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Isolation of a CENTRORADIALIS/TERMINAL FLOWER1 homolog in saffron (Crocus sativus L.): characterization and expression analysis

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Abstract

Genes in the phosphatidyl-ethanolamine-binding protein (PEBP) family are instrumental in regulating the fate of meristems and flowering time. To investigate the role of these genes in the monocotyledonous plant Crocus (Crocus sativus L), an industrially important crop cultivated for its nutritional and medicinal properties, we have cloned and characterized a CENTRORADIALIS/TERMINAL FLOWER1 (CEN/TFL1) like gene, named CsatCEN/TFL1-like, the first reported CEN/TFL1 gene characterized from such a perennial geophyte. Sequence analysis revealed that CsatCEN/TFL1 shows high similarity to its homologous PEBP family genes CEN/TFL1, FT and MFT from a variety of plant species and maintains the same exon/intron organization. Phylogenetic analysis of the CsatCEN/TFL1 amino acid sequence confirmed that the isolated sequences belong to the CEN/TFL1 clade of the PEBP family. CsatCEN/TFL1 transcripts could be detected in corms, flower and flower organs but not in leaves. An alternative spliced transcript was also detected in the flower. Comparison of expression levels of CsatCEN/TFL1 and its alternative spliced transcript in wild type flower and a double flower mutant showed no significant differences. Overexpression of CsatCEN/TFL1 transcript in Arabidopsis tfl1 plants reversed the phenotype of early flowering and terminal flowering of the tfl1 plants to a normal one. Computational analysis of the obtained promoter sequences revealed, next to common binding motifs in CEN/TFL1-like genes as well as other flowering gene promoters, the presence of two CArG binding sites indicative of control of CEN/TFL1 by MADS-box transcription factors involved in crocus flowering and flower organ formation.

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Abbreviations

CEN:

CENTRORADIALIS

PEBP:

Phosphatidylethanolamine binding protein

TFL1:

TERMINAL FLOWER1

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Acknowledgments

We thank Yiannis Patsios for his help in collecting plant material in the field and Dr. Jonathan Rhodes for critically reviewing the manuscript. Continuous support for the Institute of Agrobiotechnology/CERTH from the General Secretariat of Research and Technology of Greece is also acknowledged.

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Author notes

  1. Apostolos Kalivas

    Present address: Cotton and Industrial Plants Institute, National Agricultural Research Foundation, Sindos, 574 00, Greece

Authors and Affiliations

  1. Institute of Agrobiotechnology, CERTH, Thermi, 570 01, Greece

    Athanasios Tsaftaris, Konstantinos Pasentsis, Apostolos Kalivas, Sofia Michailidou, Panagiotis Madesis & Anagnostis Argiriou

  2. Department of Genetics and Plant Breeding, AUTh, Thessaloniki, 540 06, Greece

    Athanasios Tsaftaris

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  1. Athanasios Tsaftaris
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Correspondence to Athanasios Tsaftaris.

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11033_2012_1634_MOESM1_ESM.jpg

Figure S1 (A) Genomic organization of CEN/TFL1-like gene and promoter analysis. Bindings elements are indicated boxed with different colors. Grey boxes with numbers inside indicate the four exons and their respective size. TSS: Transcription Start Site. (B) Promoter sequence. Predicted bindings sites are grey boxed. Putative TATA box in underlined. ATG start codon is in bold. (C). Comparison of the frequencies of the putative binding sites found in CEN/TFL1 and FT promoters from rice and Arabidopsis. (D) Schematic representation of Crocus, Rice and Arabidopsis promoters. Putative binding sites are indicated in different color and shapes as in (C) (JPEG 1711 kb)

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Tsaftaris, A., Pasentsis, K., Kalivas, A. et al. Isolation of a CENTRORADIALIS/TERMINAL FLOWER1 homolog in saffron (Crocus sativus L.): characterization and expression analysis. Mol Biol Rep 39, 7899–7910 (2012). https://doi.org/10.1007/s11033-012-1634-8

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