# A novel method for measuring heat injury in leaves provides insights into the sequence of processes of heat injury development

## TL;DR

A new method using DSC helps measure heat injury in plant leaves and reveals the sequence of damage processes caused by high temperatures.

## Contribution

A novel DSC method for detecting heat-induced molecular changes in intact leaves is introduced.

## Key findings

- DSC provides consistent and repeatable heat thresholds in leaves.
- Heat injury starts with membrane leakage and progresses to protein denaturation.
- DSC complements classical methods by detecting molecular-level damage.

## Abstract

Global warming is currently occurring at a rapid rate and is having a particularly severe impact on plants, which, as sessile organisms, have a limited ability to escape high temperatures. This requires a better understanding of the thermal limits for different plant species and a better understanding of the processes involved in the development of heat injury in plant leaves. Heat injury results from multiple processes and occurs at the molecular level, involving increased membrane fluidity, lipid peroxidation, and protein aggregation and denaturation.

We have tested whether the DSC method allows the detection of heat-induced denaturation and aggregation of molecules in intact leaves. During controlled heating a consistent and repeatable pattern was observed in the DSC plot, from which critical heat thresholds could be derived. These critical temperatures were in good agreement with the temperatures determined using classical methods and also clearly mark the thermal limits of molecular structures. The advantage of the DCS method is the precise, rapid and easy detection of heat thresholds. Finally, taken all thresholds together, we can draw a better image of the sequence of events associated with heat injury in plant leaves: heat injury begins with membrane leakage and continues with protein denaturation and aggregation at high (sublethal, lethal) temperatures.

Since heat injury results from multiple processes, a holistic understanding requires the acquisition of parameters indicative of different processes. The presented DSC method, which allows the detection of denaturation and aggregation of cellular compounds, therefore complements well the classical methods that reflect photosynthetic impairment and whole leaf tissue damage. The new simple and rapid method requires only a minimal amount of leaf material and allows rapid collection of data on damaging temperatures for different plants, which is particularly important in the face of rapidly progressing climatic changes.

The online version contains supplementary material available at 10.1186/s13007-025-01404-6.