Molecular Pathogens 2024, Vol.15, No.5, 246-254 http://microbescipublisher.com/index.php/mp 249 Figure 1 Experimental evolution of TALEs (Adopted from Teper and Wang, 2021) Image caption: A: RVD repeat arrays of PthA4 (XACb0065) and dTALEs used in experimental evolution test. The nucleotide sequence of the effector-binding element of CsLOB1 from sweet orange (Citrus sinensis) is represented at the bottom. “Adapted” column indicates whether the dTALE variant was able to adapt in the duration of the experiment. B: Schematic representation of the experimental evolution workflow (Adopted from Teper and Wang, 2021) 5 Diagnostic and Detection Techniques for TAL Effector Activity 5.1 Molecular tools for detecting TAL effector targets Molecular tools have been developed to identify and characterize TAL effector targets in host plants. One such approach involves the use of single molecule real-time (SMRT) sequencing, which allows for the accurate assembly of TAL effector genes despite their repetitive nature. This method has been successfully applied to identify TAL effector genes in various Xanthomonas strains, revealing dynamic genome structures and complex relationships among TAL effector genes (Wilkins et al., 2015; Zlobin et al., 2021). Genome-wide cloning pipelines have been established to facilitate the rapid identification of TAL effector genes, enabling the prediction of potential host targets for further functional analysis (Yu et al., 2015). 5.2 Bioinformatics approaches for TAL effector prediction Bioinformatics tools play a crucial role in predicting TAL effector binding sites and their corresponding target genes. The TAL effector-DNA recognition code has been utilized to predict effector binding elements (EBEs) in the promoters of upregulated genes, aiding in the identification of new TAL effector targets. Specialized software suites like QueTAL have been developed to classify and compare TAL effectors functionally and phylogenetically. QueTAL includes tools such as DisTAL and FuncTAL, which transform TAL effector sequences into coded
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