Identification and characterisation of CYP75A31, a new flavonoid 3'5'-hydroxylase, isolated from Solanum lycopersicum

BMC Plant Biology

Research Article

Kristine M Olsen¹  Cathrine Lillo¹  Rune Slimestad²  Frédéric Bourgaud³  Hélène Jugdé³  Romain Larbat³  Alain Hehn³ 

[1] Centre for Organelle Research, Faculty of Science and Technology, University of Stavanger, N-4036, Stavanger, Norway;PlantChem, Saerheim Research Center, N-4353, Klepp stasjon, Norway;UMR 1121 Nancy Université (INPL)-INRA Agronomie et Environnement (Nancy/Colmar), IFR110, 2, av. de la Forêt de Haye, 54505, Vandoeuvre-lès-Nancy, France;

关键词: Flavonoid;    Rutin;    Naringenin;    Anthocyanin Synthesis;    Dihydroquercetin;   

DOI  :  10.1186/1471-2229-10-21

 received in 2009-07-15, accepted in 2010-02-03,  发布年份 2010

来源: Springer

【 摘 要 】

BackgroundUnderstanding the regulation of the flavonoid pathway is important for maximising the nutritional value of crop plants and possibly enhancing their resistance towards pathogens. The flavonoid 3'5'-hydroxylase (F3'5'H) enzyme functions at an important branch point between flavonol and anthocyanin synthesis, as is evident from studies in petunia (Petunia hybrida), and potato (Solanum tuberosum). The present work involves the identification and characterisation of a F3'5'H gene from tomato (Solanum lycopersicum), and the examination of its putative role in flavonoid metabolism.ResultsThe cloned and sequenced tomato F3'5'H gene was named CYP75A31. The gene was inserted into the pYeDP60 expression vector and the corresponding protein produced in yeast for functional characterisation. Several putative substrates for F3'5'H were tested in vitro using enzyme assays on microsome preparations. The results showed that two hydroxylation steps occurred. Expression of the CYP75A31 gene was also tested in vivo, in various parts of the vegetative tomato plant, along with other key genes of the flavonoid pathway using real-time PCR. A clear response to nitrogen depletion was shown for CYP75A31 and all other genes tested. The content of rutin and kaempferol-3-rutinoside was found to increase as a response to nitrogen depletion in most parts of the plant, however the growth conditions used in this study did not lead to accumulation of anthocyanins.ConclusionsCYP75A31 (NCBI accession number GQ904194), encodes a flavonoid 3'5'-hydroxylase, which accepts flavones, flavanones, dihydroflavonols and flavonols as substrates. The expression of the CYP75A31 gene was found to increase in response to nitrogen deprivation, in accordance with other genes in the phenylpropanoid pathway, as expected for a gene involved in flavonoid metabolism.

【 授权许可】

CC BY   
© Olsen et al; licensee BioMed Central Ltd. 2010