Construction of a High Throughput Salt-Specific RNAi Library for Creeping Bentgrass (Agrostis stolonifera L.)

RNA interference (RNAi) constructs containing inverted repeats can effectively silence target genes based on sequence homology in plants. To generate a plant mutant population with many genes silenced, however, would require construction of numerous individual RNAi constructs, which is not practical. We used restriction enzyme-mediated method to convert a salt-specific cDNA library from creeping bentgrass (Agrotis stolonifera L.) into a population of inverted repeats (IR), each of which potentially represents a unique gene/cDNA insert (known or unknown). This IR population is being cloned into a binary vector that will be used to transform creeping bentgrass. Our goal is to discover genes that contribute to salt tolerance by generating a population of mutants, each of which may carry a silenced salt-specific gene. Our approach is particularly suitable for polyploid plant species in which multiple alleles of a gene often exist and in which genome sequence information is not available.