Terpenoid Biosynthesis Pathway Research Articles

Quantitative Trait Locus Mapping and Candidate Gene Analysis of the Contents of Three Tanshinone Components in Salvia miltiorrhiza Bunge.

Tanshinones are abietane diterpenoid quinone compounds with diverse biological activities and pharmacological effects found in Salvia miltiorrhiza. Leveraging the high-density genetic map established through our prior research endeavors, we conducted a quantitative trait locus (QTL) analysis pertaining to the concentrations of three major tanshinone components, cryptotanshinone, tanshinone I, and tanshinone IIA, in S. miltiorrhiza. This extensive investigation was conducted across three distinct planting environments, ultimately identifying a comprehensive repertoire of 27 discernible QTLs. These QTLs were mapped onto four distinct linkage groups (LG), namely LG1, LG5, LG6, and LG7, which explained 3.11%-37.85% phenotypic variation. Candidate genes were projected based on consistent QTLs detected for each active ingredient in three environments. Nineteen putative candidate genes involved in the regulation of tanshinone biosynthesis were identified. These genes participate in primary metabolic and multiple branching terpenoid biosynthesis pathways, forming a complex regulatory network. Our findings have the potential to offer novel insights into advancing the understanding of the regulatory mechanisms governing tanshinone biosynthesis. Furthermore, these results establish crucial groundwork for gene discovery, marker-assisted selection breeding, and map-based cloning of functional genes associated with tanshinone content in S. miltiorrhiza.

Widely targeted volatilomics and transcriptome analyses reveal the differences in volatile organic components in differently shaped Amomum tsao-ko fruits

BackgroundAmomum tsao-ko is an important aromatic crop used in medicines and food. It can be categorized into three main types based on the fruit shape: long (L), oval (O), and round (R). However, limited information is available on the volatile substances present in differently shaped A. tsao-ko fruits. This study investigated the characteristics and biosynthesis of volatile organic compounds (VOCs) in fresh and dried A. tsao-ko fruits of different shapes using widely targeted volatilomics and transcriptome analyses.ResultsIn total, 978 VOCs, primarily terpenoids, esters, and heterocyclic compounds, were detected. The number of differentially accumulated volatile organic compounds (DAVOCs) in dried fruits of various shapes was significantly higher than that in fresh fruits, with terpenoids, esters, and heterocyclic compounds accounting for approximately 50% of the total DAVOCs. Notably, α-phellandrene, identified as a shared differential accumulated terpenoid across various fruit shapes, was detected in both fresh and dried fruits. Through transcriptome analysis, 40 candidate genes implicated in the terpenoid biosynthesis pathway were screened. An integrated analysis of the metabolome and transcriptome revealed that the structural genes HMGR-2, TPS7, TPS5-10, TPS21-3, TPS21-5, TPS21-6, TPS21-7, and TPS21-9, along with 81 transcription factors (including 17 NACs, 16 MYBs, 16 AP2/ERFs, 13 WRKYs, 13 bHLHs, and 6 bZIPs), co-regulate the biosynthesis of volatile terpenoids.ConclusionsThis study expands our understanding of the volatile metabolism profile of A. tsao-ko and provides a solid foundation for future investigations of the mechanisms governing fruit quality.

Transcriptome Analyses Reveal Differences in the Metabolic Pathways of the Essential Oil Principal Components of Different Cinnamomum Chemotypes

The genus Cinnamomum exhibits a rich variety of chemotypes and is an economically important essential oil (EO)-producing plant belonging to the family Lauraceae. Here, we aimed to explore the potential differences in the terpenoid (the principal components of EOs) biosynthesis pathways of different chemotypes at the molecular level in four Cinnamomum species—C. camphora var. linaloolifera, C. kanehirae, C. longipaniculatum, and C. micranthum. Gas chromatography–mass spectrometry (GC-MS) was employed to elucidate the discrepancies in the chemical profiles and compositions of leaf EO terpenoids among the four Cinnamomum species. The results revealed significant variations in leaf EO yields. The main constituents of the leaf EOs from C. camphora var. linaloolifera and C. kanehirae were the acyclic monoterpene linalool, and those of C. longipaniculatum and C. micranthum were the monoterpene eucalyptol and the sesquiterpene β-caryophyllene, respectively. Furthermore, a comparative transcriptome analysis of the leaves from the four Cinnamomum species revealed that differentially expressed genes (DEGs) were significantly enriched in terpene-related entries. Specifically, 42 and 24 DEGs were significantly enriched to the mevalonate (MVA)/2-methylerythritol 4-phosphate (MEP) pathways and terpene synthase (TPS) activity, respectively. Most genes encoding proteins involved in the terpenoid precursor MVA and MEP pathways exhibited differential expression across the four species, which correlated with the distinct terpenoid profiles observed in their leaf EOs. Four acyclic monoterpene linalool synthase genes—Maker00024100, Maker00014813, Maker00014818, and Maker00018424—were highly expressed in C. camphora var. linaloolifera and C. kanehirae. A monoterpene eucalyptol synthesis gene, Maker00001509, was highly expressed in C. longipaniculatum, and a sesquiterpene β-stigmasterol synthesis gene, Maker00005791, was highly expressed in C. micranthum. These expression levels were subsequently validated through quantitative real-time polymerase chain reaction (qRT-PCR). In conclusion, the combined results of the GC-MS and transcriptome analyses revealed a strong correlation between the metabolite content of the EOs and gene expression. This research contributes to a better understanding of the differences in terpene accumulation in various chemotypes of Cinnamomum at the molecular and mechanistic levels, laying a solid foundation for the cultivation of an ideal Cinnamomum variety.

Pangenome Identification and Analysis of Terpene Synthase Gene Family Members in Gossypium.

Terpene synthases (TPSs), key gatekeepers in the biosynthesis of herbivore-induced terpenes, are pivotal in the diversity of terpene chemotypes across and within plant species. Here, we constructed a gene-based pangenome of the Gossypium genus by integrating the genomes of 17 diploid and 10 tetraploid species. Within this pangenome, 208 TPS syntelog groups (SGs) were identified, comprising 2 core SGs (TPS5 and TPS42) present in all 27 analyzed genomes, 6 softcore SGs (TPS11, TPS12, TPS13, TPS35, TPS37, and TPS47) found in 25 to 26 genomes, 131 dispensable SGs identified in 2 to 24 genomes, and 69 private SGs exclusive to a single genome. The mutational load analysis of these identified TPS genes across 216 cotton accessions revealed a great number of splicing variants and complex splicing patterns. The nonsynonymous/synonymous Ka/Ks value for all 52 analyzed TPS SGs was less than one, indicating that these genes were subject to purifying selection. Of 208 TPS SGs encompassing 1795 genes, 362 genes derived from 102 SGs were identified as atypical and truncated. The structural analysis of TPS genes revealed that gene truncation is a major mechanism contributing to the formation of atypical genes. An integrated analysis of three RNA-seq datasets from cotton plants subjected to herbivore infestation highlighted nine upregulated TPSs, which included six previously characterized TPSs in G. hirsutum (AD1_TPS10, AD1_TPS12, AD1_TPS40, AD1_TPS42, AD1_TPS89, and AD1_TPS104), two private TPSs (AD1_TPS100 and AD2_TPS125), and one atypical TPS (AD2_TPS41). Also, a TPS-associated coexpression module of eight genes involved in the terpenoid biosynthesis pathway was identified in the transcriptomic data of herbivore-infested G. hirsutum. These findings will help us understand the contributions of TPS family members to interspecific terpene chemotypes within Gossypium and offer valuable resources for breeding insect-resistant cotton cultivars.

Multi-omics strategy to reveal the numbing taste of Zanthoxylum armatum DC. at different developmental stages

Green Sichuan pepper (Zanthoxylum armatum DC.) holds important development and utilization values in the food and pharmaceutical industries due to the special pungent and numbing taste of their fruits. Interactions among genes, metabolites and proteins are activated at the molecular level during the development of Z. armatum fruits. In this study, five key stages of Z. armatum development were characterized using the transcriptome, metabolome and proteome, and a comprehensive analysis of the numbing taste was performed. Transcriptome enrichment analyses showed that the differentially expressed genes were mainly associated with flavonoid biosynthesis and tyrosine metabolism. The results also revealed that there were significant differences in the accumulation of metabolites such as fatty acids, tyrosine metabolism, terpenoids, and alkaloids in Z. armatum at different developmental stages. Proteome enrichment analyses indicated that these proteins were mainly involved in flavonoid biosynthesis, and terpenoid backbone biosynthesis. Integrated analysis of transcriptome, metabolome and proteome reveals that different developmental stages of Z. armatum were associated with flavonoid biosynthesis, fatty acids and terpenoid biosynthesis pathways, contributing to its numbing taste and medicinal value. These findings provide novel insights into the numbing taste of Z. armatum, and are expected to inform optimal harvesting times for enhanced economic benefits.

Dynamic DNA methylation modification in catechins and terpenoids biosynthesis during tea plant (Camellia sinensis) leaf development

DNA methylation plays important roles in regulating gene expression during development. However, little is known about the influence of DNA methylation on secondary metabolism during leaf development in the tea plant (Camellia sinensis). In this study, we combined the methylome, transcriptome, and metabolome to investigate the dynamic changes in DNA methylation and its potential regulatory roles in secondary metabolite biosynthesis. In this study, the level of genomic DNA methylation increased as leaf development progressed from tender to old leaf. It additionally exhibited a similar distribution across the genomic background at the two distinct developmental stages studied. Notably, integrated analysis of transcriptomic and methylomic data showed that DNA hypermethylation primarily occurred in genes of the phenylpropanoid, flavonoid, and terpenoid biosynthesis pathways. The effect of methylation on transcription of these secondary metabolite biosynthesis genes was dependent on the location of methylation (i.e., in the promoter, gene or intergenic regions) and the sequence context (i.e., CpG, CHG, or CHH). Changes in the content of catechins and terpenoids were consistent with the changes in gene transcription and the methylation state of structural genes, such as serine carboxypeptidase-like acyltransferases 1A (SCPL1A), leucoanthocyanidin reductase (LAR), and nerolidol synthase (NES). Our study provides valuable information for dissecting the effects of DNA methylation on regulation of genes involved in secondary metabolism during tea leaf development.

Molecular Regulatory Mechanisms Affecting Fruit Aroma.

Aroma, an important quality characteristic of plant fruits, is produced by volatile organic compounds (VOCs), mainly terpenes, aldehydes, alcohols, esters, ketones, and other secondary metabolites, in plant cells. There are significant differences in the VOC profile of various fruits. The main pathways involved in the synthesis of VOCs are the terpenoid, phenylalanine, and fatty acid biosynthesis pathways, which involve several key enzyme-encoding genes, transcription factors (TFs), and epigenetic factors. This paper reviews the main synthetic pathways of the main volatile components in fruit, summarizes studies on the regulation of aroma formation by key genes and TFs, summarizes the factors affecting the fruit aroma formation, describes relevant studies on the improvement of fruit flavor quality, and finally proposes potential challenges and prospects for future research directions. This study provides a theoretical basis for the further precise control of fruit aroma quality and variety improvement.

Comparative transcriptome analysis reveals genes involved in trichome development and metabolism in tobacco

BackgroundThe glandular trichomes of tobacco (Nicotiana tabacum) can efficiently produce secondary metabolites. They act as natural bioreactors, and their natural products function to protect plants against insect-pests and pathogens and are also components of industrial chemicals. To clarify the molecular mechanisms of tobacco glandular trichome development and secondary metabolic regulation, glandular trichomes and glandless trichomes, as well as other different developmental tissues, were used for RNA sequencing and analysis.ResultsBy comparing glandless and glandular trichomes with other tissues, we obtained differentially expressed genes. They were obviously enriched in KEGG pathways, such as cutin, suberine, and wax biosynthesis, flavonoid and isoflavonoid biosynthesis, terpenoid biosynthesis, and plant–pathogen interaction. In particular, the expression levels of genes related to the terpenoid, flavonoid, and wax biosynthesis pathway mainly showed down-regulation in glandless trichomes, implying that they lack the capability to synthesize certain exudate compounds. Among the differentially expressed genes, 234 transcription factors were found, including AP2-ERFs, MYBs, bHLHs, WRKYs, Homeoboxes (HD-ZIP), and C2H2-ZFs. These transcription factor and genes that highly expressed in trichomes or specially expressed in GT or GLT. Following the overexpression of R2R3-MYB transcription factor Nitab4.5_0011760g0030.1 in tobacco, an increase in the number of branched glandular trichomes was observed.ConclusionsOur data provide comprehensive gene expression information at the transcriptional level and an understanding of the regulatory pathways involved in glandular trichome development and secondary metabolism.

Transcriptome analysis unravels differential genes involved in essential oil content in callus and tissue culture seedlings of Lavandula angustifolia

Lavender (Lavandula angustifolia Mill.) essential oil is of high medicinal and economic importance. The preliminary results of this study found that lavender tissue culture seedlings contained 2.1% essential oil, which was 21 times that of callus tissue. However, the mechanisms involved in its synthesis are unclear. This study used transcriptome sequencing of L. angustifolia callus and tissue culture seedlings to compare changes in differentially expressed genes (DEGs) and metabolic pathways. Transcriptome analysis revealed 2394 DEGs in callus and tissue culture seedlings. Several DEGs were enriched in plant hormone signal transduction, as well as the biosynthesis of terpenoids and other secondary metabolites. In the terpenoid biosynthesis pathway, 93 genes were differentially expressed, and four receptors protein genes (BRI1) of brassinosteroid (BR) hormone were significantly differentially expressed. This study provides deeper insights into regulatory genes involved in terpenoid biosynthesis and BR signal transduction in L. angustifolia, contributing to the genetic improvement of L. angustifolia.

The dynamics of N6-methyladenine RNA modification in resistant and susceptible rice varieties responding to rice stem borer damage.

N6-methyladenosine (m6A) is the most prevalent modification in cellular RNA which orchestrates diverse physiological and pathological processes during stress response. However, the differential m6A modifications that cope with herbivore stress in resistant and susceptible crop varieties remain unclear. Here, we found that rice stem borer (RSB) larvae grew better on indica rice (e.g., MH63, IR64, Nanjing 11) than on japonica rice varieties (e.g., Nipponbare, Zhonghua 11, Xiushui 11). Then, transcriptome-wide m6A profiling of representative resistant (Nipponbare) and susceptible (MH63) rice varieties were performed using a nanopore direct RNA sequencing approach, to reveal variety-specific m6A modifications against RSB. Upon RSB infestation, m6A methylation occurred in actively expressed genes in Nipponbare and MH63, but the number of methylation sites decreased across rice chromosomes. Integrative analysis showed that m6A methylation levels were closely associated with transcriptional regulation. Genes involved in herbivorous resistance related to mitogen-activated protein kinase, jasmonic acid (JA), and terpenoid biosynthesis pathways, as well as JA-mediated trypsin protease inhibitors, were heavily methylated by m6A, and their expression was more pronounced in RSB-infested Nipponbare than in RSB-infested MH63, which may have contributed to RSB resistance in Nipponbare. Therefore, dynamics of m6A modifications act as the main regulatory strategy for expression of genes involved in plant-insect interactions, which is attributed to differential responses of resistant and susceptible rice varieties to RSB infestation. These findings could contribute to developing molecular breeding strategies for controlling herbivorous pests.

Cold-stress induced metabolomic and transcriptomic changes in leaves of three mango varieties with different cold tolerance

BackgroundMango (Mangifera indica L.) is grown in Hainan, Guangdong, Yunnan, Sichuan, and Fujian provinces and Guanxi autonomous region of China. However, trees growing in these areas suffer severe cold stress during winter, which affects the yield. To this regard, data on global metabolome and transcriptome profiles of leaves are limited. Here, we used combined metabolome and transcriptome analyses of leaves of three mango cultivars with different cold stress tolerance, i.e. Jinhuang (J)—tolerant, Tainung (T) and Guiremang No. 82 (G)—susceptible, after 24 (LF), 48 (MF) and 72 (HF) hours of cold.ResultsA total of 1,323 metabolites belonging to 12 compound classes were detected. Of these, amino acids and derivatives, nucleotides and derivatives, and lipids accumulated in higher quantities after cold stress exposure in the three cultivars. Notably, Jinhuang leaves showed increasing accumulation trends of flavonoids, terpenoids, lignans and coumarins, and alkaloids with exposure time. Among the phytohormones, jasmonic acid and abscisic acid levels decreased, while N6-isopentenyladenine increased with cold stress time. Transcriptome analysis led to the identification of 22,526 differentially expressed genes. Many genes enriched in photosynthesis, antenna proteins, flavonoid, terpenoid (di- and sesquiterpenoids) and alkaloid biosynthesis pathways were upregulated in Jihuang leaves. Moreover, expression changes related to phytohormones, MAPK (including calcium and H2O2), and the ICE-CBF-COR signalling cascade indicate involvement of these pathways in cold stress responses.ConclusionCold stress tolerance in mango leaves is associated with regulation of primary and secondary metabolite biosynthesis pathways. Jasmonic acid, abscisic acid, and cytokinins are potential regulators of cold stress responses in mango leaves.

Exogenous methyl jasmonate treatment induced the transcriptional responses and accumulation of volatile terpenoids in Oenanthe javanica (Blume) DC

Water dropwort is favored by consumers for its unique flavor and medicinal value. Terpenoids were identified as the main volatile compounds related to its flavor. In this study, water dropwort was treated with different concentrations of exogenous methyl jasmonate (MeJA). The contents of volatile terpenoids were determined under various MeJA treatments. The results indicated that 0.1 mM of MeJA most effectively promoted the biosynthesis of flavor-related terpenoids in water dropwort. Terpinolene accounted the highest proportion among terpene compounds in water dropwort. The contents of jasmonates in water dropwort were also increased after exogenous MeJA treatments. Transcriptome analysis indicated that DEGs involved in the terpenoid biosynthesis pathway were upregulated. The TPS family was identified from water dropwort, and the expression levels of Oj0473630, Oj0287510 and Oj0240400 genes in TPS-b subfamily were consistent with the changes of terpene contents under MeJA treatments. Oj0473630 was cloned from the water dropwort and designated as OjTPS3, which is predicted to be related to the biosynthesis of terpinolene in water dropwort. Subcellular localization indicated that OjTPS3 protein was localized in chloroplast. Protein purification and enzyme activity of OjTPS3 protein were conducted. The results showed that the purified OjTPS3 protein catalyzed the biosynthesis of terpinolene by using geranyl diphosphate (GPP) as substrate in vitro. This study will facilitate to further understand the molecular mechanism of terpenoid biosynthesis and provide a strategy to improve the flavor of water dropwort.

Studies on Differentially Expressed Genes of Tetrastigma hemsleyanum and Its Antitumor Activity

Objective: To analyze highly expressed genes in Tetrastigma hemsleyanum roots and the in vitro antitumor effects of extracts from different parts of this plant. Methods: The expression profiles of the roots, stems, and leaves of T hemsleyanum were analyzed via high-throughput sequencing technology with the Illumina Genome Analyzer platform. Then, gene ontology (GO) functional and pathway enrichment analyses of the significantly differentially expressed genes were performed. The inhibition rates of the extracts from different plant parts were determined through the CCK-8 method. Results: Genes related to catalytic activity and cytochrome P450 were upregulated in roots compared with leaves. The genes that we screened that were expressed at high levels in the roots may play an important role in the accumulation of active ingredients. GO functional enrichment analysis revealed that there were substantial differences in genes related to metabolic processes in roots compared with those in stems, and KEGG analysis revealed significant differences in genes related to the zeatin biosynthesis and diterpenoid biosynthesis pathways. Moreover, the expression of genes related to catalytic activity, ion binding, and hydrolase activity differed significantly between the roots and leaves, and KEGG analysis revealed significant differences in the expression of genes related to terpenoid biosynthesis and flavonoid biosynthesis pathways. Quantitative PCR indicated that Thhg was highly expressed in roots. The total triterpene content was also high in the tissue. The CCK-8 assays showed that the root ethanol extract had the highest antitumor activity. Conclusion: The genes we screened that were expressed at high levels in the roots may play an important role in the accumulation of active ingredients. Importantly, stem and leaf ethanol extracts also showed certain antitumor effects on human HCC Huh-7 and Hep3B cells in vitro and may also have medicinal value.

Genome sequencing and functional analysis of a multipurpose medicinal herb Tinospora cordifolia (Giloy).

Tinospora cordifolia (Willd.) Hook.f. & Thomson, also known as Giloy, is among the most important medicinal plants that have numerous therapeutic applications in human health due to the production of a diverse array of secondary metabolites. To gain genomic insights into the medicinal properties of T. cordifolia, the genome sequencing was carried out using 10× Genomics linked read and Nanopore long-read technologies. The draft genome assembly of T. cordifolia was comprised of 1.01 Gbp, which is the genome sequenced from the plant family Menispermaceae. We also performed the genome size estimation for T. cordifolia, which was found to be 1.13 Gbp. The deep sequencing of transcriptome from the leaf tissue was also performed. The genome and transcriptome assemblies were used to construct the gene set, resulting in 17,245 coding gene sequences. Further, the phylogenetic position of T. cordifolia was also positioned as basal eudicot by constructing a genome-wide phylogenetic tree using multiple species. Further, a comprehensive comparative evolutionary analysis of gene families contraction/expansion and multiple signatures of adaptive evolution was performed. The genes involved in benzyl iso-quinoline alkaloid, terpenoid, lignin and flavonoid biosynthesis pathways were found with signatures of adaptive evolution. These evolutionary adaptations in genes provide genomic insights into the presence of diverse medicinal properties of this plant. The genes involved in the common symbiosis signalling pathway associated with endosymbiosis (Arbuscular Mycorrhiza) were found to be adaptively evolved. The genes involved in adventitious root formation, peroxisome biogenesis, biosynthesis of phytohormones, and tolerance against abiotic and biotic stresses were also found to be adaptively evolved in T. cordifolia.

The reference genome sequence of Artemisia argyi provides insights into secondary metabolism biosynthesis.

Artemisia argyi, a perennial herb of the genus Artemisia in the family Asteraceae, holds significant importance in Chinese traditional medicine, referred to as "Aicao". Here, we report a high-quality reference genome of Artemisia argyi L. cv. beiai, with a genome size up to 4.15 Gb and a contig N50 of 508.96 Kb, produced with third-generation Nanopore sequencing technology. We predicted 147,248 protein-coding genes, with approximately 68.86% of the assembled sequences comprising repetitive elements, primarily long terminal repeat retrotransposons(LTRs). Comparative genomics analysis shows that A. argyi has the highest number of specific gene families with 5121, and much more families with four or more members than the other 6 plant species, which is consistent with its more expanded gene families and fewer contracted gene families. Furthermore, through transcriptome sequencing of A. argyi in response to exogenous MeJA treatment, we have elucidated acquired regulatory insights into MeJA's impact on the phenylpropanoid, flavonoid, and terpenoid biosynthesis pathways of A. argyi. The whole-genome information obtained in this study serves as a valuable resource for delving deeper into the cultivation and molecular breeding of A. argyi. Moreover, it holds promise for enhancing genome assemblies across other members of the Asteraceae family. The identification of key genes establishes a solid groundwork for developing new varieties of Artemisia with elevated concentrations of active compounds.

Antimicrobial mechanism of Limosilactobacillus fermentum SHY10 metabolites against pickle film-producing strain by metabolomic and transcriptomic analysis

The antimicrobial mechanism of Limosilactobacillus fermentum SHY10 metabolites on the pickle film-producing strain Candida parapsilosis B7 was investigated in this study. The results showed that SHY10 metabolites mainly consisted of peptides, alkaloids, nucleic acids, hormones and transmitters, and sterol lipids. Peptides and alkaloids accounted respectively for 15.5% and 22.7% of the total. The SHY10 metabolites induced pits on the cell membrane and death of film-producing strain, which were evidenced by scanning electron microscope and fluorescence. Metabolomic results showed that SHY10 metabolites had the most significant effect on the purine metabolic pathway of Candida parapsilosis B7. Transcriptome results showed that 505 genes of Candida parapsilosis B7 related to peroxisome, α-linolenic acid metabolism, ubiquinone and other terpenoid quinone biosynthesis were remarkably changed with the presence of SHY10 metabolites. The expression of PEX 3, PEX 12, ACOX 1 and wrbA determined by qRT-PCR were consistent with the transcriptome results. In summary, the SHY10 metabolites changed the expression of PEX 3, PEX 12, ACOX 1 and wrbA and affected peroxisome, α-linolenic acid metabolism, ubiquinone and other terpenoid quinone biosynthesis pathways, and eventually led to the death of Candida parapsilosis B7.

Preharvest application of MeJA enhancing the quality of postharvest grape berries via regulating terpenes biosynthesis and phenylpropanoid metabolisms

Methyl jasmonate (MeJA) is an important phytohormone that regulates the development of grape, but the effect and underpin mechanism of its preharvest application on secondary metabolites accumulation in postharvest grape berries are still unclear. In this study, the transcriptome profiles combined with metabolic components analysis were used to determine the effect of preharvest MeJA application on the quality formation of postharvest rose-flavor table grape Shine Muscat. The results indicated that preharvest MeJA treatment had no significant effect on TSS content, but had a down-regulation effect on the accumulation of reducing sugar and titratable acid in the berries. The content of chlorophylls and carotenoids in treated berries was significantly higher than that of the control. Many phenolic components, such as trans-ferulic acid, resveratrol, quercetin, and kaempferol, were sensitive to MeJA and their contents were also significantly higher than that of the control under MeJA treatments during the shelf life. Compared with other volatile aroma components, terpenoid components were more sensitive to preharvest MeJA signals, the content of which presented an overall upward trend with increasing MeJA concentration and prolonging storage time. Furthermore, most of the differentially expressed genes in the general phenylpropanoid pathway and terpenoid biosynthesis pathway were up-regulated responding to MeJA signals. The most upregulated regulatory factors, such as VvWRKY72, VvMYB24, and VvWRI1, may be involved in MeJA signal transduction and regulation. Preharvest MeJA may be an effective technique for enhancing the quality of postharvest Shine Muscat grape berries, with its positive effect on enhancing the characteristic aroma and nutritional components.

Evolutionary flexibility and rigidity in the bacterial methylerythritol phosphate (MEP) pathway.

Terpenoids are a diverse class of compounds with wide-ranging uses including as industrial solvents, pharmaceuticals, and fragrances. Efforts to produce terpenoids sustainably by engineering microbes for fermentation are ongoing, but industrial production still largely relies on nonrenewable sources. The methylerythritol phosphate (MEP) pathway generates terpenoid precursor molecules and includes the enzyme Dxs and two iron-sulfur cluster enzymes: IspG and IspH. IspG and IspH are rate limiting-enzymes of the MEP pathway but are challenging for metabolic engineering because they require iron-sulfur cluster biogenesis and an ongoing supply of reducing equivalents to function. Therefore, identifying novel alternatives to IspG and IspH has been an on-going effort to aid in metabolic engineering of terpenoid biosynthesis. We report here an analysis of the evolutionary diversity of terpenoid biosynthesis strategies as a resource for exploration of alternative terpenoid biosynthesis pathways. Using comparative genomics, we surveyed a database of 4,400 diverse bacterial species and found that some may have evolved alternatives to the first enzyme in the pathway, Dxs making it evolutionarily flexible. In contrast, we found that IspG and IspH are evolutionarily rigid because we could not identify any species that appear to have enzymatic routes that circumvent these enzymes. The ever-growing repository of sequenced bacterial genomes has great potential to provide metabolic engineers with alternative metabolic pathway solutions. With the current state of knowledge, we found that enzymes IspG and IspH are evolutionarily indispensable which informs both metabolic engineering efforts and our understanding of the evolution of terpenoid biosynthesis pathways.

Deciphering the key pathway for triterpenoid biosynthesis in Azadirachta indica A. Juss.: a comprehensive review of omics studies in nature's pharmacy.

Since ancient times, Azadirachta indica, or Neem, has been a well-known species of plant that produces a broad range of bioactive terpenoid chemicals that are involved in a variety of biological functions. Understanding the molecular mechanisms that are responsible for the biosynthesis and control of terpenoid synthesis is majorly dependent on successfully identifying the genes that are involved in their production. This review provides an overview of the recent developments concerning the identification of genes in A. indica that are responsible for the production of terpenoids. Numerous candidate genes encoding enzymes that are involved in the terpenoid biosynthesis pathway have been found through the use of transcriptomic and genomic techniques. These candidate genes include those that are responsible for the precursor synthesis, cyclization, and modification of terpenoid molecules. In addition, cutting-edge omics technologies, such as metabolomics and proteomics, have helped to shed light on the intricate regulatory networks that govern terpenoid biosynthesis. These networks are responsible for the production of terpenoids. The identification and characterization of genes involved in terpenoid biosynthesis in A. indica presents potential opportunities for genetic engineering and metabolic engineering strategies targeted at boosting terpenoid production as well as discovering novel bioactive chemicals.

Macroevolution of floral scent chemistry across radiations of male euglossine bee-pollinated plantsMacroevolución de olores florales a través de radiaciones de plantas polinizadas por abejas euglosinas machosMacroevolução dos voláteis florais em radiações de plantas polinizadas por machos de abelhas Euglossini.

Floral volatiles play key roles as signaling agents that mediate interactions between plants and animals. Despite their importance, few studies have investigated broad patterns of volatile variation across groups of plants that share pollinators, particularly in a phylogenetic context. The "perfume flowers," Neotropical plant species exhibiting exclusive pollination by male euglossine bees in search of chemical rewards, present an intriguing system to investigate these patterns due to the unique function of their chemical phenotypes as both signaling agents and rewards. We leverage recently developed phylogenies and knowledge of biosynthesis, along with decades of chemical ecology research, to characterize axes of variation in the chemistry of perfume flowers, as well as understand their evolution at finer taxonomic scales. We detect pervasive chemical convergence, with many species across families exhibiting similar volatile phenotypes. Scent profiles of most species are dominated by compounds of either the phenylpropanoid or terpenoid biosynthesis pathways, while terpenoid compounds drive more subtle axes of variation. We find recapitulation of these patterns within two independent radiations of perfume flower orchids, in which we further detect evidence for the rapid evolution of divergent floral chemistries, consistent with the putative importance of scent in the process of adaptation and speciation.