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CRISPR/Cas9 genome-editing applied to MdPGT1 in apple results in reduced foliar phloridzin without impacting plant growth
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SYSNO ASEP 0568825 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title CRISPR/Cas9 genome-editing applied to MdPGT1 in apple results in reduced foliar phloridzin without impacting plant growth Author(s) Miranda, S. (IT)
Piazza, S. (IT)
Nuzzo, F. (IT)
Li, M. (CN)
Lagrèze, J. (IT)
Mithöfer, A. (DE)
Cestaro, A. (IT)
Tarkowská, Danuše (UEB-Q) RID, ORCID
Espley, R. (NZ)
Dare, A. (NZ)
Malnoy, M. (IT)
Martens, S. (IT)Number of authors 12 Source Title Plant Journal. - : Wiley - ISSN 0960-7412
Roč. 113, č. 1 (2023), s. 92-105Number of pages 14 s. Language eng - English Country GB - United Kingdom Keywords dihydrochalcones ; genome-editing ; glycosyltransferase ; Malus × domestica ; phloridzin ; phytohormones OECD category Biochemical research methods R&D Projects EF16_019/0000738 GA MŠMT - Ministry of Education, Youth and Sports (MEYS) Method of publishing Open access Institutional support UEB-Q - RVO:61389030 UT WOS 000896772700001 EID SCOPUS 85143980424 DOI 10.1111/tpj.16036 Annotation Phloridzin is the most abundant polyphenolic compound in apple (Malus × domestica Borkh.), which results from the action of a key phloretin-specific UDP-2′-O-glucosyltransferase (MdPGT1). Here, we simultaneously assessed the effects of targeting MdPGT1 by conventional transgenesis and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9)-mediated genome editing. To this end, we conducted transcriptomic and metabolic analyses of MdPGT1 RNA interference knockdown and genome-edited lines. Knockdown lines exhibited characteristic impairment of plant growth and leaf morphology, whereas genome-edited lines exhibited normal growth despite reduced foliar phloridzin. RNA-sequencing analysis identified a common core of regulated genes, involved in phenylpropanoid and flavonoid pathways. However, we identified genes and processes differentially modulated in stunted and genome-edited lines, including key transcription factors and genes involved in phytohormone signalling. Therefore, we conducted a phytohormone profiling to obtain insight into their role in the phenotypes observed. We found that salicylic and jasmonic acid were increased in dwarf lines, whereas auxin and ABA showed no correlation with the growth phenotype. Furthermore, bioactive brassinosteroids were commonly up-regulated, whereas gibberellin GA4 was distinctively altered, showing a sharp decrease in RNA interference knockdown lines. Expression analysis by reverse transcriptase-quantitative polymerase chain reaction expression analysis further confirmed transcriptional regulation of key factors involved in brassinosteroid and gibberellin interaction. These findings suggest that a differential modulation of phytohormones may be involved in the contrasting effects on growth following phloridzin reduction. The present study also illustrates how CRISPR/Cas9 genome editing can be applied to dissect the contribution of genes involved in phloridzin biosynthesis in apple. Workplace Institute of Experimental Botany Contact David Klier, knihovna@ueb.cas.cz, Tel.: 220 390 469 Year of Publishing 2023 Electronic address https://doi.org/10.1111/tpj.16036
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