Identification of a novel low-temperature-response element in the promoter of the barley (Hordeum vulgare L) gene blt101.1

Two winter barley (Hordeum vulgare L. cv. Igri) genomic clones, λgblt101.1 and λgblt101.2, encoding the blt101 gene family, were isolated from a genomic library. Deletion analysis of the blt101.1 promoter, using transient β-glucuronidase (GUS) reporter expression assays, indicated that it contains a...

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Bibliographic Details
Published in:	Planta Vol. 213; no. 5; pp. 770 - 780
Main Authors:	Brown, A.P.C., Dunn, M.A., Goddard, N.J., Hughes, M.A.
Format:	Journal Article
Language:	English
Published:	Berlin Springer-Verlag 01-09-2001 Springer
Subjects:	Adaptation to environment and cultivation conditions Adaptation, Physiological Agronomy. Soil science and plant productions Barley Biological and medical sciences Chromatin Chromatin - metabolism Cloning, Molecular Cold Temperature Deoxyribonuclease I - metabolism DNA DNA probes Fundamental and applied biological sciences. Psychology Gene Expression Genes Genetics and breeding of economic plants Genomic Library Glucuronidase - genetics Glucuronidase - metabolism Hordeum - genetics Hordeum - metabolism Low temperature Micrococcal Nuclease - metabolism Molecular and cellular biology Molecular genetics Oligonucleotide probes Plant Proteins - genetics Plant Proteins - metabolism Polymerase chain reaction Promoter regions Promoter Regions, Genetic Recombinant Fusion Proteins - genetics Recombinant Fusion Proteins - metabolism Reporter genes Sequence Deletion Transcription. Transcription factor. Splicing. Rna processing Varietal selection. Specialized plant breeding, plant breeding aims Monocotyledones Chromatin Hordeum vulgare Transcription promoter Cold resistance DNA binding protein Environmental factor Gene expression Structure function relationship Cereal crop Regulation(control) Gene Gramineae Angiospermae Molecular rearrangement Regulatory sequence Spermatophyta Intraspecific comparison Molecular cloning Low temperature
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Summary:	Two winter barley (Hordeum vulgare L. cv. Igri) genomic clones, λgblt101.1 and λgblt101.2, encoding the blt101 gene family, were isolated from a genomic library. Deletion analysis of the blt101.1 promoter, using transient β-glucuronidase (GUS) reporter expression assays, indicated that it contains at least three regulatory regions. A 107-bp region between nucleotides -168 and -275 with respect to the translation initiation codon, confers high -level GUS reporter expression at low temperature and contains a sequence (designated CR1) that is highly conserved in equivalent positions within the promoters of both members of the blt101 gene family. A 10-bp motif contained within CR1 binds proteins present in nuclear extracts from both control and low-temperature-treated barley tissue. Loss-of-function experiments, using transient-expression analysis, confirmed that this motif acts as a previously unreported low-temperature-responsive element. Nuclease sensitivity analysis of intact chromatin indicated that the blt101.1 promoter becomes more susceptible to DNase and micrococcal nuclease at low temperature, consistent with chromatin reorganisation upon transcriptional induction. It is proposed that both the 10-bp motif and chromatin reorganisation are involved in the regulation of blt101.1 at low temperature. This is the first detailed analysis of a low-temperature-specific plant promoter and identifies a novel low-temperature-response element.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0032-0935 1432-2048
DOI:	10.1007/s004250100549