*4.5. Construction and Stress Treatment of Transgenic A. thaliana*

The *IlAP2* sequence was inserted into the pCAMBIA1305 vector with 35S promoter using the ClonExpress II One Step Cloning Kit (Vazyme), then the 35S:*IlAP2* plasmid was transformed into *A. tumefaciens GV3101*. *A. thaliana* Columbia was used for transformation by tidbit infection, and homozygous T3 transgenic *A. thaliana* lines were identified with 1/2 MS medium containing selective antibiotic hygromycin (50 mg/L). The expression of *I1AP2* in WT and transgenic *A. thaliana* was confirmed by agarose gel electrophoresis with PCR products amplified using IlAP2-ORF primers and cDNA from WT and transgenic *A. thaliana*. Total RNA from WT and transgenic *A. thaliana* under Cd stress was extracted using Plant RNeasy Mini Kit (Vazyme), and cDNA was generated using PrimeScript® RT kit (Takara). Three-day-old WT and transgenic *A. thaliana* seedlings were selected and transplanted to 1/2 MS medium containing 50 μM CdCl2. The root length and fresh weight of seedlings were measured after 7 days of growth.
