A frameshift in the coding region of a novel tomato class I basic chitinase gene makes it a pseudogene with a functional wound-responsive promoter.
A putative class I basic chitinase gene, assigned as psiBCH, was cloned from a tomato breeding line NC 24E. The gene contains a coding region with two introns. The predicted psiBCH open reading frame (ORF) is 971 bp and exhibits 81-88% identity at the nucleotide level with known class I basic chitinase genes from the Solanaceae family. However, the presence of a stop codon caused by a frameshift in the ORF of psiBCH makes it unusual among the other class I plant basic chitinases. This stop codon might be involved in the lower accumulation of fully spliced psiBCH RNA caused by nonsense-mediated decay (NMD), which is an RNA surveillance system universally found in eukaryotes. Sequence analysis of the 1883-bp 5'-flanking region of the psiBCH gene revealed the presence of potential wound-response promoter elements. To study the transcriptional regulation of the psiBCH gene, its 5'-flanking region containing the putative promoter was fused to the gus reporter gene and introduced into the tobacco genome via Agrobacterium tumefaciens-mediated transformation. Transgenic plants were functionally assayed for beta-glucuronidase activity. The psiBCH promoter drives the reporter gene expression in response to wounding stimuli. psiBCH promoter-GUS analysis indicates that wound-response of the tobacco transgene was rapid and localized in the wounded area following mechanical wounding. Therefore, our results suggest that the psiBCH promoter can provide targeted expression of genes, such as protease inhibitors in response to pest attack.
Baykal
Ulku
U
Department of Entomology and Plant Pathology, 209 Life Sciences Building, Auburn University, Auburn AL 36849, USA. ubaykal@rci.rutgers.edu
Moyne
Anne-Laure
AL
Tuzun
Sadik
S
D016960:Agrobacterium tumefaciens; D001483:Base Sequence; D002688:Chitinases / Q000235:genetics*; D018388:Codon, Terminator / Q000235:genetics; D005091:Exons / Q000235:genetics; D016368:Frameshift Mutation; D018506:Gene Expression Regulation, Plant / Q000235:genetics*; D007438:Introns / Q000235:genetics; D018551:Solanum lycopersicum / Q000201:enzymology / Q000235:genetics*; D008969:Molecular Sequence Data; D010935:Plant Diseases / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics*; D011544:Pseudogenes / Q000235:genetics*; D020871:RNA Stability / Q000235:genetics; D018749:RNA, Plant / Q000235:genetics; D020218:Response Elements / Q000235:genetics; D014158:Transcription, Genetic / Q000235:genetics
D018388:Codon, Terminator; D010940:Plant Proteins; D018749:RNA, Plant; D002688:Chitinases; C119825:chitinase C