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Journal of Heredity Advance Access published online on October 27, 2009
Journal of Heredity, doi:10.1093/jhered/esp082
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© The American Genetic Association. 2009. All rights reserved. For permissions, please email: journals.permissions@oxfordjournals.org.

Brief Communication

Isolation and Phylogenetic Footprinting Analysis of the 5'-Regulatory Region of the Floral Homeotic Gene OrcPI from Orchis italica (Orchidaceae)

Serena Aceto, Carmela Cantone, Pasquale Chiaiese, Gianluca Ruotolo, Maria Sica, and Luciano Gaudio

From the Department of Biological Sciences, Section of Genetics and Molecular Biology, University of Naples Federico II, via Mezzocannone 8, Napoli 80134, Italy (Aceto, Cantone, Sica, and Gaudio) and the Department of Soil, Plant and Environmental Sciences, University of Naples Federico II, Portici, Naples 80055, Italy (Chiaiese and Ruotolo)

Address correspondence to Serena Aceto at the address above, or e-mail: serena.aceto{at}unina.it.

The nucleotide sequences of regulatory elements from homologous genes can be strongly divergent. Phylogenetic footprinting, a comparative analysis of noncoding regions, can detect putative transcription factor binding sites (TFBSs) shared among the regulatory regions of 2 or more homologous genes. These conserved motifs have the potential to serve the same regulatory function in distantly related taxa. We isolated the 5'-noncoding region of the OrcPI gene, a MADS-box transcription factor involved in flower development in Orchis italica, using the thermal asymmetric interlaced polymerase chain reaction technique. This region (comprising 1352 bp) induced transient β-glucuronidase expression in the petal tissue of white Rosa hybrida flowers and represents the 5'-regulatory sequence of the OrcPI gene. Phylogenetic footprinting analysis detected conserved regions within the 5'-regulatory sequence of OrcPI and the homologous regions of Oryza sativa, Lilium regale, and Arabidopsis thaliana. Some of these sequences are known TFBSs described in databases of plant regulatory elements. Nucleotide sequence data reported are available in the DDBJ/EMBL/GenBank databases under the following accession numbers: AF198055 [GenBank] promoter region of the PISTILLATA (PI) gene of A. thaliana; AB094985 [GenBank] cDNA of OrcPI (PI/GLOBOSA [PI/GLO] homologue) of O. italica; AB378089 [GenBank] 5'-regulatory region of the OrcPI gene of O. italica; AP008211 [GenBank] putative promoter region of OSMADS2 (PI/GLO homologue) of O. sativa; AP008207 [GenBank] putative promoter region of OSMADS4 (PI/GLO homologue) of O. sativa; and AB158292 [GenBank] putative promoter region of the PI/GLO homologue of L. regale.

Key Words: MADS box • orchids • OrcPI • phylogenetic footprinting • 5'-regulatory sequence

Corresponding Editor: John Burke

Received February 23, 2009
Revised August 24, 2009
Accepted September 3, 2009


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