Efficient Rutin and Quercetin Biosynthesis through Flavonoids-Related Gene Expression in Fagopyrum tataricum Gaertn. Hairy Root Cultures with UV-B Irradiation

Abstract

Transformed hairy roots had been efficiently induced from the seedlings of Fagopyrum tataricum Gaertn. due to the infection of Agrobacterium rhizogenes. Hairy roots were able to display active elongation with high root branching in 1/2 MS medium without growth regulators. The stable introduction of rolB and aux1 genes of A. rhizogenes WT strain 15834 into F. tataricum plants was confirmed by PCR analysis. Besides, the absence of virD gene confirmed hairy root was bacteria-free. After six different media and different sources of concentration were tested, the culturing of TB7 hairy root line in 1/2 MS liquid medium supplemented with 30 g l-1 sucrose for 20 days resulted in a maximal biomass accumulation (13.5 g l-1 fresh weight, 1.78 g l-1 dry weight) and rutin content (0.85 mg g-1). The suspension culture of hairy roots led to a 45-fold biomass increase and a 4.11-fold rutin content increase in comparison with the suspension culture of non-transformed roots. The transformation frequency was enhanced through preculturing for 2 days followed by infection for 20 min. The UV-B stress treatment of hairy roots resulted in a striking increase of rutin and quercetin production. Furthermore, the hairy root lines of TB3, TB7, and TB28 were chosen to study the specific effects of UV-B on flavonoid accumulation and flavonoid biosynthetic gene expression by qRT-PCR. This study has demonstrated that the UV-B radiation was an effective elicitor that dramatically changed in the transcript abundance of ftpAL, FtCHI, FtCHS, FtF3H, and FtFLS-1 in F. tataricum hairy roots.

Gene expression profile indicates involvement of uniconazole in Coix lachryma‐jobi L. seedlings at low temperature

“…UNZ primarily inhibited gene expression in relation to diterpenoid biosynthesis metabolic pathways. It was evident that uniconazole could down-regulate the genes coding ent-kaurene oxidase involving in the GA3 metabolism, which were consistent with UNZ catalyzing the oxidation of ent-kaurene to ent-kaurenoic acid in gibberellin biosynthesis (Izumi, Kamiya, Sakyrai, Oahio, & Takahashi 1985 (Huang et al, 2016) and this phenomenon was also observed in our study (Table 1).…”

“…Meanwhile, brassinosteroid-6-oxidase (CYP85A2) related to the synthesis of brassinosteroid was also annotated and up-regulated, which can enhance dry weight of aboveground and belowground biomass of plants (Huang et al, 2016) and this phenomenon was also observed in our study (Table 1). Red arrows indicate positive regulation, and blue arrows indicate negative regulation caffeoyl-CoA O-methyltransferase related to the synthesis of flavonol were annotated and up-regulated.…”

Effects of light on in vitro fiber development and flavonoid biosynthesis in green cotton (Gossypium hirsutum)

“…The biosynthesis of flavonoid and the expression levels of related genes are regulated by phytochrome, cryptochrome, phototropin, and UV-B photoreceptors [43,44]. For example, flavonoid biosynthesis was suppressed by increasing the amount of far-red light of phytochrom [45], whereas UV-A / blue light induction of the flavonoids biosynthesis was mediated principally by cryptochrome [44], in addition, the biosynthesis of flavonoids and the expression levels of related genes are induced most effectively by UV-B light [46,47]. In fact, a white light source that contains all these wavelengths is usually used to induce flavonoid biosynthesis.…”

FtSAD2 and FtJAZ1 regulate activity of the FtMYB11 transcription repressor of the phenylpropanoid pathway in Fagopyrum tataricum

“…The root material was extracted twice in dry ice/cold methanol for 24 h at 4°C. The rutin content was analyzed by high-performance liquid chromatography (HPLC) from triplicate independent extractions for each line as described previously (Huang et al, 2016).…”