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Gentiana macrophylla is one of Chinese herbal medicines in which 4 kinds of iridoids or secoiridoids, such as loganic acid, sweroside, swertiamarin, and gentiopicroside, are identified as the dominant medicinal secondary metabolites. WRKY, as a large family of transcription factors (TFs), plays an important role in the synthesis of secondary metabolites in plants. Therefore, WRKY genes involved in the biosynthesis of secoiridoids in G. macrophylla were systematically studied. First, a comprehensive genome-wide analysis was performed, and 42 GmWRKY genes were identified, which were unevenly distributed in 12 chromosomes. Accordingly, gene structure, collinearity, sequence alignment, phylogenetic, conserved motif and promoter analyses were performed, and the GmWRKY proteins were divided into three subfamilies based on phylogenetic and multiple sequence alignment analyses. Moreover, the enzyme-encoding genes of the secoiridoid biosynthesis pathway and their promoters were then analysed, and the contents of the four secoiridoids were determined in different tissues. Accordingly, correlation analysis was performed using Pearson′s correlation coefficient to construct WRKY gene-enzyme-encoding genes and WRKY gene–metabolite networks. Meanwhile, G. macrophylla seedlings were treated with methyl jasmonate (MeJA) to detect the dynamic change trend of GmWRKYs , biosynthetic genes, and medicinal ingredient accumulation. Thus, a total of 12 GmWRKYs were identified to be involved in the biosynthesis of secoiridoids, of which 8 ( GmWRKY1 , 6 , 12 , 17 , 33 , 34 , 38 and 39 ) were found to regulate the synthesis of gentiopicroside, and 4 ( GmWRKY7 , 14 , 26 and 41 ) were found to regulate the synthesis of loganic acid. Taken together, this study systematically identified WRKY transcription factors related to the biosynthesis of secoiridoids in G. macrophylla , which could be used as a cue for further investigation of WRKY gene functions in secondary metabolite accumulation.

Panax notoginseng is a highly valued perennial medicinal herb plant in Yunnan Province, China, and the taproots are the main medicinal parts that are rich in active substances of P. notoginseng saponins. The main purpose of this study is to uncover the physiological and molecular mechanism of Panax notoginseng saponin accumulation triggered by methyl jasmonate (MeJA) under arbuscular mycorrhizal fungi (AMF) by determining physiological indices, high-throughput sequencing and correlation analysis. Physiological results showed that the biomass and saponin contents of P. notoginseng , the concentrations of jasmonic acids (JAs) and the key enzyme activities involved in notoginsenoside biosynthesis significantly increased under AMF or MeJA, but the interactive treatment of AMF and MeJA weakened the effect of AMF, suggesting that a high concentration of endogenous JA have inhibitory effect. Transcriptome sequencing results indicated that differential expressed genes (DEGs) involved in notoginsenoside and JA biosynthesis were significantly enriched in response to AMF induction, e.g., upregulated genes of diphosphocytidyl-2-C-methyl-d-erythritol kinases ( ISPEs ), cytochrome P450 monooxygenases ( CYP450s ) _ and glycosyltransferases ( GTs ), while treatments AMF-MeJA and salicylhydroxamic acid (SHAM) decreased the abundance of these DEGs. Interestingly, a high correlation presented between any two of saponin contents, key enzyme activities and expression levels of DEGs. Taken together, the inoculation of AMF can improve the growth and saponin accumulation of P. notoginseng by strengthening the activities of key enzymes and the expression levels of encoding genes, in which the JA regulatory pathway is a key link. This study provides references for implementing ecological planting of P. notoginseng , improving saponin accumulation and illustrating the biosynthesis mechanism.

The Amaryllidaceae alkaloid galanthamine (Gal) in Lycoris longituba is a secondary metabolite that has been used to treat Alzheimer’s disease. Plant secondary metabolism is affected by methyl jasmonate (MeJA) exposure, although the regulatory mechanisms of MeJA on L. longituba seedlings remains largely unknown. In the present study, 75, 150, and 300 μM MeJA were used as treatments on L. longituba seedlings for 7, 14, 21, and 28 days, while 0 μM MeJA was used as the control (MJ-0). The effect of exogenous MeJA on Gal synthesis in L. longituba was then investigated using transcriptomic sequencing and metabolite profiling via GC-MS and LC-MS analysis. Galanthamine (Gal), lycorine (Lyc), and lycoramine (Lycm) abundances were 2. 71-, 2. 01-, and 2.85-fold higher in 75 μM MeJA (MJ-75) treatment plants compared to MJ-0 treatment plants after 7 days of cultivation. Transcriptomic analysis further showed that MJ-75 treatment significantly induced the expression of norbelladine synthase (NBS) and norbelladine 4′-O-methyltransferase (OMT), which are involved in the Gal biosynthesis pathway. In addition, increased expression was observed in MJ-75 treatment plants for genes in the JA synthesis and JA signaling pathways including those of allene oxide cyclase (AOC), 12-oxo-phytodienoic acid reductase (OPR), jasmonic acid amino acid synthase (JAR), and transcription factor MYC. The L. longituba tyrosine decarboxylase (LlTYDC) enzyme was identified and proposed to be involved in the Gal biosynthetic pathway. Metabolomics results demonstrated that the accumulation of Amaryllidaceae alkaloids, and especially alkaloids in the Gal biosynthesis pathway, could be induced by MJ-75 treatment. Interestingly, metabolites in the JA synthesis pathway were also affected by MeJA treatment. Overall, this multi-omics study suggests that both the JA synthesis/JA signaling and Gal biosynthesis pathways were affected by exogenous MeJA treatment. This comprehensive study of gene expression and metabolite contents can help us better understand the molecular mechanisms underlying MeJA-mediated Gal biosynthesis in L. longituba .

Ginseng ( Panax ginseng ) is a globally renowned medicinal plant. The primary active compounds in ginseng are ginsenosides, which have been shown to possess preventive and therapeutic properties against a range of ailments. WNK (with no lysine) kinases is a subfamily of serine/threonine protein kinases. Members of the WNK gene family play vital roles in the regulation of plant growth, development, and biological processes. Several studies have shown that jasmine methyl ester treatment can increase the content of protopanaxatriol-type ginsenosides in adventitious ginseng roots. However, there are no reports on the effects of WNK in ginseng under methyl jasmonate (MeJA) treatment. In this study, we conducted a detailed screening, identification, and systematic research on the WNK gene family in ginseng, including phylogenetic relationships, gene structure, chromosomal distribution, GO functional classification, cis-regulatory elements, co-expression network analysis, and gene expression pattern analysis. Furthermore, we identified candidate genes of PgWNK that could significantly respond to methyl jasmonate regulation and conducted an in-depth study of their expression patterns to explore the impact of these candidate genes on the variation in triterpenoid ginsenoside content. We identified 32 WNK genes in ginseng that were unevenly distributed across 11 of the 24 chromosomes in the ginseng genome. GO functional annotation and enrichment analysis of the PgWNK genes revealed their involvement in numerous biological processes, including ginseng growth, development, and biosynthesis of secondary metabolites. Through expression network analysis, we further understand that there are complex interactions among members of the PgWNK gene family. In addition, 17 non-biological stress response components related to hormone response, growth, and development were predicted in the promoter area of the PgWNK gene. At the same time, we studied the gene expression of PgWNK after MeJA treatment and found that the expression related to ginsenoside biosynthesis was significantly increased as the regulation time was prolonged. Following MeJA regulation, the content of five protopanaxatriol-type ginsenosides (Re, Rf, Rg1, Rg2, and Rh1) increased owing to the adjustment of PgWNK05 gene expression. Therefore, our results provide an important theoretical foundation for understanding the molecular characteristics, genetic evolution, genetic functions, expression patterns, and non-biological coercion of the WNK gene family in ginseng. The underlying research on the biological synthesis of ginsenosides provides a foundation and theoretical support.

Ginseng ( Panax ginseng C. A. Mey.) is an important and valuable medicinal plant species used in traditional Chinese medicine, and its metabolite ginsenoside is the primary active ingredient. The FAR1/FHY3 gene family members play critical roles in plant growth and development as well as participate in a variety of physiological processes, including plant development and signaling of hormones. Studies have indicated that methyl jasmonate treatment of ginseng adventitious roots resulted in a significant increase in the content of protopanaxadiol ginsenosides. Therefore, it is highly significant to screen the FAR1/FHY3 gene family members in ginseng and preliminarily investigate their expression patterns in response to methyl jasmonic acid signaling. In this study, we screened and identified the FAR1/FHY3 family genes in the ginseng transcriptome databases. And then, we analyzed their gene structure and phylogeny, chromosomal localization and expression patterns, and promoter cis-acting elements, and made GO functional annotations on the members of this family. After that, we treated the ginseng adventitious roots with 200 mM methyl jasmonate and investigated the trend of the expression of four genes containing the largest number of methyl jasmonate cis-acting elements at different treatment times. All four genes were able to respond to methyl jasmonate, the most significant change was in the PgFAR40 gene. This study provides data support for subsequent studies of this family member in ginseng and provides experimental reference for subsequent validation of the function of this family member under methyl jasmonic acid signaling.

The large pine weevil ( Hylobius abietis ) is a major regeneration pest in commercial forestry. Pesticide application has historically been the preferred control method, but pesticides are now being phased out in several countries for environmental reasons. There is, thus, a need for alternative plant protection strategies. We applied methyl jasmonate (MeJA), salicylic acid (SA) or oxalic acid (OxA) on the stem of 2-year-old Norway spruce ( Picea abies ) plants to determine effects on inducible defenses and plant growth. Anatomical examination of stem cross-sections 9 weeks after application of 100 mM MeJA revealed massive formation of traumatic resin ducts and greatly reduced sapwood growth. Application of high concentrations of SA or OxA (500 and 200 mM, respectively) induced much weaker physiological responses than 100 mM MeJA. All three treatments reduced plant height growth significantly, but the reduction was larger for MeJA (~55%) than for SA and OxA (34-35%). Lower MeJA concentrations (5-50 mM) induced comparable traumatic resin duct formation as the high MeJA concentration but caused moderate (and non-significant) reductions in plant growth. Two-year-old spruce plants treated with 100 mM MeJA showed reduced mortality after exposure to pine weevils in the field, and this enhanced resistance-effect was statistically significant for three years after treatment.

The dried roots of Scutellaria baicalensis are important traditional Chinese medicine used to treat liver and lung inflammation. An anomalous structure, hollowed root, was discovered in perennial cultivated Scutellaria baicalensis . The presence of the hollow may change the contents of bioactive metabolites, such as baicalein, and other 4’-hydroxyflavones in Scutellaria baicalensis roots, but the relationship between the hollowed root and bioactive metabolite contents is poorly understood. In this study, we identified the anatomical structure of the hollowed root and detected differentially accumulating flavonoid metabolites and enzymes related to 4’-hydroxyflavone biosynthesis in 3-year-old roots with a hollow. We confirmed that methyl jasmonate (MeJA) induced the accumulation of 4’-hydroxyflavones and the expression of enzymes related to 4’-hydroxyflavone biosynthesis in hydroponically cultured Scutellaria baicalensis roots. The development of the hollowed root were divided into 4 stages. The 4’-hydroxyflavone contents and expression of enzymes related to 4’-hydroxyflavone biosynthesis increased synchronously with the content of MeJA during the development of hollowed root. Pathogen and programed-cell-death related genes were induced during hollowed root development. Taken together, our results provide novel insight into the importance of MeJA in the development of hollowed root and the accumulation of 4’-hydroxyflavones in Scutellaria baicalensis roots. Our results suggest that a pathogen and senescence are the two major causes for the development of hollowed root in Scutellaria baicalensis roots.

Background Methyl jasmonate (MeJA), which has been identified as a lipid-derived stress hormone, mediates plant resistance to biotic/abiotic stress. Understanding MeJA-induced plant defense provides insight into how they responding to environmental stimuli. Result In this work, the dynamic network analysis method was used to quantitatively identify the tipping point of growth-to-defense transition and detect the associated genes. As a result, 146 genes were detected as dynamic network biomarker (DNB) members and the critical defense transition was identified based on dense time-series RNA-seq data of MeJA-treated Arabidopsis thaliana . The GO functional analysis showed that these DNB genes were significantly enriched in defense terms. The network analysis between DNB genes and differentially expressed genes showed that the hub genes including SYP121, SYP122, WRKY33 and MPK11 play a vital role in plant growth-to-defense transition. Conclusions Based on the dynamic network analysis of MeJA-induced plant resistance, we provide an important guideline for understanding the growth-to-defense transition of plants’ response to environment stimuli. This study also provides a database with the key genes of plant defense induced by MeJA.

Haplophyllum virgatum var. virgatum is an Iranian endemic plant. Its only natural habitat is the Geno region in the Hormozgan province of Iran. This plant’s limited distribution and special habitat conditions prompted us to investigate the possibility of its micropropagation and different biological aspects. In this study, while evaluating seed germination and seedling production of H. virgatum in two Murashige and Skoog (MS) and Gamborg (B5) culture media, shoot multiplication and rooting of single-node shoot explants of this plant were also investigated in the MS culture medium containing different concentrations of growth regulators. Flavonoids are part of the medicinal metabolites of different Haplophyllum species. Here, the flavonoid elicitation in the proliferated shoots of H. virgatum under 0, 100, and 150 µM methyl jasmonate (MeJA) treatments was done. The results demonstrated that the MS culture medium was more suitable than B5 for seed germination and natural growth of seedlings. Moreover, for H. virgatum shoot multiplication, one mg/L kinetin was more appropriate compared to other concentrations of this cytokinin (0.5, 1.5, 2, 2.5, and 3 mg/L) and the corresponding concentrations of benzyl amino purine. Rooting was achieved only in a limited number of shoots, so it needs further investigation. Meanwhile, MeJA increased total phenolic, total flavonoid, and rutin (the main flavonoid of Rutaceae) contents in the proliferated shoots. This research provided new information about the micropropagation of H. virgatum and flavonoid elicitation by MeJA in this plant.Key messageFor shoot multiplication of Haplophyllum virgatum, MS medium containing one mg/L kinetin showed the best results. Methyl jasmonate increased the phenolic and flavonoid contents of its proliferated shoots.

Panax ginseng is a well-known medicinal plant with several pharmacological uses in China. The trihelix family transcription factors, also known as GT factors, can be involved in the regulation of growth and developmental processes in plants. There have been no in-depth reports or systematic studies about the trihelix transcription factor in ginseng. In this study, the structure, chromosomal localization, gene duplication, phylogeny, functional differentiation, expression patterns and coexpression interactions of trihelix transcripts were analysed using bioinformatics methods based on the ginseng transcriptome database. Thirty-two trihelix transcription factor genes were identified in ginseng, and these genes were alternatively spliced to obtain 218 transcripts. These transcripts were unevenly distributed on different chromosomes of ginseng, and phylogenetic analysis classified the PgGT transcripts into five subgroups. Gene Ontology (GO) analysis classified PgGT transcripts into eight functional subclasses, indicating that they are functionally diverse. The expression pattern analysis of 218 PgGT transcripts revealed that their expression was tissue-specific and spatiotemporally-specific in 14 different tissues of 4-year-old ginseng, 4 different ages of ginseng roots, and 42 farmers’ cultivars of 4-year-old ginseng roots. Despite the differences in the expression patterns of these transcripts, coexpression network analysis revealed that these transcripts could be expressed synergistically in ginseng. In addition, two randomly selected PgGT transcripts in each of the five different subfamilies were subjected to methyl jasmonate treatment at different times, and PgGT was able to respond to the regulation of methy1 jasmonate. These results provide a theoretical basis and gene resources for an in-depth study of the function of trihelix genes in other plants.