The Arabidopsis HSP18.2 promoter/GUS gene fusion in transgenic Arabidopsis plants: a powerful tool for the isolation of regulatory mutants of the heat‐shock response

A detailed study of the expression of the promoter of the HSP18.2 gene from Arabidopsis fused to the bacterial gene for β-glucuronidase (GUS) in transgenic Arabidopsis plants is described. High levels of GUS activity were induced in all organs of transformants except for seeds during heat shock. The optimum temperature for expression of GUS in Arabidopsis was 35°C regardless of the plant growth temperature. Heat shock of 40°C did not induce any detectable levels of GUS activity. Pre-incubation at 35°C was found to have a protective effect on the induction of GUS activity at 40°C. GUS activity was also increased in response to a gradual increase in temperature. Histochemical analysis revealed that basal levels of GUS activity were induced in the vascular tissue of leaves and sepals, as well as at the tips of carpels, at the normal growth temperature. Heat treatment of a limited part of the plant tissue did not appear to cause systemic induction of GUS activity. To extend the analysis of the plant heat-shock response, we attempted to screen mutations in genes involved in the regulation of the induction of heat-shock protein (HSP) genes, using the GUS gene as a selection marker in transgenic Arabidopsis plants, and the results of this analysis are described.

[1]  S. Naito,et al.  Sugar-Dependent Expression of the CHS-A Gene for Chalcone Synthase from Petunia in Transgenic Arabidopsis. , 1991, Plant physiology.

[2]  W. Gurley,et al.  Transcriptional regulation of the heat-shock response: a plant perspective. , 1991, Biochemistry.

[3]  E. Vierling.  The Roles of Heat Shock Proteins in Plants , 1991 .

[4]  K. Scharf,et al.  Three tomato genes code for heat stress transcription factors with a region of remarkable homology to the DNA‐binding domain of the yeast HSF. , 1990, The EMBO journal.

[5]  W. Gurley,et al.  In Vitro Interaction of Nuclear Proteins with the Promoter of Soybean Heat Shock Gene Gmhsp17.5E. , 1990, Plant physiology.

[6]  C. Dumas,et al.  Influence of Temperature Stress on in Vitro Fertilization and Heat Shock Protein Synthesis in Maize (Zea mays L.) Reproductive Tissues. , 1990, Plant physiology.

[7]  J. Bol,et al.  Plant Pathogenesis-Related Proteins Induced by Virus Infection , 1990 .

[8]  G. Wullems,et al.  Heat Shock Proteins and Survival of Germinating Pollen of Lilium longiflorum and Nicotiana tabacum , 1989 .

[9]  E. Vierling,et al.  The major low-molecular-weight heat shock protein in chloroplasts shows antigenic conservation among diverse higher plant species , 1989, Molecular and cellular biology.

[10]  S. Lindquist,et al.  Characterization of an HSP70 Cognate Gene Family in Arabidopsis. , 1988, Plant physiology.

[11]  J. Amin,et al.  Key features of heat shock regulatory elements , 1988, Molecular and cellular biology.

[12]  E S Lander,et al.  Restriction fragment length polymorphism linkage map for Arabidopsis thaliana. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[13]  F. Schöffl.  Genetic engineering strategies for manipulation of the heat shock response , 1988 .

[14]  H. Xiao,et al.  Germline transformation used to define key features of heat-shock response elements. , 1988, Science.

[15]  J. Parker-Thornburg,et al.  Mutations that induce the heat shock response of Drosophila , 1987, Cell.

[16]  M. Bevan,et al.  GUS fusions: beta‐glucuronidase as a sensitive and versatile gene fusion marker in higher plants. , 1987, The EMBO journal.

[17]  M. Bevan,et al.  The Expression of Heat-Shock Genes in Higher Plants , 1986 .

[18]  R. Horsch,et al.  Transformation of Arabidopsis thaliana with Agrobacterium tumefaciens , 1986, Science.

[19]  J. L. Key,et al.  Presence of Heat Shock mRNAs in Field Crown Soybeans. , 1985, Plant physiology.

[20]  J. P. Mascarenhas,et al.  High temperature-induced thermotolerance in pollen tubes of tradescantia and heat-shock proteins. , 1985, Plant physiology.

[21]  M. Bevan,et al.  Binary Agrobacterium vectors for plant transformation. , 1984, Nucleic acids research.

[22]  D. Melton,et al.  Efficient in vitro synthesis of biologically active RNA and RNA hybridization probes from plasmids containing a bacteriophage SP6 promoter. , 1984, Nucleic acids research.

[23]  K. Scharf,et al.  Synthesis, modification and structural binding of heat-shock proteins in tomato cell cultures. , 1984, European journal of biochemistry.

[24]  J. K. Roberts,et al.  Acquisition of Thermotolerance in Soybean Seedlings : Synthesis and Accumulation of Heat Shock Proteins and their Cellular Localization. , 1984, Plant physiology.

[25]  J. Sambrook,et al.  Molecular Cloning: A Laboratory Manual , 2001 .

[26]  J. Brewbaker,et al.  THE ESSENTIAL ROLE OF CALCIUM ION IN POLLEN GERMINATION AND POLLEN TUBE GROWTH , 1963 .