Abstract
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.
Highlights
Fresh cut roses are among the most popular ornamental plants, generating billions of dollars in trade annually and accounting for more than one-third of the total cut flower industry worldwide[1]
We developed a detached petal disc assay (DPDA) that enabled us to quantitatively evaluate B. cinerea resistance in rose flowers
The results of the DPDA in resistant and susceptible cultivars matched their resistance abilities observed in the field, suggesting that the DPDA is a reliable method for quantifying resistance against B. cinerea in rose flowers
Summary
Fresh cut roses are among the most popular ornamental plants, generating billions of dollars in trade annually and accounting for more than one-third of the total cut flower industry worldwide[1]. The supply of cut roses is heavily reliant on the logistics of long-distance transportation, as cut roses are produced mainly in developing countries with low labor costs and suitable climates, such as Kenya, Colombia, Ecuador, and China, whereas the consumer market is mainly in the developed countries of North America, Europe, and Japan. Botrytis cinerea causes the most severe postharvest losses. B. cinerea infection leads to necrotic lesions on petals, and symptoms develop rapidly during postharvest transport, during which flowers are packed in boxes with high relative humidity[5]
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