Comparative analysis of stress-induced calcium signals in the crop species barley and the model plant Arabidopsis thaliana

Abstract

Background: Plants are continuously exposed to changing environmental conditions and biotic attacks that affect plant growth. In crops, the inability to respond appropriately to stress has strong detrimental effects on agricultural production and yield. Ca²⁺ signalling plays a fundamental role in the response of plants to most abiotic and biotic stresses. However, research on stimulus-specific Ca²⁺ signals has mostly been pursued in Arabidopsis thaliana, while in other species these events are little investigated .

Results: In this study, we introduced the Ca²⁺ reporter-encoding gene APOAEQUORIN into the crop species barley (Hordeum vulgare). Measurements of the dynamic changes in [Ca²⁺]_cyt in response to various stimuli such as NaCl, mannitol, H₂O₂, and flagellin 22 (flg22) revealed the occurrence of dose- as well as tissue-dependent [Ca²⁺]_cyt transients. Moreover, the Ca²⁺ signatures were unique for each stimulus, suggesting the involvement of different Ca²⁺ signalling components in the corresponding stress response. Alongside, the barley Ca²⁺ signatures were compared to those produced by the phylogenetically distant model plant Arabidopsis. Notable differences in temporal kinetics and dose responses were observed, implying species-specific differences in stress response mechanisms. The plasma membrane Ca²⁺ channel blocker La³⁺ strongly inhibited the [Ca²⁺]_cyt response to all tested stimuli, indicating a critical role of extracellular Ca²⁺ in the induction of stress-associated Ca²⁺ signatures in barley. Moreover, by analysing spatio-temporal dynamics of the [Ca²⁺]_cyt transients along the developmental gradient of the barley leaf blade we demonstrate that different parts of the barley leaf show quantitative differences in [Ca²⁺]_cyt transients in response to NaCl and H₂O₂. There were only marginal differences in the response to flg22, indicative of developmental stage-dependent Ca²⁺ responses specifically to NaCl and H₂O₂.

Conclusion: This study reveals tissue-specific Ca²⁺ signals with stimulus-specific kinetics in the crop species barley, as well as quantitative differences along the barley leaf blade. A number of notable differences to the model plants Arabidopsis may be linked to different stimulus sensitivity. These transgenic barley reporter lines thus present a valuable tool to further analyse mechanisms of Ca²⁺ signalling in this crop and to gain insights into the variation of Ca²⁺-dependent stress responses between stress-susceptible and -resistant species.

Keywords: Aequorin; Arabidopsis thaliana; Ca2+ signalling; Ca2+ signature; Drought stress; Hordeum vulgare; Oxidative stress; Salt stress; barley; flagellin 22.