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Fresh-cut roses ( Rosa hybrida ) are one of the most important ornamental crops worldwide, with annual trade in the billions of dollars. Gray mold disease caused by the pathogen  Botrytis cinerea  is the most serious fungal threat to cut roses, causing extensive postharvest losses. In this study, we optimized a detached petal disc assay (DPDA) for artificial  B. cinerea  inoculation and quantification of disease symptoms in rose petals. Furthermore, as the identification of rose genes involved in  B. cinerea  resistance could provide useful genetic and genomic resources, we devised a virus-induced gene silencing (VIGS) procedure for the functional analysis of  B. cinerea  resistance genes in rose petals. We used  RhPR10.1  as a reporter of silencing efficiency and found that the rose cultivar ‘Samantha’ showed the greatest decrease in  RhPR10.1  expression among the cultivars tested. To determine whether jasmonic acid and ethylene are required for  B. cinerea  resistance in rose petals, we used VIGS to silence the expression of  RhLOX5  and  RhEIN3  (encoding a jasmonic acid biosynthesis pathway protein and an ethylene regulatory protein, respectively) and found that petal susceptibility to  B. cinerea  was affected. Finally, a VIGS screen of  B. cinerea -induced rose transcription factors demonstrated the potential benefits of this method for the high-throughput identification of gene function in  B. cinerea  resistance. Collectively, our data show that the combination of the DPDA and VIGS is a reliable and high-throughput method for studying  B. cinerea  resistance in rose.

A detached petal disc assay and virus-induced gene silencing facilitate the study of Botrytis cinerea resistance in rose flowers