Physiology and Molecular Biology of Plants

, Volume 24, Issue 4, pp 591–604 | Cite as

Reproductive sink enhanced drought induced senescence in wheat fertile line is associated with loss of antioxidant competence compared to its CMS line

  • Vimal Kumar Semwal
  • Renu Khanna-ChopraEmail author
Research Article


Reproductive sinks regulate monocarpic senescence in wheat as desinking delayed flag leaf senescence under irrigated condition. In this study, wheat cv. HW 2041 and its isonuclear male sterile line (CMS) were subjected to post-anthesis water deficit stress to understand the association between sink strength, senescence and drought response in relation to oxidative stress and antioxidant defense at cellular and sub-cellular level. CMS plants maintained better water relations and exhibited delayed onset and progression of flag leaf senescence in terms of green leaf area, chlorophyll and protein content than fertile plants under water deficit stress (WDS). Delayed senescence in CMS plants under water deficit stress was associated with less reactive oxygen species generation, lower damage to membranes and better antioxidant defense both in terms of antioxidant enzyme activities and metabolite content compared to fertile plants. Expression of some senescence associated genes (SAGs) such as WRKY transcription factor (WRKY53), glutamine synthetase1 (GS1), wheat cysteine protease (WCP2) and wheat serine protease (WSP) was lower while catalse 2 (CAT2) transcript levels were higher in the CMS plants compared to HW2041 during senescence under water deficit stress. Antioxidant defense in chloroplasts was better in CMS line under water deficit stress compared to HW2041. This is the first report showing that reproductive sink enhanced drought induced senescence in flag leaf of wheat fertile line is associated with higher oxidative stress and damage and loss of antioxidant competence compared to its sterile line under water deficit stress. Higher expression of some SAGs and decline in superoxide dismutase and ascorbate peroxidase activity in the chloroplasts also contributed to the accelerated senescence in fertile line compared to its CMS line under WDS.


Reproductive sink Senescence Oxidative stress Antioxidant defense Drought resistance Senescence associated genes 



Ascorbic acid reduced


Ascorbate peroxidase




Ascorbic acid oxidized


Days after anthesis


Glutamine synthetase


Glutathione reduced


Glutathione oxidized


Reactive oxygen species


Superoxide dismutase


Senescence associated genes


Wheat cysteine protease


Wheat serine protease


Water deficit stress



Dr. (Mrs.) R. Khanna-Chopra was awarded Emeritus Scientist Scheme by Council of Scientific and Industrial Research India which supported the present research. VKS thanks Council of Scientific and Industrial Research India for Research Associate fellowship.

Compliance with ethical standards

Conflict of interest

Dr. Khanna-Chopra has nothing to disclose.

Supplementary material

12298_2018_549_MOESM1_ESM.tif (21.1 mb)
Supplementary Fig. 1. Effect of water deficit stress on the expression of senescence associated genes WRKY53, GS1, WCP2, WSP and CAT2 genes in flag leaf of wheat cv. HW2041 and its CMS line during monocarpic senescence under water deficit stress (TIFF 21636 kb)


  1. Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126CrossRefPubMedGoogle Scholar
  2. Aggarwal PK, Sinha SK (1987) Performance of wheat and triticale varieties in a variable soil water environment. IV. Yield components and their association with grain yield. Field Crops Res 17:45–53CrossRefGoogle Scholar
  3. Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–399CrossRefPubMedGoogle Scholar
  4. Araus JL, Slafer GA, Reynolds MP, Royo C (2002) Plant breeding and drought in C3 cereals: what should we breed for? Ann Bot 200289:925–940CrossRefGoogle Scholar
  5. Asada K (1984) Chloroplasts: formation of active oxygen and its scavenging. Methods Enzymol 105:422–429CrossRefGoogle Scholar
  6. Barrs HD, Weatherley PE (1962) A re-examination of the relative turgidity technique for estimating water deficits in leaves. Aust J Biol Sci 15:413–428CrossRefGoogle Scholar
  7. Bartoli CG, Gómez F, Martínez DE, Guiamet JJ (2004) Mitochondria are the main target for oxidative damage in leaves of wheat (Triticum aestivum L.). J Exp Bot 55:1663–1669CrossRefPubMedGoogle Scholar
  8. Bauer D, Biehler K, Fock H, Carrayol E, Hirel B, Migge A, Becker TW (1997) A role for cytosolic glutamine synthetase in the remobilization of leaf nitrogen during water stress in tomato. Physiol Plant 99:241–248CrossRefGoogle Scholar
  9. Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assay and an assay applicable to acrylamide gels. Anal Biochem 44:276–287CrossRefPubMedGoogle Scholar
  10. Biswas AK, Mandal SK (1986) Monocarpic senescence in wheat: influence of sterile glumes and ear. Physiol Plant 67:431–434CrossRefGoogle Scholar
  11. Blum A, Mayer J, Golan G (1988) The effect of grain number (sink size) on source activity and its water relations in wheat. J Exp Bot 39:106–114CrossRefGoogle Scholar
  12. Borrell AK, Mullet JE, George-Jaeggli B, van Oosterom EJ, Hamme GL, Klein PE, Jordan DR (2014) Drought adaptation of stay-green cereals associated with canopy development, leaf anatomy, root growth and water uptake. J Exp Bot 65:6251–6263CrossRefPubMedPubMedCentralGoogle Scholar
  13. Buchanan-Wollaston V, Page T, Harrison E, Breeze E, Lim PO, Nam HG, Lin JF, Wu SH, Swidzinski J, Ishizaki K et al (2005) Comparative transcriptome analysis reveals significant differences in gene expression and signaling pathways between developmental and dark/starvation-induced senescence in Arabidopsis. Plant J 42:567–585CrossRefPubMedGoogle Scholar
  14. Casano LM, Martin M, Sabater B (1994) Sensitivity of superoxide dismutase transcript levels and activities to oxidative stress is lower in mature-senescent than in young barley leaves. Plant Physiol 106:1033–1039CrossRefPubMedPubMedCentralGoogle Scholar
  15. Casano LM, Gómez LD, Lascano HR, González CA, Trippi VS (1997) Inactivation and degradation of CuZn-SOD by active oxygen species in wheat chloroplasts exposed to photooxidative stress. Plant Cell Physiol 38:433–440CrossRefPubMedGoogle Scholar
  16. Christopher JT, Christopher MJ, Borrell AK, Fletcher S, Chenu K (2016) Stay-green traits to improve wheat adaptation in well-watered and water limited environments. J Exp Bot 67:5159–5172CrossRefPubMedPubMedCentralGoogle Scholar
  17. De Simone V, Soccio M, Borrelli GM, Pastore D, Trono D (2014) Stay-green trait-antioxidant status interrelationship in durum wheat (Triticum durum) flag leaf during post-flowering. J Plant Res 127:159–171CrossRefPubMedGoogle Scholar
  18. Derkx AP, Orford S, Griffiths S, Foulkes MJ, Hawkesford MJ (2012) Identification of differentially senescing mutants of wheat and impacts on yield, biomass and nitrogen partitioning. J Integr Plant Biol 54:555–566CrossRefPubMedGoogle Scholar
  19. Foyer CH, Noctor G (2005) Redox homeostasis and antioxidant signaling: a metabolic interface between stress perception and physiological responses. Plant Cell 17:1866–1875CrossRefPubMedPubMedCentralGoogle Scholar
  20. Gregersen PL, Holm PB (2007) Transcriptome analysis of senescence in the flag leaf of wheat (Triticum aestivum L.). Plant Biotechnol J 5:192–206CrossRefPubMedGoogle Scholar
  21. Gregersen PL, Culetic A, Boschian L, Krupinska K (2013) Plant senescence and crop productivity. Plant Mol Biol 82(6):603–622CrossRefPubMedGoogle Scholar
  22. Guo Y, Gan SS (2014) Translational researches on leaf senescence for enhancing plant productivity and quality. J Exp Bot 65:3901–3913CrossRefPubMedGoogle Scholar
  23. Hameed A, Bibi N, Akhter J, Iqbal N (2011) Differential changes in antioxidants, proteases, and lipid peroxidation in flag leaves of wheat genotypes under different levels of water deficit conditions. Plant Physiol Biochem 49:178–185CrossRefPubMedGoogle Scholar
  24. Hatch MD (1978) A simple spectrophotometric assay for fumarate hydratase in crude tissue extracts. Anal Biochem 85:271–275CrossRefPubMedGoogle Scholar
  25. Havé M, Leitao L, Bagard M, Castell JF, Repellin A (2015) Protein carbonylation during natural leaf senescence in winter wheat, as probed by fluorescein-5-thiosemicarbazide. Plant Biol 17:973–979CrossRefPubMedGoogle Scholar
  26. Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts: I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198CrossRefPubMedGoogle Scholar
  27. Hörtensteiner S, Feller U (2002) Nitrogen metabolism and remobilization during senescence. J Exp Bot 53:927–937CrossRefPubMedGoogle Scholar
  28. Huseynova IM (2012) Photosynthetic characteristics and enzymatic antioxidant capacity of leaves from wheat cultivars exposed to drought. Biochem Biophys Acta 1817:1516–1523PubMedGoogle Scholar
  29. Jiménez A, Hernández JA, Pastori G, del Río LA, Sevilla F (1998) Role of the ascorbate–glutathione cycle of mitochondria and peroxisomes in the senescence of pea leaves. Plant Physiol 118:1327–1335CrossRefPubMedPubMedCentralGoogle Scholar
  30. Joshi AK, Kumari M, Singh VP, Reddy CM, Kumar S, Rane J, Chand R (2007) Stay green trait: variation, inheritance and its association with spot blotch resistance in spring wheat (Triticum aestivum L.). Euphytica 153:59–71CrossRefGoogle Scholar
  31. Kato M, Kobayashi K, Ogiso E, Yokoo M (2004) Photosynthesis and dry-matter production during ripening stage in a female-sterile line of rice. Plant Prod Sci 7:184–188CrossRefGoogle Scholar
  32. Khanna-Chopra R (2012) Leaf senescence and abiotic stresses share reactive oxygen species-mediated chloroplast degradation. Protoplasma 249:469–481CrossRefPubMedGoogle Scholar
  33. Khanna-Chopra R, Chauhan S (2015) Wheat cultivars differing in heat tolerance show a differential response to oxidative stress during monocarpic senescence under high temperature stress. Protoplasma 252:1241–1251CrossRefPubMedGoogle Scholar
  34. Khanna-Chopra R, Selote DS (2007) Acclimation to drought stress generates oxidative stress tolerance in drought-resistant than-susceptible wheat cultivar under field conditions. Environ Exp Bot 60:276–283CrossRefGoogle Scholar
  35. Khanna-Chopra R, Sinha SK (1988) Enhancement of drought-induced senescence by the reproductive sink in fertile lines of wheat and sorghum. Ann Bot 61:649–653CrossRefGoogle Scholar
  36. Khanna-Chopra R, Nutan KK, Pareek A (2013) Regulation of leaf senescence: role of reactive oxygen species. In: Biswal B, Krupinska K, Biswal UC (eds) Plastid development in leaves during growth and senescence. Springer, Dordrecht, pp 393–416CrossRefGoogle Scholar
  37. Koide K, Ishihara K (1992) Effects of ear removal on photosynthesis of the flag leaf during grain filling in wheat. Jpn J Crop Sci 61:659–667CrossRefGoogle Scholar
  38. Kristensen BK, Askerlund P, Bykov NV, Egsgaard H, Møller IM (2004) Identification of oxidized proteins in the matrix of rice leaf mitochondria by immunoprecipitation and two-dimensional liquid chromatography-tandem mass spectrometry. Phytochemistry 65:1839–1851CrossRefPubMedGoogle Scholar
  39. Lee S, Seo PJ, Lee HJ, Park CM (2012) A NAC transcription factor NTL4 promotes reactive oxygen species production during drought-induced leaf senescence in Arabidopsis. Plant J 70:831–844CrossRefPubMedGoogle Scholar
  40. Levine RL, Williams JA, Stadtman ER, Shacter E (1994) Carbonyl assays for determination of oxidatively modified proteins. Methods Enzymol 233:346–357CrossRefPubMedGoogle Scholar
  41. Li Y, Wang M, Zhang F, Xu Y, Chen X, Qin X, Wen X (2016) Effect of post-silking drought on nitrogen partitioning and gene expression patterns of glutamine synthetase and asparagine synthetase in two maize (Zea mays L.) varieties. Plant Physiol Biochem 102:62–69CrossRefPubMedGoogle Scholar
  42. Lichtenthaler HK (1987) Chlorophylls and carotenoids: pigments of photosynthetic biomembranes. Methods Enzymol 148:350–382CrossRefGoogle Scholar
  43. Lilley RMC, Fitzgerald MP, Rienits KG, Walker DA (1975) Criteria of intactness and the photosynthetic activity of spinach chloroplast preparations. New Phytol 75:1–10CrossRefGoogle Scholar
  44. Lim PO, Kim HJ, Nam HG (2007) Leaf senescence. Ann Rev Plant Biol 58:115–136CrossRefGoogle Scholar
  45. Loggini B, Scartazza A, Brugnoli E, Navari-Izzo F (1999) Antioxidant defense system, pigment composition and photosynthetic efficiency in two wheat subjected to drought. Plant Physiol 119:1091–1099CrossRefPubMedPubMedCentralGoogle Scholar
  46. Lowry OH, Rosenbrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin-phenol reagent. J Biol Chem 193:265–275PubMedGoogle Scholar
  47. MacKown CT, Van Sanford DA, Zhang N (1992) Wheat vegetative nitrogen compositional changes in response to reduced reproductive sink strength. Plant Physiol 99:1469–1474CrossRefPubMedPubMedCentralGoogle Scholar
  48. Merewitz EB, Gianfagna T, Huang B (2011) Protein accumulation in leaves and roots associated with improved drought tolerance in creeping bentgrass expressing an ipt gene for cytokinin synthesis. J Exp Bot 62:5311–5333CrossRefPubMedPubMedCentralGoogle Scholar
  49. Miao Y, Laun T, Zimmermann P, Zentgraf U (2004) Targets of WRKY53 transcription factor and its role during leaf senescence in Arabidopsis. Plant Mol Biol 55:853–867CrossRefPubMedGoogle Scholar
  50. Møller IM, Jensen PE, Hansson A (2007) Oxidative modifications to cellular components in plants. Annu Rev Plant Biol 58:459–481CrossRefPubMedGoogle Scholar
  51. Munné-Bosch S, Alegre L (2002) Plant aging increases oxidative stress in chloroplasts. Planta 214:608–615CrossRefPubMedGoogle Scholar
  52. Munné-Bosch S, Jubany-Marí T, Alegre L (2001) Drought-induced senescence is characterized by a loss of antioxidant defences in chloroplasts. Plant Cell Environ 24:1319–1327CrossRefGoogle Scholar
  53. Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate specific peroxidase in spinach chloroplasts. Plant Cell Physiol 22:867–880Google Scholar
  54. Navari-Izzo F, Quartacci MF, Pinzino C, Vecchia FD, Sgherri CLM (1998) Thylakoid bound and stromal anti-oxidative enzymes in wheat treated with excess copper. Physiol Plant 104:630–638CrossRefGoogle Scholar
  55. Noctor G, Mhamdi A, Foyer CH (2014) The roles of reactive oxygen metabolism in drought: not so cut and dried. Plant Physiol 164:1636–1648CrossRefPubMedPubMedCentralGoogle Scholar
  56. Noh YS, Amasino RM (1999) Identification of a promoter region responsible for the senescence-specific expression of SAG12. Plant Mol Biol 41:181–194CrossRefPubMedGoogle Scholar
  57. Nooden LD (1988) Whole plant senescence. In: Nooden LD, Leopold AC (eds) Senescence and aging in plants. Academic Press, San Diego, pp 391–439Google Scholar
  58. Palma JM, Jiménez A, Sandalio LM, Corpas FJ, Lundqvist M, Gómez M, del Río LA (2006) Antioxidative enzymes from chloroplasts, mitochondria, and peroxisomes during leaf senescence of nodulated pea plants. J Exp Bot 57:1747–1758CrossRefPubMedGoogle Scholar
  59. Panchuk II, Zentgraf U, Volkov RA (2005) Expression of the APX gene family during leaf senescence of Arabidopsis thaliana. Planta 222:926–932CrossRefPubMedGoogle Scholar
  60. Pic E, de la Serve BT, Tardieu F, Turc O (2002) Leaf senescence induced by mild water deficit follows the same sequence of macroscopic, biochemical, and molecular events as monocarpic senescence in pea. Plant Physiol 128:236–246CrossRefPubMedPubMedCentralGoogle Scholar
  61. Prochazkova D, Wilhelmova N (2007) Leaf senescence and activities of the antioxidant enzymes. Biol Plant 51:401–406CrossRefGoogle Scholar
  62. Qin G, Meng X, Wang Q, Tian S (2009) Oxidative damage of mitochondrial proteins contributes to fruit senescence: a redox proteomics analysis. J Proteome Res 8:2449–2462CrossRefPubMedGoogle Scholar
  63. Rivero RM, Kojima M, Gepstein A, Sakakibara H, Mittler R, Gepstein S, Blumwald E (2007) Delayed leaf senescence induces extreme drought tolerance in a flowering plant. PNAS USA 104:19631–19636CrossRefPubMedGoogle Scholar
  64. Rosenwasser S, Rot I, Sollner E, Meyer AJ, SmithY Leviatan N, Fluhr R, Friedman H (2011) Organelles contribute differentially to ROS-related events during extended darkness. Plant Physiol 156:185–201CrossRefPubMedPubMedCentralGoogle Scholar
  65. Schaedle M, Bassham A (1977) Chloroplast glutathione reductase. Plant Physiol 53:1011–1012CrossRefGoogle Scholar
  66. Scholander PF, Hammel HT, Hemmingsen EA, Bradstreet ED (1964) Hydrostatic pressure and osmotic potential in leaves of mangroves and some other plants. PNAS USA 52:119–125CrossRefPubMedGoogle Scholar
  67. Selote DS, Khanna-Chopra R (2006) Drought acclimation confers oxidative stress tolerance by inducing co-ordinated antioxidant defense at cellular and subcellular level in leaves of wheat seedlings. Physiol Plant 127:494–506CrossRefGoogle Scholar
  68. Semwal VK, Singh B, Khanna-Chopra R (2014) Delayed expression of SAGs correlates with longevity in CMS wheat plants compared to its fertile plants. Physiol Mol Biol Plants 20:191–199CrossRefPubMedPubMedCentralGoogle Scholar
  69. Shao H, Wang H, Tang X (2015) NAC transcription factors in plants multiple abiotic stress responses: progress and prospects. Front Plant Sci 6:902. CrossRefPubMedPubMedCentralGoogle Scholar
  70. Shinozaki K, Yamaguchi-Shinozaki K (2007) Gene networks involved in drought stress response and tolerance. J Exp Bot 58:221–227CrossRefPubMedGoogle Scholar
  71. Simova-Stoilova L, Demirevska K, Petrova T, Tsenov N, Feller U (2009) Antioxidative protection and proteolytic activity in tolerant and sensitive wheat (Triticum aestivum L.) varieties subjected to long-term field drought. Plant Growth Regul 58:107–117CrossRefGoogle Scholar
  72. Smakowska E, Czarna M, Janska H (2014) Mitochondrial ATP-dependent proteases in protection against accumulation of carbonylated proteins. Mitochondrion 19:245–251CrossRefPubMedGoogle Scholar
  73. Srivalli B, Khanna-Chopra R (2001) Induction of new isoforms of superoxide dismutase and catalase enzymes in the flag leaf of wheat during monocarpic senescence. Biochem Biophys Res Commun 288:1037–1042CrossRefPubMedGoogle Scholar
  74. Srivalli B, Khanna-Chopra R (2004) The developing reproductive sink induces oxidative stress to mediate nitrogen mobilization during monocarpic senescence in wheat. Biochem Biophys Res Commun 325:198–202CrossRefPubMedGoogle Scholar
  75. Srivalli S, Khanna-Chopra R (2009) Delayed wheat flag leaf senescence due to removal of spikelets is associated with increased activities of leaf antioxidant enzymes, reduced glutathione/oxidized glutathione ratio and oxidative damage to mitochondrial proteins. Plant Physiol Biochem 47:663–670CrossRefPubMedGoogle Scholar
  76. Thomas H, Howarth CJ (2000) Five ways to stay green. J Exp Bot 51:329–337CrossRefPubMedGoogle Scholar
  77. Thomas H, Ougham H (2014) The stay-green trait. J Exp Bot 65:3889–3900CrossRefPubMedGoogle Scholar
  78. Tian FX, Gong JF, Wang GP, Wang GK, Fan ZY, Wang W (2012) Improved drought resistance in a wheat stay-green mutant tasg1 under field conditions. Biol Plant 56:509–515CrossRefGoogle Scholar
  79. Tian FX, Gong J, Zhang J, Zhang M, Wang G, Li A, Wang W (2013) Enhanced stability of thylakoid membrane proteins and antioxidant competence contribute to drought stress resistance in the tasg1 wheat stay-green mutant. J Exp Bot 64:1509–1520CrossRefPubMedPubMedCentralGoogle Scholar
  80. Uauy C, Brevis JC, Dubcovsky J (2006) The high grain protein content gene Gpc-B1 accelerates senescence and has pleiotropic effects on protein content in wheat. J Exp Bot 57:2785–2794CrossRefPubMedGoogle Scholar
  81. Veljovic-Jovanovic S, Noctor G, Foyer CH (2002) Are leaf hydrogen peroxide concentrations commonly overestimated? The potential influence of artefactual interference by tissue phenolics and ascorbate. Plant Physiol Biochem 40:501–507CrossRefGoogle Scholar
  82. Verma V, Foulkes MJ, Worland AJ, Sylvester-Bradley R, Caligari PDS, Snape JW (2004) Mapping quantitative trait loci for flag leaf senescence as a yield determinant in winter wheat under optimal and drought-stressed environments. Euphytica 135:255–263CrossRefGoogle Scholar
  83. Wang SY, Jiao HJ, Faust M (1991) Changes in ascorbate, glutathione, and related enzyme activities during thidiazuron-induced bud break of apple. Physiol Plant 82:231–236CrossRefGoogle Scholar
  84. Weaver LM, Gan S, Quirino B, Amasino RM (1998) A comparison of the expression patterns of several senescence-associated genes in response to stress and hormone treatment. Plant Mol Biol 37:455–469CrossRefPubMedGoogle Scholar
  85. Willekens H, Chamnongpol S, Davey M, Schraudner M, Langebartels C, Van Montagu M, Inze D, Van Camp W (1997) Catalase is a sink for H2O2 and is indispensable for stress defence in C3 plants. EMBO J 16:4806–4816CrossRefPubMedPubMedCentralGoogle Scholar
  86. Woo HR, Kim HJ, Nam HG, Lim PO (2013) Plant leaf senescence and death—regulation by multiple layers of control and implications for aging in general. J Cell Sci 126:4823–4833CrossRefPubMedGoogle Scholar
  87. Wu A, Allu AD, Garapati P, Siddiqui H, Dortay H, Zanor MI, Asensi-Fabado MA, Munné-Bosch S, Antonio C, Tohge T, Fernie AR (2012) JUNGBRUNNEN1, a reactive oxygen species-responsive NAC transcription factor, regulates longevity in Arabidopsis. Plant Cell 24:482–506CrossRefPubMedPubMedCentralGoogle Scholar
  88. Zhao Y, Chan Z, Gao J, Xing L, Cao M, Yu C, Gong Y, Zhu JK (2016) ABA receptor PYL9 promotes drought resistance and leaf senescence. PNAS USA 113:1949–1954CrossRefPubMedGoogle Scholar
  89. Zimmermann P, Orendi G, Heinlein C, Zentgraf U (2006) Senescence specific regulation of catalases in Arabidopsis thaliana (L.) Heynh. Plant Cell Environ 29:1049–1060CrossRefPubMedGoogle Scholar

Copyright information

© Prof. H.S. Srivastava Foundation for Science and Society 2018

Authors and Affiliations

  1. 1.Stress Physiology Lab, Water Technology CentreIndian Agricultural Research InstituteNew DelhiIndia
  2. 2.Africa Rice Center (AfricaRice), C/O IITAIbadanNigeria

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