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Maize terpene synthase 8 (ZmTPS8) produces a blend of sesquiterpenes and contributes to defense against pests and pathogens.

Maize (Zea mays) produces terpenoid-based chemical defenses through a large family of terpene synthases, but the contributions of individual enzymes to specific compounds and stress resistance remain difficult to predict. Maize terpene synthase 8 (ZmTPS8) produces multiple sesquiterpenes in heterologous systems, but its in planta function remains unknown. We integrated a metabolite genome-wide association study (mGWAS), CRISPR/Cas9 generated tps8 loss-of-function mutants, metabolite profiling, and biotic stress assays to define ZmTPS8's role in terpene synthesis and biotic stress responses. The mGWAS identified ZmTPS8 as the primary locus associated with herbivore-induced emission of the sesquiterpene volatile germacrene D. Consistently, ZmTPS8 expression was induced by foliar and root herbivory, and tps8 mutants exhibited reduced emission of germacrene D, α-copaene, and δ-cadinene during Spodoptera frugiperda feeding. Loss of ZmTPS8 increased S. frugiperda larval growth but did not affect the belowground herbivore Diabrotica virgifera virgifera. ZmTPS8 also contributed to resistance against sugarcane mosaic virus, and the fungal pathogen Fusarium verticillioides, affecting terpenoid profiles, global metabolism, and fungal toxin production, but had no impact on Cochliobolus heterostrophus or Pythium spp. susceptibility. Together, these results demonstrate that ZmTPS8 contributes to maize defense in a threat-dependent manner, shaping volatile emissions and defense outcomes.

Zea mays

The homeostasis of β-alanine is key for Arabidopsis reproductive growth and development.

β-Alanine, an abundant non-proteinogenic amino acid, acts as a precursor for coenzyme A and plays a role in various stress responses. However, a comprehensive understanding of its metabolism in plants remains incomplete. Previous metabolic genome-wide association studies (mGWAS) identified ALANINE:GLYOXYLATE AMINOTRANSFERASE2 (AGT2, AT4G39660) linked to β-alanine levels in Arabidopsis under normal conditions. In this study, we aimed to deepen our insights into β-alanine regulation by conducting mGWAS under two contrasting environmental conditions: control (12 h photoperiod, 21°C, 150 μmol m-2 sec-1) and stress (harvested after 1820 min at 32°C and darkness). We identified two highly significant quantitative trait loci (QTL) for β-alanine, including the AGT2 locus associated in both environments and ALDEHYDE DEHYDROGENASE6B2 (ALDH6B2, AT2G14170) associated only under stress conditions. A coexpression-correlation network revealed that the regulatory pathway involving β-alanine levels, AGT2, and ALDH6B2 connects the branched chained amino acid (BCAA) degradation through the propionate pathway. Metabolic profiles of AGT2 overexpression (OE) and knock-out (KO) lines (agt2) across various organs and developmental stages established the critical role of AGT2 in β-alanine metabolism. This work underscores the importance of β-alanine homeostasis for proper growth and development in Arabidopsis.

Arabidopsis

Comparative Genome-Wide Association Studies of Metabolites and Grain-Related Traits in Common Wheat.

The metabolome is highly diverse and the closest layer to phenotype; therefore, it is commonly regarded as a bridge between the genome and phenome in plants. Here, we performed large-scale metabolome analysis using liquid chromatography-tandem mass spectrometry (LC-MS/MS) and 33 grain-related traits in a diverse panel of natural accessions and a recombinant inbred line (RIL) population. We identified a new network of 2286 associations between 947 metabolites and 33 grain-related traits. Systematic integration of metabolic genome-wide association study (mGWAS) and metabolic quantitative trait locus (mQTL) analyses identified 33 566 significant single-nucleotide polymorphisms (SNPs) and 3128 mQTL. Thirteen annotated metabolites co-localized within a physical interval on 7A. Integration of metabolite-based and phenotype-based GWAS and QTL revealed an overlapped region for gibberellin A4 (GA4) content and grain roundness on 4A. Phenotyping of an ethyl methanesulfonate (EMS)-induced mutant confirmed the role of TaSDR in regulating GA4 content and grain morphology. These findings provide novel insights into the metabolic pathways influencing key grain-related traits and advance our understanding of the complex molecular mechanisms regulating grain metabolites and phenotypes in wheat. The identified metabolic markers and candidate genes provide valuable targets for molecular breeding programs aimed at improving wheat yield and quality.

QTL

A multifunctional sesquiterpene synthase integrates with cytochrome P450s to reinforce the terpenoid defense network in maize.

Terpenoids, the largest and most structurally diverse class of plant natural products, play essential roles in maize defense and ecological interactions. In this study, we identified and functionally characterized a sesquiterpenoid-based defense pathway in maize centered on α-santalenoic acid, a pathogen-inducible sesquiterpenoid antibiotic. Using a combination of metabolite-based genome-wide association studies (mGWAS), linkage mapping, and heterologous expression assays, we identified ZmTPS9 as a multiproduct terpene synthase that primarily produces α-santalene and β-bisabolene. Sequence analysis and site-directed mutagenesis revealed that threonine at position 413 is critical for enzyme activity, with its deletion resulting in a complete loss of enzyme activity. The sesquiterpene hydrocarbons produced by ZmTPS9 are further oxidized by three cytochrome P450 monooxygenases, ZmCYP71Z16, ZmCYP71Z18, and ZmCYP71Z19, to yield antimicrobial metabolites including α-santalenoic acid, zealexin D1 (ZD1), and zealexin D2 (ZD2). Together, these findings demonstrate a convergent biosynthetic strategy in maize, where multiproduct terpene synthases and promiscuous P450s collaboratively generate a flexible and robust terpenoid defense network.

Zea mays

Metabolome-based genome-wide association study provides genetic insights into the andrographolide accumulation in Andrographis paniculata.

Andrographis paniculata is a distinctive medicinal plant that produces andrographolide-related metabolites, a class of diterpenoid compounds with potent anti-inflammatory activities. To elucidate the genetic mechanisms underlying the biosynthesis of these compounds, we perform comprehensive metabolic profiling and whole-genome resequencing on a natural population of A. paniculata. Population structure analysis reveals four distinct subgroups characterized by low intra-group genetic diversity but significant inter-group differentiation. Through metabolome-based genome-wide association study, we identify a significant locus associated with 14-deoxyandrographolide content. This locus harbors the candidate gene ApNB-ARC25 (CXN00004106), which encodes an NB-ARC domain-containing resistance protein. Functional characterization using virus-induced gene silencing shows that silencing of ApNB-ARC25 significantly reduces andrographolide accumulation and downregulates expressions of key genes in the andrographolide biosynthetic pathway. Heterologous overexpression of ApNB-ARC25 in rice not only improves resistance to blast disease but also enhances diterpenoid phytoalexin production. Our findings reveal that ApNB-ARC25 promotes diterpenoid accumulation and andrographolide biosynthesis by upregulating key genes involved in terpenoid backbone formation and diterpenoid synthesis. This work not only expands the functional understanding of the ApNB-ARC gene family but also provides a genetic resource for enhancing valuable compound accumulation in medicinal plants, offering important insights into the molecular regulation of medicinal metabolite biosynthesis.

Diterpenes