Abstract
The developmental transition of the meristem from vegetative to reproductive growth is controlled by the cyclic alternation of light and darkness in photoperiodic plants. Photoperiod is perceived in the leaves or cotyledons, where a flower-inducing signal is produced and transmitted to the apex. To begin to understand the molecular basis of the photoperiodic induction of flowering, we investigated changes in gene expression at the level of mRNA abundance that occur in association with dark induction of flowering in the short-day species Pharbitis nil. Several cDNAs were isolated that corresponded to mRNAs whose abundance is altered after the transition to darkness. The pattern of increase in mRNA levels corresponding to one cDNA clone, PN1, showed a dark-induced maximum at 8 h of darkness, whereas a second clone, PN9, showed a dark-induced accumulation of mRNA with peak levels at 12 to 16 h of darkness. When plants were held in continuous darkness, both PN1 and PN9 exhibited rhythmic patterns of mRNA accumulation with an approximate circadian periodicity, suggesting that their expression is under the control of an endogenous clock. The observed pattern of expression of PN1 and PN9 in cotyledon tissue was unusual in that darkness rather than light promoted mRNA accumulation, which is a temporal pattern of expression distinct from that of several other Pharbitis genes, including Cab, PsaG, and actin, whose mRNAs were most prevalent or equally prevalent in the light. Brief illumination of an inductive dark period by a red light night break strongly inhibited the accumulation of both PN1 and PN9 mRNA. The expression of both PN1 and PN9 was spatially regulated in that mRNA transcripts were detected in the cotyledons and stems, but not the roots, of photoperiodically competent seedlings. Both PN1 and PN9 appeared to be present as single-copy genes in the Pharbitis genome. Sequence analysis has not determined the identity of these genes. Overall, the accumulation of mRNAs corresponding to both PN1 and PN9 closely paralleled the process of photoperiodic floral induction in P. nil, but a clear involvement with this process cannot be established from our findings because of the difficulty of separating photoperiodic events from other light-regulated processes, especially those involved in photosynthesis, such as Cab gene expression. These results identify the products of circadian-regulated genes in photoreceptive tissue of P. nil and support the concept that circadian-regulated gene expression interacting with darkness may be involved in the regulation of photoperiodically controlled physiological processes, including flower induction.
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- Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bowman J. L., Smyth D. R., Meyerowitz E. M. Genes directing flower development in Arabidopsis. Plant Cell. 1989 Jan;1(1):37–52. doi: 10.1105/tpc.1.1.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
- Galletti C., Battaglini P. P., Aicardi G. 'Real-motion' cells in visual area V2 of behaving macaque monkeys. Exp Brain Res. 1988;69(2):279–288. doi: 10.1007/BF00247573. [DOI] [PubMed] [Google Scholar]
- Giuliano G., Hoffman N. E., Ko K., Scolnik P. A., Cashmore A. R. A light-entrained circadian clock controls transcription of several plant genes. EMBO J. 1988 Dec 1;7(12):3635–3642. doi: 10.1002/j.1460-2075.1988.tb03244.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lincoln J. E., Cordes S., Read E., Fischer R. L. Regulation of gene expression by ethylene during Lycopersicon esculentum (tomato) fruit development. Proc Natl Acad Sci U S A. 1987 May;84(9):2793–2797. doi: 10.1073/pnas.84.9.2793. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Melzer S., Majewski D. M., Apel K. Early Changes in Gene Expression during the Transition from Vegetative to Generative Growth in the Long-Day Plant Sinapis alba. Plant Cell. 1990 Oct;2(10):953–961. doi: 10.1105/tpc.2.10.953. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Millar A. J., Kay S. A. Circadian Control of cab Gene Transcription and mRNA Accumulation in Arabidopsis. Plant Cell. 1991 May;3(5):541–550. doi: 10.1105/tpc.3.5.541. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Nagao R. T., Kimpel J. A., Key J. L. Molecular and cellular biology of the heat-shock response. Adv Genet. 1990;28:235–274. doi: 10.1016/s0065-2660(08)60528-3. [DOI] [PubMed] [Google Scholar]
- Nair P. P. Nutrients and the human genome: new frontiers for the next century. FASEB J. 1993 Apr 1;7(6):501–502. doi: 10.1096/fasebj.7.6.8472888. [DOI] [PubMed] [Google Scholar]
- Piechulla B., Gruissem W. Diurnal mRNA fluctuations of nuclear and plastid genes in developing tomato fruits. EMBO J. 1987 Dec 1;6(12):3593–3599. doi: 10.1002/j.1460-2075.1987.tb02690.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Piechulla B., Riesselmann S. Effect of Temperature Alterations on the Diurnal Expression Pattern of the Chlorophyll a/b Binding Proteins in Tomato Seedlings. Plant Physiol. 1990 Dec;94(4):1903–1906. doi: 10.1104/pp.94.4.1903. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rosbash M., Hall J. C. The molecular biology of circadian rhythms. Neuron. 1989 Oct;3(4):387–398. doi: 10.1016/0896-6273(89)90199-2. [DOI] [PubMed] [Google Scholar]
- Salisbury F. B. Photoperiodism. Hortic Rev (Am Soc Hortic Sci) 1985;4:66–105. [PubMed] [Google Scholar]
- Takimoto A., Hamner K. C. Studies on Red Light Interruption in Relation to Timing Mechanisms Involved in the Photoperiodic Response of Pharbitis nil. Plant Physiol. 1965 Sep;40(5):852–854. doi: 10.1104/pp.40.5.852. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Taylor W. C. Transcriptional regulation by a circadian rhythm. Plant Cell. 1989 Feb;1(2):259–264. doi: 10.1105/tpc.1.2.259. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Thomas P. S. Hybridization of denatured RNA transferred or dotted nitrocellulose paper. Methods Enzymol. 1983;100:255–266. doi: 10.1016/0076-6879(83)00060-9. [DOI] [PubMed] [Google Scholar]
- Tomlinson A., Kimmel B. E., Rubin G. M. rough, a Drosophila homeobox gene required in photoreceptors R2 and R5 for inductive interactions in the developing eye. Cell. 1988 Dec 2;55(5):771–784. doi: 10.1016/0092-8674(88)90133-x. [DOI] [PubMed] [Google Scholar]
- Weigel D., Jürgens G., Küttner F., Seifert E., Jäckle H. The homeotic gene fork head encodes a nuclear protein and is expressed in the terminal regions of the Drosophila embryo. Cell. 1989 May 19;57(4):645–658. doi: 10.1016/0092-8674(89)90133-5. [DOI] [PubMed] [Google Scholar]
- Zheng C. C., Bui A. Q., O'Neill S. D. Abundance of an mRNA encoding a high mobility group DNA-binding protein is regulated by light and an endogenous rhythm. Plant Mol Biol. 1993 Nov;23(4):813–823. doi: 10.1007/BF00021536. [DOI] [PubMed] [Google Scholar]