Determination of leaf heat resistance: comparative investigation of chlorophyll fluorescence changes and tissue necrosis methods.

Heat tolerance limits for a variety of vascular plant leaves were determined both with the conventional post-culture necrosis method and by measurements of the heat-induced increase in chlorophyll fluorescence (F-T curves). The reliability of the fluorescence test was improved with the addition of far-red background light which counteracts dark reduction of the Photosystem II acceptor pool by heat-stimulated endogenous electron donors. This was of particular importance in the case of xeromorphic leaves in which the diffusion barrier for oxygen is high. A satisfactory correlation was found between T L50, the temperature at which a 30 min exposure results in 50% necrotic leaf area following post culture, and the critical temperature, T c ,the temperature at which the dark fluorescence level begins to increase during slow heating of a leaf sample at a rate of 0.7 K min-1, in the fluorescence test. The correlation can be described by a linear function, T L50=1.12 T c -5.37,with a correlation coefficient, r=0.87. Maximal deviation of the regression line from the line T L50=T c was 1.2 K, with 22 determinations for leaves with widely varying heat tolerance limits. This shows that heat-induced fluorescence changes within the thylakoid membrane may be connected with the irreversible leaf tissue damage which occurs following prolonged exposure to high temperature. On the basis of the heat dosage equation of Lepeschkin, a more general expression can be obtained which allows calculation of the accumulated heat dosage under the experimental conditions of the standard fluorescence test (slow heating, 0.7 K min-1). Such calculations reveal that for a given species the 'fraction of critical dosage' begins to increase, i.e. accumulating heat reaches an injurious level, at a temperature which approximately coincides both with T L50, obtained with the necrosis method, and with T c ,the critical temperature derived from the fluorescence test. Hence, the increase in fraction of critical dosage and the rise in chlorophyll fluorescence seem to concur. It is concluded that the fluorescence assay provides a rapid and reliable means of determining the heat tolerance limit of leaf tissue.