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.
Highlights
Pisum sativum has been a model organism in plant research for more than a century; it has lagged behind other plants in becoming the model organism to study root symbioses, i.e., those resulting in nodulation and mycorrhiza-formation
Pea Transformation Proof-of-concept In our hands, the best method to produce transformed composite pea plants (Figures 1 and 2) relied on the infection of 10 day-old pea seedlings using Fibrgro® cubes imbibed with A. rhizogenes suspension as in Collier et al [24]
Our rationale was that the complementation of these mutants with the wild-type SYM10 gene should reverse their nodulation phenotype and the presence of nodules should serve as the ultimate scoring for transformation success
Summary
Pisum sativum (pea) has been a model organism in plant research for more than a century; it has lagged behind other plants in becoming the model organism to study root symbioses, i.e., those resulting in nodulation and mycorrhiza-formation Reasons for this stem from the fact that pea is a large plant which requires a large area for growth and has a relatively long life-cycle (~ 90 days). It has a large genome (about 4 × 109 base pairs [1]) still to be sequenced, and many molecular techniques, especially those of transformation, are difficult to use [1] to study the function of the genes involved in the nodulation process. Pea production is expected to increase further as farming practises are focusing on agricultural sustainability, with farmers once again utilizing the benefits of crop rotation to decrease fertilizer applications.
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