Monitoring the expression pattern of 1300 Arabidopsis genes under drought and cold stresses by using a full-length cDNA microarray.
Full-length cDNAs are essential for functional analysis of plant genes. Using the biotinylated CAP trapper method, we constructed full-length Arabidopsis cDNA libraries from plants in different conditions, such as drought-treated, cold-treated, or unstressed plants, and at various developmental stages from germination to mature seed. We prepared a cDNA microarray using approximately 1300 full-length Arabidopsis cDNAs to identify drought- and cold-inducible genes and target genes of DREB1A/CBF3, a transcription factor that controls stress-inducible gene expression. In total, 44 and 19 cDNAs for drought- and cold-inducible genes, respectively, were isolated, 30 and 10 of which were novel stress-inducible genes that have not been reported as drought- or cold-inducible genes previously. Twelve stress-inducible genes were identified as target stress-inducible genes of DREB1A, and six of them were novel. On the basis of RNA gel blot and microarray analyses, the six genes were identified as novel drought- and cold-inducible genes that are controlled by DREB1A. Eleven DREB1A target genes whose genomic sequences have been registered in the GenBank database contained the dehydration-responsive element (DRE) or DRE-related CCGAC core motif in their promoter regions. These results show that our full-length cDNA microarray is a useful material with which to analyze the expression pattern of Arabidopsis genes under drought and cold stresses, to identify target genes of stress-related transcription factors, and to identify potential cis-acting DNA elements by combining the expression data with the genomic sequence data.
Seki
M
M
Plant Mutation Exploration Team, Plant Functional Genomics Research Group, RIKEN Genomic Sciences Center, 3-1-1 Koyadai, Tsukuba 305-0074, Japan.
Narusaka
M
M
Abe
H
H
Kasuga
M
M
Yamaguchi-Shinozaki
K
K
Carninci
P
P
Hayashizaki
Y
Y
Shinozaki
K
K
D017360:Arabidopsis / Q000235:genetics*; D029681:Arabidopsis Proteins; D001483:Base Sequence; D003080:Cold Temperature; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics*; D005456:Fluorescent Dyes; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant / Q000502:physiology; D009693:Nucleic Acid Hybridization; D020411:Oligonucleotide Array Sequence Analysis; D014157:Transcription Factors / Q000502:physiology; D014867:Water
D029681:Arabidopsis Proteins; D017931:DNA Primers; D018076:DNA, Complementary; C114578:DREB1A protein, Arabidopsis; D005456:Fluorescent Dyes; D014157:Transcription Factors; D014867:Water