Leaf Water Loss and Proline Assay

The water loss rate was estimated according to the methods described by Wei et al. [46 (link)]. Leaves of the seedlings in the same position were detached and their fresh weight was measured. Then, the detached leaves were exposed in the air to induce dehydration. Desiccated weight was assessed at 1 h, 2 h and 3 h, respectively. Water loss rate (%) = (fresh weight – desiccated weight)/FW × 100. Proline content was determined following the proline assay kit (A107-1-1, Nanjing Jiancheng Bioengineering Institute, Nanjing, China), with homogenized 0.1 mg of homogenized fresh leaves. Absorbance values were measured at 520 nm.