Base Editing in Poplar Through an Agrobacterium-Mediated Transformation Method.

Area of Science:

• Plant Biotechnology
• Genome Editing
• Molecular Biology

Background:

• CRISPR-Cas9 has enabled gene editing in plants, but previous tree studies focused on NHEJ-induced indels.
• Cytosine base editors (CBEs) and adenine base editors (ABEs) allow precise base conversions (C-to-T, A-to-G).
• Base editing systems are recently established in trees, offering new avenues for genetic modification.

Purpose of the Study:

• To establish a robust protocol for precise base editing in poplar using CBEs and ABEs.
• To facilitate functional genomic studies and genetic trait improvement in trees.
• To provide a detailed methodology for Agrobacterium-mediated transformation with base editors.

Main Methods:

• Preparation of T-DNA vectors encoding two efficient CBEs (PmCDA1-BE3, A3A/Y130F-BE3) and one efficient ABE (ABE8e).
• Optimization of Agrobacterium-mediated transformation protocol for efficient T-DNA delivery in poplar.
• Application of base editors for targeted C-to-T and A-to-G base substitutions.

Main Results:

• Demonstrated successful implementation of CBEs and ABEs in poplar.
• Established a reliable protocol for base editor delivery and transformation in trees.
• Showcased the potential for introducing specific amino acid changes and regulatory element edits.

Conclusions:

• Precise base editing using CBEs and ABEs is feasible and effective in poplar.
• The developed protocol offers significant potential for advancing functional genomics and crop improvement in trees.
• This work expands the genome editing toolkit available for woody plant species.