Agrobacterium-Mediated Transient Expression in Sorghum Leaves for Accelerating Functional Genomics and Genome Editing Studie

The Science

Researchers aimed to develop a transient gene expression system in sorghum to efficiently screen promoter-gene constructs.

• Develop an Agrobacterium-mediated transient gene expression assay with intact sorghum leaves using green fluorescent protein as marker.
• Test genome editing capability using sgRNA to GFP protein in sorghum.

The Impact

• In planta Agrobacterium-mediated transient expression of transgenes is achievable in sorghum leaves
• High reproducibility, simplicity, rapidity, and feasibility to transform large constructs, which can directly be used for stable transformation.
• This method can be used for subcellular localization studies and physiological assays.

Summary

Sorghum is one of the most recalcitrant species for transformation.Considering the time and effort required for stable transformation in sorghum, establishing a transient system to screen the efficiency and full functionality of vector constructs is highly desirable.  Here, researchers report an Agrobacterium-mediated transient transformation assay with intact sorghum leaves using green fluorescent protein as marker. It also provides a good monocot alternative to tobacco and protoplast assays with a direct, native and more reliable system for testing single guide RNA (sgRNA) expression construct efficiency. Given the simplicity and ease of transformation, high reproducibility, and ability to test large constructs, this method can be widely adopted to speed up functional genomic and genome editing studies.

References

Sharma, R., Y. Liang, M. Y. Lee, V. R. Pidatala, J. C. Mortimer, H. V. Scheller. 2020. “Agrobacterium-mediated Transient Transformation of Sorghum Leaves for Accelerating Functional Genomics and Genome Editing Studies,” BMC Research Notes 13, 116. DOI:10.1186/s13104-020-04968-9.