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

Xuan Huang,Dengfeng Xie,Yangyang Zhao,Xue Jiang,Jingwen Yao,Ziqin Xu

doi:10.3389/fpls.2016.00063

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.

Highlights

As a significant food and medicinal species, tartary buckwheat (Fagopyrum tataricum Gaertn.; family Polygonaceae) is grown and used in the mountainous regions of Southwest China (Sichuan province), Northern India, Bhutan, and Nepal (Fabjan et al, 2003)
We have established an efficient protocol for A. rhizogenesmediated genetic transformation of tartary buckwheat (F. tataricum), and applied PCR analysis in the detection of hairy roots
Hairy roots cultured in 1/2 MS liquid medium supplemented with 30 g l−1 sucrose grew faster than normal roots under standard liquid culture conditions, and had higher rutin content

Summary

Introduction

As a significant food and medicinal species, tartary buckwheat (Fagopyrum tataricum Gaertn.; family Polygonaceae) is grown and used in the mountainous regions of Southwest China (Sichuan province), Northern India, Bhutan, and Nepal (Fabjan et al, 2003). The plant contains numerous functional components, including flavonoids, phenolic compounds, phytosterols, fagopyrins, Production and Gene Expression in F. tataricum d-chiro-inositol, and thiamin-binding proteins, which play essential role in antioxidant, hypocholesterolemic, and antidiabetic effects (Krkošková and Mrázová, 2005; Jiang et al, 2007; Tomotake et al, 2007; Yang and Ren, 2008; Yao et al, 2008; Qin et al, 2013). The major functional components of F. tartaricum, such as rutin, quercetin, orientin, vitexin and kaempferol, had been demonstrated to be flavonoids In comparison with Fagopyrum esculentum (common buckwheat), as a source of dietary rutin and quercetin, F. tartaricum had higher contents of flavonoids and other phenolic compounds. There had been increasing researches focusing on tartary buckwheat in recent years due to its remarkable health benefits associated with health

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