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Molecular Mechanisms Regulating Carbohydrate Metabolism During Lolium perenne Regrowth Vary in Response to Nitrogen and Gibberellin Supply


The promoting effects of both high nitrogen (N) and exogenous gibberellin (GA) supply on regrowth of Lolium perenne have been widely reported. The mobilisation of carbohydrate reserves in response to N is a critical mechanism for promoting plant regrowth. However, our knowledge about GA regulation of carbohydrate metabolism remains limited. Here, we analysed the effects of both N and exogenous GA on the molecular mechanisms controlling perennial ryegrass regrowth and investigated the similarities and differences. Our analyses show that both high N and exogenous GA supply lead to a decline in the accumulation of carbohydrate reserves, but the regulatory mechanisms responsible for this decline varied between N and GA supply. The effects of elevated N were mainly through declining fructan biosynthesis which results in improving photosynthate use efficiency to promote plant regrowth, whereas the application of exogenous GA resulted in an increase in the hydrolytic activities of fructan exohydrolase and invertases capable of cleaving reserved carbohydrates to release energy sources for plant regrowth.

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  1. Ali A, Kafkafi U, Yamaguchi I, Sugimoto Y, Inanag S (1996) Effects of low root temperature on sap flow rate, soluble carbohydrates nitrite contents and on cytokinin and gibberellin levels in root xylem exudate of sand-grown tomato. J Plant Nutr 19:619–634

  2. Ayele BT, Ozga JA, Reinecke DM (2006) Regulation of GA biosynthesis genes during germination and young seedling growth of pea (Pisum sativum L.) J Plant Growth Regul 25:219–232

  3. Bouton S, Leydesher MT, Meyer C, Truing HN (2002) Role of gibberellins and the RGA and GAI genes in controlling nitrate assimilation in Arabidopsis thaliana. Plant Physiol Biochem 40:939–947

  4. Cai Y, Shao L, Li X, Liu G (2016) Chen S (2016) Gibberellin stimulates regrowth after defoliation of sheepgrass (Leymus chinensis) by regulating expression of fructan-related genes. J Plant Res 129:935–944

  5. Cairns AL, Gallagher JA (2004) Absence of turnover and futile cycling of sucrose in leaves of Lolium temulentum L.: implications for metabolic compartmentation. Planta 219:836–846

  6. Conaghan P, O’Kiely P, Halling MA, O’Mara FP, Nesheim L (2012) Yield and quality response of perennial ryegrass selected for high concentration of water-soluble carbohydrate to nitrogen application rate. Crop Sci 52:2839–2851

  7. Davies CS (2000) Strategy differences of two potato species in response to nitrogen starvation. Do plants have a genetic switch for nitrogen signalling? Plant Cell Environ 23:759–765

  8. de Roover L, Vandenbranden K, Van Laere A, Van de Ende W (2000) Drought induces fructan synthesis and 1-SST (sucrose: sucrose fructosyltransferase) in roots and leaves of Cichorium seedlings (Cichorium intybus L.) Planta 210:808–814

  9. de Visser R, Vianden H, Schnyder H (1997) Kinetics and relative significance of remobilized and current C and N incorporation in leaf and root growth zones of Lolium perenne after defoliation: assessment by 13C and 15N steady-state labelling. Plant Cell Environ 20:37–46

  10. Edwards GR, Parsons AJ, Rasmussen S, Bryant RH (2007) High sugar ryegrasses for livestock systems in New Zealand. Proc N Z Grassl Assoc 69:161–171

  11. Gandi AP, Naik MS (1974) Role of roots, hormones and light in the synthesis of nitrate reductase and nitrite reductase in rice seedlings. FEBS Lett 40:343–345

  12. Gocal GF, Poole AT, Gubler F, Watts RJ, Blundell C, King RW (1999) Long-day up-regulation of a GAMYB gene during Lolium Temulentum inflorescence formation. Plant Physiol 119:1271–1278

  13. Guerrand D, Prud’Homme MP, Boucaud J (1996) Fructan metabolism in expanding leaves, mature leaf sheaths and mature leaf blades of Lolium perenne. Fructan synthesis, fructosyltransferase and invertase activities. New Phytol 134:205–214

  14. Hedden P, Phillips AL (2000) Gibberellin metabolism: new insights revealed by genes. Trends Plant Sci 12:523–530

  15. Hisano H, Kanazawa A, Yoshida M, Humphreys MO, Iizuka M, Kitamura K, Yamada T (2008) Coordinated expression of functionally diverse fructosyltransferase genes is associated with fructan accumulation in response to low temperature in perennial ryegrass. New Phytol 178:766–780

  16. Hong YF, Ho TH, Wu CF, Ho SL, Yeh RH, Lu CA, Chen PW, Yu LC, Chao A, Yu SM (2012) Convergent starvation signals and hormone crosstalk in regulating nutrient mobilization upon germination in cereals. Plant Cell 24:2857–2873

  17. Hunt MG, Rasmussen S, Newton PCD, Parsons AJ, Newman JA (2005) Near-term impacts of elevated CO2, nitrogen and fungal endophyte-infection on Lolium perenne L.: growth, chemical composition and alkaloid production. Plant Cell Environ 28:1345–1354

  18. Jang SW, Hamayun M, Sohn EY, Shin DH, Kim KU, Lee BH, Lee IJ (2008) Effect of elevated nitrogen levels on endogenous gibberellin and jasmonic acid contents of three rice (Oryza sativa L) cultivars. J Plant Nutr Soil Sci 171:181–186

  19. Jermyn MA (1956) A new method for determining ketohexoses in the presence of aldohexoses. Nature 177:38–39

  20. Johnson X, Lidgett A, Chalmers J, Guthridge K, Jones E, Cummings N, Spangenberg G (2003) Isolation and characterisation of an invertase cDNA from perennial ryegrass (Lolium perenne). J Plant Physiol 160:903–911

  21. Kaneko M, Itoh H, Inukai Y, Sakamoto T, Ueguchi-Tanaka M, Ashikari M, Matsuoka M (2003) Where do gibberellin biosynthesis and gibberellin signalling occur in rice plants? Plant J 35:104–115

  22. Kaufman PB, Ghoshen NS, LaCroix JD, Soni SL, Ikuma H (1973) Regulation of invertase levels in Avena stem segments by gibberellic acid, sucrose, glucose, and fructose. Plant Physiol 52:221–228

  23. Kavanová M, Lattanzi FA, Schnyder H (2008) Nitrogen deficiency inhabits leaf blade growth in Lolium perenne by increasing cell cycle duration and decreasing mitotic and post-mitotic growth rates. Plant Cell Environ 31:103–113

  24. Keating T, O’Kiely P (2000) Comparison of old permanent grassland, Lolium perenne and Lolium multiflorum swards grown for silage. 3. Effects of varying fertiliser nitrogen application rate. Ir J Agric Food Res 39:35–53

  25. Kingston-Smith AH, Walker RP, Pollock CJ (1999) Invertase in leaves: conundrum or control point? J Exp Bot 50:735–743

  26. Kozlowska M, Rybus-Zajac M, Stachowiak J, Janowska B (2007) Changes in carbohydrate contents of Zantedeschia leaves under gibberellin-stimulated flowering. Acta Physiol Plant 29:27–32

  27. Krauss A, Marschner H (1982) Influence of nitrogen nutrition, day length and temperature on contents of gibberellic and abscisic acid and on tuberizations in potato plants. Potato Res 25:13–21

  28. Lattanzi FA, Ostler U, Wild M, Morvan-Bertrand A, Decau ML, Lehmeier CA, Meuriot F, Prud’homme MP, Schaüfele R, Schnyder H (2012) Fluxes in central carbohydrate metabolism of source leaves in a fructan-storing C3 grass: rapid turnover and futile cycling of sucrose in continuous light under contrasted nitrogen nutrition status. J Exp Bot 63:2363–2375

  29. Lee JM, Sathish P, Donaghy DJ, Roche JR (2011) Impact of defoliation severity on photosynthesis, carbon metabolism and transport gene expression in perennial ryegrass. Funct Plant Biol 38:808–817

  30. Lehmeier CA, Lattanzi FA, Schaüfele R, Schnyder H (2010) Nitrogen deficiency increases the residence time of respiratory carbon in the respiratory substrate supply system of perennial ryegrass. Plant Cell Environ 33:76–87

  31. Lidgett A, Jennings K, Johnson X, Guthridge K, Jones E, Spangenberg G (2002) Isolation and characterisation of a fructosyltransferase gene from perennial ryegrass (Lolium perenne). J Plant Physiol 159:1037–1043

  32. Liu Q, Jones CS, Parsons A, Xue H, Rasmussen S (2015) Does gibberellin biosynthesis play a critical role in the growth of Lolium perenne? Evidence from a transcriptional analysis of gibberellin and carbohydrate metabolic genes after defoliation. Front Plant Sci 6:944. https://doi.org/10.3389/fpls.2015.00944

  33. Mathew C, Hofmann WA, Osborne MA (2009) Pasture response to gibberellins: a review and recommendations. N Z J Agric Res 52:213–225

  34. MacMillan CP, Blundell CA, King RW (2005) Flowering of the grass Lolium perenne: effects of verbalization and long days on gibberellin biosynthesis and signalling. Plant Physiol 138:1794–1806

  35. Martin DN, Proebsting WM, Parks TD, Dougherty WG, Lange T, Lewis MJ, Gaskin P, Hedden P (1996) Feed-back regulation of gibberellin biosynthesis and gene expression in Pisum sativum L. Planta 200:159–166

  36. Marx S, Nösberger J, Frehner M (1997) Seasonal variation of fructan-β-fructosidase (FEH) activity and characterization of a β-(2–1)-linkage specific FEH from tubers of Jerusalem artichoke (Helianthus tuberosus). New Phytol 135:267–277

  37. Matsushita A, Furumoto T, Ishida S, Takahashi Y (2007) AGF1, an AY-hook protein, is necessary for the negative feedback of AtGA3ox1 encoding GA3-oxidase. Plant Physiol 143:1152–1162

  38. McIntyre CL, Casu RE, Rattey A, Dreccer MF, Kam JW, van Herwaarden AF, Shorter R, Xue GP (2011) Linked gene networks involved in nitrogen and carbon metabolism and levels of water-soluble carbohydrate accumulation in wheat stems. Funct Integr Genomics 11:585–597

  39. Middleton A, Úbeda-Tomás S, Griffiths J, Holman T et al (2012) Mathematical modelling elucidates the role of transcriptional feedback in gibberellin signalling. Proc Natl Acad Sci USA 109:7571–7576

  40. Ministry for the Environment (2013) Ministry for the Environment New Zealand's greenhouse gas inventory 1990–2011. https://www.mfe.govt.nz/publications/climate/greenhouse-gas-inventory-2013/index.html.

  41. Morcuende R, Kostadinova P, Pèrez P, Martìn del Molino IM, Martìnez-Carrasco R (2004) Nitrate is a negative signal for fructan synthesis, and the fructosyltransferase-inducing trehalose inhibits nitrogen and carbon assimilation in excised barley leaves. New Phytol 161:749–759

  42. Morris DA, Arthur ED (1985) Effects of gibberellic acid on patterns of carbohydrate distribution and acid invertase activity in Phaseolus vulgaris. Physiol Plant 65:257–262

  43. Morvan A, Challe G, Prud’homme MP, Le Saos J, Boucaud J (1997) Rise of fructan exohydrolase activity in stubble of Lolium perenne after defoliation is decreased by uniconazole, an inhibitor of the biosynthesis of gibberellins. New Phytol 136:81–88

  44. Morvan-Bertrand A, Boucaud J, Prud’homme MP (1999) Influence of initial levels of carbohydrates, fructans, nitrogen, and soluble proteins on regrowth of Lolium perenne L. cv. Bravo following defoliation. J Exp Bot 50:1817–1826

  45. Morvan-Bertrand A, Ernstsen A, Lindgárd B, Koshioka M, Le Saos J, Boucaud J, Prud’homme MP, Junttila O (2001) Endogenous gibberellins in Lolium perenne and influence of defoliation on their contents in elongating leaf bases and in leaf sheaths. Physiol Plant 111:225–231

  46. Ogawa M, Kusano T, Katsumi M, Sano H (2000) Rice gibberellin-insensitive gene homolog, OsGAi, encodes a nuclear-localized protein capable of gene activation at transcriptional level. Gene 245:21–29

  47. Olszewski N, Sun TP, Gubler F (2002) Gibberellin signaling: biosynthesis, catabolism, and response pathways. Plant Cell 14:S61–80

  48. Parsons AJ, Rasmussen S, Liu Q, Xue H, Ball C, Shaw C (2013) Plant growth—resource or strategy limited: insights from responses to gibberellin. Grass Forage Sci 68:577–588

  49. Peng J, Carol P, Richards DE, King KE, Cowling RJ, Murphy GP, Harberd NP (1997) The Arabidopsis GAI gene defines a signalling pathway that negatively regulates gibberellin responses. Gene Dev 11:3194–3205

  50. Pollock CJ, Cairns AJ (1991) Fructan metabolism in grasses and cereals. Annu Rev Plant Physiol Plant Mol Biol 42:77–101

  51. Prud'homme MP, Gonzalez B, Billard JP, Boucaud J (1992) Carbohydrate content, fructan and sucrose enzyme activities in roots, stubble and leaves of ryegrass (Lolium perenne L.) as affected by source/sink modification after cutting. J Plant Physiol 40:282–291

  52. Ranwala AP, Miller WB (2008) Gibberellin-mediated changes in carbohydrate metabolism during flower stalk elongation in tulips. Plant Growth Regul 55:241–248

  53. Rasmussen S, Parsons AJ, Basset S, Christensen MJ, Hume DE, Johnson LJ, Johnson RD, Simpson WR, Stacke C, Viosey CR, Xue H, Newman JA (2007) High nitrogen supply and carbohydrate content reduce fungal endophyte and alkaloid concentration in Lolium perenne. New Phytol 173:787–797

  54. Rasmussen S, Parsons AJ, Xue H, Liu Q, Jones CS, Ryan GD, Newman JA (2014) Transcript profiling of fructan biosynthetic pathway genes reveals association of a specific fructosyltransferase isoform with the high sugar trait in Lolium perenne. J Plant Physiol 171:475–485

  55. Ruuska SA, Lewis DC, Kennedy G, Furbank RT, Jenkins CLD, Tabe LM (2008) Large scale transcriptome analysis of the effects of nitrogen nutrition on accumulation of stem carbohydrate reserves in reproductive stage wheat. Plant Mol Biol 66:15–32

  56. Schnyder H, de Visser R (1999) Fluxes of reserve-derived and currently assimilated carbon and nitrogen in perennial ryegrass recovering from defoliation. The regrowing tiller and its component functionally distinct zones. Plant Physiol 119:1423–1435

  57. Stitt M, Krapp A (1999) The interaction between elevated carbon dioxide and nitrogen nutrition: the physiological and molecular background. Plant Cell Environ 22:583–621

  58. Sturm A (1999) Invertases. Primary structures, functions, and roles in plant development and sucrose partitioning. Plant Physiol 121:1–8

  59. Thomas SG, Phillips AL, Hedden P (1999) Molecular cloning and functional expression of gibberellin 2-oxidases, multifunctional enzymes involved in gibberellin deactivation. Proc Natl Acad Sci USA 96:4698–4703

  60. Truong HN, Caboche M, Daniel-Vedele F (1997) Sequence and characterization of two Arabidopsis thaliana cDNAs isolated by functional complementation of a yeast gln3 gdh1 mutant. FEBS Lett 410:213–218

  61. van den Ende W, Clerens S, Vergauwen R, Va Riet L, Van Laere A, Yoshida M, Kawakami A (2003) Fructan 1-exohydrolase. β-(2,1)-trimmers during graminan biosynthesis in stems of wheat? Purification, characterization, mass mapping and cloning of two fructan 1-exohydrolase isoforms. Plant Physiol 131:621–631

  62. van den Ende W, De Roover J, Van Laere A (1999) Effect of nitrogen concentration on fructan and fructan metabolizing enzymes in young chicory plants (Cichorium intybus). Physiol Plant 105:2–8

  63. van Rossum MH, Bryant RH, Edwards GR (2013) Response of simple grass-white clover and multi-species pastures to gibberellic acid or nitrogen fertiliser in autumn. Proc N Z Grassl Assoc 75:145–150

  64. Wang C, Tillberg JE (1996) Effects of nitrogen deficiency on accumulation of fructan and fructan metabolizing enzyme activities in sink and source leaves of barley (Hordeum vulgare). Physiol Plant 5:17–37

  65. Wang C, van den Ende W, Tillberg JE (2000) Fructan accumulation induced by nitrogen deficiency in barley leaves correlates with the level of sucrose: fructan 6-fructosyltransferase mRNA. Planta 211:701–707

  66. Wu LL, Mitchell JP, Cohn NS, Kaufman PB (1993) Gibberellin (GA3) enhances cell wall invertase activity and mRNA levels in elongating dwarf pea (Pisum sativum) shoots. Int J Plant Sci 154:280–289

  67. Xu YL, Li L, Gage DA, Zeevaart AD (1999) Feedback regulation of GA5 expression and metabolic engineering of gibberellin levels in Arabidopsis. Plant Cell 11:927–935

  68. Yamaguchi J (1998) Analysis of embryo-specific α-amylase using isolated mature rice embryos. Breed Sci 48:365–370

  69. Yamaguchi S (2008) Gibberellin metabolism and its regulation. Annu Rev Plant Biol 59:225–251

  70. Yamamoto S, Mino Y (1987) Effect of sugar level on phleinase induction in stem base of orchargrass after defoliation. Physiol Plant 69:456–460

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This work was supported by the New Zealand Agricultural Greenhouse Gas Research Centre and conducted at AgResearch Grasslands, Palmerston North, New Zealand. The authors acknowledge Daniel Bastias (AgResearch) for critically reviewing the manuscript and Dongwen Luo (AgResearch) and Briar Davies (internship) for statistical advice and technical assistance.

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Correspondence to Chris S. Jones.

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Liu, Q., Rasmussen, S., Johnson, L.J. et al. Molecular Mechanisms Regulating Carbohydrate Metabolism During Lolium perenne Regrowth Vary in Response to Nitrogen and Gibberellin Supply. J Plant Growth Regul (2020). https://doi.org/10.1007/s00344-020-10070-y

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  • Perennial ryegrass
  • Regrowth
  • Fructans
  • Exogenous gibberellin
  • Nitrogen
  • Gene expression