Generating Wheat Transgenic Lines

The coding sequence of TaCAMTA1b-B.1 was amplified from leaf cDNA using the specific primers TaCAMTA1b-B.1-F and TaCAMTA1b-B.1-R to generate overexpression lines. Following sequence verification, the full-length CDS was introduced to the expression vector pUBI-GFP using an adaptor containing KpnI (NEB, Beijing, China) and BamHI (NEB, Beijing, China) digestion sites. RNAi lines were generated by cloning a 184 bp fragment amplified from TaCAMTA1b-B.1 cDNA into the pCUB vector through BamHI restriction sites in the sense orientation and SacI (NEB, Beijing, China) restriction sites in the antisense orientation. The resulting plasmids were transformed into Agrobacterium strain EHA105 and subsequently introduced into the wheat cultivar Jingdong18 (J18). Agrobacterium-mediated transformation to obtain transgenic plants following the protocol described by Wang et al. [65 (link),66 (link)]. The primers used for vector construction are listed in Table S3.