International Journal of Molecular Evolution and Biodiversity, 2025, Vol.15, No.2, 99-110 http://ecoevopublisher.com/index.php/ijmeb 103 In addition, in the process of analyzing the relationship between environmental adaptability and metabolic expression, the study found that the expression levels of genes such as terpene synthase (TPS) and cytochrome P450 monooxygenase in aromatic Lindera aggregata populations were significantly upregulated. The high expression of these synthases not only plays a core role in the accumulation of metabolites, but also further confirms their key regulatory functions in the biosynthesis of volatile oils through transgenic functional verification methods (including overexpression and RNA interference) (Gu et al., 2010). The clear positioning and functional verification of this type of genes provide targeted resource support for future molecular breeding and high-efficiency variety improvement, and also lay a molecular foundation for improving the medicinal quality of Linderae odorifera. 4.2 Screening of genes related to stress resistance Lindera aggregata exhibits excellent environmental adaptability, and its internal mechanism originates from a complex stress resistance molecular network. Transcriptome analysis identified multiple key gene families involved in abiotic stress response: dehydrins maintain cell membrane integrity to resist drought damage, heat shock proteins ensure protein homeostasis, and aquaporins precisely regulate water transport (Ye and Li, 2019). These functional units work together to form a systemic defense system for Lindera aggregata to cope with adversity. The study of Lindera aggregata population in Qingchuan revealed the dynamic regulatory characteristics of stress signaling pathways. The core components of the ABA signaling pathway, SnRK2 kinase and MYB transcription factor, were significantly upregulated at the beginning of stress (Nakamura et al., 2021). This discovery not only clarifies the molecular mechanism of Lindera aggregata's stress resistance, but also provides an important target for molecular marker-assisted breeding. In-depth analysis of the mechanism of action of these regulatory factors will accelerate the breeding of new varieties with multiple resistances. 4.3 Gene function validation technologies The establishment of a gene function verification system is the core link in clarifying the mechanism of action of candidate genes. RT-qPCR combined with RNA-Seq technology can accurately analyze the dynamic expression characteristics of Lindera aggregata genes in different tissues, developmental stages and stress conditions, providing key data support for revealing its regulatory network (Shi et al., 2024a). The innovative study by Shi et al. (2024b) systematically analyzed the structural characteristics of the mitochondrial genome of Lindera aggregata by integrating multi-omics data, including the precise positioning of functional elements and the distribution of repetitive sequences (Figure 2). This achievement not only improves the genome annotation information of this species, but also provides a new perspective for understanding its genetic evolution mechanism. The integrated application of multi-omics technologies is pushing the functional genomics research of Lindera aggregatainto a new stage. At the same time, the rapid development of gene editing technology has provided a new breakthrough for functional verification. The CRISPR/Cas9 system has been successfully applied to the functional analysis of key genes in Linderae obesa. Taking terpene synthase as an example, after specific knockout of this gene, the content of plant volatile oil decreased significantly, which clearly proved its core regulatory role in the secondary metabolic pathway (Liu et al., 2021). This type of direct correlation verification of "genetic modification-phenotypic response" has greatly improved the accuracy and practical value of functional gene analysis. 4.4 Candidate gene association analysis Establishing the association between genotype and phenotype is an important basis for molecular breeding. Whole-genome association analysis revealed a key finding: the polymorphism of terpene synthase genes was significantly correlated with the content of volatile oils. The presence of specific alleles is often accompanied by an increase in the content of medicinal ingredients, which provides a precise target for molecular marker-assisted breeding (Xiong et al., 2020).
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