Large-scale gene silencing by RNA interference (RNAi) offers the possibility to address gene function in eukaryotic organisms at a depth unprecedented until recently. Although genome-wide RNAi approaches are being carried out in organisms like @entity2199 , @entity238 spp. or @entity1 after the corresponding tools had been developed, knock-down of only single or a few genes by RNAi has been reported in plants thus far. Here, we present a method for high-throughput, transient-induced gene silencing (TIGS) by RNAi in @entity2247 epidermal cells that is based on biolistic transgene delivery. This method will be useful to address gene function of shoot epidermis resulting in cell-autonomous phenotypes such as resistance or susceptibility to the powdery-mildew fungus Blumeria graminis @entity10327 . hordei. Gene function in epidermal cell elongation, stomata regulation, or UV resistance might be addressed as well. Libraries of RNAi constructs can be built up by a new, cost-efficient method that combines highly efficient ligation and recombination by the Gateway cloning system. This method allows cloning of any blunt-ended DNA fragment without the need of adaptor sequences. The final RNAi destination vector was found to direct highly efficient RNAi, as reflected by complete knock-down of a cotransformed green fluorescent protein reporter gene as well as by complete phenolcopy of the recessive loss-of-function mlo resistance gene. By using this method, a role of the t-SNARE protein HvSNAP34 in three types of durable, race-nonspecific resistance was observed.

A high-throughput gene-silencing system for the functional assessment of defense-related genes in XXXX epidermal cells.

@entity1 :: ('9606', 'Species') :: ['human'] @entity10327 :: ('239', 'Species') :: ['f. sp'] @entity2199 :: ('6239', 'Species') :: ['Caenorhabditis elegans'] @entity2247 :: ('4513', 'Species') :: ['barley'] @entity238 :: ('7227', 'Species') :: ['Drosophila']
@entity2247 :: (4513,Species) :: ['barley']
