Rapid delivery of Cas9 gene into the tomato cv. ‘Heinz 1706’ through an optimized Agrobacterium-mediated transformation procedure

Solanum lycopersicum ‘Heinz 1706’ is a pioneer model cultivar for tomato research, whose whole genome sequence valuable genomics studies available. Nevertheless, genetic transformation procedure this has not yet been reported. Meanwhile, various editing technologies such as transfection of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated (Cas) ribonucleoprotein complexes into cells are in the limelight. Utilizing Cas9-expressing genotype possessing reference can simplify verification an off-target effect, resolve economic cost Cas9 endonuclease preparation, and avoid complex assembly process together with single-guide RNA (sgRNA) approach. Thus, study was designed to generate lines by establishing Agrobacterium tumefaciens-mediated (ATMT) procedure. Here, we report rapid reproducible finetuning factors: A. tumefaciens strain, pre-culture co-culture durations, proper combination phytohormones at each step, supplementation acetosyringone, shooting/rooting method. Particularly, through eluding subculture simultaneously inducing shoot elongation rooting from leaf cluster, achieved duration three months recovering transgenic plants expressing Cas9. The presence gene its stable expression were confirmed PCR qRT-PCR analyses, integrated T0 plant stably transmitted T1 progeny. Therefore, anticipate that our appears ease conventional ATMT 1706’, created ultimately useful via unilateral sgRNA protoplasts.