What Is Stress? Dose-Response Effects in Commonly Used in Vitro Stress Assays

TLDR

In vitro stress assays are widely used to study plant responses to abiotic stress, yet most studies employ very high stress levels and assess outcomes such as germination, survival, or visual symptoms. The authors aim to develop tools that enable the study of mild stress effects in Arabidopsis. They show that conventional metrics and transcript markers only respond to severe stress, while shoot growth exhibits a highly specific, dose‑dependent response to common stressors, making it a sensitive phenotype, though caution is advised when using mannitol.

Abstract

In vitro stress assays are commonly used to study the responses of plants to abiotic stress and to assess stress tolerance. A literature review reveals that most studies use very high stress levels and measure criteria such as germination, plant survival, or the development of visual symptoms such as bleaching. However, we show that these parameters are indicators of very severe stress, and such studies thus only provide incomplete information about stress sensitivity in Arabidopsis (Arabidopsis thaliana). Similarly, transcript analysis revealed that typical stress markers are only induced at high stress levels in young seedlings. Therefore, tools are needed to study the effects of mild stress. We found that the commonly used stress-inducing agents mannitol, sorbitol, NaCl, and hydrogen peroxide impact shoot growth in a highly specific and dose-dependent way. Therefore, shoot growth is a sensitive, relevant, and easily measured phenotype to assess stress tolerance over a wide range of stress levels. Finally, our data suggest that care should be taken when using mannitol as an osmoticum.