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Open Access Methodology

Reproducible hairy root transformation and spot-inoculation methods to study root symbioses of pea

Scott R Clemow1, Lindsey Clairmont1, Lene H Madsen2 and Frédérique C Guinel1*

Author Affiliations

1 Department of Biology, Wilfrid Laurier University, 75 University Avenue W., Waterloo, N2L 3C5, Ontario, Canada

2 Department of Molecular Biology, Centre for Carbohydrate Recognition and Signalling, Aarhus University, Gustav Wields Vej 10, Aarhus C -8000 Denmark

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Plant Methods 2011, 7:46  doi:10.1186/1746-4811-7-46

Published: 15 December 2011

Abstract

Pea has lagged behind other model legumes in the molecular study of nodulation and mycorrhizae-formation because of the difficulty to transform its roots and its poor growth on agar plates. Here we describe for pea 1) a transformation technique which permits the complementation of two known non-nodulating pea mutants, 2) a rhizobial inoculation method which allows the study of early cellular events giving rise to nodule primordia, and 3) a targeted fungal inoculation method which allows us to study short segments of mycorrhizal roots assured to be infected. These tools are certain to advance our knowledge of pea root symbioses.

Keywords:
Composite plants; gene complementation; mycorrhizae; nodulation; pea mutants; targeted spot-inoculation; transformation