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GrainGenes Reference Report: PMB-47-353

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Reference	PMB-47-353
Title	Analysis of barley chloroplast psbD light-responsive promoter elements in transplastomic tobacco
Journal	Plant Molecular Biology
Year	2001
Volume	47
Pages	353-366
Author	Thum KE [ Show all 6 ]
Abstract	Summary: The plastid gene psbD encodes D2, a photosystem II reaction center chlorophyll-binding protein. psbD is transcribed from a conserved chloroplast promoter that is activated by blue, white, or UV-A light. In this study, various forms of the barley (Hordeum vulgare L.) chloroplast psbD-LRP were fused to the uidA reporter gene and introduced into the tobacco (Nicotiana tabacum L.) plastid genome through homologous recombination. Primer extension analysis of transcripts from the psbD-LRP-uidA construct showed that the barley psbD-LRP was activated in tobacco by blue or white light. Transcription from this construct was also regulated by circadian cycling indicating that the barley psbD-LRP could respond to light modulated regulatory pathways in tobacco. Mutation of the psbD-LRP prokaryotic -10 promoter element reduced transcription to very low levels in all light regimes. In contrast, mutation of a prokaryotic -35 promoter element had no effect on transcription from the psbD-LRP. Deletion or mutation of an upstream activating element, the AAG-box (-36 to -64), also reduced transcription from the construct to very low levels. In contrast, deletion of the upstream PGT-box (-71 to -100) did not alter promoter activation by blue light, or responsiveness to circadian cycling. These in vivo studies confirm the importance of the psbD-LRP -10 promoter element and AAG-box in light regulation and demonstrate that these elements are sufficient to mediate circadian cycling of the barley psbD promoter
Keyword	[ Hide all but 1 of 45 ] activation barley beta-glucuronidase binding proteins blue light chlorophyll chloroplast chloroplast dna circadian rhythm deletion element extension gene gene expression genetic transformation genome homologous recombination hordeum hordeum vulgare in vivo introduced light light regulation light-responsive promoter mediate messenger rna mutation nicotiana nicotiana tabacum photosystem ii plastid primer extension promoter protein recombination regulation reporter responsiveness tobacco transcript transcription transgenic plant uida vivo white light

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