Assays to Assess Freeze Injury of Satsuma Mandarin

Abstract

Containerized `Owari' satsuma mandarin (Citrus unshiu Marc.) on Poncirus trifoliata `Flying Dragon' rootstock were exposed to one of two acclimation regimes (cold acclimated and unacclimated) and frozen in a computer-controlled freezer to five different low temperatures. Whole plant survival was measured and compared to the results of four leaf and stem injury assays. Acclimating plants in growth chambers at 20 °C day and 10 °C night for 14 days, followed by 15 °C day and 4 °C night for 14 to 21 days resulted in an 81% and 80% increase in leaf and stem survival, respectively, when frozen to a low of -8 °C. Electrolyte leakage and phenolic leakage assays effectively detected changes in percent leaf survival, but the TTC stain assay, using leaf disks, did not. Stem survival was best predicted by the TTC assay, using the phloem as the indicator tissue for survival. Electrolyte leakage and phenolic leakage were also reliable assays for predicting stem survival, although survival percentages were different at the same electrolyte leakage values reported in other studies. The callus growth assay accurately predicted survival for cold acclimated satsuma mandarin stems only. Chemical name used: triphenyl tetrazolium chloride (TTC).