全 文 ：Received 18 Jun. 2003 Accepted 10 Sept. 2003
Supported by the State Key Basic Research and Development Plan of China (G19990117003) and the Hi-Tech Research and Development
(863) Program of China (2001AA222131, 2002AA2Z1001, 2001AA212121).
* Contribute equally with the first author.
** Author for correspondence. E-mail: .
http://www.chineseplantscience.com
Acta Botanica Sinica
植 物 学 报 2004, 46 (5): 573-577
Transgenic Analysis of a Salt-inhibited OsZFP1 Gene from Rice
KONG Jin, CAO Wan-Hong*, ZHANG Jin-Song, CHEN Shou-Yi **
(Plant Biotechnology Laboratory, Institute of Genetics and Developmental Biology, The Chinese
Academy of Sciences, Beijing 100101, China)
Abstract : OsZFP1 (Oryza sativa zinc finger protein No.1) gene encodes a protein containing three
putative Cys2/Cys2-type zinc-finger domains, and is negatively regulated by salt stress. The plant expres-
sion vector carrying the OsZFP1 gene was constructed under the control of 35S promoter in pCAMBIA1300,
and then transformed into Arabidopsis plants and rice calli to overexpress OsZFP1 gene. Both the transgenic
Arabidopsis plants and the transgenic rice calli were more sensitive to salt treatment than the wild type.
This result implies that OsZFP1 gene may function as a repressor, which probably inhibits the expression
of some salt-inducible genes. The transgenic Arabidopsis plants also bolted later than wild type plants upon
ABA treatment, suggesting that the function of the OsZFP1 gene may be regulated by ABA.
Key words: OsZFP1 gene; transgenic plant; salt stress; rice
High salinity is one of the most severe abiotic stress in
agriculture. Many researches have been performed on the
gene expressions under salt stress, and a number o f salt-
induced genes have been isolated and studied in different
plant species (Claes et al., 1990; Lippuner et al, 1996; Zhang
et al., 1999; Li and Chen, 1999; 2000). However, only a few
down-regulated genes by abiotic stress have been charac-
terized (Sheng et al ., 1991; Yu et a l., 1996; Li and Chen,
2001).
We are interes ted in the regulation of gene expression
by salt stress in plants and have isolated a series of salt-
responsive genes, e.g. SAMDC from rice (Li and Chen, 2000),
NTHK1 from tobacco (Zhang et al., 2001; Xie et al., 2003),
AhCMO and BADH from Atrip lex hortensis (Shen et a l.,
2002). An OsZFP1 gene (AF171223) is also isolated from
rice by differential display in our laboratory (Li and Chen,
2001). This gene encodes a putative zinc-finger protein
containing th ree pu tat ive Cys2/Cys 2-type zinc-finger
domains. Prev ious studies (Li and Chen, 2001) showed
that the OsZFP1 transcripts were more abundant in rice
shoots than in the roots, and the level was down-regulated
by salt stres s. The expression of OsZFP1 in rice shoots
was also significantly repressed by exogenous application
of abscisic acid (ABA) (Li and Chen, 2001). However, de-
tailed functions of OsZFP1 remain unclear.
Genetic transformation is a useful technique in improv-
ing plant stres s-tolerance as well as in studying the func-
t ion of s tres s-related genes (Dubouzet et a l., 2003).
However, the report about the funct ional research o f
salt-stress down-regulated genes via transgenic approach
has not been found yet.
In the present study , OsZFP1 gene was transformed
into Arabidopsis plants and rice calli to study its roles. We
found that the transgenic plants or tissues were more sen-
sit ive to s alt stress. The function of OsZFP1 was also
discussed.
1 Materials and Methods
1.1 Construction of the plant expression vector for
OsZFP1
The full length of OsZFP1 cDNA including 3-UTR in
pGEM-T easy vector (Promega, USA) was completely di-
ges ted by SacⅠ and SacⅡ, and ligated into the vector
pBluescript-SK. The fragment containing OsZFP1 cDNA
was obtained by BamHⅠ and SacⅠ digestion , and then
inserted downstream of the 35S promoter in pCAMBIA 1300,
res ulting in the plant expression vector pCAMBIA35S:
OsZFP1 (Fig.1).
1.2 Transformation of Arabidopsis plants and rice calli
The pCAMBIA35S:OsZFP1 plasmid was int roduced
into Agrobacterium AGL1 for Arabidopsis transformation.
The Columbia type o f Arabidopsis was us ed fo r
transformation, and the transformation was carried out by
vacuum infiltration technique. The seeds were harvested
and s elected on MS medium con tain ing 50 mg/L
hygromycin. The hygromycin resis tant plan ts were re-
moved to s oil for growth and the seeds were harves ted.
This procedure was repeated until no segregation occurred



KONG Jin et al.: Transgenic Analysis of a Salt-inhibited OsZFP1 Gene from Rice 577
(Managing editor: ZHAO Li-Hui)
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