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Gene up-regulation by DNA demethylation in 35S-gshI-transgenic poplars (Populus x canescens)

Gyulai, Gábor and Tóth, Z. and Bittsánszky, András and Szabó, Zoltán and Gullner, Gábor and Kiss, J. and Kőmíves, Tamás and Heszky, László (2008) Gene up-regulation by DNA demethylation in 35S-gshI-transgenic poplars (Populus x canescens). In: Genetically Modified Plants: New Research Trends. Nova Science Publishers Inc., Hauppauge, pp. 173-191. ISBN 978-1-60456-696-3

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Abstract

Gene expression levels of transgene 35S-gshI (γ-glutamylcysteine synthetase) cloned from E. coli, and the endogenous gene gsh1 of poplar (Populus x canescens) were upregulated by the DNA demethylating agent DHAC (5,6-dihydro-5'-azacytidine hydrochloride) (10-4 M for 7 days) in aseptic leaf discs cultures. Two 35S-gshI-transgenic (6lgl and 11ggs) and wild type (WT) poplar clones were used. The efficiency of gene upregulation was also analyzed under herbicide paraquat stress (4 x 10-7 M). Levels of gshI-mRNA and gsh1-mRNA were determined by RT-qPCR (reverse transcriptase quantitative PCR) after cDNA synthesis. For internal control, the constitutively expressed housekeeping poplar genes α-tubulin and actin were used, and the 2−HHCt method was applied for data analysis. In long term DHAC treatment (21 days), a morphogenetic response of de novo root development was observed on leaf discs in a wide concentration range of DHAC (10-8 to 10-6 M). Adventitious shoots (11ggs clone) also emerged from leaf discs after a combined treatment with DHAC (10-4 M) and paraquat (10-7 M). Shoots were dissected, rooted and transplanted in glass houses for further analyses for phytoremediation capacity. Since DNA methylation patterns are inherited (epigenetic memory), these poplar plants with increased gene expression levels of both transgene 35S-gshI and endogenous gene gsh1 provide novel plant sources for in situ application.

Item Type: Book Section
Subjects: Q Science / természettudomány > QH Natural history / természetrajz > QH301 Biology / biológia > QH3015 Molecular biology / molekuláris biológia
Q Science / természettudomány > QK Botany / növénytan
Depositing User: Erika Bilicsi
Date Deposited: 08 Jul 2013 13:05
Last Modified: 08 Jul 2013 13:05
URI: http://real.mtak.hu/id/eprint/5838

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