Visualization of resistance responses in Phaseolus vulgaris using reporter tagged clones of Bean common mosaic virus

Reporter tagged virus clones can provide detailed information on virus–host interactions. In Phaseolus vulgaris (bean), four recessive and one dominant gene are known to control infection by strains of the potyvirus species Bean common mosaic virus (BCMV). To study the interactions between BCMV and bean genotypes with different resistance gene combinations, an infectious clone of the strain RU1 was tagged with the UidA gene encoding β-glucuronidase (GUS). The clone was agroinoculated to bean genotypes with different combinations of the resistance genes bc-u, bc-1, bc-2, bc-3 and I. In situ histochemical GUS assays showed new details of the resistance responses, which were previously analysed by immunological methods and symptom descriptions. In some instances GUS assays suggested that resistance breaking strains appeared at single foci in uninoculated leaves. To allow recovery of resistance breaking strains for further studies, BCMV RU1 was tagged with the sequence encoding green fluorescent protein (GFP), which was visualized directly without destruction of the tissue. In this paper we present details of the construction of the infectious clone and discuss its application in studies of BCMV resistance in bean.

► We constructed GUS and GFP tagged cDNA clones of bean common mosaic virus.
► The cDNA clones were adapted for agroinoculation.
► Infectivity was tested on bean lines with various combinations of resistance genes.