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First published online 15 February 2005
doi: 10.1242/jcs.01685

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Large-scale chromatin decondensation induced in a developmentally activated transgene locus

¹ Department of Cell and Developmental Biology, John Innes Centre, Norwich Research Park, Colney Lane, Norwich, NR4 7UH, UK
² Centro de Estudios Fotosintéticos y Bioquímicos (CEFOBI), CONICET-UNR-F-LILLO, Suipacha 531, S202LRK Rosario, Argentina
³ Fraunhofer Institute for Molecular Biology and Applied Ecology, IME, Grafschaft, Auf dem Aberg 1, 57392 Schmallenberg, Germany
⁴ Institute of Molecular Biotechnology, BiologieVII, RWTH Aachen, Worringerweg 1, 52074 Aachen, Germany

^* Author for correspondence (e-mail: peter.shaw{at}bbsrc.ac.uk)

Accepted 9 December 2004

The high molecular weight (HMW) glutenin-encoding genes in wheat are developmentally activated in the endosperm at about 8 days after anthesis. We have investigated the physical changes that occur in these genes in two transgenic lines containing about 20 and 50 copies each of the HMW glutenin genes together with their promoters. Using fluorescence in-situ hybridisation (FISH) and confocal imaging, we demonstrate that, in non-expressing tissue, each transgene locus consists of one or two highly condensed sites, which decondense into many foci upon activation of transcription in endosperm nuclei. Initiation of transcription can precede decondensation but not vice versa. We show that, in one of the lines, cytoplasmic transcript levels are high after onset of transcription but disappear by 14 days after anthesis, whereas small interfering RNAs, which indicate post-transcriptional gene silencing (PTGS), are detected at this stage. However, the transcript levels remain high at the transcription sites, most of the transgene copies are transcriptionally active and transcriptional activity in the nucleus ceases only with cell death at the end of endosperm development.

Key words: Chromatin Decondensation, PTGS, HMW glutenin, Nuclear transcript

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