Functional characterization of nine critical genes encoding rate-limiting enzymes in the flavonoid biosynthesis of the medicinal herb Grona styracifolia.

Grona styracifolia is a photophilous legume that contains abundant flavonoids with multiple pharmacological activities, which is used to cure urethral and biliary calculus in China for thousands of years. The authentication of the rate-limiting enzymes involved in the flavonoids biosynthesis pathway enabled a better understanding of the molecular aspect of quality formation and modulation of this medicinal herb. In this study, the chemical distribution characteristics and content of flavonoids in different tissues of Grona styracifolia were analyzed using ultraperormance liquid chromatography coupled with Q-TOF mass spectrometry and showed that active flavonoids were primarily synthesized and stored in the leaves. Subsequently, RNA sequencing (RNA-seq)-based transcriptome profiling of the different tissues revealed that the flavonoids biosynthesis in the leaves was the most active. Meanwhile, 27 full-length transcripts inferred encoding vital enzymes involved in the flavonoids biosynthesis were preliminarily excavated. Finally, four CHSs, four CHIs, and one FNSII were successfully characterized by heterologous expression, which involved in three rate-limiting steps of the flavonoid biosynthetic pathway. In conclusion, these results laid a foundation for further investigation of the molecular mechanism of the biosynthesis and modulation of active flavonoids in Grona styracifolia.

Functional characterization of four mono-terpene synthases (TPSs) provided insight into the biosynthesis of volatile monoterpenes in the medicinal herb Blumea balsamifera.

Blumea balsamifera, a wooden plant belonging to the family Asteraceae, is a medicinal herb with anticancer, antiviral, and multiple pharmacological effects, which are believed to be caused by its essential oil. The essential oil from B. balsamifera is comprised of mono- and sesqui-terpenes as the majority. Unfortunately, this plant has been facing the challenge of resource shortage, which could be effectively alleviated by biological engineering. Therefore, the identification of key elements involved in the biosynthesis of active ingredients becomes an indispensable prerequisite. In this study, candidate genes encoding monoterpene synthase were screened by transcriptome sequencing combined with metabolomics profiling in the roots, stems, and leaves of B. balsamifera. Then, these candidates were successfully cloned and verified by heterologous expression and in vitro enzyme activity assays. As a result, six candidate BbTPS genes were isolated from B. balsamifera, of which three encoded single-product monoterpene synthases and one encoded a multi-product monoterpene synthase. Among them, BbTPS1, BbTPS3, and BbTPS4 could catalyze the formation of D-limonene, α-phellandrene, and L-borneol, respectively. Meanwhile, BbTPS5 functioned in catalyzing GPP into terpinol, ß-phellandrene, ß-myrcene, D-limonene, and 2-carene in vitro. In general, our results provided important elements for the synthetic biology of volatile terpenes in B. balsamifera, which laid a foundation for subsequent heterologous production of these terpenoids through metabolic engineering and increasing their yield, as well as promoting sustainable development and utilization of B. balsamifera. Supplementary Information: The online version contains supplementary material available at 10.1007/s12298-023-01306-8.

Integrated UPLC-MS, network pharmacology, and intestinal flora analysis to determine the treatment effect of Qiangzhi decoction on comorbid Tourette syndrome and RRTI.

Background: Tourette syndrome (TS) is a complex neurodevelopmental disorder characterized by vocal and motor tics. Recurrent respiratory tract infection (RRTI), a commonly occurring disease in childhood, correlates with recurrent and severe course of tic symptoms. Qiangzhi decoction (QZD) is a traditional Chinese medicine that can alleviate TS symptoms while reducing the recurrence of RRTI. However, the mechanism of QZD on TS and RRTI remains unclear. This study aimed to determine the treatment effect of QZD on comorbid TS and RRTI by integrating ultrahigh-performance liquid chromatography mass spectrometry (UPLC-MS), network pharmacology, and intestinal flora analysis. Methods: The components of QZD were first identified by UPLC-quadrupole (Q)-orbitrap-MS/MS. The mechanism of QZD on comorbid RRTI and TS was investigated by a series of network pharmacological methods, including target prediction and bioinformatics analysis. Finally, a comorbid TS and RRTI rat model was established by intraperitoneal injection of 3,3-iminodipropionitrile (IDPN), cyclophosphamide (CTX), and lipopolysaccharide (LPS). Alteration of gut microbiota in the alleviation of TS and RRTI by QZD was investigated via intestinal flora analysis. Results: The results of UPLC-Q-orbitrap-MS/MS showed that QZD had 96 types of chemical components. The network pharmacology results demonstrated that targets of QZD involved in the treatment of TS and RRTI involved 1045 biological processes (BPs), 109 cellular components (CCs), and 133 molecular functions (MFs), including synaptic and transsynaptic signaling, chemical synaptic transmission, neurotransmitter receptor activity, G protein-coupled amine receptor activity, and serotonin receptor activity, among others. Firmicutes, Bacteroidetes, Coprococcus, and Lachnospiraceae played crucial roles in gut microbiota of a QZD-treated comorbid TS and RRTI model. Conclusions: Our results revealed QZD provided a multicomponent, multitarget, and multipathway synergistic treatment of comorbid TS and RRTI.

Metabolic stimulation-elicited transcriptional responses and biosynthesis of acylated triterpenoids precursors in the medicinal plant Helicteres angustifolia.

BACKGROUND: Helicteres angustifolia has long been used in Chinese traditional medicine. It has multiple pharmacological benefits, including anti-inflammatory, anti-viral and anti-tumor effects. Its main active chemicals include betulinic acid, oleanolic acid, helicteric acid, helicterilic acid, and other triterpenoid saponins. It is worth noting that some acylated triterpenoids, such as helicteric acid and helicterilic acid, are characteristic components of Helicteres and are relatively rare among other plants. However, reliance on natural plants as the only sources of these is not enough to meet the market requirement. Therefore, the engineering of its metabolic pathway is of high research value for enhancing the production of secondary metabolites. Unfortunately, there are few studies on the biosynthetic pathways of triterpenoids in H. angustifolia, hindering its further investigation. RESULTS: Here, the RNAs of different groups treated by metabolic stimulation were sequenced with an Illumina high-throughput sequencing platform, resulting in 121 gigabases of data. A total of 424,824 unigenes were obtained after the trimming and assembly of the raw data, and 22,430 unigenes were determined to be differentially expressed. In addition, three oxidosqualene cyclases (OSCs) and four Cytochrome P450 (CYP450s) were screened, of which one OSC (HaOSC1) and one CYP450 (HaCYPi3) achieved functional verification, suggesting that they could catalyze the production of lupeol and oleanolic acid, respectively. CONCLUSION: In general, the transcriptomic data of H. angustifolia was first reported and analyzed to study functional genes. Three OSCs, four CYP450s and three acyltransferases were screened out as candidate genes to perform further functional verification, which demonstrated that HaOSC1 and HaCYPi3 encode for lupeol synthase and ß-amyrin oxidase, which produce corresponding products of lupeol and oleanolic acid, respectively. Their successful identification revealed pivotal steps in the biosynthesis of acylated triterpenoids precursors, which laid a foundation for further study on acylated triterpenoids. Overall, these results shed light on the regulation of acylated triterpenoids biosynthesis.

Effects of dance therapy on non-motor symptoms in patients with Parkinson's disease: a systematic review and meta-analysis.

BACKGROUND: Parkinson's disease (PD) represents the second most common neurodegenerative disease. OBJECTIVE: To evaluate the effects of dance therapy (DT) aimed at improving non-motor symptoms in PD. METHODS: Studies were performed through PubMed, Web of Science, The Cochrane Library, Embase, and Science Direct from inception to October 27, 2021. The data were screened independently by two reviewers, and the quality of the papers was assessed using the Cochrane manual. The included studies were randomized controlled trials and quasi-randomized controlled trials, reporting random-effects standardized mean differences, and 95% confidence intervals as the effect size. I2 statistics were used to assess heterogeneity. The main outcomes included the Montreal Cognitive Assessment Scale (MOCA), Baker Depression Scale (BDI), Parkinson's Fatigue Scale (FPS-16), and Apathy Scale (AS). RevMan 5.3 software was integrated for meta-analysis. RESULTS: Nine literatures were analyzed for the meta-analysis with a total of 307 patients. Random effects showed that DT significantly improved cognitive of PD (MD = 1.50, 95% CI [0.52, 2.48], P = 0.0003; I2 = 51%). However, this meta-analysis demonstrated that dance therapy had no significance for improving depression (MD = - 1.33, 95% CI [- 4.11, 1.45], P = 0.35; I2 = 79%), fatigue (MD = 0.26, 95% CI [- 0.31, 0.83], P = 0.37; I2 = 0%), and apathy (MD = 0.07, 95% CI [- 2.55, 2.69], P = 0.96; I2 = 50%). CONCLUSION: The meta-analysis suggests that dance can improve cognitive function in PD.

Chitin combined with selenium reduced nitrogen loss in soil and improved nitrogen uptake efficiency in Guanxi pomelo orchard.

Nitrogen cycling in soil, which associated with microbes, plays an important role in plant growth. Irrational application of nitrogen fertilizer could disrupt the structure of soil microbial community, thus inhibiting the uptake of nitrogen by plants and increasing nitrogen leaching in soil. Field and pot leaching experiments with the combined application of chitin fertilizer and selenium (Se) were carried out in order to develop an approach to improve the efficiency of nitrogen fertilizer utilization and reduce runoff by nitrogen loss in orchards of Guanxi pomelo in Fujian Province. Our results showed that application of chitin fertilizer combined with Se to the soil with reduced nitrogen and phosphate fertilizer (nitrogen fertilizer decreased by 25% and phosphate fertilizer decreased by 50%) could significantly increase the fruit yield, vitamin C and solid-acid ratio in the fruit. The application of chitin fertilizer and Se can not only lead to the increase of total nitrogen content in plant leaves but also the alkali-hydrolyzable nitrogen content in soil which enhancing soil nitrogen supplying capacity. It has been found that the adding the chitin fertilizer and Se into soil can significantly affect the structure and functional categories microbial communities and its activities. This is directly evidenced by the findings that the expression level of several groups of N metabolism and transporting related genes (i.e. amoAB and nxrA in nitrification, narG, nirK, norBC, and nosZ in denitrification, nirD, narH in dissimilatory nitrogen reduction, and ureC in ammoniation) has been drastically up-regulated. Our results indicate this strategy for reducing N and P input while maintaining and improving plant performance by supplementing with micronutrient Se and chitin fertilizer can increase the fruit yield and improve the quality of Guanxi pomelo through improving fertilizer use efficiency.

Enhancement and improvement of selenium in soil to the resistance of rape stem against Sclerotinia sclerotiorum and the inhibition of dissolved organic matter derived from rape straw on mycelium.

Sclerotinia stem rot (SSR), caused by Sclerotinia sclerotiorum (S. sclerotiorum), one of the most destructive diseases in many crops including Brassica napus L. The extensive use of fungicides to control S. sclerotiorum caused severe damage to the environment in the long term. Increasing study reported that selenium (Se) is a beneficial element for plant by promoting growth and enhancing disease resistance. In this study, it was found that Se in soil shortened lesion length by 19.14% on rape stem infected with S. sclerotiorum. While resistance mechanism of rape stem against S. sclerotiorum remains unknown. Transcriptomic analysis of rape stem was performed and the results indicated that genes related to antifungal pathways were up-regulated. Moreover, metabonomic analysis was carried out to study the inhibitive effect of the dissolved organic matter derived from rape straw with Se pretreatment in soil (RSDOMSe) on S. sclerotiorum mycelium, results showed that RSDOMSe caused severe damage to energy metabolism of mycelium. Further study indicated that RSDOMSe decreased the pathogenicity of mycelium on rape leaves significantly, and enhanced content of chlorophyII, carotenoids, OD phenol and activities of phenylalanine ammonia-lyase (PAL), polyphenol oxidase (PPO) in rape leaves, which suggested that RSDOMSe plays a positive role in regulating oxidative stress responses of plant when infected with S. sclerotiorum. In addition, when compared with dimcthachlon (DIM) treatment alone, DIM combined with RSDOMSe resulted in higher inhibition on mycelial growth of S. sclerotiorum (the inhibition ratio of nearly 60%). Results in this study suggested that Se enhanced the resistance of rape stem against S. sclerotiorum because of the up-regulated genes related to antifungal pathways, and RSDOMSe improved the mycelial growth inhibition and decreased the pathogenicity of mycelium on rape leaves. Overall, Se as well as Se-enrich byproducts, possessed great potential to be developed as ecological fungicides for controlling S. sclerotiorum.