, Volume 227, Issue 5, pp 1063–1077

Carbohydrate mobilization and gene regulatory profile in the pseudobulb of Oncidium orchid during the flowering process

  • Chih-Yu Wang
  • Chung-Yi Chiou
  • Heng-Long Wang
  • Ramanarayan Krishnamurthy
  • Shripathi Venkatagiri
  • Jun Tan
  • Kai-Wun Yeh
Original Article


The pseudobulb of Oncidium orchid is a storage organ for supplying water, minerals and carbohydrates to the developing inflorescence. Different patterns of mannan, starch and pectin metabolism were observed in the pseudobulb of three developmental stages by histochemical staining and high performance anion exchange chromatographic (HPAEC) analysis. Copious pectin was strongly stained by ruthenium red in young pseudobulbs demonstrating that mannan and pectin were preferentially accumulated in the young pseudobulb sink at inflorescence pre-initiation stage. Concomitant with the emergence of the inflorescence, mannan and pectin decreased gradually and converted to starch. The starch, synthesized at the inflorescence developing stage, was eventually degraded at the floral development stage. A systematic survey on the subtractive EST (expression sequence tag) library of pseudobulb in the inflorescence pre-initiation stage revealed the presence of five groups of gene homologues related to sucrose, mannan, starch, pectin and other carbohydrate metabolism. The transcriptional level of 13 relevant genes related to carbohydrate metabolism was characterized from pseudobulbs of three different developmental stages. The specific activities of the enzymes encoded by these genes were also assayed. The expression profiles of these genes show that the transcriptional levels largely correlated with the enzyme activities, which were associated with the respective carbohydrate pools. These results demonstrated a novel functional profile of polysaccharide mobilization pathway as well as their relevant gene expression in the pseudobulb of Oncidium orchid during the flowering process.


Carbohydrate metabolism EST (expressed sequence tags) Inflorescence Oncidium Pseudobulb 



High performance anion exchange chromatography


Expression sequence tags


  1. Baier M, Hemmann G, Holman R, Corke F, Card R, Smith C, Rook F, Bevan MW(2004) Characterization of mutants in Arabidopsis showing increased sugar-specific gene expression, growth, and developmental responses. Plant Physiol 134:81–91PubMedCrossRefGoogle Scholar
  2. Beck E, Ziegler P (1989) Biosynthesis and degradation of starch in higher plants. Annu Rev Plant Physiol Plant Mol Biol 40:95–117CrossRefGoogle Scholar
  3. Bernier G, Havelange A, Houssa C, Petitjean A, Lejeune P (1993) Physiological signals that induce flowering. Plant Cell 5:1147–1155PubMedCrossRefGoogle Scholar
  4. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  5. Buckeridge MS, Santos HP, Tiné MAS (2000) Mobilization of storage cell wall polysaccharides in seeds. Plant Physiol Biochem 38:141–156CrossRefGoogle Scholar
  6. Caissard J-C, Meekijjironenroj A, Baudino S, Anstett M-C (2004) Localization and emission of pollinator attractant on whole leaves of Chamaerops humilis (Arecaceae). Am J Bot 91:1190–1199CrossRefGoogle Scholar
  7. Chang S, Puryear J, Cairney J (1993) A simple and efficient method for isolating RNA from pine trees. Plant Mol Biol Rep 11:113–116CrossRefGoogle Scholar
  8. Corbesier L, Lejeune P, Bernier G (1998) The role of carbohydrates in the induction of flowering in Arabidopsis thaliana: comparison between the wild type and a starchless mutant. Planta 206:131–137PubMedCrossRefGoogle Scholar
  9. Creelman RA, Mullet JE (1997) Oligosaccharins, brassinolides and jasmonates: nontraditional regulators of plant growth, development, and gene expression. Plant Cell 9:1211–1223PubMedCrossRefGoogle Scholar
  10. Denyer K, Barber M, Edwards E.A, Smith AM, Wang TL (1997) Two isoforms of the GBSS 1 class of granule-bound starch synthase are differentially expressed in pea plants (Pisum sativum L.). Plant Cell Environ 20:1566–1572CrossRefGoogle Scholar
  11. Dreier LP, Hter JJ, Ruffner HP (1998) Invertase activity, grape berry development and cell compartmentation. Plant Physiol Biochem 36:865–872CrossRefGoogle Scholar
  12. Dumville JC, Fry SC (2000) Uronic acid-containing oligosaccharins: Their biosynthesis, degradation and signaling roles in non-deseased plant tissues. Plant Physiol Biochem 38:125–140CrossRefGoogle Scholar
  13. Dumville JC, Fry SC (2003) Solubilization of tomato fruit pectins by ascorbate: a possible non-enzymic mechanism of fruit softening. Planta 217:951–961PubMedCrossRefGoogle Scholar
  14. Etzler ME (1998) Oligosaccharide signaling of plant cells. J Cell Biochem Suppl 31:123–128CrossRefGoogle Scholar
  15. Fiske CH, Subbarow Y (1925) The colorimetric determination of phosphorous. J Biol Chem 66:375–400Google Scholar
  16. Gana JA, Kalengamaliro NE, Cunningham SM, Volenec JJ (1998) Expression of β-amylase from alfalfa taproots. Plant Physiol 118:495–1505CrossRefGoogle Scholar
  17. Geiger DR (1987) Understanding interactions of source and sink regions of plants. Plant Physiol Biochem 25:659–666Google Scholar
  18. Hew CS, Yong JWH (1994) Growth and photosynthesis of Oncidium ‘Goldiana’. J Hortic Sci 69:809–819Google Scholar
  19. Hew CS, Ng CKY (1996) Changes in mineral and carbohydrate content in pseudobulbs of C3 epiphytic orchid hybrid Oncidium Goldiana at different growth stages. Lindleyana 11:125–134Google Scholar
  20. Iwai H, Kikuchi A, Kobayashi T, Kamada H, Satoh S (1999) High levels of non-methylesterified pectins and low levels of peripherally located pectins in loosely attached non-embryogenic callus of carrot. Plant Cell Rep 18:561–566CrossRefGoogle Scholar
  21. Marolda CL, Valvano MA (1993) Identification, expression, and DNA sequence of the GDP-mannose biosynthesis genes encoded by the rfb gene cluster of strain VW187 (Escherichia coli 07:K1). J Bacteriol 175:148–158PubMedGoogle Scholar
  22. Marraccini P, John Rogers W, Allard C, Andre ML, Caillet V, Lacoste N, Lausanne F, Michaux S (2001) Molecular and biochemical characterization of endo-β-mannanases from germinating coffee (coffea arabica) grains. Planta 213:296–308PubMedCrossRefGoogle Scholar
  23. Mulimani VH, Prashanth SJ (2002) Investigating plant galactomannans. Biochem Mol Biol Educ 30:101–103CrossRefGoogle Scholar
  24. Ng C, Hew C (1996) Pathway of phleom loading in the C3 tropical orchid hybrid Oncidium Goldiana. J Exp Bot 47:1935–1939CrossRefGoogle Scholar
  25. Ni Y, Turner D, Yates KM, Tizard I (2004) Isolation and characterization of structural components of Aloe vera L. leaf pulp. Int Immunopharm 4:1745–1755CrossRefGoogle Scholar
  26. Rook F, Hadingham SA, Li YH, Bevan MW (2006) Sugar and ABA response pathways and the control of gene expression. Plant Cell Environ 29:426–434PubMedCrossRefGoogle Scholar
  27. Salehuzzhaman SNIM, Jacobson E, Visser RGF (1994) Expression patterns of 2 starch biosynthetic genes in in-vitro cultured cassava plants and their induction by sugar. Plant Sci 98:53–62CrossRefGoogle Scholar
  28. Smirnoff N, Conklin PL, Loewus FA (2001) Biosynthesis of ascorbic acid in plants: a renaissance. Annu Rev Plant Physiol Plant Mol Biol 52:437–467PubMedCrossRefGoogle Scholar
  29. Sobry S, Havelange A, Liners F, Van Cutsem P (2005) Immunolocalization of homogalacturonans in the apex of the long-day plant Sinapis alba at floral transition. The pectin content drops dramatically in the first hours of this transition. Physiol Plant 123:339–347CrossRefGoogle Scholar
  30. Stancato GC, Mazzafera P, Buckeridge MS (2001) Effect of a drought period on the mobilization of non-structural carbohydrates, photosynthetic efficiency and water status in an epiphytic orchid. Plant Physiol Biochem 39:1009–1016CrossRefGoogle Scholar
  31. Stolle-Smits T, Beekhuizen JG, Kok MTC, Pijnenburg M, Recourt K, Derksen J Voragen AHJ (1999) Changes in cell wall polysaccharides of green bean pods during development. Plant Physiol 121:363–372PubMedCrossRefGoogle Scholar
  32. Sung SS, Sheih WJ, Geiger DR, Black CC (1994) Growth, sucrose synthase, and invertase activities of developing Phaseolus vulgaris L. fruits. Plant Cell Environ 17:419–426CrossRefGoogle Scholar
  33. Talaga P, Vialle S, Moreau M (2002) Development of a high-performance anion-exchange chromatography with pulsed-amperometric detection-based quantification assay for pneumococcal polysaccharides and conjugates. Vaccine 20:2474–2484PubMedCrossRefGoogle Scholar
  34. Tan J, Wang HL, Yeh KW (2005) Analysis of organ-specific, expressed genes in Oncidium orchid by subtractive expressed sequence tags library. Biotech Lett 27:1517–1528CrossRefGoogle Scholar
  35. Thomas BR, Rodriguez RL (1994) Metabolite signals regulate gene expression and source/sink relations in cereal seedlings. Plant Physiol 106:1235–1239PubMedGoogle Scholar
  36. Tymowska-Lalanne Z, Kreis M (1988) Expression of the Arabidopsis thaliana invertase gene family. Planta 207:259–265CrossRefGoogle Scholar
  37. Van Oosten JJ, Besford RT (1994) Sugar feeding mimics effects 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
  38. Wang HL, Chung JD, Yeh KW (2003) Changes in carbohydrate and free amino acid pools in current pseudobulb of Oncidium Gower Ramsey during inflorescence development. J Agric Assoc China 4:476–488Google Scholar
  39. Wang HL, Yeh KW, Chen PR, Chang CH, Chen JM, Khoo KH (2006) Isolation and characterization of a pure mannan from Oncidium cv. Gower Ramsey. Current pseudobulb during initial inflorescence development. Biosci Biotechnol Biochem 70:551–553PubMedCrossRefGoogle Scholar
  40. Wang Q, Monroe J, Sjolund RD (1995) Identification and characterization of phloem-specific β-amylase. Plant Physiol 109:743–750PubMedCrossRefGoogle Scholar
  41. Western TL, Burn J, Tan WL, Skinner DJ, Martin-McCaffrey L, Moffatt BA, Haughn GW (2001) Isolation and characterization of mutants defective in seed coat mucilage secretory cell development in Arabidopsis. Plant Physiol 127:998–1011PubMedCrossRefGoogle Scholar
  42. Willats WG, McCartney L, Knox JP (2001) In-situ analysis of pectic polysaccharides in seed nucilage and at the root surface of Arabidopsis thaliana. Planta 213:37–44PubMedCrossRefGoogle Scholar
  43. Yong JWH, Hew CS (1995) The importance of photoassimilate contribution from current shoots and connected back shoots to inflorescence size in the thin-leaved sympodial orchid Oncidium Goldiana. Int J Plant Sci 156:450–459CrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2007

Authors and Affiliations

  • Chih-Yu Wang
    • 1
  • Chung-Yi Chiou
    • 1
  • Heng-Long Wang
    • 2
  • Ramanarayan Krishnamurthy
    • 1
  • Shripathi Venkatagiri
    • 4
  • Jun Tan
    • 1
    • 3
  • Kai-Wun Yeh
    • 1
  1. 1.Institute of Plant BiologyNational Taiwan UniversityTaipei, 106Taiwan
  2. 2.Department of Life ScienceNational University of KaohsiungKaohsiungTaiwan
  3. 3.College of BioinformaticsChongqing University of Posts and TelecommunicationsChongqingChina
  4. 4.Department of BotanyKarnatak UniversityDharwadIndia

Personalised recommendations