Biologia plantarum 62:80-88, 2018 | DOI: 10.1007/s10535-017-0739-2

Knockout mutants of Arabidopsis thaliana β-galactosidase. Modifications in the cell wall saccharides and enzymatic activities

M. Moneo-Sánchez1, L. Izquierdo1, I. Martín1, J. Hernández-Nistal2, L. Albornos1, B. Dopico1, E. Labrador1,*
1 Dpto de Botánica y Fisiología Vegetal, Instituto Hispano Luso de Investigaciones Agrarias, Universidad de Salamanca, Campus Miguel de Unamuno, Salamanca, Spain
2 Dpto de Biología Funcional, Universidad de Santiago de Compostela, Campus de Lugo, Lugo, Spain

This work studied the six β-galactosidases (BGALs) of the subfamily a1 of Arabidopsis, that have been proposed to play important roles in the cell wall remodelling during plant development, although their precise functions are still unknown. Knockout mutants bgal1, bgal2, bgal3, bgal4, bgal5, and bgal12 of Arabidopsis and their wild type (WT) plants were analysed to determine their morphology and composition of their cell walls. The gas chromatography and the Fourier transform infrared spectroscopy revealed differences between the mutants and their WT such as in the proportions of glucose, galactose, or xylose in bgal2 and bgal4 and in cell walls polysaccharides in bgal1, bgal3, and bgal5. However, these slight changes did not result in morphological variations during plant development. None of the mutant seedlings displayed a clear reduction in β(1,4)-galactan content, analysed by immunolocalization. The absence of significant phenotypic changes in the β-galactosidase subfamily a1 mutants could indicate possible β-galactosidases functional redundancy. Future studies will focus on the construction of multiple mutants that help to establish the precise function of each member of the β-galactosidase subfamily a1.

Keywords: FTIR; β(1,4)-galactan; galactose; glucose; glycosyl hydrolase family 35; immunolocalization; xylose
Subjects: knockout mutant; beta-galactosidase; sugars; gene expression; glucose; galactose; arabinose; rhamnose; xylose

Received: September 22, 2016; Revised: February 7, 2017; Accepted: March 13, 2017; Published: January 1, 2018Show citation

ACS AIP APA ASA Harvard Chicago IEEE ISO690 MLA NLM Turabian Vancouver
Moneo-Sánchez, M., Izquierdo, L., Martín, I., Hernández-Nistal, J., Albornos, L., Dopico, B., & Labrador, E. (2018). Knockout mutants of Arabidopsis thaliana β-galactosidase. Modifications in the cell wall saccharides and enzymatic activities. Biologia plantarum62(1), 80-88. doi: 10.1007/s10535-017-0739-2.
Download citation

Supplementary files

Download filebpl-201801-0008_S1.pdf

File size: 2.23 MB

References

  1. Ahn, Y.O., Zheng, M., Bevan, D.R., Esen, A., Shiu, S.H., Benson, J., Peng, H.P., Miller, J.T., Cheng, C.L., Poulton, J.E., Shih, M.C.: Functional genomic analysis of Arabidopsis thaliana glycoside hydrolase family 35. - Phytochemistry 68: 1510-1520, 2007. Go to original source...
  2. Albersheim, P., Nevins, D.J., English, P.D.: A method for the analysis of sugars in plant cell wall polysaccharides by gas liquid chromatography. - Carbohydr. Res. 5: 340-345, 1967. Go to original source...
  3. Albornos, L., Martín, I., Pérez, P., Marcos, R., Dopico, B., Labrador, E.: Promoter activities of genes encoding β-galactosidases from Arabidopsis a1 subfamily. - Plant Physiol. Biochem. 60: 223-232, 2012. Go to original source...
  4. Alonso, J.M., Stepanova, A.N., Leisse, T.J., Kim, C.J., Chen, H., Shinn, P., Stevenson, D.K., Zimmerman, J., Barajas, P., Cheuk, R., Gadrinab, C., Heller, C., Jeske, A., Koesema, E., Meyers, C.C., Parker, H., Prednis, L., Ansari, Y., Choy, N., Deen, H., Geralt, M., Hazari, N., Hom, E., Karnes, M., Mulholland, C., Ndubaku, R., Schmidt, I., Guzman, P., Aguilar-Henonin, L., Schmid, M., Weigel, D., Carter, D.E., Marchand, T., Risseeuw, E., Brogden, D., Zeko, A., Crosby, W.L., Berry, C.C., Ecker, J.R.: Genome-wide insertional mutagenesis of Arabidopsis thaliana. - Science 301: 653-657, 2003. Go to original source...
  5. Alonso-Simón, A., García-Angulo, P., Mélida, H., Encina, A., Álvarez, J.M., Acebes, J.L.: The use of FTIR spectroscopy to monitor modifications in plant cell wall architecture caused by cellulose biosynthesis inhibitors. - Plant Signal. Behav. 6: 1104-1110, 2011. Go to original source...
  6. Balasubramaniam, S., Lee, H.C., Lazan, H., Othman, R., Ali, Z.M.: Purification and properties of a beta-galactosidase from carambola fruit with significant activity towards cell wall polysaccharides. - Phytochemistry 66: 153-163, 2005. Go to original source...
  7. Bradford, M.N.: Rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. - Anal. Biochem. 72: 248-254, 1976. Go to original source...
  8. Černá, M., Barros, A.S., Nunes, A., Rocha, S.M., Delgadillo, I., Čopíková, J., Coimbra, M.A.: Use of FT-IR spectroscopy as a tool for the analysis of polysaccharide food additives. - Carbohydr. Polymer. 51: 383-389, 2003.
  9. Coimbra, M.A., Barros, A., Rutledge, D.N., Delgadillo, I.: FTIR spectroscopy as a tool for the analysis of olive pulp cell wall polysaccharide extracts. - Carbohydr. Res. 317: 145-154, 1999. Go to original source...
  10. Desprez, T., Juraniec, M., Crowell, E.F., Jouy, H., Pochylova, Z., Parcy, F., Höfte, H., Gonneau, M., Vernhettes, S.: Organization of cellulose synthase complexes involved in primary cell wall synthesis in Arabidopsis thaliana. - Proc. nat. Acad. Sci. USA 104: 15572-15577, 2007. Go to original source...
  11. Dokken, K.M., Davis, L.C.: Infrared monitoring of dinitrotoluenes in sunflower and maize roots. - J. Environ. Qual. 40: 719-730, 2011. Go to original source...
  12. Dopico, B., Nicolás, G., Labrador, E.: Partial purification of cell wall β-galactosidases from Cicer arietinum epicotyls. Relationship with cell wall autolytic processes. - Physiol. Plant. 75: 458-464, 1989. Go to original source...
  13. Esteban, R., Dopico, B., Muñoz, F.J., Romo, S., Martín, I., Labrador, E.: Cloning of a Cicer arietinum β-galactosidase with pectin-degrading function. - Plant Cell Physiol. 44: 718-725, 2003. Go to original source...
  14. Esteban, R., Labrador, E., Dopico, B.: A family of β-galactosidase cDNAs related to development of vegetative tissue in Cicer arietinum. - Plant Sci. 168: 457-466, 2005. Go to original source...
  15. Gantulga, D., Ahn, Y.O., Zhou, C., Battogtokh, D., Bevan, D.R., Winkel, B.S.J., Esen, A.: Comparative characterrization of the Arabidopsis subfamily a1 β-galactosidases. - Phytochemistry 70: 1999-2009, 2009. Go to original source...
  16. Hara, Y., Yokoyama, R., Osakabe, K., Toki, S., Nishitani, K.: Function of xyloglucan endotransglucosylase/hydrolases in rice. - Ann. Bot. 114: 1309-1318, 2014. Go to original source...
  17. Hernández-Nistal, J., Martín, I., Dopico, B., Labrador, E.: Coordinated action of β-galactosidases in the cell wall of embryonic axes during chickpea germination and seedling growth. - Plant Biol. 16: 404-410, 2014. Go to original source...
  18. Jamet, E., Canut, H., Boudart, G., Pont-Lezica, R.F.: Cell wall proteins, a new insight through proteomics. - Trends Plant Sci. 11: 33-39, 2006. Go to original source...
  19. Jones, L., Seymour, G.B., Knox, J.P.: Localization of pectic galactan in tomato cell walls using a monoclonal antibody specific to (1-4)-β-D-galactan. - Plant Physiol. 113: 1505-1412, 1997. Go to original source...
  20. Kačuráková, M., Capek, P., Sasinková, V., Wellner, N., Ebringerová, A.: FTIR study of plant cell wall model compounds: pectic polysaccharides and hemicelluloses. - Carbohydr. Polymer. 43: 195-203, 2000. Go to original source...
  21. Kačuráková, M., Wilson, R.H.: Developments in mid-infrared FT-IR spectroscopy of selected carbohydrates. - Carbohydr. Polymer. 44: 291-303, 2001. Go to original source...
  22. Kotake, T., Dina, S., Konishi, T., Kaneko, S., Igarashi, K., Samejima, M., Watanabe, Y., Kimura, K., Tsumuraya, Y.: Molecular cloning of a β-galactosidase from radish that specifically hydrolyzes β-(1-3)-and β-(1-6)-galactosyl residues of arabinogalactan protein. - Plant Physiol. 138: 1563-1576, 2005. Go to original source...
  23. Lazan, H., Ng, S.Y., Goh, L.Y., Ali, Z.M.: Papaya β-galacto-sidase/galactanase isoforms in differential cell wall hydrolysis and fruit softening during ripening. - Plant Physiol. Biochem. 42: 847-853, 2004. Go to original source...
  24. Li, S.C., Han, J.W., Chen, K.C., Chen, C.S.: Purification and characterization of iso-forms of beta-galactosidases in mung bean seedlings. - Phytochemistry 57: 349-359, 2001. Go to original source...
  25. Martín, I., Hernández-Nistal, J., Albornos, L., Labrador, E., Dopico, B.: βIII-Gal is involved in galactan reduction during phloem element differentiation in chickpea stems. - Plant Cell Physiol. 54: 960-970, 2013. Go to original source...
  26. Martín, I., Jiménez, T., Esteban, R., Dopico, B., Labrador, E.: Immunolocalization of a cell wall β-galactosidase reveals its developmentally regulated expression in Cicer arietinum and its relationship to vascular tissue. - J. Plant Growth Regul. 27: 181-191, 2008. Go to original source...
  27. Martín, I., Jiménez, T., Hernández-Nistal, J., Dopico, B., Labrador, E.: The βI-galactosidase of Cicer arietinum is located in thickened cell walls such as those of collenchyma, sclerenchyma and vascular tissue. - Plant Biol. 13: 777-783, 2011. Go to original source...
  28. Martín, I., Jiménez, T., Hernández-Nistal, J., Labrador, E., Dopico, B.: The location of the chickpea cell wall βVgalactosidase suggests involvement in the transition between cell proliferation and cell elongation. - J. Plant Growth Regul. 28: 1-11, 2009. Go to original source...
  29. Moneo-Sánchez, M., Izquierdo, L., Martín, I., Labrador, E. Dopico, B.: Subcellular location of Arabidopsis thaliana subfamily a1 β-galactosidases and developmental regulation of transcript levels of their coding genes. - Plant Physiol. Biochem. 109: 137-145, 2016. Go to original source...
  30. Morris, V.J., Ring, S.G., MacDougall, A.J., Wilson, R.H.: Biophysical characterisation of plant cell walls. - In: Rose, J. (ed.): The Plant Cell Wall. Annual Plant Reviews. Pp. 55-91. Blackwell Publishing, Oxford 2003. Go to original source...
  31. Murashige, T., Skoog, F.: A revised medium for rapid growth and bioassays with tobacco tissue cultures. - Physiol. Plant. 15: 473-497, 1962. Go to original source...
  32. Osato, Y., Yokoyama, R., Nishitani, K.: A principal role for AtXTH18 in Arabidopsis thaliana root growth, a functional analysis using RNAi plants. - J. Plant Res. 119: 153-162, 2006. Go to original source...
  33. Persson, S., Paredez, A., Carroll, A., Palsdottir, H., Doblin, M., Poindexter, P., Khitrov, N., Auer, M., Somerville, C.R.: Genetic evidence for three unique components in primary cell-wall cellulose synthase complexes in Arabidopsis. - Proc. nat. Acad. Sci. USA 104: 15566-15571, 2007. Go to original source...
  34. Sessions, A., Burke, E., Presting, G., Aux, G., McElver, J., Patton, D., Dietrich, B., Ho, P., Bacwaden, J., Ko, C., Clarke, J.D., Cotton, D., Bullis, D., Snell, J., Miguel, T., Hutchison, D., Kimmerly, B., Mitzel, T., Katagiri, F., Glazebrook, J., Law, M., Goff, S.A.: A high-throughput Arabidopsis reverse genetics system. - Plant Cell 14: 2985-2994, 2002. Go to original source...
  35. Smith, L.D., Gross, K.C.: A family of at least seven β-galactosidase genes is expressed during tomato fruit development. - Plant Physiol. 123: 1173-1183, 2000. Go to original source...
  36. Szymanska-Chargot, M., Zdunek, A.: Use of FT-IR spectra and PCA to the bulk characterization of cell wall residues of fruits and vegetables along a fraction process. - Food Biophys. 8: 29-42, 2013. Go to original source...
  37. Verbelen, J.P., Vissenberg, K.: The Expanding Cell. Plant Cell Monographs. Vol. 5. Springer, New York 2007. Go to original source...
  38. Vincken, J.P., Schols, H.A., Oomen, R.J.F.J., Beldman G., Visser R.G.F., Voragen A.G.J. Pectin-the hairy thing. - In: Voragen, A.G.J., Schols, H., Visser, R. (ed.): Advances in Pectin and Pectinase Research. Pp. 47-59. Kluwer Academic Publishers, Dordrecht 2003. Go to original source...
  39. Wilson, R.H., Smith, A.C., Kačuráková, M., Saunders, P.K., Wellner, N., Waldron, K.W.: The mechanical properties and molecular dynamics of plant cell wall polysaccharides studied by Fourier-transform infrared spectroscopy. - Plant Physiol. 124: 397-405, 2000. Go to original source...