Abstract
Precise cellular localization of the GUS stain is notoriously difficult inArabidopsis seeds. Here we report an improved protocol for the clearing of seeds after GUS staining. Incubation in ethanol-acetic acid (EtAc) and Hoyer’s medium allows reliable cellular localization of the GUS, even in seeds from late developmental stages. This method also leads to the staining of nucleoli in the endosperm and embryo, facilitating nuclear counts in endosperm development.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Beeckman T and Engler G (1994) Clearing of histochemically stained plant tissue. Plant Mol Biol Rep 12: 37–42.
Berleth T and Jurgens G (1993) The role of the monopterous gene in organising the basal body region of the Arabidopsis embryo, Development 118: 575–587.
Boisnard-Lorig C, Colon-Carmona A, Bauch M, Hodge S, Doerner P, Bancharel E, Dumas C, Haseloff J, and Berger F (2001) Dynamic analyses of the expression of the HISTONE::YFP fusion protein in Arabidopsis show that syncytial endosperm is divided in mitotic domains. Plant Cell 13: 495–509.
Brown RC, Lemmon BE, Nguyen H, and Olsen OA (1999) Development of endosperm in Arabidopsis thaliana. Sex Plant Reprod 12: 32–42.
Jefferson RA, Kavanagh TA, and Bevan MW (1987) GUS fusions: b-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6: 3901–3907.
Liu CM and Meinke D (1998) The titan mutants of Arabidopsis are disrupted in mitosis and cell cycle control during seed development. Plant J 16: 21–31.
Luo M, Bilodeau P, Dennis ES, Peacock WJ, and Chaudhury A (2000) Expression and parent-of-origin effects for FIS2, MEA, and FIE in the endosperm and embryo of developing Arabidopsis seeds. Proc Natl Acad Sci USA 97: 10637–10624.
Mandal A, Sandgren M, Holmström KO, Gallois P, and Palva ET (1995) Identification of Arabidopsis thaliana sequences responsive to low temperature and abscisic acid by T-DNA tagging and in-vivo gene fusion. Plant Mol Biol Rep 13: 243–254.
Martin T, Wohner R-V, Hummel S, Willmitzer L, and Frommer WB (1992) The GUS reporter system as a tool to study plant gene expression. In: GUS Protocols: Using the GUS Gene as a Reporter of Gene Expression. Academic Press, Inc., pp 23–43.
Motzfeldt-Laane M and Lie T (1992) Overflatestudier av ugjennomsiktige objekter. In: Håndbok i mikroskopi og framstilling av preparater. Universitetsforlaget, Oslo, pp 173–174.
Stangeland B, Salehian Z, Aalen RB, Mandal A, and Olsen O-A (2001) Promoter trapping of seven genes expressed in Arabidopsis endosperm. 12th International Conference on Arabidopsis Research, June 23–27, Madison, pp 534.
Sørensen MB, Chaudhury AM, Robert H, Bancharel E, and Berger F (2001) Polycomb group genes control pattern formation in plant seed. Current Biology 11: 277–281.
Topping JF, Agyeman F, Henricot B, and Lindsey K (1994) Identification of molecular markers of embryogenesis in Arabidopsis thaliana by promoter trapping. Plant J 5: 895–903.
Vielle-Calzada J-P, Baskar R, and Grossniklaus U (2000) Delayed activation of the paternal genome during seed development. Nature 404: 91–94.
Author information
Authors and Affiliations
Corresponding author
Additional information
An erratum to this article is available at http://dx.doi.org/10.1007/BF02782473.
Rights and permissions
About this article
Cite this article
Stangeland, B., Salehian, Z. An improved clearing method for GUS assay inArabidopsis endosperm and seeds. Plant Mol Biol Rep 20, 107–114 (2002). https://doi.org/10.1007/BF02799426
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF02799426