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| Cloning and Analysis of Promoter Sequence of Rx4 Gene for Resistance to Bacterial Spot Race T3 in Solanum pimpinellifolium Accession PI128216 |
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Abstract Promoters of resistance genes play important roles on interactions between the pathogen of Xanthomonas and the host plant. Bacterial spot caused by Xanthomonas is a disease that severely threats tomato production. Current data suggest that the recognition of the pathogen race T3 to the resistance gene Rx4 might be associated with the promoter of the gene. Therefore, a 2 149 bp upstream sequence of the resistance gene Rx4 was obtained from the genomic DNA of Solanum pimpinellifolium accession PI128216 by PCR amplification in this study. Comparing to the database of Plant Cis-acting Regulatory DNA Elements (PLACE) revealed that the sequence contained the basic core promoter elements, gibberellin, abscisic acid, and ethylene related cis-elements, dehydration, salt, and cold response-related cis-elements, large number of pathogen-related transcription factors such as WRKY and MYB binding elements, and resistance-related elements W-box, G-box. A series deletion fragments of the promoter sequence were fused with the GUS gene and transiently expressed in tomato (Nicotiana benthamiana) cotyledon and tobacco leaves. Quantitative analysis of GUS activity indicated that the activity was very weak using a 297 bp fragment from -338 to -41. The remaining three fragments of 511 bp from -552 to -41, 1 244 bp from -1 285 to -41, and 733 bp from -1 285 to -552 showed increased activities but the differences were not remarkably. These results suggested that the region from -1 285 to -338 had an important role on promoter activity of the Rx4 gene. This finding provides useful information for further investigating the promoter types as well as the interaction between the resistance gene Rx4 and the race T3 pathogen of bacterial spot in tomato.
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Received: 11 December 2014
Published: 13 May 2015
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