Tolerance of Triploid Hybrids of White Poplar ‘Beilinxiongzhu 1’ to Genetic Transformation Screening Agents In Vitro

Abstract

Genetic transformation of forest trees is essential for validating gene functions and breeding new varieties through molecular means. Appropriate selective pressure is critical for creating an effective screening system. ‘Beilinxiongzhu 1’ sensitivity testing showed that the critical tolerance concentrations for hygromycin (Hyg), kanamycin (Kan), and glyphosate (PPT) in leaf explants were 2.0 mg/L, 20 mg/L, and 1.0 mg/L, respectively. Among the physiological indicators, soluble sugar content, soluble protein content, and endogenous hormone levels were identified as key markers of the effects of the different antibiotic treatments. Transcriptome analysis showed that Hyg treatment resulted in a large number of differentially expressed genes (DEGs) involved in leaf cell wall synthesis and glucose metabolism. Under Kan treatment, the DEGs were associated with pathways such as ribosome biosynthesis and histone packaging in eukaryotes. Under PPT treatment, significant DEGs were related to ABC transporters. DEGs common to all three antibiotics were involved in glutathione metabolism pathways. A weighted gene co-expression network analysis identified TRXH2, H3.2, H2B, GST, U71K1, and CHS as key genes in response to antibiotic stress. By elucidating the physiological and molecular mechanisms by which different antibiotics affect leaf sprouting, our study serves as a reference for research into the genetic transformation of poplar leaves.