, Volume 2, Issue 1, pp 39-52
Date: 01 Feb 2006

Analysis of gene expression during bud burst initiation in Norway spruce via ESTs from subtracted cDNA libraries

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Abstract

We have made and partially sequenced two subtracted cDNA libraries, one representing genes predominantly expressed in a tree from an early-flushing family of Norway spruce (early-flushing library; EFL) and the second from a late flushing family (late flushing library; LFL), during 4 weeks before bud burst. In the EFL, expressed sequence tags (ESTs) encoding proteins of the photosynthetic apparatus and energy metabolism and proteins related to stress (abiotic and biotic) and senescence were abundant. ESTs encoding metallothionein-like and histone proteins as well as transcription factors were abundant in the LFL. We used quantitative real-time reverse transcription polymerase chain reaction to study the expression patterns of 25 chosen genes and observed that the highest levels of activity for most genes were present when plants were still ecodormant. The results indicate that the late flushing is not a result of a delay in gene activity, but is rather associated with an active transcriptional process. Accordingly, certain metabolic processes may be turned on in order to prevent premature flushing. We discuss the putative role of the studied genes in regulation of bud burst timing. Among the candidate genes found, the most interesting ones were the DNA-binding proteins, water-stress-related genes and metallothioneins. Expression patterns of some genes involved in chemical modification of DNA and histones indicate that epigenetic factors are involved in the timing of bud burst. In the obtained transcriptomes, we could not find genes commonly believed to be involved in dormancy and bud set regulation (PHY, CRY, ABI etc.) in angiosperm plants.