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. 2021 Dec 28;11(1):90.
doi: 10.3390/plants11010090.

Expression Analysis of Phenylpropanoid Pathway Genes and Metabolomic Analysis of Phenylpropanoid Compounds in Adventitious, Hairy, and Seedling Roots of Tartary Buckwheat

Minsol Choi  1 , Ramaraj Sathasivam  1 , Bao Van Nguyen  2 , Nam Il Park  3 , Sun-Hee Woo  4 , Sang Un Park  1   2
Affiliations

Expression Analysis of Phenylpropanoid Pathway Genes and Metabolomic Analysis of Phenylpropanoid Compounds in Adventitious, Hairy, and Seedling Roots of Tartary Buckwheat

Minsol Choi et al. Plants (Basel). .

Abstract

Tartary buckwheat (Fagopyrum tataricum) is an important crop that belongs to the Polygonaceae family, whose roots have received considerable attention due to the presence of compounds with high nutritional and medicinal value. In this study, we aimed to develop an efficient protocol for the culture of adventitious (ARs) and hairy (HRs) roots on a half-strength Schenk and Hildebrandt (SH) medium containing different concentrations of the auxins, α-naphthaleneacetic acid (NAA), indole-3-butyric acid (IBA), and indole-3-acetic acid (IAA). The highest percentage of root induction (91.67%) was achieved with 0.5 mg/L IAA, whereas the greatest number of roots was found in 1 mg/L IAA. In contrast, 0.1 mg/L IBA returned the longest roots. As expected, HRs were obtained from in vitro leaf explants infected with Agrobacterium rhizogenes R1000. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis of 11 phenolic pathway genes revealed that five genes (FtPAL, FtC3H, FtHQT, FtCHS, and FtANS) were highly expressed in HRs, whereas only four (FtC4H, FtFLS2, FtDFR, and FtANR), and three (Ft4CL, FtCHI, and FtF3H) were recognized in the ARs and seedling roots (SRs), respectively. HPLC analysis of phenolic compounds in different root cultures showed that the majority of the phenolic compounds (both individual and total) were significantly accumulated in the HRs. Principal component analysis (PCA) identified differences among the three root types, whereby HRs were separated from ARs and SRs based on the amount of phenolic compounds present. Analysis of the metabolic pathway revealed that among the identified metabolites, the 3, 2, and 1 pathways were associated with flavonoid, flavone and flavonol, and phenylpropanoid biosynthesis, respectively. Hierarchical clustering analysis and the heat map showed that the different root cultures presented unique metabolites.

Keywords: Agrobacterium rhizogenes; Fagopyrum tataricum; auxins; buckwheat; phenolic compounds; root culture.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Proposed phenylpropanoid biosynthetic pathway in F. tataricum. The enzyme involved in each step is shown in red. The phenylpropanoid compounds measured in this study are shown in black and bold. Asterisks denote the genes used for gene expression analysis. PAL, phenylalanine ammonia-lyase; C4H, cinnamic acid 4-hydroxylase; 4CL, 4-coumaric acid: CoA ligase; HCT, cinnamoyl-CoA shikimate/quinate transferase; HQT, hydroxycinnamoyl CoA:quinate hydroxycinnamoyl transferase; C3H, p-coumaroylester 3-hydroxylase; CHS, chalcone synthase; CHI, chalcone isomerase; F3H, flavanone 3-hydroxylase; FLS, flavonol synthase; F3′H, flavonol 3′ hydroxylase; F3′5′H, flavonoid 3′,5′-hydroxylase; DFR, dihydroflavonol 4-Reductase; LAR, leucocyanidin reductase; ANR, anthocyanidin reductase; ANS, anthocyanidin synthase; FGS, flavan-3-O-gallate synthase; 3 GT, flavonoid-3-O-glucosyltransferase; RT, flavonoid 3-O-glucoside-6-O-rhamnosyltransferase. The pathway scheme is adapted and modified from Li et al. [12].
Figure 2
Figure 2
Growth of different types of root cultures of F. tataricum in a flask. Values followed by different letters within a column indicate a significant difference (p < 0.05) between areas for that parameter using DMRT (n ≥ 5, mean ± SD).
Figure 3
Figure 3
The gene expression profiles of the adventitious root (AR), hairy root (HR), and seedling root (SR) of F. tataricum. Values followed by different letters within a column indicate a significant difference (p < 0.05) between areas for that parameter using DMRT (n ≥ 5, mean ± SD).
Figure 4
Figure 4
Score and loading plot of (A) PCA model and (B) PLS-DA model obtained from the metabolites detected in different root cultures of F. tataricum.
Figure 5
Figure 5
The main components separating the different types of root cultures of F. tataricum are based on the VIP in PLS-DA.
Figure 6
Figure 6
Identified metabolites and their pathway impact on different root cultures of F. tataricum.
Figure 7
Figure 7
Heat map representing the change in the concentration of relative metabolites in root cultures of F. tataricum.
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References

    1. Fabjan N., Rode J., Košir I.J., Wang Z., Zhang Z., Kreft I. Tartary buckwheat (Fagopyrum tataricum Gaertn.) as a source of dietary rutin and quercitrin. J. Agric. Food Chem. 2003;51:6452–6455. doi: 10.1021/jf034543e. - DOI - PubMed
    1. Park N.I., Li X., Suzuki T., Kim S.-J., Woo S.-H., Park C.H., Park S.U. Differential expression of anthocyanin biosynthetic genes and anthocyanin accumulation in tartary buckwheat cultivars ‘Hokkai T8’and ‘Hokkai T10’. J. Agric. Food Chem. 2011;59:2356–2361. doi: 10.1021/jf200020b. - DOI - PubMed
    1. Zhu F. Chemical composition and health effects of Tartary buckwheat. Food Chem. 2016;203:231–245. doi: 10.1016/j.foodchem.2016.02.050. - DOI - PubMed
    1. Guillon S., Trémouillaux-Guiller J., Pati P.K., Gantet P. Hairy roots: A powerful tool for plant biotechnological advances. In: Ramawat K.G., Merillon J.M., editors. Bioactive Molecules and Medicinal Plants. Springer; Berlin/Heidelberg, Germany: 2008. pp. 271–283.
    1. Bhatia S., Bera T., Dahiya R., Bera T., Bhatia S., Bera T. Classical and nonclassical techniques for secondary metabolite production in plant cell culture. In: Bhatia S., Sharma K., Dahiya R., Bera T., editors. Modern Applications of Plant Biotechnology in Pharmaceutical Sciences. Elsevier; Amsterdam, The Netherlands: 2015. pp. 231–291.