Computational Molecular Biology 2025, Vol.15, No.2, 84-90 http://bioscipublisher.com/index.php/cmb 87 In addition to jasmonic acid, hormones such as abscisic acid (ABA) and ethylene (ET) are also involved in the regulation of secondary metabolites. Studies have found that some transcription factors of the AP2/ERF family are activated under the regulation of these hormones, thereby affecting the synthesis of metabolites including alkaloids (Xie et al., 2019). Therefore, the synergistic relationship between hormones and transcription factors enables motherwort to flexibly adjust the accumulation of metabolites to better adapt to environmental changes (Zhang et al., 2007). 4.3 The regulatory effect of environmental stress on the accumulation of Leonurus japonicus alkaloids The changes in natural conditions also profoundly affect the accumulation of alkaloids in motherwort. Whether it is extreme temperature, insufficient water, or excessive soil salinity, these stresses will change the hormone levels in plants, thereby triggering a series of stress-related transcription factors and further affecting metabolic pathways. For instance, the AP2/ERF family is considered to have a potential connection with alkaloid synthesis due to its significant role in adverse response (He et al., 2018). What's more interesting is that environmental stress often does not act alone but exerts its influence through interaction with hormone signaling pathways. For instance, jasmonic acid-related transcription factors have been proven to regulate the synthesis of secondary metabolites under adverse conditions, and this mechanism may also exist in motherwort (Mertens et al., 2016). Therefore, this regulatory network can be understood as the way plants "self-regulate" in complex environments. It not only ensures the production of alkaloids but also enhances the defense and survival capabilities of the plants. 5 Molecular Research Methods of Leonurus japonicus Alkaloids 5.1 Application of transcriptomics and metabolomics in the research of Leonurus japonicus alkaloids Transcriptomics and metabolomics are two indispensable tools when exploring the formation and regulation of alkaloids in Leonurus japonicus. Take the transcriptome as an example. It can help researchers identify key enzymes related to metabolic pathways in the vast amount of genetic information. The research on diterpene synthase is a typical case. This type of enzyme is closely related to the formation of complex diterpene structures. The functional analysis not only enables us to see the diversity of plant synthetic capabilities, but also provides important clues for understanding the synthetic pathways of alkaloids (Zhu, 2024). Through the analysis of gene expression profiles in the transcriptome, the regulatory network of alkaloid synthesis can be gradually outlined. Meanwhile, metabolomics offers another perspective. It directly approaches from the metabolite level and uses methods such as LC-MS/MS and HPLC-DAD to conduct a comprehensive detection of the chemical components in Leonurus japonicus extract. In this way, not only can the presence of multiple alkaloids be confirmed, but also other secondary metabolites such as flavonoids and phenolic acids can be discovered simultaneously. When the data of the transcriptome and metabolome are combined, researchers can match gene expression with specific metabolites, thus forming a more complete picture. This comprehensive analysis is crucial for further understanding the metabolic mechanism of Leonurus japonicus and its potential pharmacological value (Shang et al., 2014; Li et al., 2019). 5.2 Application of CRISPR technology in the research of aloids from mothergrass With the development of gene editing technology, CRISPR has become a powerful means for studying the molecular mechanism of motherwort. Although the related applications are still relatively limited at present, their potential cannot be ignored. Through this technology, researchers can precisely knock out or modify genes related to alkaloid synthesis, thereby regulating the yield of specific alkaloids. For example, the editing of certain key enzymes or regulatory genes may enable the optimization of the medicinal activity of Leonurus japonicus. CRISPR also provides a direct way to verify gene functions. Candidate genes previously screened out through transcriptome studies, such as those encoding diterpene synthase, can be used to detect their specific roles in alkaloid synthesis through gene editing (Wang et al., 2022). This method can not only deepen our understanding of the genetic basis of alkaloid synthesis, but also open up a new direction for improving motherwort and breeding varieties with higher medicinal value (Rong et al., 2022).
RkJQdWJsaXNoZXIy MjQ4ODYzNA==