Functional Characterization of Water-Deficit Stress Responsive Genes Using RNAi

In response to water-deficit stress, plants alter expression of thousands of genes and as a result, cellular, physiological, and biochemical processes are modified. Understanding the functional role of water-deficit stress-responsive genes is important in order to develop stress-tolerant plants. RNA interference (RNAi) technology is one of the potential reverse genetics tool for assessing the functional significance of these genes. We describe here the protocols for developing stable gene knockdown lines for stress-induced genes using RNAi. In addition, stress imposition method that allows plants to experience gradual acclimation stress is enumerated. Further, precautions that should be taken while developing RNAi lines and during stress imposition are discussed.