Loading calcium fluorescent probes into protoplasts to detect calcium in the flesh tissue cells of Malus domestica.

ABSTRACT: Cytosolic Ca²⁺ plays a key role in signal transduction in plants. Calcium imaging is the most common approach to studying dynamic changes in the cytoplasmic Ca²⁺ content. Here, we used mature 'Fuji' apples (Malus pumila Mill.) to obtain viable protoplasts from flesh tissue cells by enzymatic hydrolysis; then, three small-molecule fluorescent probes (fluo-8/AM, fluo-4/AM, and rhod-2/AM) were loaded into the protoplasts. All three Ca²⁺ fluorescent probes successfully entered the cytoplasm but did not enter the vacuole. Both the Ca²⁺ chelator (EGTA) and Ca²⁺ channel blocker (La³⁺) reduced the fluorescence intensity in the cytoplasm. The calcium ionophore A23187 increased the fluorescence intensity in the cytoplasm at 5 µ

SUBMITTER: Qiu L 

PROVIDER: S-EPMC7261807 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature