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Aquilaria sinensis, a plant belonging to the genus Aquilaria of the Thymelaeaceae family, can produce agarwood when exposed to natural factors (such as lightning strikes, fire, and insect bite) or artificial factors (such as cuts, holes, and inoculation of fungi). MADS is a transcription factor that plays a significant role in plant growth and development and stress response. However, its function in A. sinensis remains unclear. To elucidate the stress response of MADS-box genes in A. sinensis after injury, this study utilized transcriptomic data derived from A. sinensis to expression heatmaps and screen for differentially expressed genes, leading to the identification and cloning of a MADS-box encoding gene, designated AsMADS12, which exhibited significant differential expression. Comprehensive analyses were subsequently conducted on the domain architecture, physicochemical properties, phylogenetic relationships, and predicted secondary and tertiary structures of AsMADS12. The expression dynamics of the AsMADS12 at various time points after methyl jasmonate (MeJA) treatment were assessed via real-time quantitative PCR (RT-qPCR). Furthermore, subcellular localization of the AsMADS12 was determined by transiently expressing in onion epidermal cells (Agrobacterium-mediated infiltration) and observing fluorescence under a confocal laser scanning microscope. Sequence and evolutionary analyses revealed that AsMADS12 possessed a complete open reading frame (ORF) of 678 bp, encoding a protein of 255 amino acids. The predicted molecular weight of the encoded protein was 25.28 kDa. AsMADS12 contained a typical MEF2-like MADS domain and was classified as a MIKC-type transcription factor. The secondary structure of the protein was mainly composed of 52.00% α-helix, 11.56% extended chain, and 36.44% random coil. Phylogenetic cluster analysis revealed that AsMADS12 was clustered in the same branch as the MADS of Citrus sinensis and Citrus trifoliata. The subcellular localization experiment showed that the AsMADS12 protein was located in the nucleus. The real-time fluorescence quantitative results indicated that methyl jasmonate treatment could induce the expression of AsMADS12, and the expression level was the highest at 24 hours after treatment. It was speculated that AsMADS12 might play a potential role in the formation of agarwood in A. sinensis. This study completed the cloning of the gene for the first time and conducted preliminary exploration, laying the foundation for further research on whether it participates in the formation of agarwood and its role in biotic or abiotic stress responses, thereby providing a theoretical basis for understanding the molecular regulatory mechanism of agarwood formation in A sinensis.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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