An efficient system for Agrobacterium-mediated transient transformation in Pinus tabuliformis

Area of Science:

• Plant Biotechnology
• Conifer Genomics
• Functional Genomics

Background:

• Conifer functional genomics are challenging due to large genomes and long growth periods.
• Heterologous expression in Arabidopsis is limited by evolutionary divergence between gymnosperms and angiosperms.
• Efficient genetic transformation protocols are crucial for Pinus tabuliformis gene function studies.

Purpose of the Study:

• To establish a highly efficient transient gene expression system for Pinus tabuliformis.
• To optimize Agrobacterium-mediated transformation for conifer species.
• To enable rapid gene function identification in Pinus tabuliformis.

Main Methods:

• Agrobacterium tumefaciens-mediated transient transformation.
• Optimization of co-cultivation parameters including bacterial strain, optical density, acetosyringone concentration, and duration.
• Use of a β-glucuronidase (GUS) reporter gene for efficiency assessment.

Main Results:

• Achieved a highly efficient transient expression system for Pinus tabuliformis.
• Determined optimal conditions yielding 70.1% transformation efficiency.
• Demonstrated GUS staining 24 hours post-infection, indicating rapid gene expression.

Conclusions:

• A simple, fast, and resilient Agrobacterium-mediated transformation system for Pinus tabuliformis was developed.
• This protocol enhances high-level expression of target genes.
• The system is expected to improve transformation efficiency in other conifer species.