Tropical Plant Biology

, Volume 1, Issue 2, pp 142–158 | Cite as

Differential Expression of Genes in the Leaves of Sugarcane in Response to Sugar Accumulation



In C4 sugarcane (Saccharum spp. hybrids), photosynthetic activity has been shown to be regulated by the demand for carbon from sink tissues. There is evidence, from other plant species, that sink-limitation of photosynthesis is facilitated by sugar-signaling mechanisms in the leaf that affect photosynthesis through regulation of gene expression. In this work, we manipulated leaf sugar levels by cold-girdling leaves (5°C) for 80 h to examine the mechanisms whereby leaf sugar accumulation affects photosynthetic activity and assess whether signaling mechanisms reported for other species operate in sugarcane. During this time, sucrose and hexose concentrations above the girdle increased by 77% and 81%, respectively. Conversely, leaf photosynthetic activity (A) and electron transport rates (ETR) decreased by 66% and 54%, respectively. Quantitative expression profiling by means of an Affymetrix GeneChip Sugarcane Genome Array was used to identify genes responsive to cold-girdling (56 h). A number of genes (74) involved in primary and secondary metabolic pathways were identified as being differentially expressed. Decreased expression of genes related to photosynthesis and increased expression of genes involved in assimilate partitioning, cell wall synthesis, phosphate metabolism and stress were observed. Furthermore four probe sets homologous to trehalose 6-phosphate phosphatase (TPP; EC and trehalose 6-phosphate synthase (TPS; EC were up- and down-regulated, respectively, indicating a possible role for trehalose 6-phosphate (T6P) as a putative sugar-sensor in sugarcane leaves.


Cold-girdling Expression profiling Genes Leaf Photosynthesis Sugar Sugarcane Trehalose 



The authors are grateful for funding provided by the South African Sugarcane Research Institute, SA Sugar Association Trust Fund for Education and the National Research Foundation.

Supplementary material

12042_2008_9013_MOESM1_ESM.xls (126 kb)
Supplementary Table 1 List of probe sets differentially expressed during the cold-girdling treatment. Putative identity was assigned using the BLASTX function within the National Centre of Biotechnological Information (NCBI) GenBank database ( Where E values are absent, probe sets homology was matched to those assigned by Casu et al. [12]. Fold changes indicate statistical significance values (P < 0.05) as determined by ANOVA (n = 4) (XLS 128 KB)
12042_2008_9013_Fig1_ESM.gif (496 kb)
Fig. 1

Normalised gene expression profile comparison between cold-girdled (56 h) and control leaves. Up- and down-regulation of genes in controls (n = 4) is seen in red and blue, respectively, whereas the reverse applies for the cold-girdled leaves. Genes that remain unaffected by the treatment are depicted in yellow (GIF 2.24 MB)

12042_2008_9013_Fig1_ESM.tif (2.2 mb)
Supplementary Fig. 1 High resolution image file (TIFF 495 kb)


  1. 1.
    Allison JCS, Williams HT, Pammenter NW (1997) Effect of specific leaf nitrogen on photosynthesis of sugarcane. Ann Appl Biol 63:135–144CrossRefGoogle Scholar
  2. 2.
    Amaya A, Cock JH, Hernandez A, Irvine J (1995) Bioligía. In: Casselett C, Torres J, Isaacs C (eds) El cultivo de la Caňa en la zona azucarera de Colombia. Cenicaňa, Cali, Colombia, pp 31–62Google Scholar
  3. 3.
    Altschul SF, Madden TL, Schaffer AA, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402PubMedCrossRefGoogle Scholar
  4. 4.
    Arruda P (2001) Sugarcane transcriptome. A landmark in plant genomics in the tropics. Genet Mol Biol 24:1–4CrossRefGoogle Scholar
  5. 5.
    Basu PS, Sharma A, Garg ID, Sukumaran NP (1999) Tuber sink modifies photosynthetic response in potato under water stress. Environ Exp Bot 42:25–29CrossRefGoogle Scholar
  6. 6.
    Bläsing OE, Gibon Y, Günther M, Höhne M, Morcuende R, Osuna D, Thimm O, Usadel B, Scheible WR, Stitt M (2005) Sugars and circadian regulation make major contributions to the global regulation of diurnal gene expression in Arabidopsis. The Plant Cell 17:3257–3281PubMedCrossRefGoogle Scholar
  7. 7.
    Bugos RC, Chiang VL, Zhang XH, Campbell ER, Podila GK, Campbell WH (1995) RNA isolation from plant tissues recalcitrant to extraction in guanidine. Biotechniques 19:734–737PubMedGoogle Scholar
  8. 8.
    Bull TA, Tovey DA (1974) Aspects of modelling sugar cane growth by computer simulation. Proc Int Soc Sugarcane Technol 165:1021–1032Google Scholar
  9. 9.
    Carson DL, Huckett BI, Botha FC (2002) Sugarcane ESTs differentially expressed in immature and maturing internodal tissue. Plant Sci 162:289–300CrossRefGoogle Scholar
  10. 10.
    Casu RE, Dimmock CM, Chapman SC, Grof CPL, McIntyre CL, Bonnett GD, Manners JM (2004) Identification of differentially expressed transcripts from maturing stem of sugarcane by in silico analysis of stem expressed sequence tags and gene expression profiling. Plant Mol Biol 54:503–517PubMedCrossRefGoogle Scholar
  11. 11.
    Casu RE, Grof CPL, Rae AL, McIntyre CL, Dimmock CM, Manners JM (2003) Identification of a novel sugar transporter homologue strongly expressed in maturing stem vascular tissues in sugarcane by expressed sequence tag and microarray analysis. Plant Mol Biol 52:371–386PubMedCrossRefGoogle Scholar
  12. 12.
    Casu RE, Jarmey J, Bonnett G, Manners J (2007) Identification of transcripts associated with cell wall metabolism and development in the stem of sugarcane by Affymetrix GeneChip Sugarcane Genome Array expression profiling. Funct Integr Genomics 7:153–167PubMedCrossRefGoogle Scholar
  13. 13.
    Ciereszko I, Johnsson H, Hurry V, Kleczkowski LA (2001) Phosphate status affects the gene expression, protein content and enzymatic activity of UDP-glucose pyrophosphorylase in wild-type and pho mutants of Arabidopsis. Planta 212:598–605PubMedCrossRefGoogle Scholar
  14. 14.
    Davies C, Robinson SP (2000) Differential screening indicates a dramatic change in mRNA profiles during grape berry ripening. Cloning and characterization of cDNAs encoding putative cell wall and stress response genes. Plant Physiol 122:803–812PubMedCrossRefGoogle Scholar
  15. 15.
    Du YC, Nose A, Kondo A, Wasano K (2000) Diurnal changes in photosynthesis in sugarcane leaves. II. Enzyme activities and metabolite levels relating to sucrose and starch metabolism. Plant Prod Sci 3:9–16CrossRefGoogle Scholar
  16. 16.
    Eastmond PJ, Li Y, Graham IA (2003) Is trehalose-6-phosphate a regulator of sugar metabolism in plants? J Exp Bot 54:533–537PubMedCrossRefGoogle Scholar
  17. 17.
    Eastmond PJ, van Dijken AJ, Spielman M, Kerr A, Tissier AF, Dickinson HG, Jones JD, Smeekens SC, Graham IA (2002) Trehalose-6-phosphate synthase 1, which catalyses the first step in trehalose synthesis, is essential for Arabidopsis embryo maturation. Plant J 29:225–235PubMedCrossRefGoogle Scholar
  18. 18.
    Edwards GE, Baker NR (1993) Can CO2 assimilation in maize leaves be predicted accurately from chlorophyll fluorescence analysis? Photosynth Res 37:89–102CrossRefGoogle Scholar
  19. 19.
    Ehness R, Ecker M, Godt DE, Roitsch T (1997) Glucose and stress independently regulate source and sink metabolism and defense mechanisms via signal transduction pathways involving protein phosphorylation. Plant Cell 9:1825–1841PubMedCrossRefGoogle Scholar
  20. 20.
    Franck N, Vaast P, Genard M, Dauzat J (2006) Soluble sugars mediate sink feedback down-regulation of leaf photosynthesis in field-grown Coffea arabica. Tree Physiol 26:517–525PubMedGoogle Scholar
  21. 21.
    Franco-Zorrilla JM, Martin AC, Leyva A, Paz-Ares J (2005) Interaction between phosphate-starvation, sugar, and cytokinin signaling in Arabidopsis and the roles of cytokinin receptors CRE1/AHK4 and AHK3. Plant Physiol 138:847–857PubMedCrossRefGoogle Scholar
  22. 22.
    Gibon Y, Blaesing OE, Hannemann J, Carillo P, Höhne M, Hendriks JHM, Palacios N, Cross J, Selbig J, Stitt M (2006) A robot-based platform to measure multiple enzyme activities in Arabidopsis using a set of cycling assays: comparison of changes of enzyme activities and transcript levels during diurnal cycles and in prolonged darkness. Plant Cell 16:3304–3325CrossRefGoogle Scholar
  23. 23.
    Grof C, Campbell J (2001) Sugarcane sucrose metabolism: scope for molecular manipulation. Aust J Plant Physiol 28:1–12Google Scholar
  24. 24.
    Goldschmidt EE, Huber SC (2001) Regulation of photosynthesis by end-product accumulation in leaves of plants storing starch, sucrose, and hexose sugars. Plant Physiol 99:1443–1448Google Scholar
  25. 25.
    Gutiérrez-Miceli FA, Morales-Torres R, de Jesús Espinosa-Castañeda Y, Rincón-Rosales R, Mentes-Molina J, Oliva-Llaven MA, Dendooven L (2004) Effects of partial defoliation on sucrose accumulation, enzyme activity and agronomic parameters in sugar cane (Saccharum spp.). J Agron Crop Sci 190:256–261CrossRefGoogle Scholar
  26. 26.
    Hartt CE, Burr GO (1967) Factors affecting photosynthesis in sugarcane. Proc Int Soc Sugarcane Technol 12:590–609Google Scholar
  27. 27.
    Huckett BA, Botha FC (1995) Stability and potential use of RAPD markers in a sugarcane genealogy. Euphytica 86:117–125CrossRefGoogle Scholar
  28. 28.
    Iglesias DJ, Lliso I, Tadeo FR, Talon M (2002) Regulation of photosynthesis through source: sink imbalance in citrus is mediated by carbohydrate content in leaves. Physiol Plant 116:563–572CrossRefGoogle Scholar
  29. 29.
    Ingelbrecht IL, Mandelbaum CI, Mirkov TE (1998) Highly sensitive northern hybridization using a rapid protocol for downward alkaline blotting of RNA. BioTechniques 25:420–425PubMedGoogle Scholar
  30. 30.
    Irizarry RA, Bolstad BM, Collin F, Cope LM, Hobbs B, Speed TP (2003) Summaries of Affymetrix GeneChip probe level data. Nucleic Acid Res 4(e15):1–8Google Scholar
  31. 31.
    Jackson PA (2005) Breeding for improved sugar content in sugarcane. Field Crops Res 92:277–290CrossRefGoogle Scholar
  32. 32.
    Jones MG, Outlaw WH, Lowry OH (1977) Enzymic assay of 10−7 to 10−14 moles of sucrose in plant tissues. Plant Physiol 60:379–383PubMedCrossRefGoogle Scholar
  33. 33.
    Kalt-Torres W, Kerr PS, Usuda H, Huber SC (1987) Diurnal changes in maize leaf photosynthesis. Plant Physiol 83:283–288PubMedGoogle Scholar
  34. 34.
    Krapp A, Hofman B, Schäfer C, Stitt M (1993) Regulation of the expression of rbcS and other photosynthetic genes by carbohydrates: mechanism for the sink regulation of photosynthesis? Plant J 3:817–828CrossRefGoogle Scholar
  35. 35.
    Krapp A, Quick WP, Stitt W (1991) Ribulose-1,5-bisphosphate carboxylase-oxygenase, other Calvin cycle enzymes and chlorophyll decrease when glucose is supplied to mature spinach leaves via the transcription stream. Planta 186:58–59CrossRefGoogle Scholar
  36. 36.
    Krapp A, Stitt M (1995) An evaluation of direct and indirect mechanisms for the “sink regulation” of photosynthesis in spinach: changes in gas exchange, carbohydrates, metabolites, enzyme activities and steady state transcript levels after cold-girdling leaves. Planta 195:313–323CrossRefGoogle Scholar
  37. 37.
    Kolbe A, Tiessen A, Schluepmann H, Paul M, Ulrich S, Geigenberger P (2005) Trehalose 6-phosphate regulates starch synthesis via post-translational redox activation of ADP-glucose pyrophosphorylase. Proc Natl Acad Sci USA 102:11118–11123PubMedCrossRefGoogle Scholar
  38. 38.
    Lawlor DW (1987) Photosynthesis: metabolism, control and physiology. Longman, Harlow, UKGoogle Scholar
  39. 39.
    Lee JM, Williams ME, Tingey SV, Rafalski AJ (2002) DNA array profiling of gene expression changes during maize embryo development. Funct Integr Genomics 2:13–27PubMedCrossRefGoogle Scholar
  40. 40.
    Leibbrandt NB, Snyman SJ (2003) Stability of gene expression and agronomic performance of a transgenic herbicide-resistant sugarcane line in South Africa. Crop Sci 43:671–677Google Scholar
  41. 41.
    Lloyd JC, Zakhleniuk OV (2004) Responses of primary and secondary metabolism to sugar accumulation revealed by microarray expression analysis of the Arabidopsis mutant, pho3. J Exp Bot 55:1221–1230PubMedCrossRefGoogle Scholar
  42. 42.
    Lunn JE, Furbank RT (1999) Sucrose biosynthesis in C4 plants. New Phytol 143:221–237CrossRefGoogle Scholar
  43. 43.
    Lunn JE, Feil R, Hendriks JHM, Gibon Y, Morcuende Osuna D, Scheible WR, Carillo P, Hajirezaei MR, Stitt M (2006) Sugar-induced increases in trehalose 6-phosphate are correlated with redox activation of ADPglucose pyrophosphorylase and higher rates of starch synthesis in Arabidopsis thaliana. Biochem J 397:139–148PubMedCrossRefGoogle Scholar
  44. 44.
    Ma H, Albert HA, Paull R, Moore PH (2000) Metabolic engineering of invertase activities in different subcellular compartments affects sucrose accumulation in sugarcane. Aust J Plant Physiol 27:1021–1030Google Scholar
  45. 45.
    Masclaux-Daubresse C, Purdy S, Lemaitre T, Pourtau N, Taconnat L, Renou JP, Wingler A (2007) Genetic variation suggests interaction between cold acclimation and metabolic regulation of leaf senescence. Plant Physiol 143:434–446PubMedCrossRefGoogle Scholar
  46. 46.
    Maxwell K, Johnson GN (2000) Chlorophyll fluorescence—a practical guide. J Exp Bot 51:659–668PubMedCrossRefGoogle Scholar
  47. 47.
    McCormick AJ, Cramer MD, Watt DA (2006) Sink strength regulates photosynthesis in sugarcane. New Phytol 171:759–770PubMedCrossRefGoogle Scholar
  48. 48.
    McCormick AJ, Cramer MD, Watt DA (2008) Changes in photosynthetic rates and gene expression of leaves during a source–sink perturbation in sugarcane. Ann Bot 101:89–102PubMedCrossRefGoogle Scholar
  49. 49.
    McCormick AJ, Cramer MD, Watt DA (2008) Regulation of photosynthesis by sugars in sugarcane leaves. J Plant Physiol. doi: 10.1016/j.jplph.2008.01.008
  50. 50.
    Minchin PEH, Thorpe MR, Farrar JF, Koroleva OA (2002) Source–sink coupling in young barley plants and control of phloem loading. J Exp Bot 53:1671–1676PubMedCrossRefGoogle Scholar
  51. 51.
    Müller R, Morant M, Jarmer H, Nilsson L, Nielsen TH (2007) Genome-wide analysis of the Arabidopsis transcriptome reveals interaction of phosphate and sugar metabolism. Plant Physiol 143:156–171PubMedCrossRefGoogle Scholar
  52. 52.
    Nielsen TH, Krapp A, Röber-Schwarz U, Stitt M (1998) The sugar-mediated regulation of genes encoding the small subunit of Rubisco and the regulatory subunit of ADP glucose pyrophosphorylase is modified by phosphate and nitrogen. Plant Cell Environ 21:443–454CrossRefGoogle Scholar
  53. 53.
    Paul MJ (2007) Trehalose 6-phosphate. Curr Opin Plant Biol 10:303–309PubMedCrossRefGoogle Scholar
  54. 54.
    Paul MJ, Driscoll SP (1997) Sugar repression of photosynthesis: the role of carbohydrates in signalling nitrogen deficiency through source:sink imbalance. Plant Cell Environ 20:110–116CrossRefGoogle Scholar
  55. 55.
    Paul MJ, Foyer CH (2001) Sink regulation of photosynthesis. J Exp Bot 52:1381–1400CrossRefGoogle Scholar
  56. 56.
    Paul MJ, Pellny TK (2003) Carbon metabolite feedback regulation of leaf photosynthesis and development. J Exp Bot 54:539–547PubMedCrossRefGoogle Scholar
  57. 57.
    Paul MJ, Pellny TK, Goddijn IJM (2001) Enhancing photosynthesis with sugar signals. Trends Plant Sci 6:197–200PubMedCrossRefGoogle Scholar
  58. 58.
    Pego JV, Kortsee AJ, Huijser C, Smeekens SCM (2000) Photosynthesis, sugars and the regulation of gene expression. J Exp Bot 51:407–416PubMedCrossRefGoogle Scholar
  59. 59.
    Pellny TK, Ghannoum O, Conroy JP, Schluepmann H, Smeekens S, Andralojc J, Krause KP, Goddijn O, Paul MJ (2004) Genetic modification of photosynthesis with E. coli genes for trehalose synthesis. Plant Biotechnol 2:71–82CrossRefGoogle Scholar
  60. 60.
    Pieters AJ, Paul MJ, Lawlor DW (2001) Low sink demand limits photosynthesis under Pi deficiency. J Exp Bot 52:1083–1091PubMedCrossRefGoogle Scholar
  61. 61.
    Prioul JL, Reyss A (1988) Rapid variations in the content of the RNA of the small subunit of ribulose-1,5-bisphosphate carboxylase of mature tobacco leaves in response to localized changes in light quantity. Relationships between the activity and quantity of the enzyme. Planta 174:488–494CrossRefGoogle Scholar
  62. 62.
    Ramon M, Rolland F (2007) Plant development: introducing trehalose metabolism. Trends Plant Sci 12:185–188PubMedCrossRefGoogle Scholar
  63. 63.
    Rodermel S, Haley J, Jiang CZ, Tsai CH, Bogorad L (1996) A mechanism for intergenomic integration: abundance of ribulose bisphosphate carboxylase small-subunit protein influences the translation of the large-subunit mRNA. Proc Natl Acad Sci USA 93:3881–3885PubMedCrossRefGoogle Scholar
  64. 64.
    Roitsch T, Balibrea ME, Hofmann M, Proeis R, Sinna AK (2003) Extracellular invertase: key metabolic enzyme and PR protein. J Exp Bot 54:513–524PubMedCrossRefGoogle Scholar
  65. 65.
    Rolland F, Baena-Gonzalez E, Sheen J (2006) Sugar sensing and signaling in plants: conserved and novel mechanisms. Annu Rev Plant Biol 57:675–709PubMedCrossRefGoogle Scholar
  66. 66.
    Rolland F, Moore B, Sheen J (2002) Sugar sensing and signaling in plants. Plant Cell 14:185–205Google Scholar
  67. 67.
    Sawers RJH, Liu P, Anufrikova K, Hwang JT, Brutnell TP (2007) A multi-treatment experimental system to examine photosynthetic differentiation in the maize leaf. BMC Genomics 8:1–13CrossRefGoogle Scholar
  68. 68.
    Schäfer C, Simper H, Hofmann B (1992) Glucose feeding results in co-ordinated changes of chlorophyll content, ribulose-1,5-bisphosphate carboxylase/oxygenase activity and photosynthetic potential in photoautotrophic suspension-cultured cells of Chenopodium rubrum. Plant Cell Environ 15:343–350CrossRefGoogle Scholar
  69. 69.
    Schluepmann H, van Dijken A, Smeekens S, Paul M (2003) Trehalose-6-phosphate is indispensable for carbohydrate utilization and growth in Arabidopsis thaliana. Proc Natl Acad Sci USA 100:6849–6854PubMedCrossRefGoogle Scholar
  70. 70.
    Smeekens S (2000) Sugar-induced signal transduction in plants. Annu Rev Plant Physiol Plant Mol Biol 51:49–81PubMedCrossRefGoogle Scholar
  71. 71.
    Solfanelli C, Poggi A, Loreti E, Alpi A, Perata P (2006) Sucrose-specific induction of the anthocyanin biosynthetic pathway in Arabidopsis. Plant Physiol 147:637–646CrossRefGoogle Scholar
  72. 72.
    Stitt M, Quick WP (1989) Photosynthetic carbon partitioning: its regulation and possibilities for manipulation. Physiol Plant 77:633–641CrossRefGoogle Scholar
  73. 73.
    Stitt M, Gibon Y, Lunn JE, Piques M (2007) Multilevel genomics analysis of carbon signalling during low carbon availability: coordinating the supply and utilization of carbon in a fluctuating environment. Funct Plant Biol 34:526–549CrossRefGoogle Scholar
  74. 74.
    Tiessen A, Prescha K, Branscheid A, Palacios N, McKibbin R, Halford NG, Geigenberger P (2003) Evidence that SNF1-related kinase and hexokinase are involved in separate sugar-signalling pathways modulating post-translational redox activation of ADP-glucose pyrophosphorylase in potato tubers. Plant J 35:490–500PubMedCrossRefGoogle Scholar
  75. 75.
    Toroser D, Plaut Z, Huber SC (2000) Regulation of a plant SNF1-related protein kinase by glucose-6-phosphate. Plant Physiol 123:403–411PubMedCrossRefGoogle Scholar
  76. 76.
    Van Oosten JJ, Besford RT (1994) Sugar feeding mimics effect of acclimation to high CO2: rapid downregulation of RuBisCO small subunit transcripts, but not of the large subunit transcripts. J Plant Physiol 143:306–312Google Scholar
  77. 77.
    Watt DA, McCormick AJ, Govender C, Carson DL, Cramer MD, Huckett BI, Botha FC (2005) Increasing the utility of genomics in unraveling sucrose accumulation. Field Crops Res 92:149–158CrossRefGoogle Scholar
  78. 78.
    Williams LE, Lemoine R, Sauer N (2000) Sugar transporters in higher plants—a diversity of roles and complex regulation. Trends Plant Sci 5:283–290PubMedCrossRefGoogle Scholar
  79. 79.
    Wu L, Birch RG (2007) Doubled sugar content in sugarcane plants modified to produce a sucrose isomer. Plant Biotechnol J 5:109–117PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  • A. J. McCormick
    • 1
    • 2
    • 3
  • M. D. Cramer
    • 4
    • 5
  • D. A. Watt
    • 1
    • 3
  1. 1.Crop Biology Resource CentreSouth African Sugarcane Research Institute (SASRI)Mt. EdgecombeSouth Africa
  2. 2.Department of Plant SciencesUniversity of OxfordOxfordUK
  3. 3.School of Biological and Conservation SciencesUniversity of KwaZulu-NatalDurbanSouth Africa
  4. 4.Botany DepartmentUniversity of Cape TownCape TownSouth Africa
  5. 5.School of Plant Biology, Faculty of Natural and Agricultural SciencesThe University of Western AustraliaCrawleyAustralia

Personalised recommendations