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Additional file 1: Table S1. Primer information and sequences.

Among the most critical processes in simulating terrestrial ecosystem performance is the regulatory role of sto- mata in carbon and water cycles. Compared with field measurements, the changes in stomatal slope caused by the biophysical environment provide a simple but effective synthetic framework for studying climate-related carbon and water cycling, due to its sensitivity to CO2, vapor pressure deficit, and photosynthesis. It is also crucial in un- derstanding the effects of climate change on photosynthesis and water use efficiency. Endeavored by numerous scholastic efforts, stomatal conductance models have been improved based on experimental, semi-experimental, and mechanical processes. However, the underlying biological mechanisms and the dynamics of key parameters in these models remain unexplored, especially regarding the changes in stomatal slope. By improving the under- s tanding of the stomata's regulatory role, we reduced the uncertainty of stomatal conductance simulation. We then synthesized the recent developments and lessons in optimal stomatal behavior theory to simulate stomatal con- ductance and included an introduction to widely used stomatal conductance models and parameters, the main fac- tors influencing stomatal slopes, and applications of the mechanical stomatal conductance models in different ecosystems. Based on our literature review, we proposed that future research is needed on the optimal stomatal