RESUMEN
Biosynthesis of Amaryllidaceae alkaloids (AA) starts with the condensation of tyramine with 3,4-dihydroxybenzaldehyde. The latter derives from the phenylpropanoid pathway that involves modifications of trans-cinnamic acid, p-coumaric acid, caffeic acid, and possibly 4-hydroxybenzaldehyde, all potentially catalyzed by hydroxylase enzymes. Leveraging bioinformatics, molecular biology techniques, and cell biology tools, this research identifies and characterizes key enzymes from the phenylpropanoid pathway in Leucojum aestivum. Notably, we focused our work on trans-cinnamate 4-hydroxylase (LaeC4H) and p-coumaroyl shikimate/quinate 3'-hydroxylase (LaeC3'H), two key cytochrome P450 enzymes, and on the ascorbate peroxidase/4-coumarate 3-hydroxylase (LaeAPX/C3H). Although LaeAPX/C3H consumed p-coumaric acid, it did not result in the production of caffeic acid. Yeasts expressing LaeC4H converted trans-cinnamate to p-coumaric acid, whereas LaeC3'H catalyzed specifically the 3-hydroxylation of p-coumaroyl shikimate, rather than of free p-coumaric acid or 4-hydroxybenzaldehyde. In vivo assays conducted in planta in this study provided further evidence for the contribution of these enzymes to the phenylpropanoid pathway. Both enzymes demonstrated typical endoplasmic reticulum membrane localization in Nicotiana benthamiana adding spatial context to their functions. Tissue-specific gene expression analysis revealed roots as hotspots for phenylpropanoid-related transcripts and bulbs as hubs for AA biosynthetic genes, aligning with the highest AAs concentration. This investigation adds valuable insights into the phenylpropanoid pathway within Amaryllidaceae, laying the foundation for the development of sustainable production platforms for AAs and other bioactive compounds with diverse applications.
Asunto(s)
Alcaloides de Amaryllidaceae , Proteínas de Plantas , Transcinamato 4-Monooxigenasa , Proteínas de Plantas/metabolismo , Proteínas de Plantas/genética , Transcinamato 4-Monooxigenasa/metabolismo , Transcinamato 4-Monooxigenasa/genética , Alcaloides de Amaryllidaceae/metabolismo , Sistema Enzimático del Citocromo P-450/metabolismo , Sistema Enzimático del Citocromo P-450/genética , Ácidos Cumáricos/metabolismo , Regulación de la Expresión Génica de las PlantasRESUMEN
BACKGROUND: Expansins are cell wall proteins loosening plant cell in pH-dependent manner. This study aimed to investigate the role of AtEXPA18 in different morphological, physiological, and cellular responses of transgenic tobacco plants to moderate and severe drought stress. METHODS AND RESULTS: Previously synthesized AtEXPA18 gene construct was successfully transferred to the tobacco plants through an agrobacterium-mediate transformation system. Upon obtaining the second generation, tobacco transgenic plants were confirmed by conventional polymerase chain reaction (PCR) technique alongside reverse transcription PCR (RT-PCR) using specific primers. Under drought stress, the transgenic lines showed remarkable growth and significantly improved based on morphological traits such as height and stem diameter, leaf area, leaf number, root dry weight, and Abscisic acid levels of leaves compared control plants. As a result, the Cytokinin content of transgenic plants has increased under severe stress levels. Notably, the area's expansion for abaxial epidermal cells under the microscope confirmed in transgene cells compared with the -transgene cells. CONCLUSION: These results, altogether, could support the AtEXPA18 gene implication in cell expansion and improving tolerance capacity of transgenic crops under drought stress.
Asunto(s)
Proteínas de Arabidopsis/metabolismo , Tolerancia a la Sal/genética , Estrés Fisiológico/genética , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Clorofila/metabolismo , Sequías , Expresión Génica/genética , Regulación de la Expresión Génica de las Plantas/genética , Hojas de la Planta/genética , Proteínas de Plantas/genética , Plantas Modificadas Genéticamente/genética , Estrés Fisiológico/fisiología , Nicotiana/genéticaRESUMEN
The medicinal sage plant (Salvia spp.), belonging to Lamiaceae family, is one of the most important medicinal and aromatic plants. The members of this genus are globally known due to its antimicrobial, antioxidant, astringent, spasmolytic, antihidrotic and specific sensorial properties. In this study, we investigated the potential impact of water deficit on transcript abundance, and essential oil composition of five major metabolites, i.e. 1-8 cineole, α-ß-thujone, camphor, and borneol in three genotypes of Salvia spp. Results showed that relative expression of three genes and their corresponding metabolites increased together at three stages under drought condition, but the CS gene transcript decreased independently from 1,8-cineole in garden sage. Furthermore, borneol changed differently compared to the BS gene expression in control and drought treatment plants of S. reuterana (Yasuj). The competitive synthesis of ß-thujone, and α-thujone by SS gene were demonstrated in S. officinalis and Yasuj ecotype of S. reuterana; whereas, no change was observed for Urmia ecotype of S. reuterana. There was no precursor shortage to synthesis of borneol and camphor in garden sage; however increasing the BS led to high production of borneol and low camphor in S. reuterana under drought stress. As a mechanism, secondary metabolites enable the plants to cope with unfavorable conditions, but genetic differences might affect the quantity and quality of these compounds.
