On the induction of injury in tomato under continuous light: circadian asynchrony as the main triggering factor
Aaron I. Velez-Ramirez A B E H , Gabriela Dünner-Planella A F , Dick Vreugdenhil B C , Frank F. Millenaar D G and Wim van Ieperen A
+ Author Affiliations
- Author Affiliations
A Horticulture and Product Physiology, Wageningen University, PO Box 630, 6700 AP Wageningen, The Netherlands.
B Laboratory of Plant Physiology, Wageningen University, PO Box 658, 6700 AR Wageningen, The Netherlands.
C Centre for Bio Systems Genomics, PO Box 98, 6700 AB Wageningen, The Netherlands.
D Monsanto Holland B.V., PO Box 1050, 2660 BB Bergschenhoek, The Netherlands.
E Present address: Laboratory of Functional Plant Biology, Ghent University, K.L. Ledeganckstraat 35, 9000 Ghent, Belgium.
F Present address: Sociedad Agricola Quilin Ltda., Avenuenida Vespucio Sur 4100, Santiago de Chile, Chile.
G Present address: Bayer CropScience Vegetable Seeds, PO Box 4005, 6080 AA Haelen, The Netherlands.
H Corresponding author. Email: ai.velezramirez@gmail.com
Functional Plant Biology 44(6) 597-611 https://doi.org/10.1071/FP16285
Submitted: 10 August 2016 Accepted: 23 February 2017 Published: 3 May 2017
Abstract
Unlike other species, when tomato plants (Solanum lycopersicum L.) are deprived of at least 8 h of darkness per day, they develop a potentially lethal injury. In an effort to understand why continuous light (CL) is injurious to tomato, we tested five factors, which potentially could be responsible for triggering the injury in CL-grown tomato: (i) differences in the light spectral distribution between sunlight and artificial light, (ii) continuous light signalling, (iii) continuous supply of light for photosynthesis, (iv) continuous photo-oxidative pressure and (v) circadian asynchrony – a mismatch between the internal circadian clock frequency and the external light/dark cycles. Our results strongly suggest that continuous-light-induced injury does not result from the unnatural spectral distribution of artificial light nor from the continuity of light per se. Instead, circadian asynchrony seems to be the main factor inducing the CL-induced injury, but the mechanism is not by the earlier hypothesised circadian pattern in sensitivity for photoinhibition. Here, however, we show for the first time diurnal fluctuations in sensitivity to photoinhibition during normal photoperiods. Similarly, we also report for the first time diurnal and circadian rhythms in the maximum quantum efficiency of PSII (Fv/Fm) and the parameter F0.
Additional keywords: circadian regulation, light-induced signalling, photodamage, photoinhibition, photoperiod, quantum yield.
References
Arthur JM (1936) Plant growth in continuous illumination. In ‘Biological effects of radiation. Vol. 2’. (Ed. BM Duggar) pp. 715–725. (McGraw-Hill Book Company: New York)Arthur JM, Guthrie JD, Newell JM (1930) Some effects of artificial climates on the growth and chemical composition of plants. American Journal of Botany 17, 416–482.
| Some effects of artificial climates on the growth and chemical composition of plants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaA3cXjslGktQ%3D%3D&md5=5dba61627414a35ca071bf0a70586f61CAS |
Bae G, Choi G (2008) Decoding of light signals by plant phytochromes and their interacting proteins. Annual Review of Plant Biology 59, 281–311.
| Decoding of light signals by plant phytochromes and their interacting proteins.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXntFaqsLY%3D&md5=5ad69b3360bd1e5ab696a0ec752fb662CAS |
Baker NR (2008) Chlorophyll fluorescence: a probe of photosynthesis in vivo. Annual Review of Plant Biology 59, 89–113.
| Chlorophyll fluorescence: a probe of photosynthesis in vivo.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXntFaqsL8%3D&md5=2e931356ff6efeac700a0aa61c516f11CAS |
Barajas-López J de D, Serrato AJ, Cazalis R, Meyer Y, Chueca A, Reichheld JP, Sahrawy M (2011) Circadian regulation of chloroplastic f and m thioredoxins through control of the CCA1 transcription factor. Journal of Experimental Botany 62, 2039–2051.
| Circadian regulation of chloroplastic f and m thioredoxins through control of the CCA1 transcription factor.Crossref | GoogleScholarGoogle Scholar |
Casal JJ (2013) Photoreceptor signaling networks in plant responses to shade. Annual Review of Plant Biology 64, 403–427.
| Photoreceptor signaling networks in plant responses to shade.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3sXosFSktb4%3D&md5=d89bf18f16776ee1923196688aaa35d5CAS |
Casal JJ, Sánchez RA, Botto JF (1998) Modes of action of phytochromes. Journal of Experimental Botany 49, 127–138.
Christie JM (2007) Phototropin blue-light receptors. Annual Review of Plant Biology 58, 21–45.
| Phototropin blue-light receptors.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXnsVahsrc%3D&md5=eff094243c476ff054569ac9be4aae1eCAS |
Cornelissen G (2014) Cosinor-based rhythmometry. Theoretical Biology & Medical Modelling 11, 16
| Cosinor-based rhythmometry.Crossref | GoogleScholarGoogle Scholar |
Cushman KE, Tibbitts TW (1998) The role of ethylene in the development of constant-light Injury of potato and tomato. Journal of the American Society for Horticultural Science 123, 239–245.
Darrow GM (1933) Tomatoes, berries and other crops under continuous light in Alaska. Science 78, 370
| Tomatoes, berries and other crops under continuous light in Alaska.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BC3cvnsleksg%3D%3D&md5=310d32f57504fe2a6f834544da6e55ecCAS |
Daskaloff C, Ognjanova A (1965) Das verhalten von Lycopersycon esculentum Mill., L. racemigerum Lange und L. hirsutum Humb. et Bonpl. gegenüber dauerbelichtung. Zeitschrift für Pflanzenzüchtung 54, 169–181.
Demers DA, Gosselin A (2002) Growing greenhouse tomato and sweet pepper under supplemental lighting: optimal photoperiod, negative effects of long photoperiod and their causes. Acta Horticulturae 83–88.
| Growing greenhouse tomato and sweet pepper under supplemental lighting: optimal photoperiod, negative effects of long photoperiod and their causes.Crossref | GoogleScholarGoogle Scholar |
Demers DA, Dorais M, Wien CH, Gosselin A (1998) Effects of supplemental light duration on greenhouse tomato (Lycopersicon esculentum Mill.) plants and fruit yields. Scientia Horticulturae 74, 295–306.
| Effects of supplemental light duration on greenhouse tomato (Lycopersicon esculentum Mill.) plants and fruit yields.Crossref | GoogleScholarGoogle Scholar |
Dorais M, Carpentier R, Yelle S, Gosselin A (1995) Adaptability of tomato and pepper leaves to changes in photoperiod: effects on the composition and function of the thylakoid membrane. Physiologia Plantarum 94, 692–700.
| Adaptability of tomato and pepper leaves to changes in photoperiod: effects on the composition and function of the thylakoid membrane.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2MXpt1KqtL0%3D&md5=fb1cce80c43071d6b5995021f3213bffCAS |
Dorais M, Yelle S, Gosselin A (1996) Influence of extended photoperiod on photosynthate partitioning and export in tomato and pepper plants. New Zealand Journal of Crop and Horticultural Science 24, 29–37.
| Influence of extended photoperiod on photosynthate partitioning and export in tomato and pepper plants.Crossref | GoogleScholarGoogle Scholar |
Facella P, Lopez L, Carbone F, Galbraith DW, Giuliano G, Perrotta G (2008) Diurnal and circadian rhythms in the tomato transcriptome and their modulation by cryptochrome photoreceptors. PLoS One 3, e2798
| Diurnal and circadian rhythms in the tomato transcriptome and their modulation by cryptochrome photoreceptors.Crossref | GoogleScholarGoogle Scholar |
Folta KM, Kaufman LS (2003) Phototropin 1 is required for high-fluence blue-light-mediated mRNA destabilization. Plant Molecular Biology 51, 609–618.
| Phototropin 1 is required for high-fluence blue-light-mediated mRNA destabilization.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3sXhtFCqur8%3D&md5=afa17e4593416b00c3f062677c4c4c79CAS |
Globig S, Rosen I, Janes HW (1997) Continuous light effects on photosynthesis and carbon metabolism in tomato. Acta Horticulturae 141–152.
| Continuous light effects on photosynthesis and carbon metabolism in tomato.Crossref | GoogleScholarGoogle Scholar |
Haque MS, Kjaer KH, Rosenqvist E, Ottosen C-O (2015) Continuous light increases growth, daily carbon gain, antioxidants and alters carbohydrate metabolism in a cultivated and a wild tomato species. Frontiers in Plant Science 6, 522
| Continuous light increases growth, daily carbon gain, antioxidants and alters carbohydrate metabolism in a cultivated and a wild tomato species.Crossref | GoogleScholarGoogle Scholar |
Highkin HR, Hanson JB (1954) Possible interaction between light-dark cycles and endogenous daily rhythms on the growth of tomato plants. Plant Physiology 29, 301–302.
| Possible interaction between light-dark cycles and endogenous daily rhythms on the growth of tomato plants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaG2cXmsFSitg%3D%3D&md5=a12072b244a86115a9b9f7c2db0ee432CAS |
Hillman WS (1956) Injury of tomato plants by continuous light and unfavorable photoperiodic cycles. American Journal of Botany 43, 89–96.
| Injury of tomato plants by continuous light and unfavorable photoperiodic cycles.Crossref | GoogleScholarGoogle Scholar |
Hogewoning SW, Douwstra P, Trouwborst G, van Ieperen W, Harbinson J (2010a) An artificial solar spectrum substantially alters plant development compared with usual climate room irradiance spectra. Journal of Experimental Botany 61, 1267–1276.
| An artificial solar spectrum substantially alters plant development compared with usual climate room irradiance spectra.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXjt1Kgu7g%3D&md5=2764bf3ea6e99acf6c9106b1c993437cCAS |
Hogewoning SW, Trouwborst G, Maljaars H, Poorter H, van Ieperen W, Harbinson J (2010b) Blue light dose–responses of leaf photosynthesis, morphology, and chemical composition of Cucumis sativus grown under different combinations of red and blue light. Journal of Experimental Botany 61, 3107–3117.
| Blue light dose–responses of leaf photosynthesis, morphology, and chemical composition of Cucumis sativus grown under different combinations of red and blue light.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3cXotVWktrY%3D&md5=0a58f5c66f617b635dba333352d4f9bdCAS |
Holmes MG, Fukshansky L (1979) Phytochrome photoequilibria in green leaves under polychromatic radiation: a theoretical approach. Plant, Cell & Environment 2, 59–65.
| Phytochrome photoequilibria in green leaves under polychromatic radiation: a theoretical approach.Crossref | GoogleScholarGoogle Scholar |
Jiao Y, Lau OS, Deng XW (2007) Light-regulated transcriptional networks in higher plants. Nature Reviews. Genetics 8, 217–230.
| Light-regulated transcriptional networks in higher plants.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXhslOgtrY%3D&md5=4a8ee935d58eb805f20424d5903da75cCAS |
Jończyk M, Sobkowiak A, Siedlecki P, Biecek P, Trzcinska-Danielewicz J, Tiuryn J, Fronk J, Sowiński P (2011) Rhythmic diel pattern of gene expression in juvenile maize leaf. PLoS One 6, e23628
| Rhythmic diel pattern of gene expression in juvenile maize leaf.Crossref | GoogleScholarGoogle Scholar |
Jones LW, Kok B (1966) Photoinhibition of chloroplast reactions. I. Kinetics and action spectra. Plant Physiology 41, 1037–1043.
| Photoinhibition of chloroplast reactions. I. Kinetics and action spectra.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaF28XkslGgtrs%3D&md5=5577fa297551aee05477d53a407fbcecCAS |
Kalaji H, Schansker G, Ladle R, Goltsev V, Bosa K, Allakhverdiev S, Brestic M, Bussotti F, Calatayud A, Dąbrowski P, Elsheery N, Ferroni L, Guidi L, Hogewoning S, Jajoo A, Misra A, Nebauer S, Pancaldi S, Penella C, Poli D, Pollastrini M, Romanowska-Duda Z, Rutkowska B, Serôdio J, Suresh K, Szulc W, Tambussi E, Yanniccari M, Zivcak M (2014) Frequently asked questions about in vivo chlorophyll fluorescence: practical issues. Photosynthesis Research 122, 121–158.
| Frequently asked questions about in vivo chlorophyll fluorescence: practical issues.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXhsVSqs73O&md5=247df091b9bf038e99325a66cae25ddfCAS |
Kristoffersen T (1963) Interactions of photoperiod and temperature in growth and development of young tomato plants (Lycopersicon esculentum Mill.). Physiologia Plantarum 16, 1–98.
Larkin RM, Ruckle ME (2008) Integration of light and plastid signals. Current Opinion in Plant Biology 11, 593–599.
| Integration of light and plastid signals.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXhsVWhsL7M&md5=41fd22f1dc1cf27fd2e426eec903ede0CAS |
Lee Gierke C (2013) ‘Analysis of rhythms using R: chronomics analysis toolkit (CAT).’ (University of Minesota: Minneapolis, MN, USA)
Li Z, Wakao S, Fischer BB, Niyogi KK (2009) Sensing and responding to excess light. Annual Review of Plant Biology 60, 239–260.
| Sensing and responding to excess light.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXntFGls7k%3D&md5=d82f211181d5d70dcb94d04f0e617082CAS |
Matsuda R, Ozawa N, Fujiwara K (2012) Effects of continuous lighting with or without a diurnal temperature difference on photosynthetic characteristics of tomato leaves. Acta Horticulturae 165–170.
| Effects of continuous lighting with or without a diurnal temperature difference on photosynthetic characteristics of tomato leaves.Crossref | GoogleScholarGoogle Scholar |
Matsuda R, Ozawa N, Fujiwara K (2014) Leaf photosynthesis, plant growth, and carbohydrate accumulation of tomato under different photoperiods and diurnal temperature differences. Scientia Horticulturae 170, 150–158.
| Leaf photosynthesis, plant growth, and carbohydrate accumulation of tomato under different photoperiods and diurnal temperature differences.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXmvV2kt74%3D&md5=370b6caf231cec1bc50765a15258ceefCAS |
Matsuda R, Yamano T, Murakami K, Fujiwara K (2016) Effects of spectral distribution and photosynthetic photon flux density for overnight LED light irradiation on tomato seedling growth and leaf injury. Scientia Horticulturae 198, 363–369.
| Effects of spectral distribution and photosynthetic photon flux density for overnight LED light irradiation on tomato seedling growth and leaf injury.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2MXitVGhtbjN&md5=08f757d3ddbc7bba8039a181ceeb7edcCAS |
Morrow RC, Tibbitts TW (1988) Evidence for involvement of phytochrome in tumor development on plants. Plant Physiology 88, 1110–1114.
| Evidence for involvement of phytochrome in tumor development on plants.Crossref | GoogleScholarGoogle Scholar | 1:STN:280:DC%2BD3MnlsF2ksw%3D%3D&md5=c5ab6df9d85fce07d2a93805d9c8321dCAS |
Müller NA, Wijnen CL, Srinivasan A, Ryngajllo M, Ofner I, Lin T, Ranjan A, West D, Maloof JN, Sinha NR, Huang S, Zamir D, Jiménez-Gómez JM (2016) Domestication selected for deceleration of the circadian clock in cultivated tomato. Nature Genetics 48, 89–93.
| Domestication selected for deceleration of the circadian clock in cultivated tomato.Crossref | GoogleScholarGoogle Scholar |
Murage EN, Watashiro N, Masuda M (1997) Influence of light quality, PPFD and temperature on leaf chlorosis of eggplants grown under continuous illumination. Scientia Horticulturae 68, 73–82.
| Influence of light quality, PPFD and temperature on leaf chlorosis of eggplants grown under continuous illumination.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaK2sXislOgur8%3D&md5=57e5578f346f2e7189dd4b7d22ee9331CAS |
Niwa Y, Yamashino T, Mizuno T (2009) The circadian clock regulates the photoperiodic response of hypocotyl elongation through a coincidence mechanism in Arabidopsis thaliana. Plant & Cell Physiology 50, 838–854.
| The circadian clock regulates the photoperiodic response of hypocotyl elongation through a coincidence mechanism in Arabidopsis thaliana.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1MXkslKgsrs%3D&md5=d6bfe114ad0845c9ea14cab41648caddCAS |
Nozue K, Covington MF, Duek PD, Lorrain S, Fankhauser C, Harmer SL, Maloof JN (2007) Rhythmic growth explained by coincidence between internal and external cues. Nature 448, 358–361.
| Rhythmic growth explained by coincidence between internal and external cues.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD2sXnvVeqsLY%3D&md5=22edf3c9b64487f14da06a509d785c88CAS |
R Core Team (2013) ‘R: a language and environment for statistical computing.’ (R Foundation for Statistical Computing: Vienna, Austria)
Refinetti R, Cornélissen G, Halberg F (2007) Procedures for numerical analysis of circadian rhythms. Biological Rhythm Research 38, 275–325.
| Procedures for numerical analysis of circadian rhythms.Crossref | GoogleScholarGoogle Scholar |
Rockwell NC, Su Y-S, Lagarias JC (2006) Phytochrome structure and signaling mechanisms. Annual Review of Plant Biology 57, 837–858.
| Phytochrome structure and signaling mechanisms.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD28XosVKhtL0%3D&md5=ac5bddaec8c578dc62960cdcbadc8e12CAS |
Ruckle ME, DeMarco SM, Larkin RM (2007) Plastid signals remodel light signaling networks and are essential for efficient chloroplast biogenesis in Arabidopsis. The Plant Cell 19, 3944–3960.
| Plastid signals remodel light signaling networks and are essential for efficient chloroplast biogenesis in Arabidopsis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD1cXhvF2ht70%3D&md5=9ae9291bd35615f9b6c6a0450e0d8a5aCAS |
Sager JC, Smith WO, Edwards JL, Cyr KL (1988) Photosynthetic efficiency and phytochrome photoequilibria determination using spectral data. Transactions of the ASAE. American Society of Agricultural Engineers 31, 1882–1889.
| Photosynthetic efficiency and phytochrome photoequilibria determination using spectral data.Crossref | GoogleScholarGoogle Scholar |
Schaffer R, Landgraf J, Accerbi M, Simon V, Larson M, Wisman E (2001) Microarray analysis of diurnal and circadian-regulated genes in Arabidopsis. The Plant Cell 13, 113–123.
| Microarray analysis of diurnal and circadian-regulated genes in Arabidopsis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXjslCrtrk%3D&md5=99425ec685f50e4c080d75253305d5eaCAS |
Schneider CA, Rasband WS, Eliceiri KW (2012) NIH Image to ImageJ: 25 years of image analysis. Nature Methods 9, 671–675.
| NIH Image to ImageJ: 25 years of image analysis.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC38XhtVKntb7P&md5=d0e047c13797abd8a794b8b1e00e9cedCAS |
Siemens CW (1879) On the influence of electric light upon vegetation, and on certain physical principles involved. Proceedings of the Royal Society of London 30, 210–219.
| On the influence of electric light upon vegetation, and on certain physical principles involved.Crossref | GoogleScholarGoogle Scholar |
Sorek M, Yacobi YZ, Roopin M, Berman-Frank I, Levy O (2013) Photosynthetic circadian rhythmicity patterns of Symbiodium, the coral endosymbiotic algae. Proceedings of the Royal Society of London. Series B, Biological Sciences 280, 20131012
| Photosynthetic circadian rhythmicity patterns of Symbiodium, the coral endosymbiotic algae.Crossref | GoogleScholarGoogle Scholar |
Sysoeva MI, Shibaeva TG, Sherudilo EG, Ikkonen EN (2012) Control of continuous irradiation injury on tomato plants with a temperature drop. Acta Horticulturae 956, 283–289.
| Control of continuous irradiation injury on tomato plants with a temperature drop.Crossref | GoogleScholarGoogle Scholar |
Trouwborst G, Hogewoning SW, van Kooten O, Harbinson J, van Ieperen W (2016) Plasticity of photosynthesis after the ‘red light syndrome’ in cucumber. Environmental and Experimental Botany 121, 75–82.
| Plasticity of photosynthesis after the ‘red light syndrome’ in cucumber.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2MXosFKktb8%3D&md5=7cd689ce876faaa9255c309b77750b2fCAS |
Velez-Ramirez AI (2014) Continuous light on tomato: from gene to yield. PhD thesis, Graduate School Experimental Plant Sciences, Wageningen University, Wageningen, The Netherlands.
Velez-Ramirez AI, van Ieperen W, Vreugdenhil D, Millenaar FF (2011) Plants under continuous light. Trends in Plant Science 16, 310–318.
| Plants under continuous light.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC3MXnsVyrs70%3D&md5=bc0023955ba8367ce6e31f0bb20b3f3fCAS |
Velez-Ramirez AI, van Ieperen W, Vreugdenhil D, van Poppel PMJA, Heuvelink E, Millenaar FF (2014) A single locus confers tolerance to continuous light and allows substantial yield increase in tomato. Nature Communications 5, 4549
| A single locus confers tolerance to continuous light and allows substantial yield increase in tomato.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXitVWgt7%2FP&md5=72c752adb5c49d08fd0b2bd6733fea76CAS |
Velez-Ramirez AI, van Ieperen W, Vreugdenhil D, Millenaar FF (2015) Continuous-light-tolerance in tomato is graft-transferable. Planta 241, 285–290.
| Continuous-light-tolerance in tomato is graft-transferable.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BC2cXhvFGjurrF&md5=121d2268a86412aef032d08b09f79057CAS |
Withrow AP, Withrow RB (1949) Photoperiodic chlorosis in tomato. Plant Physiology 24, 657–663.
| Photoperiodic chlorosis in tomato.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DyaG3cXhvVClsA%3D%3D&md5=46c988cda578f003a5aee8307ab44501CAS |
Xu Y, Johnson CH (2001) A clock- and light-regulated gene that links the circadian oscillator to LHCB gene expression. The Plant Cell 13, 1411–1426.
| A clock- and light-regulated gene that links the circadian oscillator to LHCB gene expression.Crossref | GoogleScholarGoogle Scholar | 1:CAS:528:DC%2BD3MXkvVakt7o%3D&md5=3e56e4d23598847e84edf4381bdb6e0aCAS |
