Search

• TyrAT, PPO, TDC, and SOMT are important in alkaloid synthesis of L. aggregate. • N-methyllaurotetanine accumulates exclusively in roots of L. aggregata. • Isoquinoline alkaloids of L. aggregata accumulate preferentially in roots. • Taproots of L. aggregata can be valuable raw materials for pharmaceutical industry. Lindera aggregata (L. aggregata) is a wild shrub growing in the forests of Southeast Asia, whose main bioactive constituents are isoquinoline alkaloids. They are widely used in food and pharmaceutical industries. The studies on the metabolites and biosynthesis pathways in L. aggregata remain poorly understood. Nine isoquinoline alkaloid compounds were identified by UPLC Triple TOF-MS/MS in this study. Except for N-methyllaurotetanine, most isoquinoline alkaloid compounds were widely distributed in various parts of L. aggregata and accumulated preferentially in roots than in leaves. Transcriptome data showed that several isoquinoline alkaloid biosynthetic genes, such as TyrAT, PPO, TDC, and SOMT, were identified to play important roles in generating differential metabolites in roots and leaves of L. aggregata. Concentration-dependent analgesic effects and toxic effects of them were demonstrated in zebrafish experiments, and the overall ranking was JRAL > TRAL > LAL. The results of this study would provide useful information for the synthesis mechanisms of isoquinoline alkaloids in L. aggregata , and provide valuable information for the application of traditional non-medicinal parts of L. aggregata in food and pharmaceutical industries. [ABSTRACT FROM AUTHOR]

The traditional Chinese herb Rubus chingii Hu (R. chingii) is widely used in clinical practice due to its beneficial effects. Flavonoids are the important class of pharmacological substances in R. chingii , however, the molecular mechanism underlying the differences in active flavonoid contents in R. chingii at different developmental stages remain poorly understood. In this experiment, we selected four developmental stages (GG, GY, YR, RR) of R. chingii as the research material. We studied the untargeted and targeted metabolic profiles of flavonoids in different periods of R. chingii , combining full-length and comparative transcriptome analyses. Functional analyses were conducted on genes implicated in flavonoid differences. GG and RR displayed relatively higher and lower contents of flavonols, flavones, flavanols, flavanones, and isoflavonoid, respectively. RNA-seq analyses showed structural genes such as RcPAL, RcC4H, Rc4CL, RcCHS, RcCHI, RcF3H, RcF3′H, and RcFLS in flavonoid biosynthesis pathway were upregulated in GG, which were essential for the accumulation flavanones, flavones, and flavonols (effective components). qRT-PCR analyses investigated that six structural genes RcCHI, RcF3H, 2 RcCHS, and 2 Rc4CL, two TFs RcMYB308 and RcMYB123 had a consistent expression pattern with which in transcriptome. Also, an interaction network showed that the RcMYB308 could positively regulate Ka3R, Qu, Qu3G, AS, Hy, Ti through RcF3H. Furthermore, Subcellular localization analysis revealed that RcMYB308 was localization to the nucleus. In tobacco, RcMYB308 was overexpressed, resulting in higher flavonoids, RcF3H, RcF3′H, RcCHI, and RcFLS. RcMYB308 upregulated RcF3H in dual-luciferase assays. These results provide new insights for further understanding the molecular mechanism regulating flavonol biosynthesis in R. chingii fruit, and also provide a potential MYB regulator for molecular breeding of R. chingii. • Metabolic profiles with transcriptome analyses were performed in R. chingii. • GG and RR displayed relatively higher and lower contents of flavonoids. • Important structural genes in flavonoid biosynthesis were upregulated in GG. • RcMYB308 led to increased flavonols, RcF3H, RcF3′H, RcCHI, and RcFLS. • Dual-luciferase assays confirmed that RcMYB308 upregulated RcF3H. [ABSTRACT FROM AUTHOR]

Pseudostellariae Radix is an important traditional Chinese medicine (TCM), which is consumed commonly for its positive health effects. However, a lack of transcriptomic and genomic information hinders research on Pseudostellariae Radix . Here, high-throughput RNA sequencing (RNA-seq) was employed for the de novo assembly to analyze the transcriptome in Pseudostellariae Radix , finding significantly differentially expressed genes in this TCM from different fields based on RNA-seq and bioinformatic analysis. A total of 146,408,539 paired-end reads were generated and assembled into 89,857 unigenes with an average length of 862 bp. All of the assembly unigenes were annotated by running BLASTx and BLASTn similarity searches on the Non-redundant nucleotide database (NT), the Non-redundant protein database (NR), Swiss-Prot, Cluster of Orthologous Groups (COG), Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene Ontology (GO), and Interpro. On the basis of bioinformatic analysis and the expression profiles for Pseudostellariae Radix , 29 significantly differentially expressed genes were identified, which provides the basic information for exploring the molecular mechanisms that determine the quality of Pseudostellariae Radix from different fields. The expression levels of 29 genes were validated by real-time quantitative PCR (RT-qPCR). This is the first study to sample Pseudostellariae Radix , which provides an invaluable resource for understanding the genome of this herb. [ABSTRACT FROM AUTHOR]

Pseudostellaria heterophylla is an important traditional Chinese herbal medicine with vast clinical consumption because of its positive effects. To date, changes in the metabolite composition of P. heterophylla have been well documented; however, the molecular differences between cultivated P. heterophylla and its wild-type are still unknown. The aim of this study was to investigate differences in cultivated and wild P. heterophylla . Due to the lack of a genomic database, we used high-throughput transcriptomic and proteomic technologies to identify proteins in the herb. Isobaric tags for relative and absolute quantification (iTRAQ) MS/MS were used to detect statistically significant changes between cultivated and wild P. heterophylla . We detected 3775 proteins; 332 showed differential accumulations across two different ecotypes of P. heterophylla . 71 significant differential expressions of proteins were selected based on GO annotations, KEGG, STRING analysis, and expression level. Quantitative real-time PCR analysis confirmed the authenticity and accuracy of the proteomic analysis. The results indicated that the carbohydrate and cellular amino acid metabolisms in cultivated P. heterophylla were weaker than those in its wild-type; seven important proteins were found to regulate sucrose and amino acids. This will provide the basic information for exploring the cause of differences in secondary metabolites in different ecotypes of P. heterophylla and the protein mechanism of its quality formation. Biological significance This study combined a transcriptome, proteome, and metabolism approach for analyzing differentially expressed proteins of cultivated and wild P. heterophylla and established the relationship between significantly differentially expressed proteins and differential chemical components in non-model plants. The results of proteomic analysis provide the basic information for exploring the quality forming process, which will demonstrate, and provide guidance for, the study of effective constituents of P. heterophylla and its quality formation mechanism. [ABSTRACT FROM AUTHOR]