Yan Bai

Introduction: Tetrastigma hemsleyanum Diels et Gilg (Vitaceae) is a medicinally valuable plant typically with a whitish root cross-section. Resource surveys discovered some roots of T. hemsleyanum with a yellow-brown root cross-section and stronger antioxidant properties. However, the molecular mechanisms underlying these phenotypic differences remain poorly understood.Methods: We employed integrated metabolomic and transcriptomic analyses to compare the two root types. Key candidate genes were cloned and functionally characterized to uncover the regulatory mechanisms involved.Results: Metabolomic analysis identified significant changes in 217 metabolites between the two types, among which 57 flavonoids, such as Hesperetin-7-O-glucoside, were significantly higher in the yellow-brown roots. Transcriptomic analysis revealed 3516 differentially expressed genes, many associated with flavonoid biosynthesis. Differences in root quality were largely attributed to varied flavonoid production. Analysis of transcription factors regulating this process identified a strong correlation between MYB6 genes and flavonoids biosynthesis. Functional analysis indicated that the ThMYB6 factor of the R2R3 MYB subgroup 5 regulates flavonoid production in T. hemsleyanum.Conclusion: The ThMYB6-mediated divergence in flavonoid biosynthesis is the key factor influencing quality differences between the two phenotypes. These results establish a theoretical foundation for targeted quality improvement and offer scientific support for the medicinal development of T. hemsleyanum.

Introduction: Tetrastigma hemsleyanum is a folk and rare medicinal plant, and specifically, it is distributed in the south, China. To investigate the cumulative properties of its medicinal components, we examined the effect of UV light on flavonoid content and related enzyme activity changes in T. hemsleyanum.Methods: The leaves and tubers were treated with UV-A, UV-B and UV-C for 1 h, 1L/23D h, 3 h and 3L/21D h (D represents darkness treatment). High-performance liquid chromatography (HPLC) analysis showed that the content of many flavonoids decreased significantly during UV-A treatment, increased after UV-B and UV-C irradiation and accumulated again after darkness treatment.Results: In the root tubers of the UV-A group, naringin content in the 3L/21D h group (0.069 μg/g) was 16.30 times higher than that of 3 h group (0.0042 μg/g). The rutin content was elevated after UV irradiation but was not detected in the CK group. The test results of the enzyme-linked kit indicated that the activities of many enzymes were higher in the UV-A and UV-B irradiation groups than those in the CK group, but the results were reversed in the UV-C treatment. After darkness treatment, the activities of most enzymes were higher than those with UV irradiation alone; F3’5’H activity in the 3L/21D h group (97.25 U/L) was 1.24 times higher than that in the 3 h group (78.12 U/L) in the UV-A-treated group.Discussion: The study results suggest that appropriate UV-B and UV-C irradiation, as well as darkness supplementation, had a promotive effect on flavonoids in the leaves and root tubers of T. hemsleyanum. Additionally, UV irradiation and darkness treatment enhanced the activity of most enzymes.

Paris polyphylla Smith var. Chinensis (Franch.) Hara is a medicinal plant that belongs to the Liliaceae family. Its main components are parissaponins, which have excellent medicinal effects such as anti-inflammatory, anti-tumor, etc. Improving the quality of parissaponins through artificial directional regulation has emerged as a practice to meet medical demand and is a new research hotspot. In this paper, P. polyphylla plants were treated with UVA, UVB, and UVC, and the contents of PolyPhyllin VI (PPVI) and PolyPhyllin VII (PPVII), saponin synthase (squalene synthase, SS; cycloartenol synthase, CAS; cytochrome P450, CYP450; and glycosyl transferases, GT) activity, MDA, and the photosynthetic pigment indexes were measured and analyzed. The results showed that PPVII content increased by 32.43% with UVC treatment after 4 h (3.43 mg/g), but the PPVI and PPVII contents in the other groups decreased compared with CK (control group) and they did not return to the original level after 4 h. SS, CAS, CYP450, and GT synthases were activated in varying degrees via UV treatment and increased, respectively, by 22.93%, 10.83%, 20.15%, and 25.98%. Among them, GT, as the last of the synthetases, had a shorter response time to UVB (30 min) and UVC (15 min); the difference was sensible compared with CK. Moreover, UV had a stressing effect and promoted the rapid accumulation of MDA content (increased 17.66%, 34.53%, and 9.65%) and carotenoid (increased 7.58, 5.60, and 7.76 times) within 4 h compared to CK. UVB and UVC radiation visibly improved chlorophyll a content (42.56% and 35.45%), but UVA did not, and the change in chlorophyll b content showed no overt statistical difference. In addition, PPVI and PPVII were negatively correlated with SS, CAS, carotenoids, and MDA (p < 0.05) and positively correlated with CYP450, GT, and chlorophyll a (p < 0.05). This study provides a theoretical basis for using UV light to regulate secondary metabolism in P. polyphylla, which is of great value for production management.

Tetrastigma hemsleyanum Diels et Gilg, known as a “plant antibiotic”, possesses several attractive properties including anti-inflammatory, anti-tumor, and antioxidant effects, with its efficacy being attributed to flavonoids. However, the flavonoid biosynthesis of T. hemsleyanum has rarely been studied. In this study, we investigated the flavonoid metabolism of T. hemsleyanum through metabolome analysis and transcriptome sequencing. The metabolomic results showed differences in the flavonoids of the leaves and root tubers of T. hemsleyanum. A total of 22 flavonoids was detected, and the concentrations of most flavonoids in the leaves were higher than those in the root tubers. Transcriptome analysis revealed that differentially expressed genes (DEGs) in the leaves and root tubers were enriched in photosynthesis-antenna proteins. Pearson correlation analysis indicated that the expression levels of chalcone isomerase (CHI) and UDP-glycose flavonoid glycosyltransferase (UFGT) were highly correlated with the concentrations of most flavonoids. Further, this study found that the photosynthesis-antenna proteins essentially contributed to the difference in the flavonoids in T. hemsleyanum. The gene expressions and concentrations of the total flavonoids of leaves and root tubers in Hangzhou, Jinhua, Lishui, and Taizhou in Zhejiang Province, China, showed that CHI (CL6715.Contig1_All, Unigene19431_All, CL921.Contig4_All) and UFGT (CL11556.Contig3_All, CL11775.Contig1_All) were the potential key genes of accumulation of most flavonoids in T. hemsleyanum.

Tetrastigma hemsleyanum Diels et Gilg is a folk herb in Zhejiang Province with anti-inflammatory, antineoplastic, and anti-oxidation effects. Given its pharmacological activity, T. hemsleyanum is known as New “Zhebawei” and included in the medical insurance system of Zhejiang and other provinces. Flavonoids are the most important components of T. hemsleyanum, and their contents are mainly regulated by ultraviolet (UV) radiation. In this study, the total flavonoid contents, flavonoid monomer contents, and flavonoid synthesis related enzyme activities (phenylalanine ammonia–lyase, chalcone synthase, and chalcone isomerase), anti-oxidant enzyme activities (catalase, peroxidase, and superoxide dismutase), and biochemical indicators (malondialdehyde, free amino acid, soluble protein, and soluble sugar) in the leaves (L) and root tubers (R) of T. hemsleyanum with UV treatments were determined. Three kinds of UV radiation (UV-A, UV-B, and UV-C) and six kinds of radiation durations (15 and 30 min, 1, 2, 3, and 5 h) were used. Appropriate doses of UV-B and UV-C radiation (30 min to 3 h) induced eustress, which contributed to the accumulation of flavonoids and improve protective enzyme system activities and bioactive compound contents. Especially, certain results were observed in several special structures of the flavonoid monomer: quercetin contents in L increased by nearly 20 times, isoquercitrin contents in R increased by nearly 34 times; most of flavonoids with glycoside content, such as quercitrin (19 times), baicalin (16 times), and apigenin-7G (13 times), increased multiple times. Compared with the CK group, the flavonoid synthase activities, anti-oxidant enzyme activities, and biochemical substance contents in L and R all increased with UV treatments. This study provides a theoretical foundation for regulating flavonoids by light factors and improving the quality of T. hemsleyanum in production and medical industries.

Tetrastigma hemsleyanum is a traditional Chinese medicine herb, commonly used for its anti-inflammatory and antitumor properties. Flavonoids are the main functional constituents of T. hemsleyanum, and their production in the herb is affected by light quality. T. hemsleyanum is a shade-loving plant and is usually covered by black shade nets during cultivation. However, there are only a few studies on the effects of using color films on growth and flavonoid synthesis in T. hemsleyanum. In this study, we measured the influence of five different color films on growth indexes—sugar, soluble amino acid, soluble protein, and flavonoid content—and flavonoid-synthesizing enzyme activities in T. hemsleyanum . The films used were colorless plastic film as the control group (CK-W), red film (RF), yellow film (YF), green film (GF), and blue film (BF). BF promoted plant growth and increased yield, as evidenced by the highest growth indexes, soluble amino acid content, and chalcone isomerase (CHI) enzyme activity. RF increased the content of secondary metabolites, thereby enhancing herb quality, as evidenced by the highest phenylalanine ammonia lyase (PAL) activity and increased flavonoid content.