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A chimeric Lhcb::Nia gene: an inducible counter selection system for mutants in the phytochrome signal transduction pathway
Year:
1995
Source of publication :
Plant Molecular Biology
Authors :
Kenigsbuch, David
;
.
Volume :
27
Co-Authors:
Heimer, Y.M., Plant Adaptation Research Unit, J. Blaustein Institute for Desert Research, Ben-Gurion University of the Negev, Sede Boker, 84493, Israel
Brusslan, J.A., Department of Biology, University of California Los Angeles, CA, 90024, United States
Kenigsbuch, D., Department of Biology, University of California Los Angeles, CA, 90024, United States
Tobin, E.M., Department of Biology, University of California Los Angeles, CA, 90024, United States
Facilitators :
From page:
129
To page:
136
(
Total pages:
8
)
Abstract:
One approach towards understanding the transduction pathways of phytochromes is the selection of mutants impaired in various steps. We report here the construction of an inducible counter-selection system for such mutants employing the enzyme nitrate reductase. This enzyme can convert the benign substrate analogue chlorate to the toxic product chlorite, resulting in severe growth inhibition. An Arabidopsis thaliana nitrate reductase gene (Nia1*2) was placed under the regulation of an Arabidopsis thaliana light-harvesting chlorophyll a/b protein (Lhcb1*3) promoter that is phytochrome-responsive. The chimeric Lhcb::Nia gene was transformed into A. thaliana. Homozygous transformant lines were selected and grown in the absence of nitrate and the presence of L-glutamine, conditions that substantially inhibited the expression of the endogenous nitrate reductase genes. In darkness seedlings of the trans-formed lines were resistant to chlorate; however, when seedlings were grown with intermittent red light, increased sensitivity to chlorate was observed. This sensitivity was correlated with an increase in both Nia1*2 RNA levels and nitrate reductase activity. The resistant seedlings were clearly distinguishable from the sensitive ones based on hypocotyl length, with no overlap in this parameter between the two populations. Thus, this system should allow for the selection of mutants that are impaired in phytochrome regulation of the transcription of Lhcb genes. © 1995 Kluwer Academic Publishers.
Note:
Related Files :
arabidopsis
Arabidopsis thaliana
drug resistance
Gene
Genetics
genetic transformation
metabolism
mutation
photosynthesis
Show More
Related Content
More details
DOI :
10.1007/BF00019184
Article number:
Affiliations:
Database:
Scopus
Publication Type:
article
;
.
Language:
English
Editors' remarks:
ID:
29847
Last updated date:
02/03/2022 17:27
Creation date:
17/04/2018 00:49
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Scientific Publication
A chimeric Lhcb::Nia gene: an inducible counter selection system for mutants in the phytochrome signal transduction pathway
27
Heimer, Y.M., Plant Adaptation Research Unit, J. Blaustein Institute for Desert Research, Ben-Gurion University of the Negev, Sede Boker, 84493, Israel
Brusslan, J.A., Department of Biology, University of California Los Angeles, CA, 90024, United States
Kenigsbuch, D., Department of Biology, University of California Los Angeles, CA, 90024, United States
Tobin, E.M., Department of Biology, University of California Los Angeles, CA, 90024, United States
A chimeric Lhcb::Nia gene: an inducible counter selection system for mutants in the phytochrome signal transduction pathway
One approach towards understanding the transduction pathways of phytochromes is the selection of mutants impaired in various steps. We report here the construction of an inducible counter-selection system for such mutants employing the enzyme nitrate reductase. This enzyme can convert the benign substrate analogue chlorate to the toxic product chlorite, resulting in severe growth inhibition. An Arabidopsis thaliana nitrate reductase gene (Nia1*2) was placed under the regulation of an Arabidopsis thaliana light-harvesting chlorophyll a/b protein (Lhcb1*3) promoter that is phytochrome-responsive. The chimeric Lhcb::Nia gene was transformed into A. thaliana. Homozygous transformant lines were selected and grown in the absence of nitrate and the presence of L-glutamine, conditions that substantially inhibited the expression of the endogenous nitrate reductase genes. In darkness seedlings of the trans-formed lines were resistant to chlorate; however, when seedlings were grown with intermittent red light, increased sensitivity to chlorate was observed. This sensitivity was correlated with an increase in both Nia1*2 RNA levels and nitrate reductase activity. The resistant seedlings were clearly distinguishable from the sensitive ones based on hypocotyl length, with no overlap in this parameter between the two populations. Thus, this system should allow for the selection of mutants that are impaired in phytochrome regulation of the transcription of Lhcb genes. © 1995 Kluwer Academic Publishers.
Scientific Publication
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