The Aconitum carmichaelii F3′5′H Gene Overexpression Increases Flavonoid Accumulation in Transgenic Tobacco Plants

Abstract

Aconitum carmichaelii Debx. is a herbal species that contains many precious bioactive substances, which are alkaloids, flavonoids, steroids, and glycosides. Flavonoids, which are major secondary compounds, play an important role in maintaining redox balance in the cells of the plant body. Many flavonoids have antibacterial, antioxidant, and anticancer properties. However, studies have mainly focused on aconitine, which is a highly toxic group A poison belonging to the alkaloid group, but with little mention of flavonoids. The flavonoids in A. carmichaelii are a group of substances with high content, concentrated in leaves and flowers, including quercetin and kaempferol. F3′5′H (Flavonoid 3′5′-hydroxylase) has been identified as the key enzyme involved in the final steps of flavonoid biosynthesis in plants in general and in A. carmichaelii specifically. This study offers the first report, and demonstrates that the overexpression of the F3′5′H gene from a herbal plant, A. carmichaelii, increases flavonoid content in genetically modified tobacco plants. The A. carmichaelii gene was transformed into tobacco leaf tissue to create transgenic tobacco plants. The AcF3′5′H gene was incorporated into the tobacco genome and was expressed in four transgenic tobacco lines (T01, T03, T05, and T014). The F3′5′H content increased from 20.33% to 32.00% compared with that in non-transformed plants (P < 0.001). Therefore, the flavonoid content of four transgenic tobacco lines increased compared to the WT, from 69.23% to 122.23% (P < 0.001). The results of the successful expression of the AcF3′5′H gene in model tobacco plants are the basis for using the AcF3′5′H gene for improving flavonoid content in other medicinal plants. Thus, the AcF3′5′H gene considered in this work could be a candidate for gene technology to enhance flavonoid accumulation in plants.

Integrated transcriptome and metabolome analysis reveals antioxidant machinery in grapevine exposed to salt and alkali stress

“…In this study, the transcriptome analysis revealed that both SS and AS had an impact on the expression of genes associated with flavonoid biosynthesis ( PAL , 4CL , CHS , CHI , F3H , FLS , and F3′5′H ). Previous research has shown that overexpressing PAL , Df4CL1 , EaCHS1 , DcCHI1 , DcCHI4 , CsF3H , EkFLS , and AcF3′5′H can enhance flavonoid production in transgenic plants (Chen et al, 2015; Li et al, 2021; Mahajan & Yadav, 2014; Nguyen et al, 2021; Park et al, 2012; Wang et al, 2021; Zhu et al, 2021). Therefore, the upregulation of PAL , 4CL , CHS , CHI , F3H , FLS , and F3′5′H genes under SS, as well as the upregulation of CHS and CHI genes under AS, may contribute to increased flavonoid accumulation in grapevines, thereby enhancing their tolerance to SS and AS.…”

Cajanus platycarpus Flavonoid 3′5′ Hydroxylase_2 (CpF3′5′H_2) Confers Resistance to Helicoverpa armigera by Modulating Total Polyphenols and Flavonoids in Transgenic Tobacco

“…This information not only demonstrated the involvement of CpF3′5′H_2 in the response of the wild relative to herbivory but also corroborated with earlier studies from our group towards understanding the chemical basis of resistance response in C. platycarpus [ 11 ]. Though there have been studies demonstrating the utility of increased expression of F3′5′H in plants [ 21 , 24 , 25 ], deciphering the role of genes identified during the resistance response towards herbivory has been scarce.…”

“…Studies have demonstrated the industrial utility of F3′5′H for improving colour in ornamental flowers [ 19 , 20 ]. Additionally, the overexpression of F3′5′H has also shown increased flavonoids in many plant species [ 21 ]. Further, there have also been several studies establishing the role of other pivotal enzymes of the flavonoid pathway in the management of abiotic stresses [ 22 ].…”

Integrated Data Reveal the Flavonoid Biosynthesis Metabolic Pathway in Semen Ziziphi Spinosae Using Transcriptomic and Metabolomic

“…F3′H, and F3′5′H which belonged to the CYP450 family played an important role in regulating the biosynthesis of different types of flavonoids. Overexpression of these genes could increase the flavonoid accumulation in plants [44]. Anthocyanins were often highly modified into anthocyanin derivatives by acyltransferase, methyltransferase, and UDP-glycosyltransferase [45].…”