Co-transformation with one Agrobacterium tumefaciens strain containing two binary plasmids as a method for producing...

Area of Science:

• Plant biotechnology
• Molecular biology
• Genetics

Background:

• Traditional plant transformation often involves selectable markers that can be undesirable in final crop varieties.
• Developing methods to eliminate selectable markers post-transformation is crucial for producing marker-free transgenic plants.

Purpose of the Study:

• To investigate co-transformation as a strategy for obtaining marker-free transgenic plants.
• To assess the efficiency of gene expression and segregation from co-transformed Brassica napus and Nicotiana tabacum.

Main Methods:

• Co-cultivation of rapeseed and tobacco with a single Agrobacterium tumefaciens strain harboring two binary plasmids.
• Selection of primary transformants and analysis of transgene expression in progeny.
• Molecular analysis to determine gene insertion loci and segregation patterns.

Main Results:

• Co-transformation resulted in the expression of genes from both plasmids in approximately 50% of primary transformants.
• About 50% of co-transformed lines showed progeny expressing only one transgene, indicating independent insertion.
• Successful recovery of marker-free progeny was achieved.

Conclusions:

• Single-strain co-transformation is an effective method for plant genetic engineering.
• This approach facilitates the use of selectable markers during regeneration while allowing their removal in subsequent generations.
• Co-transformation offers a viable strategy for producing marker-free transgenic crops.