The TiO 2 -assisted photodegradation of a squarylium cyanine dye (SQ) has been examined under visible light irradiation (λ g 430 nm) by UV-vis, proton-NMR, ESR, and GC-MS spectroscopies, by peroxide assays in the presence of peroxidase and catalase enzymes, and by chemical oxygen demand (COD) methods. Significant results were obtained relevant to the mechanism(s) of TiO 2 -assisted photodegradations. The active oxygen species produced first when an aqueous TiO 2 dispersion containing SQ is irradiated by visible light is the superoxide radical anion, which is stable in methanol solvent but very unstable in aqueous media, and is readily converted to • OH radicals via formation and subsequent reduction of H 2 O 2 . The quantity of H 2 O 2 increased during the photodegradation; though expected, no organoperoxides were detected. Of import, cleavage of the cyanine CdC double bond in the SQ dye dominated over the whole degradation process yielding 1-sulfopropyl-3,3-dimethyl-5-bromoindolenium-2-one, the predominant component in the dispersion. This intermediate is not excited by visible light and degrades no further. A sequence of steps is proposed in the initiation of the TiO 2 -assisted photochemical transformation.
BEL1-and KNOTTED1-type proteins are transcription factors from the three-amino-loop-extension superclass that interact in a tandem complex to regulate the expression of target genes. In potato (Solanum tuberosum), StBEL5 and its Knox protein partner regulate tuberization by targeting genes that control growth. RNA movement assays demonstrated that StBEL5 transcripts move through the phloem to stolon tips, the site of tuber induction. StBEL5 messenger RNA originates in the leaf, and its movement to stolons is induced by a short-day photoperiod. Here, we report the movement of StBEL5 RNA to roots correlated with increased growth, changes in morphology, and accumulation of GA2-oxidase1, YUCCA1a, and ISOPENTENYL TRANSFERASE transcripts. Transcription of StBEL5 in leaves is induced by light but insensitive to photoperiod, whereas in stolon tips growing in the dark, promoter activity is enhanced by short days. The heterodimer of StBEL5 and POTH1, a KNOTTED1-type transcription factor, binds to a tandem TTGAC-TTGAC motif that is essential for regulating transcription. The discovery of an inverted tandem motif in the StBEL5 promoter with TTGAC motifs on opposite strands may explain the induction of StBEL5 promoter activity in stolon tips under short days. Using transgenic potato lines, deletion of one of the TTGAC motifs from the StBEL5 promoter results in the reduction of GUS activity in new tubers and roots. Gel-shift assays demonstrate BEL5/POTH1 binding specificity to the motifs present in the StBEL5 promoter and a double tandem motif present in the StGA2-oxidase1 promoter. These results suggest that, in addition to tuberization, the movement of StBEL5 messenger RNA regulates other aspects of vegetative development.
BEL1-like transcription factors are ubiquitous in plants and interact with KNOTTED1 types to regulate numerous developmental processes. In potato (Solanum tuberosum subsp. andigena), the BEL1-like transcription factor StBEL5 and its Knox protein partner regulate tuber formation by targeting genes that control growth. RNA detection methods and heterografting experiments demonstrated that StBEL5 transcripts are present in phloem cells and move across a graft union to localize in stolon tips, the site of tuber induction. This movement of RNA originates in leaf veins and petioles and is induced by a shortday photoperiod, regulated by the untranslated regions, and correlated with enhanced tuber production. Assays for RNA mobility suggest that both 5# and 3# untranslated regions contribute to the preferential accumulation of the StBEL5 RNA but that the 3# untranslated region may contribute more to transport from the leaf to the stem and into the stolons. Addition of the StBEL5 untranslated regions to another BEL1-like mRNA resulted in its preferential transport to stolon tips and enhanced tuber production. Transcript stability assays showed that the untranslated regions and a long-day photoperiod enhanced StBEL5 RNA stability in shoot tips. Upon fusion of the untranslated regions of StBEL5 to a b-glucuronidase marker, translation in tobacco (Nicotiana tabacum) protoplasts was repressed by those constructs containing the 3# untranslated sequence. These results demonstrate that the untranslated regions of the mRNA of StBEL5 are involved in mediating its long-distance transport, in maintaining transcript stability, and in controlling translation.
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