Proteomic analysis of rice anthers under salt stress

Salinity is a major factor that limits rice production worldwide. Rice is considered generally to be sensitive to salt stress during the reproductive stage. To determine the molecular mechanisms of salt tolerance at the reproductive stage, anther proteomic patterns for two contrasting rice genotypes IR64 (salt sensitive) and Cheriviruppu (salt tolerant) under salt stress were compared. Plants were grown in a greenhouse and salt stress (100 mM NaCl) was imposed at the booting stage. Anther samples were collected from control and salt-treated plants at the anthesis stage. The Na⁺/K⁺ ratio in IR64 anthers under salt stress was >1.7 times greater than that under control conditions, whereas no significant change was observed in Cheriviruppu. We also observed an 83% reduction in IR64 pollen viability, whereas this reduction was only 23% in Cheriviruppu. Of 454 protein spots detected reproducibly on two-dimensional electrophoresis gels, 38 showed significant changes in at least one genotype in response to stress. Using Mass spectrometry (MALDI TOF/TOF) analysis, we identified 18 protein spots that were involved in several processes that might increase plant adaptation to salt stress, such as carbohydrate/energy metabolism, anther wall remodelling and metabolism, and protein synthesis and assembly. Three isoforms of fructokinase-2 were upregulated only in Cheriviruppu under salt stress. This upregulation might result in increased starch content in pollen, which would support pollen growth and development under salt stress. The results also suggested that anther and pollen wall remodelling/metabolism proteins contribute to the tolerance of rice to salt stress.