RNA Interference-Based Gene Silencing as an Efficient Tool for Functional Genomics in Hexaploid Bread Wheat

摘要: Insertional mutagenesis and gene silencing are efficient tools for the determination of function. In contrast to gain- or loss-of-function approaches, RNA interference (RNAi)-induced can possibly silence multigene families homoeologous genes in polyploids. This is great importance functional studies hexaploid wheat (Triticum aestivum), where most present at least three copies conventional insertional not effective. We have introduced into bread double-stranded RNA-expressing constructs containing fragments encoding Phytoene Desaturase (PDS) signal transducer ethylene, Ethylene Insensitive 2 (EIN2). Transformed plants showed phenotypic changes that were stably inherited over two generations. These very similar mutant phenotypes diploid model plants. Quantitative real-time polymerase chain reaction revealed a good correlation between decreasing mRNA levels increasingly severe phenotypes. RNAi had same quantitative effect on all genes. The observed homozygous strongest reduction and, interestingly, produced around 2-fold amount small RNAs compared heterozygous suggests gene-dosage dependent. Wheat seedlings with low EIN2 ethylene insensitive. Thus, positive regulator ethylene-signaling pathway wheat, its homologs Arabidopsis (Arabidopsis thaliana) rice (Oryza sativa). Our data show results therefore represents an tool genomic polyploid wheat.