, Volume 224, Issue 4, pp 828–837

Xylem-specific and tension stress-responsive coexpression of KORRIGAN endoglucanase and three secondary wall-associated cellulose synthase genes in aspen trees

  • Suchita Bhandari
  • Takeshi Fujino
  • Shiv Thammanagowda
  • Dongyan Zhang
  • Fuyu Xu
  • Chandrashekhar P. Joshi
Original Article


In nature, angiosperm trees develop tension wood on the upper side of their leaning trunks and drooping branches. Development of tension wood is one of the straightening mechanisms by which trees counteract leaning or bending of stem and resume upward growth. Tension wood is characterized by the development of a highly crystalline cellulose-enriched gelatinous layer next to the lumen of the tension wood fibers. Thus experimental induction of tension wood provides a system to understand the process of cellulose biosynthesis in trees. Since KORRIGAN endoglucanases (KOR) appear to play an important role in cellulose biosynthesis in Arabidopsis, we cloned PtrKOR, a full-length KOR cDNA from aspen xylem. Using RT-PCR, in situ hybridization, and tissue-print assays, we show that PtrKOR gene expression is significantly elevated on the upper side of the bent aspen stem in response to tension stress while KOR expression is significantly suppressed on the opposite side experiencing compression stress. Moreover, three previously reported aspen cellulose synthase genes, namely, PtrCesA1, PtrCesA2, and PtrCesA3 that are closely associated with secondary cell wall development in the xylem cells exhibited similar tension stress-responsive behavior. Our results suggest that coexpression of these four proteins is important for the biosynthesis of highly crystalline cellulose typically present in tension wood fibers. Their simultaneous genetic manipulation may lead to industrially relevant improvement of cellulose in transgenic crops and trees.


Aspen Cellulose Cellulose synthase KORRIGAN endoglucanase Tension wood Trees 



Cellulose synthase


Korrigan endoglucanase


Degree of polymerization




  1. Brummell DA, Catala C, Lashbrook CC, Bennett AB (1997) A membrane-anchored E-type endo-1, 4-beta-glucanase is localized on Golgi and plasma membranes of higher plants. Proc Natl Acad Sci USA 94:4794–4799CrossRefPubMedGoogle Scholar
  2. del Campillo E (1999) Multiple endo-1, 4-β-d-glucanase (cellulose) genes in Arabidopsis. Curr Topics Dev Biol 46:39–61CrossRefGoogle Scholar
  3. Delmer DP (1999) Cellulose biosynthesis: exciting times for a difficult field of study. Ann Rev Plant Physiol Plant Mol Biol 50:245–276CrossRefGoogle Scholar
  4. Doblin M, Kurek I, Jacob-Wilk D, Delmer DP (2002) Cellulose biosynthesis in plants: from genes to rosettes. Plant Cell Physiol 43:1407–1420CrossRefPubMedGoogle Scholar
  5. Gardiner JC, Taylor NG, Turner SR (2003) Control of cellulose synthase complex localization in developing xylem. Plant Cell 15:1740–1748CrossRefPubMedGoogle Scholar
  6. Geisler-Lee J, Geisler M, Coutinho PM, Segerman B, Nishikubo N, Takahashi J, Aspeborg H, Djerbi S, Master E, Andersson-Gunneras S, Sundberg B, Karpinski S, Teeri TT, Kleczkowski LA, Henrissat B, Mellerowicz EJ (2006) Poplar carbohydrate-active enzymes (CAZymes). Gene identification and expression analyses. Plant Physiol 140:946–962CrossRefPubMedGoogle Scholar
  7. Joshi CP (1987a) An inspection of the domain between putative TATA box and translation start site in seventy-nine plant genes. Nucleic Acids Res 15:6643–6653CrossRefGoogle Scholar
  8. Joshi CP (1987b) Putative polyadenylation signals in nuclear genes of higher plants: a compilation and analysis. Nucleic Acids Res 15:9627–9640CrossRefGoogle Scholar
  9. Joshi CP, Bhandari S, Ranjan P, Kalluri UC, Liang X, Fujino T, Samuga A (2004) Genomics of cellulose biosynthesis in poplars. New Phytol 164:53–61CrossRefGoogle Scholar
  10. Kalluri U (2003) Comparative expression analysis of cellulose biosynthesis related genes from aspen trees. Ph. D. thesis. Michigan Technological University, HoughtonGoogle Scholar
  11. Kalluri U, Joshi CP (2003) Isolation and characterization of a new, full-length cellulose synthase cDNA from developing xylem of aspen trees. J Exp Bot 54:2187–2188CrossRefPubMedGoogle Scholar
  12. Kalluri U, Joshi CP (2004) Differential expression of two cellulose synthase genes associated with primary wall and secondary wall development in aspen trees. Planta 220:47–55CrossRefPubMedGoogle Scholar
  13. Lane DR, Wiedemeier A, Peng L, Hofte H, Vernhettes S, Desprez T, Hocart CH, Birch RJ, Baskin TI, Burn JE, Arioli T, Betzner AS, Williamson RE (2001) Temperature-sensitive alleles of rsw2 link the KORRIGAN endo-1,4-beta-glucanase to cellulose synthesis and cytokinesis in Arabidopsis. Plant Physiol 126:278–288CrossRefPubMedGoogle Scholar
  14. Libertini E, Li Y, McQueen-Mason SJ (2004) Phylogenetic analysis of the plant endo-beta-1,4-glucanase gene family. J Mol Evol 58:506–515CrossRefPubMedGoogle Scholar
  15. Master ER, Rudsander UJ, Zhou W, Henriksson H, Divne C, Denman S, Wilson DB, Teeri TT (2004) Recombinant expression and enzymatic characterization of PttCel9A, a KOR homologue from Populus tremula × tremuloides. Biochemistry 43:10080–10089CrossRefPubMedGoogle Scholar
  16. Molhoj M, Ulvskov P, Dal Degan F (2001) Characterization of a functional soluble form of a Brassica napus membrane-anchored endo-1,4-beta-glucanase heterologously expressed in Pichia pastoris. Plant Physiol 127:674–684CrossRefPubMedGoogle Scholar
  17. Molhoj M, Pagant S, Hofte H (2002) Towards understanding the role of membrane-bound endo-beta-1, 4-glucanases in cellulose biosynthesis. Plant Cell Physiol 43:1399–1406CrossRefPubMedGoogle Scholar
  18. Nicol F, His I, Jauneau A, Vernhettes S, Canut H, Hofte H (1998) A plasma membrane-bound putative endo-1, 4-beta-d-glucanase is required for normal wall assembly and cell elongation in Arabidopsis. EMBO J 17:5563–5576CrossRefPubMedGoogle Scholar
  19. Pear JR, Kawagoe Y, Schreckengost WE, Delmer DP, Stalker DM (1996) Higher plants contain homologs of the bacterial celA genes encoding the catalytic subunit of cellulose synthase. Proc Natl Acad Sci USA 93:12637–12642CrossRefPubMedGoogle Scholar
  20. Pérez-Garcia A, Pereira S, García Gutiérrez A, Cazorla F, Salema R, de Vicente A, Cánovas F (1998) Cytosolic localization in tomato mesophyll cells of a novel glutamine synthetase induced in response to bacterial infection or phosphinothricin treatment. Planta 206:426–434CrossRefGoogle Scholar
  21. Ranjan P, Kao Y, Jiang H, Joshi CP, Harding SA, Tsai C (2004) Suppression Subtractive Hybridization-mediated transcriptome analysis from multiple tissues of aspen (Populus tremuloides) trees altered in phenylpropanoid metabolism. Planta 219:694–704CrossRefPubMedGoogle Scholar
  22. Read SM, Bacic T (2002) Prime time for cellulose. Science 295:59–60CrossRefPubMedGoogle Scholar
  23. Robert S, Mouille G, Hofte H (2004) The mechanism and regulation of cellulose synthesis in primary walls: lessons from cellulose deficient Arabidopsis mutants. Cellulose 11:351–364CrossRefGoogle Scholar
  24. Sambrook J, Russell DW (2001) Molecular cloning: a laboratory manual, 3rd edn, Vol 3. Cold Spring Harbor Laboratory Press, NYGoogle Scholar
  25. Samuga A (2003) Molecular characterization of four members of cellulose synthase gene superfamily in aspen. Ph. D. thesis. Michigan Technological University, Houghton, MI, USAGoogle Scholar
  26. Samuga A, Joshi CP (2002) A new cellulose synthase gene (PtrCesA2) from aspen xylem is orthologous to Arabidopsis AtCesA7 (irx3) gene associated with secondary cell wall synthesis. Gene 296:37–44CrossRefPubMedGoogle Scholar
  27. Samuga A, Joshi CP (2004) Expression patterns of two primary cell wall-related cellulose synthase cDNAs, PtrCesA6 and PtrCesA7 from aspen. Gene 334:73–82CrossRefPubMedGoogle Scholar
  28. Sato S, Kato T, Kakegawa K, Ishii T, Liu YG, Awano T, Takabe K, Nishiyama Y, Kuga S, Sato S, Nakamura Y, Tabata S, Shibata D (2001) Role of the putative membrane-bound endo-1,4-beta-glucanase KORRIGAN in cell elongation and cellulose synthesis in Arabidopsis thaliana. Plant Cell Physiol 42:251–263CrossRefPubMedGoogle Scholar
  29. Scheible W-R, Eshed R, Richmond T, Delmer D, Somerville CR (2001) Modifications of cellulose synthase confer resistance to isoxaben and thiazolidinone herbicides in Arabidopsis ixr1 mutants. Proc Natl Acad Sci USA 98:10079–10084PubMedCrossRefGoogle Scholar
  30. Somerville C, Bauer S, Brininstool G, Facette M, Hamann T, Milne J, Osborne E, Paredez A, Persson S, Raab T, Vorwerk S, Youngs H (2004) Toward a systems approach to understanding plant cell walls. Science 306:2206–2211CrossRefPubMedGoogle Scholar
  31. Szyjanowicz PM, McKinnon I, Taylor NG, Gardiner J, Jarvis MC, Turner SR (2004) The irregular xylem2 mutant is an allele of korrigan that affects the secondary cell wall of Arabidopsis thaliana. Plant J 37:730–740CrossRefPubMedGoogle Scholar
  32. Taylor NG, Scheible W-R, Cutler S, Somerville CR, Turner SR (1999) The irregular xylem3 locus of Arabidopsis encodes a cellulose synthase required for secondary cell wall synthesis. Plant Cell 11:769–779CrossRefPubMedGoogle Scholar
  33. Taylor NG, Laurie S, Turner SR (2000) Multiple cellulose synthase catalytic subunits are required for cellulose synthesis in Arabidopsis. Plant Cell 12:2529–2540CrossRefPubMedGoogle Scholar
  34. Taylor NG, Howells RM, Huttly AK, Vickers K, Turner SR (2003) Interactions among three distinct CesA proteins essential for cellulose synthesis. Proc Natl Acad Sci USA 100:1450–1455PubMedCrossRefGoogle Scholar
  35. Timell TE (1969) The chemical composition of tension wood. Svenk Papperstidning 72:173–181Google Scholar
  36. Turner SR, Somerville CR (1997) Collapsed xylem phenotype of Arabidopsis identifies mutants deficient in cellulose deposition in the secondary cell wall. Plant Cell 9:689–701CrossRefPubMedGoogle Scholar
  37. Williamson RE, Burn JE, Hocart CH (2002) Towards the mechanism of cellulose synthesis. Trends Plant Sci 7:461–467CrossRefPubMedGoogle Scholar
  38. Wu L, Joshi CP, Chiang VL (2000) A xylem-specific cellulose synthase gene from aspen (Populus tremuloides) is responsive to mechanical stress. Plant J 22:495–502CrossRefPubMedGoogle Scholar
  39. Zuo J, Niu QW, Nishizawa N, Wu Y, Kost B, Chua NH (2000) KORRIGAN, an Arabidopsis endo-1,4-beta-glucanase, localizes to the cell plate by polarized targeting and is essential for cytokinesis. Plant Cell 12:1137–1152CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Suchita Bhandari
    • 1
    • 2
  • Takeshi Fujino
    • 1
  • Shiv Thammanagowda
    • 1
  • Dongyan Zhang
    • 1
  • Fuyu Xu
    • 1
  • Chandrashekhar P. Joshi
    • 1
  1. 1.Biotechnology Research Center, School of Forest Resources and Environmental SciencesMichigan Technological UniversityHoughtonUSA
  2. 2.BILTGurgaonIndia

Personalised recommendations