Establishment of a novel system to elucidate the mechanisms underlying light-induced ripening of strawberry fruit with an Agrobacterium-mediated RNAi technique
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Traditional methods used to study strawberry ripening-related gene function are time-consuming, and require at least 15 months from initiating the transformation experiment until the first ripe fruits are available for analysis. To accelerate data acquisition during gene function studies, we explored a transient assay method that employs an <i>Agrobacterium</i>-mediated RNAi (<i>A</i>mRNAi) technique in post-harvest strawberry fruit, <i>Fragaria</i>×<i>ananassa</i> (<i>Fa</i>) cv. Sachinoka, a Japanese cultivar. Our results showed that artificial white light induced strong expression of <i>Fa′chalcone synthase</i> (<i>Fa′CHS</i>), <i>Fa′chalcone isomerase</i> (<i>Fa′CHI</i>), and <i>Fa′flavonoid 3′-hydroxylase</i> orthologues (<i>Fa′F3′H</i>) in post-harvest fruit. <i>Fa′CHS</i> and <i>Fa′F3′H</i> function was subsequently examined by performing <i>A</i>mRNAi with post-harvest fruit. Although reduction of light-induced <i>Fa′F3′H</i> expression by <i>A</i>mRNAi resulted in no significant change in anthocyanin content, reduction of <i>Fa′CHS</i> significantly decreased anthocyanin levels, and up-regulated <i>Fa′F3′H</i> levels. Our results are consistent with previous data indicating that while <i>CHS</i> is required for anthocyanin accumulation during late stage strawberry fruit maturation, <i>Fa′F3′H</i> is not required. The novel system described here enabled gene function data to be available within 10 days of initiating the incubation period following infiltration. Therefore, we conclude our system is a valuable tool to elucidate the molecular mechanisms underlying light-induced ripening of strawberry fruit.
- Plant Biotechnology
Plant Biotechnology 29(3), 271-277, 2012-06-25
Japanese Society for Plant Cell and Molecular Biology