RNAi-mediated gene silencing reveals involvement of Arabidopsis chromatin-related genes in Agrobacterium-mediated root transformation.

Abstract

We investigated the effect of RNAi-mediated gene silencing of 109 Arabidopsis thaliana chromatin-related genes (termed "chromatin genes" hereafter) on Agrobacterium-mediated root transformation. Each of the RNAi lines contains a single- or low-copy-number insertion of a hairpin construction that silences the endogenous copy of the target gene. We used three standard transient and stable transformation assays to screen 340 independent RNAi lines, representing 109 target genes, for the rat (resistant to Agrobacterium transformation) phenotype. Transformation frequency was not affected by silencing 85 of these genes. Silencing of 24 genes resulted in either a weak or strong rat phenotype. The rat mutants fell into three general groups: (i) severely dwarfed plants exhibiting a strong rat phenotype (CHC1); (ii) developmentally normal plants showing a reduced response to three transformation assays (HAG3, HDT1, HDA15, CHR1, HAC1, HON5, HDT2, GTE2, GTE4, GTE7, HDA19, HAF1, NFA2, NFA3, SGA1, and SGB2); or (iii) varying response among the three transformation assays (DMT1, DMT2, DMT4, SDG1, SDG15, SDG22, and SDG29). A direct molecular assay indicated that SGA1, HDT1, and HDT2 are important for T-DNA integration into the host genome in Arabidopsis roots.