Characterization of an ethylene-responsive glutathione S-transferase gene cluster in carnation

• Published in: Plant molecular biology Vol. 22; no. 1; pp. 43 - 58
• Main Authors: Itzhaki, H, Woodson, W.R
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer 01-04-1993
• Subjects: Amino Acid Sequence; amino acid sequences; Base Sequence; beta-glucuronidase; Biological and medical sciences; Cloning, Molecular; corolla; Dianthus caryophyllus; Enzyme Induction - drug effects; ethylene; Ethylenes - pharmacology; exons; Fundamental and applied biological sciences. Psychology; genbank/l05915; genbank/l05916; gene expression; Gene Expression Regulation, Enzymologic; Genes, Plant - genetics; Genes. Genome; genetic regulation; genetic transformation; Glutathione Transferase - biosynthesis; Glutathione Transferase - genetics; Glutathione Transferase - isolation & purification; gst1 gene; gst2 gene; histochemistry; Histocytochemistry; introns; messenger RNA; Molecular and cellular biology; Molecular genetics; Molecular Sequence Data; multigene family; Multigene Family - genetics; nucleotide sequences; Plants - drug effects; Plants - genetics; Recombinant Proteins - biosynthesis; Recombinant Proteins - isolation & purification; Regulatory Sequences, Nucleic Acid - genetics; reporter genes; RNA Splicing; RNA, Messenger - genetics; senescence; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Sequence Homology, Nucleic Acid; structural genes; Transcription, Genetic; Transformation, Genetic; Genome organization; Senescence; Nucleotide sequence; Enzyme; Sequence alignment; Ethylene; Homology; Gene organization; Tandemly repeated sequence; Dianthus caryophyllus; Caryophyllaceae; Glutathione transferase; Dicotyledones; Angiospermae; Flower crop; Regulatory sequence; Spermatophyta; Comparative study; Aminoacid sequence
• ISSN: 0167-4412; 1573-5028
• DOI: 10.1007/BF00038994

In this paper we present the structural analysis of two tightly linked genes from the glutathione S-transferase (GST) gene family in carnation (Dianthus caryophyllus). Southern blot analysis and restriction endonuclease mapping revealed a single cloned re on of the carnation genome was highly homologous to the previously characterized ethylene-responsive GST mRNA expressed in flower petals during senescence. Nucleotide sequencing of this region revealed the presence of two tandemly arranged genes designated GST1 and GST2. Comparison of the nucleotide sequences of the cloned genomic region with the previously characterized GST cDNA clone pSR8 revealed that GST1 contains the entire transcription unit in 10 exons interrupted by 9 introns. The transcription unit of GST2 was found to be very similar to GST1 with complete conservation of intron position. In addition, the length and nucleotide sequences of the two genes' introns were highly conserved. GST2 was not completely represented by the cloned genomic region, missing the 3' portion of the transcription unit. Primer extension analysis indicated a single transcriptional start site for transcripts which accumulate in senescing carnation petals. The 5'-flanking sequences of GST1 and GST2 were compared and regions of homology and divergence identified. These upstream sequences were compared with other plant ethylene-responsive genes and GST genes and several sequence motifs of potential importance in the regulation of GST expression were identified. A chimeric gene constructed between -1457 bp of the 5' -flanking DNA of GST1 and the coding region of beta-glucuronidase was found to confer ethylene-inducible expression in flower petals following delivery of the construct into tissue by particle bombardment.