A rapid and simple vacuum infiltration system of germinated seeds for plant genetic transformation

Plant transformation methods play a crucial role in molecular breeding that enhances crop yield and quality. The genetic transformation of soybean is mainly achieved through Agrobacterium -mediated methods. To solve the problem of long cycles in plant transformation, this study established and optimized a rapid and simple transformation system, namely the vacuum infiltration system of germinated seeds, in soybean ( Glycine max (Linn.) Merr.). The expression of β-glucuronidase (GUS) in soybean was achieved using this system, which did not need sterile conditions. The results of reverse transcription quantitative PCR (RT-qPCR) analysis indicated that the GUS gene was stably expressed in T1-generation transgenic plants. Compared with traditional transformation methods, this system is rapid and simple and does not require tissue culture procedures. Moreover, to explore the CRISPR/Cas9 gene editing system in combination with vacuum infiltration of germinated seeds, we used the gene editing vector pHEE- pds to knock out the PDS gene in tobacco ( Nicotiana tabacum L.). In addition, the visual RUBY reporter gene combined with the vacuum infiltration of the cotyledons was applied to the soybean transformation, which effectively improves the screening of candidate transformants.