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PMID: 12956533 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

The promoter of the nematode resistance gene Hs1pro-1 activates a nematode-responsive and feeding site-specific gene expression in sugar beet (Beta vulgaris L.) and Arabidopsis thaliana.

Plant molecular biology ·Vol. 52 ·No. 3 ·2003-06-00 ·Pages 643-60

Thurau T, Kifle S, Jung C, Cai D

Abstract

The Hs1pro-1 gene confers resistance to the beet cyst nematode Heterodera schachtii in sugar beet (Beta vulgaris L.) on the basis of a gene-for-gene relationship. RNA-gel blot analysis revealed that the transcript of Hs1pro-1 was present in uninfected roots of resistant beet at low levels but increased by about fourfold one day after nematode infection. Treatments of plants with external stimuli including salicylic acid, jasmonic acid, gibberellic acid and abscisic acid as well as wounding or salt stress did not result in changes in the gene transcription, indicating de novo transcription of Hs1pro-1 upon nematode infection specifically. To study transcriptional regulation of Hs1pro-1 expression at the cellular level, a 3082 bp genomic fragment representing the Hs1pro-1 promoter, isolated from the YAC-DNA housing the Hs1pro-1 gene, was fused to the beta-glucuronidase reporter gene (1832prm1::GUS) and transformed into susceptible beet roots and Arabidopsis plants, respectively. Fluorometric and histochemical GUS assays on transgenic beet roots and Arabidopsis plants carrying the 1832prm1::GUS construct demonstrated that the Hs1pro-1 promoter is functional in both species and drives a nematode responsive and feeding site-specific GUS-expression. GUS activity was detected as early as at initiation of the nematode feeding sites and GUS staining was restricted to the nematode feeding sites. To delineate the regulatory domains of the Hs1pro-1 promoter, fusion genes with various 5' deletions of the Hs1pro-1 promoter and the GUS gene were constructed and analysed in transgenic beet roots as well. Cis elements responsible for feeding site-specific gene expression reside between -355 and +247 from the transcriptional initiation site of Hs1pro-1 whereas an enhancer region necessary for higher gene expression is located between -1199 and -705 of the promoter. The Hs1pro-1 promoter drives a nematode feeding site-specific GUS expression in both sugar beet and Arabidopsis suggesting a conserved mechanism of regulation of Hs1pro-1 expression in these two species.

MeSH Terms
Animals Arabidopsis/genetics,parasitology Base Sequence Beta vulgaris/genetics,parasitology Cloning, Molecular DNA, Plant/chemistry,genetics Gene Expression Regulation, Plant Glucuronidase/genetics,metabolism Immunity, Innate/genetics Membrane Proteins/genetics Molecular Sequence Data Nematoda/physiology Plant Diseases/genetics,parasitology Plant Proteins Plants, Genetically Modified Promoter Regions, Genetic/genetics Recombinant Fusion Proteins/genetics,metabolism Sequence Analysis, DNA Up-Regulation
Chemicals
DNA, Plant Hs1 pro-1 protein, Beta procumbens Membrane Proteins Plant Proteins Recombinant Fusion Proteins Glucuronidase
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Thurau Tim
Institute of Crop Science and Plant Breeding, Christian Albrechts University of Kiel, Olshausenstrasse 40, 24098 Kiel, Germany.
Kifle Sirak
Jung Christian
Cai Daguang
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Article Info
Journal
Plant molecular biology
Abbr.
Plant Mol Biol
ISSN
0167-4412
Published
2003-06-00
Pages
643-60
Language
English
Region
Netherlands
NLM ID
9106343
Subset
IM
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