RESUMEN
Secondary metabolites play multiple crucial roles in plants by modulating various regulatory networks. The biosynthesis of these compounds is unique to each species and is intricately controlled by a range of developmental and environmental factors. While light's role in certain secondary metabolites is evident, its impact on sterol biosynthesis remains unclear. Previous studies indicate that ELONGATED HYPOCOTYL5 (HY5), a bZIP transcription factor, is pivotal in skotomorphogenesis to photomorphogenesis transition. Additionally, PHYTOCHROME INTERACTING FACTORs (PIFs), bHLH transcription factors, act as negative regulators. To unveil the light-dependent regulation of the mevalonic acid (MVA) pathway, a precursor for sterol biosynthesis, mutants of light signaling components, specifically hy5-215 and the pifq quadruple mutant (pif 1,3,4, and 5), were analyzed in Arabidopsis thaliana. Gene expression analysis in wild-type and mutants implicates HY5 and PIFs in regulating sterol biosynthesis genes. DNA-protein interaction analysis confirms their interaction with key genes like AtHMGR2 in the rate-limiting pathway. Results strongly suggest HY5 and PIFs' pivotal role in light-dependent MVA pathway regulation, including the sterol biosynthetic branch, in Arabidopsis, highlighting a diverse array of light signaling components finely tuning crucial growth pathways.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico , Regulación de la Expresión Génica de las Plantas , Esteroles , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/metabolismo , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/genética , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/metabolismo , Factores de Transcripción con Motivo Hélice-Asa-Hélice Básico/genética , Esteroles/metabolismo , Esteroles/biosíntesis , Proteínas Nucleares/metabolismo , Proteínas Nucleares/genética , Mutación , Luz , Ácido Mevalónico/metabolismoRESUMEN
The Raffinose Family of Oligosaccharides (RFOs), including Galactinol, Raffinose, and Stachyose, are pivotal carbohydrates with significant roles in abiotic stress tolerance and growth within dynamic environments. Plant development is profoundly influenced by light, a major environmental signal. Despite this, the interconnections between the biosynthesis of secondary sugars and light signaling have remained unexplored. This study reveals that exposure to light induces the expression of Galactinol synthase (AtGolS1), a key enzyme in the RFO biosynthesis pathway. The light-inducible response of AtGolS1 operates downstream of ELONGATED HYPOCOTYL 5 (HY5), a central regulator in light signaling. Mutant seedlings with disrupted HY5 function (hy5-215) exhibit reduced AtGolS1 transcript accumulation compared to wild-type (WT) and HY5 overexpression seedlings. DNA-protein interaction studies demonstrate that HY5 directly binds to light-responsive cis-elements in the promoter region of AtGolS1, thereby mediating its light responsiveness. Quantification of galactinol revealed a diminished accumulation in the hy5-215 mutant compared to wild-type (WT) and HY5 overexpression seedlings. Consequently, these findings shed light on the intricate crosstalk between RFO biosynthesis and light signaling in Arabidopsis.
Asunto(s)
Proteínas de Arabidopsis , Arabidopsis , Disacáridos , Galactosiltransferasas , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/genética , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/metabolismo , Regulación de la Expresión Génica de las Plantas , Hipocótilo/metabolismo , Rafinosa/metabolismo , Plantones/genética , Plantones/metabolismoRESUMEN
The terpenoid pathway serves as an essential source of all isoprenoid precursors and metabolites that are of great pharmacological importance. The major enzymes for the synthesis of these diverse molecules is the terpene synthases (TPSs), which catalyse the final step of the synthesis of the important secondary products, the terpenes. Previous studies have reported that the various environmental factors, including light govern the synthesis of terpenoids. However, the molecular components and steps involved in the regulation of synthesis of these molecules have not been studied in detail. In this study, we report that the light regulates the expression of the members of terpene synthase gene family in Arabidopsis thaliana. We demonstrate that ELONGATED HYPOCOTYL (HY5), a basic leucine zipper transcription factor, plays a crucial role in light-mediated transcriptional regulation of terpene synthase, AtTPS03. Expression analysis using hy5-215 mutant and HY5 over-expression lines revealed that HY5 acts as a positive regulator of AtTPS03. Additionally, studies including AtTPS03 Promoter::reporter transgenic lines in wild-type and hy5-215, as well as electrophoretic mobility shift assay (EMSA), suggest an interaction of HY5 with the AtTPS03 promoter. Together, our analysis indicate the requirement for HY5 for light-mediated regulation of AtTPS03 for the terpenoid biosynthesis in Arabidopsis.
Asunto(s)
Transferasas Alquil y Aril/genética , Proteínas de Arabidopsis/genética , Arabidopsis/genética , Factores de Transcripción con Cremalleras de Leucina de Carácter Básico/genética , Regulación de la Expresión Génica de las Plantas , Luz , Mutación , Regulación hacia ArribaRESUMEN
Arsenic (As) is a ubiquitous element, which imposes threat to crops productivity and human health through contaminated food chain. As a part of detoxification mechanism, As is chelated and sequestered into the vacuoles via sulphur containing compounds glutathione (GSH) and phytochelatins (PCs). Under limiting sulphur (LS) conditions, exposure of As leads to enhanced toxic effects in plants. Therefore, it is a prerequisite to understand molecular mechanisms involved in As stress response under sulphur deficiency conditions in plants. In recent years, natural variation has been utilized to explore the genetic determinants linked to plant development and stress response. In this study, natural variation in Arabidopsis has been utilized to understand the molecular mechanisms underlying LS and As(III) stress response. Analysis of different accession of Arabidopsis led to the identification of Koz2-2 and Ri-0 as the most tolerant and sensitive accessions, respectively, towards As(III) and LS+As(III) stress. Biochemical analysis and expression profiling of the genes responsible for sulphur transport and assimilation as well as metal detoxification and accumulation revealed significantly enhanced sulphur assimilation mechanism in Koz2-2 as compared to Ri-0. Analyses suggest that genetic variation regulates differential response of accessions towards As(III) under LS condition.