Tree Genetics and Molecular Breeding 2024, Vol.14, No.5, 218-228 http://genbreedpublisher.com/index.php/tgmb 223 Figure 2PaMYB199 interacts withPaC3H17 (Adopted from Tang et al., 2019) Image caption: (a) Yeast two hybrid assays were performed using PaMYB199 fused to the DNA binding domain (BD) and PaC3H17 fused to the activation domain (AD). AH109 cells containing different plasmid combinations were grown on selective medium SD-LTHA (L, leucine; T, tryptophan; H, histidine; A, adenine) with 0 or 20 mM 3-amino-1,2,4-triazole, and stained for α-galactosidase activity. (b) In vitropull-down assays. GST-PaC3H17 and MBP-PaMYB199 proteins were immunoprecipitated with an anti-GST antibody and then immunoblotted using an anti-MBP antibody. (c) Bimolecular fluorescence complementation assay showing that PaC3H17-YFPNE and PaMYB199-YFPCE interact to form a functional yellow fluorescent protein (YFP) in the nucleus of Arabidopsis leaf protoplasts. The DNA fluorochrome 4′,6-diamidino-2-phenylindole (DAPI) was used to stain the cell nucleus (blue). Bars, 10 μm. (d) Co-immunoprecipitation assay. 35S:FLAG-PaC3H17 (GFP-FLAGas the control) and 35S:PaMYB199-HA were co-transformed into Arabidopsis leaf protoplasts. Total proteins were immunoprecipitated using an anti-HA antibody and the immunoblots were probed with an anti-FLAG antibody (Adopted from Tang et al., 2019) In Arabidopsis, the transcription factors WOX4 and KNAT1 are central to cambium development. Their interaction with other regulatory pathways, such as those involving phytohormones like auxin and cytokinin, underscores the collaborative nature of these gene networks (Zhang et al., 2019). Additionally, the role of DELLA proteins in mediating gibberellin signaling and their interaction with AUXIN RESPONSE FACTORS (ARF6 and ARF8) further illustrates the intricate regulation of cambium and xylem development (Figure 3) (Ben-Targem et al., 2021). In Betula pendula, trunk girdling experiments have shown that changes in photoassimilate levels can significantly impact cambium activity and the differentiation of xylem and phloem. Increased photoassimilates lead to enhanced phloem formation and changes in the anatomical structure of the conducting tissues, demonstrating the environmental regulation of cambium activity (Serkova et al., 2022). 6 Applications and Prospects of Gene Editing Technology in Cambium Regulation 6.1 Overview of gene editing technologies Gene editing technologies, such as CRISPR-Cas9, have revolutionized the field of plant biology by enabling precise modifications of specific genes. These technologies allow for targeted alterations in the genome,
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