In a significant stride towards enhancing genetic manipulation techniques for Bacillus strains, a team of researchers led by Wei Quan from the College of Horticulture and Plant Protection at Inner Mongolia Agricultural University has developed an efficient electrotransformation method. Published in the journal *Applied Microbiology and Biotechnology*, this study focuses on optimizing transformation conditions for three Bacillus species: *Bacillus amyloliquefaciens YN-J3*, *Bacillus velezensis JN-Y2*, and *Bacillus subtilis S-16*.
Bacillus strains are pivotal in food fermentation and plant disease control, making efficient transformation methods crucial for their genetic manipulation. The researchers employed orthogonal experiments combined with response surface analysis to fine-tune transformation parameters. Their findings revealed that the optimal conditions for *B.a YN-J3* and *B.v JN-Y2* included an OD600 of 0.70, a competent cell volume of 91 μL, a plasmid concentration of 1040 ng·μL⁻1, and a field strength of 18.1 kV·cm⁻1. For *B.s S-16*, the optimal parameters were slightly different: an OD600 of 0.71, a competent cell volume of 92 μL, a plasmid concentration of 1052 ng·μL⁻1, and a field strength of 18.2 kV·cm⁻1.
Under these optimized conditions, the transformation efficiencies soared to 22,198.33 CFU·μg⁻1 DNA for *B.a YN-J3*, 24,498.67 CFU·μg⁻1 DNA for *B.v JN-Y2*, and 23,305.00 CFU·μg⁻1 DNA for *B.s S-16*. The researchers also discovered that treating cells with 50 mg/mL glycine significantly boosted transformation efficiency by 40, 36, and 24 times for *B.a YN-J3*, *B.v JN-Y2*, and *B.s S-16*, respectively.
“We were astonished by the dramatic increase in transformation efficiency when we introduced glycine treatment,” said Wei Quan, the lead author of the study. “This finding opens up new possibilities for genetic engineering in Bacillus strains, which are vital for agricultural and industrial applications.”
The implications of this research are far-reaching for the agriculture sector. Efficient genetic manipulation of Bacillus strains can lead to the development of more robust and effective biopesticides and biofertilizers. These advancements can enhance crop protection and productivity, ultimately contributing to sustainable agriculture practices.
“This study not only provides a stable and efficient electroporation transformation system but also paves the way for future developments in genetic engineering of Bacillus strains,” added Quan. “The combination of optimized transformation conditions and glycine treatment offers a powerful tool for researchers and industry professionals alike.”
The research, published in *Applied Microbiology and Biotechnology*, represents a significant leap forward in the field of agritech, promising to revolutionize the way we approach plant disease control and food fermentation. As the agricultural industry continues to evolve, such innovations will be crucial in meeting the growing demands for sustainable and efficient food production.