The identification of homologous genes with functional overlap in forward genetic screens is $)! severely limited. Here we report the generation of over 14,000 amiRNA-expressing plants that $*! enable screens of the functionally redundant gene space in Arabidopsis. A protocol is developed %+! here for isolating robust and reproducible amiRNA-mutants. Examples of validation approaches %"! and essential controls are presented for two new amiRNA mutants that exhibit genetically %#! redundant phenotypes and circumvent double mutant lethality. In a forward genetic screen for %$! abscisic acid (ABA)-mediated inhibition of seed germination, amiRNAs that target combinations %%! of known redundant ABA receptor and SnRK2 kinase genes were rapidly isolated, providing a %&! strong proof of principle for this approach. A new ABA insensitive amiRNA line is isolated, which %'! targets three genes encoding avirulence-induced gene2-like (AIG2) genes. A thermal imaging %(! screen for plants with impaired stomatal opening in response to low CO 2 exposure led here to %)
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