A Methodological Approach to Facilitate Abscission Zone Isolation and Gene Regulation Studies of Rose.

Isolation of petal abscission zones (AZs) for molecular and biochemical studies is challenging due to their small size and the difficulty in separating them cleanly from surrounding non-AZ tissues, followed by the rapid exposure of the excised tissue to wound and dehydration stresses, post-excision. Immediate freezing of all the excised tissue is essential but practically difficult. In this chapter, we present a refined protocol for isolating petal AZ tissues from rose petals by optimizing the dissection process to minimize contamination with non-AZ cells coupled with immediate freezing to prevent stress-induced artifacts. The protocol includes careful tissue handling, precise excision, and rapid freezing, enabling the acquisition of high-quality samples for subsequent molecular and biochemical analyses. We also provide a method that facilitates the study of promoters by GUS expression through agro-injection of rose flower buds.