Triggering of a plant molecular defense mechanism: Increase in gene expression levels of transgene gshI and poplar gene gsh1 (Populus × canescens) by response to the DNA demethylating drug DHAC - An qRT-PCR analysis

A. Bittsánszky, G. Gyulai, R. P. Malone, G. Gullner, J. Kiss, M. Czakó, L. Márton, L. Heszky, T. Kömíves

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

As DNA methylation patterns are inherited (epigenetic memory) gshI transgenic poplar (Populus canescens) clones (11ggs and 6Lgl) were treated with the DNA demethylating drug DHAC (5,6-dihydro-5-azacytidine hydrochloride) at 104 M for 7 days in order to study acquired plant molecular defense mechanisms in novel plant sources. In this study, the response of relative gene expression levels of transgene gshI and poplar gene gsh1 to DHAC treatment were analyzed by qRT-PCR (quantitative reverse transcriptase PCR). High expression levels of transgene gshI were observed in the 6Lgl clone (13.5-fold increase) compared to 11ggs (1.0) sample. The expression level doubled (1.8-fold increase) in the DHAC-treated 6Lgl samples but not in the 11ggs clone (0.4-fold). Contrary to this, the relative copy number of transgene gshI in the 6Lgl clone was found to be 60% less (1.0) than in the 11ggs sample (1.6). Relative expression level of endogenous poplar gene gsh1 showed significantly higher responsiveness to DHAC-induced demethylation than the transgene gshI with the highest expression level in the untransformed WT poplar (19.7-fold increase) compared to transformed clones of 6Lgl (8.7-fold increase) and 11ggs (2.5-fold increase), respectively. Competition in the reactivation capacity between transgene gshI and poplar gsh1 of 6Lgl clone was also observed as the relative gene expression level of transgene gshI increased from a high relative expression level (13.5) up by about twofold (1.8 times) rate (to 23.7) compared to poplar gsh1 gene that increased by an 8.7 increment from a lower level (1.6 rel. expression) to 13.9.

Original languageEnglish
Pages (from-to)235-243
Number of pages9
JournalActa Phytopathologica et Entomologica Hungarica
Volume42
Issue number2
DOIs
Publication statusPublished - Dec 1 2007

Keywords

  • DNA-demethylation
  • Poplar (cottonwood)
  • Relative gene expression

ASJC Scopus subject areas

  • Plant Science
  • Insect Science

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