Agrobacterium rhizogenes-mediated transformation of Pisum sativum L. roots as a tool for studying the mycorrhizal and root nodule symbioses

Irina V Leppyanen; Anna N Kirienko; Elena A Dolgikh

doi:10.7717/peerj.6552

Agrobacterium rhizogenes-mediated transformation of Pisum sativum L. roots as a tool for studying the mycorrhizal and root nodule symbioses

PeerJ. 2019 Mar 6:7:e6552. doi: 10.7717/peerj.6552. eCollection 2019.

Authors

Irina V Leppyanen¹, Anna N Kirienko¹, Elena A Dolgikh¹

Affiliation

¹ All-Russia Research Institute for Agricultural Microbiology, Saint-Petersburg, Russia.

Abstract

In this study, we demonstrated the successful transformation of two pea (Pisum sativum L.) cultivars using Agrobacterium rhizogenes, whereby transgenic roots in the resulting composite plants showed expression of the gene encoding the green fluorescent protein. Subsequent to infection with A. rhizogenes, approximately 70%-80% of pea seedlings developed transgenic hairy roots. We found out that the transgenic roots can be efficiently nodulated by Rhizobium leguminosarum bv. viciae and infected by the arbuscular mycorrhizal (AM) fungus Rhizophagus irregularis. The morphology of nodules in the transgenic roots was found to be identical to that of nodules observed in wild-type roots, and we also observed the effective induction of markers typical of the symbiotic association with AM fungi. The convenient protocol for highly efficient A. rhizogenes-mediated transformation developed in this study would be a rapid and effective tool for investigating those genes involved in the development of the two types of symbioses found in pea plants.

Keywords: Agrobacterium rhizogenes; Composite plants; Mycorrhizae; Nodulation; Pea transformation.

Grants and funding

This work was financially supported by the Russian Science Foundation (grant 17-76-30016). The research was performed using equipment of the Core Centrum “Genomic Technologies, Proteomics and Cell Biology” in ARRIAM. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.