta 1, respectively. RNAi-based suppression of MtGbeta1 and PsGbeta1 significantly decreased the number of nodules formed, suggesting the involvement of G-protein beta subunits in symbiosis in both legumes. Analysis of composite M. truncatula plants carrying the pMtGbeta1:GUS construct showed β-glucuronidase (GUS) staining in developing nodule primordia and young nodules, consistent with data on the role of G-proteins in controlling organ development and cell proliferation. In mature nodules, GUS staining was the most intense in the meristem and invasion zone (II), while it was less prominent in the apical part of the nitrogen-fixing zone (III). Thus, MtG beta 1 may be involved in the maintenance of meristem development and regulation of the infection process during symbiosis. Protein–protein interaction studies using co-immunoprecipitation revealed the possible composition of G-protein complexes and interaction of G-protein subunits with phospholipase C (PLC), suggesting a cross-talk between G-protein- and PLC-mediated signaling pathways in these legumes. Our findings provide direct evidence regarding the role of MtG beta 1 and PsG beta 1 in symbiosis development regulation.

AB - Heterotrimeric G-proteins regulate plant growth and development as master regulators of signaling pathways. In legumes with indeterminate nodules (e.g., Medicago truncatula and Pisum sativum), the role of heterotrimeric G-proteins in symbiosis development has not been investigated extensively. Here, the involvement of heterotrimeric G-proteins in M. truncatula and P. sativum nodulation was evaluated. A genome-based search for G-protein subunit-coding genes revealed that M. truncatula and P. sativum harbored only one gene each for encoding the canonical heterotrimeric G-protein beta subunits, MtG beta 1 and PsG beta 1, respectively. RNAi-based suppression of MtGbeta1 and PsGbeta1 significantly decreased the number of nodules formed, suggesting the involvement of G-protein beta subunits in symbiosis in both legumes. Analysis of composite M. truncatula plants carrying the pMtGbeta1:GUS construct showed β-glucuronidase (GUS) staining in developing nodule primordia and young nodules, consistent with data on the role of G-proteins in controlling organ development and cell proliferation. In mature nodules, GUS staining was the most intense in the meristem and invasion zone (II), while it was less prominent in the apical part of the nitrogen-fixing zone (III). Thus, MtG beta 1 may be involved in the maintenance of meristem development and regulation of the infection process during symbiosis. Protein–protein interaction studies using co-immunoprecipitation revealed the possible composition of G-protein complexes and interaction of G-protein subunits with phospholipase C (PLC), suggesting a cross-talk between G-protein- and PLC-mediated signaling pathways in these legumes. Our findings provide direct evidence regarding the role of MtG beta 1 and PsG beta 1 in symbiosis development regulation.

KW - beta subunits

KW - co-immunoprecipitation

KW - heterotrimeric G-protein

KW - legume-rhizobial symbiosis

KW - Medicago truncatula Gaertn

KW - pea Pisum sativumL

KW - promoter-GUS fusion localization

KW - RNAi based suppression

KW - PLANT

KW - KINASE

KW - RECEPTOR

KW - CELL-PROLIFERATION

KW - PHOSPHOLIPID SIGNALING PATHWAYS

KW - pea Pisum sativum L

KW - RESPONSES

KW - NOD-FACTOR

KW - SYMBIOTIC ROOT-NODULES

KW - ARABIDOPSIS

KW - BACTERIAL ENTRY

UR - http://www.scopus.com/inward/record.url?scp=85123453225&partnerID=8YFLogxK

U2 - 10.3389/fpls.2021.808573

DO - 10.3389/fpls.2021.808573

M3 - Article

AN - SCOPUS:85123453225

VL - 12

JO - Frontiers in Plant Science

JF - Frontiers in Plant Science

SN - 1664-462X

M1 - 808573

ER -