5 research outputs found

    First genome edited poinsettias: targeted mutagenesis of flavonoid 3′-hydroxylase using CRISPR/Cas9 results in a colour shift

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    The CRISPR/Cas9 system is a remarkably promising tool for targeted gene mutagenesis, and becoming ever more popular for modification of ornamental plants. In this study we performed the knockout of flavonoid 3′-hydroxylase (F3′H) with application of CRISPR/Cas9 in the red flowering poinsettia (Euphorbia pulcherrima) cultivar ‘Christmas Eve’, in order to obtain plants with orange bract colour, which accumulate prevalently pelargonidin. F3′H is an enzyme that is necessary for formation of cyanidin type anthocyanins, which are responsible for the red colour of poinsettia bracts. Even though F3′H was not completely inactivated, the bract colour of transgenic plants changed from vivid red (RHS 45B) to vivid reddish orange (RHS 33A), and cyanidin levels decreased significantly compared with the wild type. In the genetically modified plants, an increased ratio of pelargonidin to cyanidin was observed. By cloning and expression of mutated proteins, the lack of F3′H activity was confirmed. This confirms that a loss of function mutation in the poinsettia F3′H gene is sufficient for obtaining poinsettia with orange bract colour. This is the first report of successful use of CRISPR/Cas9 for genome editing in poinsettia

    Alteration of the phenylpropanoid pathway by watercore disorder in apple (Malus x domestica)

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    Watercore is a physiological disorder of apple expressed as fluid deposition in intercellular spaces. We analysed the watercore prone cultivar \u27Fuji\u27 to establish a link between enzymatic response and fluid accumulation in the fruit. Individual phenolic content, selected enzyme activities of the phenylpropanoid pathway, sugars and organic acids were quantified in watercore affected flesh and peel. In addition, transcriptome sequencing was performed to obtain a first insight into molecular mechanisms underlying the physiological disorder. Sampled material included: peel (HP) and flesh from healthy fruit (H), peel from watercore affected fruit (WP), healthy flesh from fruit with watercore (HW) and watercore flesh (WW) from fruit with watercore. A sorbitol increase in watercore affected apple was observed not only in the flesh but also in the peel. Moreover, two phenolic groups (hydroxycinnamic acids and flavonols) were significantly affected by watercore and their content was higher in WP in comparison to HP. Dihydrochalcone content was the highest in WW flesh. This was supported by RNA-seq data, which indicated a generally increased expression of genes from the early flavonoid pathway, sorbitol metabolism and polyphenol oxidases. Among the phenylpropanoid pathway enzymes analysed, phloretin 2′-O-glycosyltransferase was the only one exhibiting altered activity
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