Tissue?￠????type?￠????specific transcriptome analysis identifies developing xylem?￠????specific promoters in poplar

摘要

Plant biotechnology offers a means to create novel phenotypes. However, commercial application of biotechnology in crop improvement programmes is severely hindered by the lack of utility promoters (or freedom to operate the existing ones) that can drive gene expression in a tissue?￠????specific or temporally controlled manner. Woody biomass is gaining popularity as a source of fermentable sugars for liquid fuel production. To improve the quantity and quality of woody biomass, developing xylem (DX)?￠????specific modification of the feedstock is highly desirable. To develop utility promoters that can drive transgene expression in a DX?￠????specific manner, we used the Affymetrix Poplar Genome Arrays to obtain tissue?￠????type?￠????specific transcriptomes from poplar stems. Subsequent bioinformatics analysis identified 37 transcripts that are specifically or strongly expressed in DX cells of poplar. After further confirmation of their DX?￠????specific expression using semi?￠????quantitative PCR, we selected four genes ( DX5 , DX8 , DX11 and DX15 ) for in vivo confirmation of their tissue?￠????specific expression in transgenic poplars. The promoter regions of the selected DX genes were isolated and fused to a ???2?￠????glucuronidase (GUS)?￠????reported gene in a binary vector. This construct was used to produce transgenic poplars via Agrobacterium ?￠????mediated transformation. The GUS expression patterns of the resulting transgenic plants showed that these promoters were active in the xylem cells at early seedling growth and had strongest expression in the developing xylem cells at later growth stages of poplar. We conclude that these DX promoters can be used as a utility promoter for DX?￠????specific biomass engineering.