Employing Biolistic Transformation to Investigate Subcellular Protein Localization and Interactions in Growing Tobacco Pollen Tubes.

The transformation of certain plant tissues, such as pollen grains, poses significant challenges when using conventional methods such as Agrobacterium-mediated or viral vector-based approaches. These tissues are often resistant to direct transformation, which complicates functional studies. This issue is further exacerbated in crop species, where stable transformation remains difficult and hinders many research projects. Consequently, transient biolistic transformation of Nicotiana tabacum (tobacco) pollen has become a valuable (heterologous) system for preliminary protein characterization, offering a rapid alternative to the lengthy process of stable transformation in crop plants. In this chapter, we present an optimized, robust protocol for the transient transformation of tobacco pollen, demonstrating its applicability to a variety of protein targets.