Flavanone 3-hydroxylase expression in Citrus paradisi and Petunia hybrida seedlings

• Published in: Phytochemistry (Oxford) Vol. 64; no. 2; pp. 435 - 444
• Main Authors: Pelt, Jennifer L., Downes, W.Andrew, Schoborg, Robert V., McIntosh, Cecilia A.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Ltd 01.09.2003; Elsevier
• Subjects: Amino Acid Sequence; amino acid sequences; biochemical pathways; Biological and medical sciences; biosynthesis; chemical structure; Citrus paradisi; Citrus paradisi - enzymology; Citrus paradisi - genetics; Citrus paradisi - growth & development; Developmental expression; developmental gene expression; developmental stages; DNA Primers - genetics; DNA, Complementary - genetics; F3H; Flavanone 3-hydroxylase; flavanone diglycoside; Flavanones - metabolism; Flavonoid; flowers; Flowers - anatomy & histology; Flowers - metabolism; Fundamental and applied biological sciences. Psychology; gene expression; Grapefruit; grapefruits; leaves; messenger RNA; Metabolism; Metabolism. Physicochemical requirements; mixed function oxidase; Mixed Function Oxygenases - biosynthesis; Mixed Function Oxygenases - genetics; Molecular Sequence Data; naringenin; naringin; nucleotide sequences; Petunia - enzymology; Petunia - genetics; Petunia - growth & development; Petunia hybrida; Plant Leaves - metabolism; Plant physiology and development; quantitative analysis; Reverse Transcriptase Polymerase Chain Reaction - methods; RNA, Messenger - biosynthesis; RNA, Messenger - genetics; RNA, Ribosomal, 18S - genetics; roots; Rutaceae; seedlings; Seedlings - enzymology; Seedlings - genetics; Seedlings - growth & development; Sequence Alignment; Sequence Homology, Amino Acid; Solanaceae; Templates, Genetic; tissue distribution; Developmental expression; Petunia hybrida; Flavanone 3-hydroxylase; Citrus paradisi; Rutaceae; Grapefruit; F3H; Solanaceae; Flavonoid; Enzyme; Naringenin 3-dioxygenase; Tissue specificity; Gene expression; Flavanone derivatives; Flavonoid: Flavanone 3-hydroxylase; Dicotyledones; Angiospermae; Spermatophyta; Developmental stage; Citrus paradist; Oxidoreductases; Biological accumulation
• ISSN: 0031-9422; 1873-3700
• DOI: 10.1016/S0031-9422(03)00341-8

Petunia hybrida and Citrus paradisi have significantly different flavonoid accumulation patterns. Petunia sp. tend to accumulate flavonol glycosides and anthocyanins while Citrus paradisi is known for its accumulation of flavanone diglycosides. One possible point of regulation of flavanone metabolism is flavanone 3-hydroxylase (F3H) expression. To test whether this is a key factor in the different flavanone usage by Petunia hybrida and Citrus paradisi, F3H mRNA expression in seedlings of different developmental stages was measured using semi-quantitative RT-PCR. Primers were designed to conserved regions of F3H and used to amplify an approximately 350 bp segment for quantitation by PhosphorImaging. Primary leaves of 32 day old grapefruit seedlings and a grapefruit flower bud had the highest levels of F3H mRNA expression. Petunia seedlings had much lower levels of F3H mRNA expression relative to grapefruit. The highest expression in petunia was in primary leaves and roots of 65 day old seedlings. These results indicate that preferential use of naringenin for production of high levels of flavanone glycosides in young grapefruit leaves cannot be attributed to decreased F3H mRNA expression. Quantification of flavanone-3-hydroxylase (F3H) mRNA in petunia and grapefruit seedlings at different developmental stages indicates that differential metabolism of naringenin into flavanone glycosides by grapefruit and flavonols/anthocyanins by petunia is not due to lower levels of F3H transcription in grapefruit tissues.