Triticeae Genomics and Genetics, 2024, Vol.15, No.1, 19-30 http://cropscipublisher.com/index.php/tgg 26 2.3 Identification and verification of candidate genes for stress tolerance trait genes In modern agricultural science, the identification and verification of candidate genes for stress tolerance trait genes are key steps to improve crops' ability to withstand stress. This process involves not only the discovery of potential stress-resistant genes but also the verification of the functions of these genes through a series of experiments. As climate change and environmental stress intensify, it is particularly important to develop crop varieties that can withstand stresses such as drought, salinity, temperature extremes, and disease. Identification of candidate genes for stress tolerance traits first requires an in-depth understanding of the molecular mechanisms of crop responses to stress. Through tissue-specific expression analysis, transcriptomics, and proteomics studies, scientists are able to identify genes whose expression levels are significantly altered under stress conditions. In addition, using comparative genomics methods, researchers can also search for conserved stress tolerance genes in different species, which may have adapted to specific environmental stresses during evolution (Oyiga et al., 2017). Subsequently, through bioinformatics tools and databases, such as NCBI, PlantGDB, and Ensembl Plants, further bioinformatics analysis of these candidate genes can be performed, including prediction of gene structure, functional domains, and the signal transduction pathways they may participate in. exploration. These analyzes help screen out the most promising candidate genes for subsequent validation. Once candidate genes are identified, the next step is to confirm the role of these genes in plant stress tolerance through functional validation. This typically involves gene overexpression or knockout (gene silencing) experiments and subsequent assessment of phenotypic, physiological and biochemical properties. Gene overexpression experiments usually use genetic transformation methods to introduce target genes into model plants (such as Arabidopsis thaliana) or target crops. These transgenic plants are then exposed to corresponding stresses under laboratory or field conditions to assess whether enhanced gene expression improves the plants' stress tolerance. In contrast, gene knockout or silencing experiments reduce or eliminate the expression of a target gene through specific technologies (such as RNA interference or CRISPR/Cas9 gene editing) and observe how this change affects the plant's response to stress. In addition, functional verification of candidate genes needs to be performed at the cellular and molecular levels. For example, by analyzing the activity of genes encoding proteins in in vitro experiments, or using fluorescence microscopy to observe the localization of proteins in cells, and exploring interactions between proteins through bimolecular fluorescence complementation experiments. These experiments help reveal how candidate genes regulate physiological and molecular responses in plants in response to external stresses. 3 Challenges and Future Prospects 3.1 Interaction between genes and environment The interaction between genes and the environment is an important topic in biology and genetics research, especially in understanding the expression and variation of biological traits. This interaction reflects the complex process of how an organism's genetic properties are affected by the environment in which it grows. In the fields of agricultural production and human health, research on the interaction between genes and the environment provides an important theoretical basis for improving crop yields, disease prevention and treatment. Gene-environment interaction refers to the process of interaction between an individual's genetic background and external environmental factors. This interaction can affect the individual's phenotype, that is, the observable traits. Specifically, a certain genetic variant may lead to different phenotypic outcomes under different environmental conditions, and vice versa, the same environmental factor may have different effects in different genetic backgrounds (Saeideh et al., 2018). In the field of plant science, the study of gene-environment interactions helps scientists understand the adaptability and yield performance of crops in different ecosystems. Under drought conditions, some crop
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