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Plant Physiol, June 2001, Vol. 126, pp. 707-716
Networking Senescence-Regulating Pathways by Using Arabidopsis
Enhancer Trap Lines1
Yuehui
He,
Weining
Tang,
Johnnie D.
Swain,
Anthony L.
Green,
Thomas P.
Jack, and
Susheng
Gan*
Plant Physiology/Biochemistry/Molecular Biology Program,
Department of Agronomy and Tobacco and Health Research Institute,
University of Kentucky, Lexington, Kentucky 40546-0236 (Y.H., W.T.,
J.D.S., A.L.G., S.G.); and Department of Biological Sciences, Dartmouth
College, Hanover, New Hampshire 03755 (T.P.J.)
The last phase of leaf development, generally referred to as leaf
senescence, is an integral part of plant development that involves
massive programmed cell death. Due to a sharp decline of photosynthetic
capacity in a leaf, senescence limits crop yield and forest plant
biomass production. However, the biochemical components and regulatory
mechanisms underlying leaf senescence are poorly characterized.
Although several approaches such as differential cDNA screening,
differential display, and cDNA subtraction have been employed to
isolate senescence-associated genes (SAGs), only a
limited number of SAGs have been identified, and
information regarding the regulation of these genes is fragmentary.
Here we report on the utilization of enhancer trap approach toward the identification and analysis of SAGs. We have developed a
sensitive large-scale screening method and have screened 1,300 Arabidopsis enhancer trap lines and have identified 147 lines in which
the reporter gene GUS ( -glucuronidase) is expressed
in senescing leaves but not in non-senescing ones. We have
systematically analyzed the regulation of -glucuronidase expression
in 125 lines (genetically, each contains single T-DNA insertion) by six
senescence-promoting factors, namely abscisic acid, ethylene, jasmonic
acid, brassinosteroid, darkness, and dehydration. This analysis not
only reveals the complexity of the regulatory circuitry but also allows
us to postulate the existence of a network of senescence-promoting
pathways. We have also cloned three SAGs from randomly
selected enhancer trap lines, demonstrating that reporter expression
pattern reflects the expression pattern of the endogenous gene.
1
This work was supported by the U.S. Department
of Agriculture National Research Initiative (grant no.
2001-35304-09994 to S.G.) and by the Tobacco and Health Research
Institute's Biotechnology Program at the University of
Kentucky (grant to S.G.). Y.H. was supported in part by the
University of Kentucky's Research Challenge Trust Fund (Plant
Sciences). J.D.S. was supported in part by the University of
Kentucky's Science Outreach Center. This is publication no. 01-06-25
of the Kentucky Agricultural Experiment Station.
*
Corresponding author; e-mail sgan{at}pop.uky.edu; fax 859-323-1077.
© 2001 American Society of Plant Physiologists
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