Functional analysis of splice variant expression of MADS AFFECTING FLOWERING 2 of Arabidopsis thaliana
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Functional analysis of splice variant expression of MADS AFFECTING FLOWERING 2 of Arabidopsis thaliana Sarah Marie Rosloski • Anandita Singh • Sathya Sheela Jali • Sureshkumar Balasubramanian Detlef Weigel • Vojislava Grbic
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Received: 14 March 2012 / Accepted: 19 October 2012 / Published online: 31 October 2012 Ó The Author(s) 2012. This article is published with open access at Springerlink.com
Abstract The MADS-AFFECTING FLOWERING 2 (MAF2) gene of Arabidopsis thaliana has been characterized as a repressor of flowering. The molecular basis of MAF2 gene function and role of alternative MAF2 transcripts in flowering time modulation is not understood. MAF2 splice variant expression was quantified in cold-acclimated plants by quantitative RT-PCR. Cold influenced the abundance of splice variants and prompted a functional study of splice forms. Individual variants were overexpressed in the Col background and were assayed for their ability to delay flowering. Overexpression of MAF2 variants 2 and 4 had limited effect on flowering time. Overexpression of MAF2 splice variant 1 resulted in early flowering and affected the expression of the endogenous MAF2 gene and its paralogues, confounding functional assessment. In the Ll-2 Arabidopsis Electronic supplementary material The online version of this article (doi:10.1007/s11103-012-9982-2) contains supplementary material, which is available to authorized users. S. M. Rosloski S. S. Jali V. Grbic (&) Department of Biology, Western University, London, ON N6A 5B8, Canada e-mail: [email protected] A. Singh S. Balasubramanian D. Weigel Department of Molecular Biology, Max-Planck Institute for Developmental Biology, 72076 Tu¨bingen, Germany Present Address: A. Singh Department of Biotechnology, Faculty of Applied Science, TERI University, 10 Institutional Area, Vasant Kunj, New Delhi 110070, India Present Address: S. Balasubramanian School of Biological Sciences, Monash University, Clayton, VIC 3800, Australia
accession, a MAF2, MAF3, MAF4 and FLC null line, MAF2 var1 was consistent in its effect on reproductive delay under ambient and reduced temperatures, indicating that it acts as a repressor of flowering. Keywords Arabidopsis thaliana Flowering time Alternative splicing Response to cold MAF2
Introduction The number of genes discovered in early genome sequencing projects was lower than anticipated considering the apparent complexity of eukaryotic organisms (Rubin et al. 2000; International Human Genome Sequencing Consortium 2001). However, a small gene set may result in complex phenotypes if genes produce multiple products with diverse functions (Pesole 2008). Alternative splicing is one of the mechanisms that can expand the functional diversity of genes (Modrek and Lee 2002; Reddy 2007). Alternative splicing operates through selection of variable donor and acceptor sites during pre-mRNA processing and can result in a suite of divergent mRNAs produced from a single gene (Sharp 1994). Alternative splicing can instigate a spectrum of functional outcomes that may cons
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