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A novel high‐throughput genetic screen for stress‐responsive mutants of Arabidopsis thaliana reveals new loci involving stress responses
Author(s) -
Rama Devi S.,
Chen Xi,
Oliver David J.,
Xiang Chengbin
Publication year - 2006
Publication title -
the plant journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.058
H-Index - 269
eISSN - 1365-313X
pISSN - 0960-7412
DOI - 10.1111/j.1365-313x.2006.02814.x
Subject(s) - mutant , biology , reporter gene , mutagenesis , arabidopsis , promoter , gene , genetics , genetic screen , arabidopsis thaliana , transposable element , rna interference , forward genetics , microbiology and biotechnology , gene expression , rna
Summary Activation sequence‐1 ( as‐1 ) cognate promoter elements are widespread in the promoters of plant defense‐related genes as well as in plant pathogen promoters, and may play important roles in the activation of defense‐related genes. The as‐1 ‐type elements are highly responsive to multiple stress stimuli such as jasmonic acid (JA), salicylic acid (SA), H 2 O 2 , xenobiotics and heavy metals, and therefore provide a unique opportunity for identifying additional signaling components and cross‐talk points in the various signaling networks. A single as‐1 ‐type cis ‐element‐driven GUS reporter Arabidopsis line responsive to JA, SA, H 2 O 2 , xenobiotics and heavy metals was constructed for mutagenesis. A large‐scale T‐DNA mutagenesis has been conducted in the reporter background, and an efficient high‐throughput mutant screen was established for isolating mutants with altered responses to the stress chemicals. A number of mutants with altered stress responses were obtained, some of which appear to identify new components in the as‐1 ‐based signal transduction pathways. We characterized a mutant (Δ8L4) with a T‐DNA insertion in the coding sequence of the gene At4g24275. The as‐1 ‐regulated gene expression and GUS reporter gene expression were altered in the Δ8L4 mutant, but there was no change in the expression of genes lacking as‐1 elements in their promoters. The phenotype observed with the Δ8L4 mutant was further verified using RNAi plants for At4g24275 (8L4‐RNAi), suggesting the feasibility of use of this high‐throughput mutant screening in isolating stress‐signaling mutants.