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.
Highlights
Pea (Pisum sativum L.) is one of the most agriculturally important legumes and is a potential target for crop improvement
We describe a convenient protocol for highly efficient A. rhizogenes-mediated transformation of P. sativum
In order to select for transformed hairy roots and quantify the efficiency of transformation, we introduced a plasmid carrying a fusion with the gusA/GFP reporter gene driven by the CaMV35S promoter and subsequently examined hairy roots and nodules for green fluorescent protein (GFP) or β-glucuronidase (GUS) staining
Summary
Pea (Pisum sativum L.) is one of the most agriculturally important legumes and is a potential target for crop improvement. In comparison with other legumes such as Lotus japonicus and Medicago truncatula (Schauser et al, 1999; Ané et al, 2002; Limpens et al, 2003; Madsen et al, 2003; Arrighi, 2006; Gonzalez-Rizzo, Crespi & Frugier, 2006; Heckmann et al, 2006; Tirichine et al, 2006; Smit et al, 2007; Floss et al, 2013; Yano et al, 2017), the technique of transformation necessary for such experiments, as well as for crop improvement, has not been widely applied for pea plants.
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