Volume 3, Issue 5, Pages 842-853 (September 2010) Unraveling the Biochemical and Molecular Networks Involved in Maize Cell Habituation to the Cellulose Biosynthesis Inhibitor Dichlobenil  Mélida Hugo , Encina Antonio , Álvarez Jesús , Acebes José Luis , Caparrós-Ruiz David   Molecular Plant  Volume 3, Issue 5, Pages 842-853 (September 2010) DOI: 10.1093/mp/ssq027 Copyright © 2010 The Authors. All rights reserved. Terms and Conditions

Figure 1 Cellulose Content of Cell Walls Isolated from Non-Habituated (NH), Habituated to 12 μM DCB (H12; Collected in the Active Growth or in the Stationary Phase), Dehabituated (DH; in the Stationary Phase), and Non-Habituated Supplemented with DCB for 5–6 d (NH/DCB) Maize Cell Cultures Analyzed by the Updegraff Method. Values are means ± S.D. of three measurements. Growth measured as relative increase in fresh weight of the cells was reduced by 38.9 and 12.4% in H12 and DH cells, respectively, with regard to NH. Molecular Plant 2010 3, 842-853DOI: (10.1093/mp/ssq027) Copyright © 2010 The Authors. All rights reserved. Terms and Conditions

Figure 2 Effect of DCB on the Growth of Non-Habituated (NH), Habituated to 12 μM DCB (H12), and Dehabituated (DH) Maize Cell Cultures Expressed as the Concentration of DCB (μM) Required to Inhibit Weight Increase by 10% (I10), 50% (I50), and 90% (I90) with Respect to Control. Molecular Plant 2010 3, 842-853DOI: (10.1093/mp/ssq027) Copyright © 2010 The Authors. All rights reserved. Terms and Conditions

Figure 3 Relative ZmCesA Gene Expression Analyzed by RT–PCR of Different Maize Cell Cultures and Culture Phases (Key as in Figure 1).↑, more mRNA accumulation than control (NH); ↓, less mRNA than control. ZmCesA4, ZmCesA9, ZmCesA10, ZmCesA11, ZmCesA12, and ZmMAP20 were not detected. ZmMAP20, microtubule-associated protein 20; ZmBk2L3, brittle stalk 2 Like 3. Molecular Plant 2010 3, 842-853DOI: (10.1093/mp/ssq027) Copyright © 2010 The Authors. All rights reserved. Terms and Conditions

Figure 4 Ferulic (A) and p-Coumaric Acid (B) Content of Cell Walls Isolated from Different Maize Cell Cultures (Key as in Figure 1) Released by Treatment with 1 M NaOH and Measured by HPLC–PAD. Values are mean ± S.D. of three measurements. Molecular Plant 2010 3, 842-853DOI: (10.1093/mp/ssq027) Copyright © 2010 The Authors. All rights reserved. Terms and Conditions

Figure 5 (A) Phenolic Biosynthetic Pathway, from Phenylalanine to Arabinoxylan Feruloylation. Dashed Arrows Indicate Putative Pathways.(B) Relative Expression of the Genes Involved in the Synthesis of Phenolic Compounds of Different Maize Cell Cultures (key as in Figure 1). ↑ more mRNA content than control; ↓, less mRNA content than control; PAL, Phenylalanine Ammonia-Lyase; C4H, Cinnamate 4-Hydroxylase; 4CL, 4-Coumarate CoA Ligase; C3H, 4-coumarate 3-hydroxylase; HCT, Hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyl Transferase, CCoAOMT, Caffeoyl-CoA O-MethylTransferase; COMT, Caffeic acid O-MethylTransferase. Molecular Plant 2010 3, 842-853DOI: (10.1093/mp/ssq027) Copyright © 2010 The Authors. All rights reserved. Terms and Conditions

Figure 6 Activity of Glutathione S-Tranferase (GST), Glutathione Reductase (GR), Guaiacol-Type Peroxidase (GPOX), Ascorbate Peroxidase (APOX), and Catalase (CAT) in Non-Habituated (NH), Habituated to DCB (H12), and Non-Habituated Supplemented with DCB (NH/DCB) Maize Cultured Cells. Values are means ± S.D. (n = 10) and are expressed as μmol min−1 mg prot−1. Molecular Plant 2010 3, 842-853DOI: (10.1093/mp/ssq027) Copyright © 2010 The Authors. All rights reserved. Terms and Conditions