Highly efficient hairy root genetic transformation and applications in citrus

Area of Science:

• Plant Biotechnology
• Genetics
• Molecular Biology

Background:

• Traditional Agrobacterium tumefaciens-mediated transformation is inefficient for woody plants like citrus.
• This inefficiency hinders gene function analysis and crop improvement.

Purpose of the Study:

• To develop a simple, universal, and highly efficient genetic transformation technology for citrus.
• To enable rapid genome editing and gene function analysis in multiple citrus genotypes.

Main Methods:

• Utilized Agrobacterium rhizogenes K599 for genetic transformation, bypassing traditional tissue culture.
• Applied the method to multiple citrus genotypes for transformation and genome editing.
• Used Agrobacterium rhizogenes for direct viral vector inoculation, facilitating VIGS and VIGS.

Main Results:

• Achieved highly efficient genetic transformation in 2-8 weeks across various citrus genotypes.
• Successfully performed genome editing using 11 plasmids, obtaining targeted knockouts.
• Demonstrated utility for obtaining transgenic root tissue and direct viral vector inoculation.

Conclusions:

• Established a rapid, efficient Agrobacterium rhizogenes-mediated transformation system for citrus.
• This technology significantly reduces time and effort for genetic studies in citrus.
• The method supports genome editing, gene overexpression, and virus-mediated gene function analysis.