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Plant Cell, Vol. 13, 61-72, January 2001, Copyright © 2001, American Society of Plant Physiologists

Monitoring the Expression Pattern of 1300 Arabidopsis Genes under Drought and Cold Stresses by Using a Full-Length cDNA Microarray

Motoaki Sekia,b, Mari Narusakaa, Hiroshi Abec, Mie Kasugac, Kazuko Yamaguchi-Shinozakic, Piero Carnincid, Yoshihide Hayashizakid, and Kazuo Shinozakia,b
a Plant Mutation Exploration Team, Plant Functional Genomics Research Group, RIKEN Genomic Sciences Center, 3-1-1 Koyadai, Tsukuba 305-0074, Japan
b Laboratory of Plant Molecular Biology, RIKEN Tsukuba Institute, 3-1-1 Koyadai, Tsukuba 305-0074, Japan
c Biological Resources Division, Japan International Research Center for Agricultural Sciences, Ministry of Agriculture, Forestry, and Fisheries, 2-1 Ohwashi, Tsukuba, Ibaraki 305-0851, Japan
d Genome Science Laboratory, RIKEN Tsukuba Institute, 3-1-1 Koyadai, Tsukuba 305-0074, Japan

Correspondence to: Kazuo Shinozaki, shinozaki{at}rtc.riken.go.jp (E-mail), 81-298-36-9060 (fax)

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 ~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.




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