A high efficient protocol for soybean root transformation by Agrobacterium rhizogenes and most stable reference genes...

Area of Science:

• Plant biotechnology
• Molecular biology
• Genetics

Background:

• Agrobacterium rhizogenes-mediated transformation is crucial for studying soybean root-pathogen interactions.
• Existing protocols for soybean transformation have limited efficiency and plant recovery rates.
• Accurate gene expression analysis via RT-qPCR requires stable reference genes.

Purpose of the Study:

• To optimize Agrobacterium rhizogenes-mediated soybean root transformation for higher efficiency.
• To identify stable reference genes for RT-qPCR analysis in transformed soybean composite plants.
• To enhance the study of soybean responses to abiotic stress and phytopathogens.

Main Methods:

• Optimization of Agrobacterium rhizogenes-mediated transformation protocol.
• Inclusion of an acclimation step involving hydroponics and specific selection processes.
• Validation of reference gene stability using RT-qPCR in composite plants, including under nematode infection.

Main Results:

• Achieved a 55% transformation efficiency, a significant improvement over existing methods.
• Demonstrated higher rates of transformed plant recovery.
• Identified GmELF1-β and GmELF1-α as the most stable reference genes for soybean gene expression studies.

Conclusions:

• The optimized protocol significantly enhances soybean transformation efficiency and plant recovery.
• GmELF1-β and GmELF1-α are validated as reliable reference genes for RT-qPCR in soybean composite plants, particularly under stress conditions.
• This advancement facilitates more robust functional genomics studies in soybean.