1 3 Plant Mol Biol (2014) 86:395–407 DOI 10.1007/s11103-014-0236-3 The pineapple AcMADS1 promoter confers high level expression in tomato and Arabidopsis flowering and fruiting tissues, but AcMADS1 does not complement the tomato LeMADS-RIN (rin) mutant Richard L. Moyle · Jonni H. Koia · Julia Vrebalov · James Giovannoni · Jose R. Botella Received: 3 December 2013 / Accepted: 4 August 2014 / Published online: 20 August 2014 © Springer Science+Business Media Dordrecht 2014 proteins. The AcMADS1 promoter directed strong expres- sion of the GUS reporter gene to fruits and developing flo- ral organs in tomato and Arabidopsis thaliana, suggesting AcMADS1 may play a role in flower development as well as fruitlet ripening. The AcMADS1 promoter provides a useful molecular tool for directing transgene expression, particularly where up-regulation in developing flowers and fruits is desirable. Keywords Ananas comosus · Fruit ripening · Promoter analysis · Beta-glucuronidase · MADS box transcription factor Introduction Fruit ripening and softening have a significant impact on fruit quality, nutritional value, harvesting costs, shelf-life, storage and ultimately human health (Klee and Giovan- noni 2011). Physiologically, fruit ripening and senescence are characterized as catabolic events that cause changes in color, texture, flavor, and pathogen susceptibility dur- ing fruit maturation. Modification in the rate of ethylene biosynthesis and respiration, decrease in the rate of plant growth, increase in ROS activity, breakdown of chloro- phyll, and degradation of cell wall and other organelle structures have also been associated with fruit ripening pro- cesses. The regulation of fruit ripening is an important agri- cultural concern and it is of interest to determine if diverse plants bearing fleshy fruits share conserved ripening regu- latory mechanisms. Fleshy fruits are generally classified into two groups according to their ripening physiologies. Climacteric fruits such as tomato, banana, and apple exhibit a burst of eth- ylene biosynthesis and an increase in respiration at the Abstract A previous EST study identified a MADS box transcription factor coding sequence, AcMADS1, that is strongly induced during non-climacteric pineapple fruit rip- ening. Phylogenetic analyses place the AcMADS1 protein in the same superclade as LeMADS-RIN, a master regulator of fruit ripening upstream of ethylene in climacteric tomato. LeMADS-RIN has been proposed to be a global ripening regulator shared among climacteric and non-climacteric species, although few functional homologs of LeMADS- RIN have been identified in non-climacteric species. AcMADS1 shares 67 % protein sequence similarity and a similar expression pattern in ripening fruits as LeMADS- RIN. However, in this study AcMADS1 was not able to complement the tomato rin mutant phenotype, indicating AcMADS1 may not be a functionally conserved homolog of LeMADS-RIN or has sufficiently diverged to be unable to act in the context of the tomato network of interacting Richard L. Moyle and Jonni H. Koia have contributed equally to this work. Electronic supplementary material The online version of this article (doi:10.1007/s11103-014-0236-3) contains supplementary material, which is available to authorized users. R. L. Moyle · J. H. Koia · J. R. Botella (*) Plant Genetic Engineering Laboratory, School of Agriculture and Food Sciences, University of Queensland, Brisbane 4072, Australia e-mail: j.botella@uq.edu.au R. L. Moyle e-mail: r.moyle1@uq.edu.au J. Vrebalov · J. Giovannoni US Department of Agriculture, Boyce Thompson Institute for Plant Research, Tower Road, Cornell University Campus, Ithaca, NY 14853, USA