Abstract
We used a transgenic Arabidopsis line expressing a translational fusion between a mitotic cyclin and the reporter gene β-glucuronidase (GUS) to investigate cell divisions in postembryonic root meristems. The fusion protein contains the cyclin destruction box (CDB) and this leads to a rapid degradation of the chimeric GUS-protein after mitosis. Hence, the staining pattern of the meristem marks dividing cells. We observed that upon germination the first cell divisions occur in epidermis cells at the junction with the hypocotyl. Moreover, the accelerated root growth on media supplemented with sucrose correlates with an increased number of dividing cells and an enlargement of the root meristematic zone. The conditional root expansion mutants porn porn 1 and procuste 1 (quill) suppress this sugar effect leading to a smaller meristematic zone. Simultaneous visualisation of the nucleus revealed that the CYCAT1:CDB:GUS expression is subcellularly localised around the nucleus. This particular staining starts at prophase and disappears after the completion of the new cell wall. In metaphase the staining invades the cytoplasm whereas in the telophase it concentrates again around the nucleus. This cell cycle-dependent distribution was used to characterise the two root specific cytokinesis mutants pleiade 1 and hyade 1. In both mutants, cells which fail to develop a complete cell wall during cytokinesis divide synchronously in further cell divisions leading to multinucleate cells. These experiments demonstrate the usefulness of the CYCAT1:CDB:GUS marker line for studying cell division of wild-type and mutants. Furthermore, this line can be used to analyse the influence of biotic and abiotic signals on the rate and spatial distribution of cell divisions.
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References
Blancaflor E B, Fasano J M and Gilroy S 1998 Mapping the functional roles of cap cells in the response of Arabidopsis primary root to gravity. Plant Physiol. 116, 213–222.
Beemster G T S and Baskin T I 1998 Analysis of cell division and elongation underlying the developmental acceleration of root growth in Arabidopsis thaliana. Plant Physiol. 116, 1515–1526.
Cheng J-C, Seeley K A and Sung Z R 1995 RML1 and RML2, Arabidopsis genes required for cell proliferation at the root tip. Plant Physiol. 107, 365–376.
Clowes F A L 1961 Apical Meristems. Blackwell, Oxford.
Desnos T, Orbovic V, Bellini C, Kronenberger J, Caboche M, Traas J and Höfte H 1996 Procustel mutants identify two distinct genetic pathways controlling hypocotyl cell elongation, respectively in dark-and light-grown Arabidopsis seedlings. Development 122, 683–693.
Doerner P W, Jorgensen J-E, You R, Steppuhn J and Lamb C J 1996 Control of root growth and development by cyclin expression. Nature 380, 520–523.
Dolan L, Janmaat K, Willemsen V, Linstead P, Poethig S, Roberts K and Scheres B 1993 Cellular organisation of the Arabidopsis root. Development 119, 71–84.
Dolan L, Duckett C M, Grierson C, Linstead P, Schneider K, Lawson E, Dean C, Poethig S and Roberts K 1994 Clonal relationships and cell patterning in the root epidermis of Arabidopsis. Development 120, 2465–2474.
Evans T, Rosenthal E T, Youngblom J, Distel D and Hunt T 1983 Cyclin: A protein specified by maternal mRNA in sea urchin eggs that is destroyed at each cleavage division. Cell 33, 389–396.
Ferreira P, Hemerly A, Engler J D, Bergounioux C, Burssens S, Van Montagu M, Engler G and Inze D 1994 Three discrete classes of Arabidopsis cyclins are expressed during different intervals of the cell cycle. Proc Natl Acad Sci USA 91, 11313–11317.
Fobert P R, Coen E S, Murphy G J and Doonan J H 1994 Patterns of cell division revealed by transcriptional regulation of genes during the cell cycle in plants. EMBO J 13, 616–24.
Genschik P, Criqui M C, Parmentier Y, Derevier A and Fleck J 1998 Cell cycle-dependent proteolysis in plants: identification of the destruction box pathway and metaphase arrest produced by the proteasome inhibitor MG132. Plant Cell 10, 2063–2075.
Hauser M-T and Benfey P N 1994 Genetic regulation of root expansion in Arabidopsis thaliana. In NATO-ASI Plant Molecular Biology Series. Eds P Puigdomenech and G Coruzzi. pp 31–40. Springer-Verlag, New York.
Hauser M-T, Morikami A and Benfey P N 1995 Conditional root expansion mutants of Arabidopsis. Development 121, 1237–1252.
Hemerly A S, Ferreira P, Engler J, Van Montagu M, Engler G and Inze D 1993. Cdc2a expression in Arabidopsis is linked with competence for cell division. Plant Cell 5, 1722–1723.
Hemerly A S, Bergounioux C, Van Montagu M, Inze D and Ferreira F 1992 Genes regulating the plant cell cycle: isolation of a mitotic like cyclin from Arabidopsis thaliana. Proc Natl Acad Sci USA 89, 3295–3299.
Hirt H, Mink M, Bögre L, Györgyey J, Jonak C, Gartner A, Dudits D and Heberle-Bors E 1992 Alfalfa cyclins: differential expression during the cell cycle and in plant organs. Plant Cell 4, 15311538.
Ivanov V B 1997 Relationship between cell proliferation and transition to elongation in plant roots. Int. J. Dev. Biol. 41, 907–915.
Kouchi H, Sekine M and Hata S 1995 Distinct classes of mitotic cyclins are differentially expressed in the soybean shoot apex during the cell cycle. Plant Cell 7, 1143–1155.
Martinez M C, Jorgensen J-E, Lawton M A, Lamb C J and Doerner P W 1992 Spatial pattern of cdc2 expression in relation to meristem activity and cell proliferation during plant development. Proc Natl Acad Sci USA 80, 7360–7364.
Meskiene I, Bögre L, Dahl M, Pieck M, Ha D T C, Swoboda I, Heberle-Bors E, Ammerer G and Hirt H 1995 CycMs3, a novel B-type alfalfa cyclin gene, is induced in the G(0)-to-G(1) transition of the cell cycle. Plant Cell 7, 759–771.
Murashige T and Skoog F 1962 A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant 15, 473–479.
Ovecka M, Ciamporova M and Hauser M-T 1996 Root-specific mutation of Arabidopsis thaliana affecting cytokinesis: production and fate of multinucleate cells. J. Computer-Assisted Microscopy 8, 277–278.
Renaudin J-P, Doonan J H, Freeman D, Hashimoto J, Hirt H, Inze D, Jacobs T, Kouchi H, Rouze P, Sauter M, Savoure A, Sorrell D A, Sundaresan V and Murray J A H 1996 Plant cyclins: a unified nomenclature for plant A-, B- and D-type cyclins based on sequence organization. Plant Mol Biol 32, 1003–1018.
Scheres B, Wolkenfeldt H, Willemsen V, Terlouw M, Lawson E, Dean C and Weisbeek P 1994 Embryonic origin of Arabidopsis primary root and root meristem initials. Development 120, 2475–2487.
Setiady Y Y, Sekine M, Hariguchi N, Yamamoto T, Kouchi H and Shinmyo A 1995 Tobacco mitotic cyclins: cloning, characterization, gene expression and functional assay. Plant J 8, 949–957.
Shaul O, Mironov V, Burssens S, Van Montagu M and Inze D 1996 Two Arabidopsis cyclin promoters mediate distinctive transcriptional oscillation in synchronized tabocco BY-2 cells. Proc Natl Acad Sci USA 93, 4868–4872.
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Hauser, MT., Bauer, E. (2001). Histochemical analysis of root meristem activity in Arabidopsis thaliana using a cyclin:GUS (β-glucuronidase) marker line. In: Gašparíková, O., Čiamporová, M., Mistrík, I., Baluška, F. (eds) Recent Advances of Plant Root Structure and Function. Developments in Plant and Soil Sciences, vol 90. Springer, Dordrecht. https://doi.org/10.1007/978-94-017-2858-4_1
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DOI: https://doi.org/10.1007/978-94-017-2858-4_1
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