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Plant Physiol, January 2000, Vol. 122, pp. 15-24
Identification by Large-Scale Screening of Phytochrome-Regulated
Genes in Etiolated Seedlings of Arabidopsis Using a Fluorescent
Differential Display Technique1
Norihito
Kuno,
Takamichi
Muramatsu,
Fumiaki
Hamazato, and
Masaki
Furuya*
Hitachi Advanced Research Laboratory, Hatoyama, Saitama 350-0395,
Japan.
Phytochrome A (PhyA)-regulated genes
in 6-d-old etiolated seedlings of Arabidopsis Landsberg
erecta were identified by fluorescent differential
display. To screen for PhyA-regulated genes, mRNA fingerprints of the
wild type and the phyA-201 mutant were compared from
samples prepared 4 h after far-red light irradiation.
Approximately 30,000 bands of cDNA were displayed by fluorescent
differential display, and 24 differentially expressed bands were
observed. Sequence analysis revealed that they represent 20 distinct
genes. Among them, 15 genes were confirmed as PhyA regulated by
northern-blot (or reverse transcription-polymerase chain reaction)
analysis. Thirteen up-regulated genes included 12 known genes that
encode nine photosynthetic proteins, two enzymes involved in the
biosynthesis of chlorophyll, one DNA damage repair/toleration-related
protein, and one unknown gene. Two down-regulated genes were identified as encoding a xyloglucan endotransglycosylase-related protein and a
novel member of the ASK protein kinase family. In the
phyA-201 mutant and the phyA-201phyB-1
double mutant, expression of all of these genes was photoreversibly up-
or down-regulated by type II phytochromes. The results indicate that
modes of photoperception differ between PhyA and PhyB, but that both
types of phytochromes have overlapping effects on the photoregulation
of gene expression.
1
This work was partly supported by grants to M.F.
from the Hitachi Advanced Research Laboratory (B2023) and the Program
for Promotion of Basic Research Activities for Innovative Biosciences.
*
Corresponding author; e-mail mfuruya{at}harl.hitachi.co.jp; fax
81-492-96-7511.
© 2000 American Society of Plant Physiologists
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