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Molecular Biology Reports

, Volume 39, Issue 4, pp 3523–3532 | Cite as

Characterization of PROFILIN genes from allotetraploid (Gossypium hirsutum) cotton and its diploid progenitors and expression analysis in cotton genotypes differing in fiber characteristics

  • Anagnostis Argiriou
  • Apostolos Kalivas
  • Georgios Michailidis
  • Athanasios Tsaftaris
Article

Abstract

The actin-binding protein profilin (PRF) plays an important role in cell growth and expansion by regulating the organization of the actin filaments. Recent studies have reported association between fiber elongation in cultivated cotton (Gossypium hirsutum) and PRF expression. In the present study, we cloned four genomic clones from allotetraploid cotton (G. hirsutum) and its putative diploid progenitors (G. arboreum and G. raimondii) designated GhPRF1_A, GhPRF1_D, GaPRF1, and GrPRF1 encoding cotton PRF and characterized their genomic structure, phylogenetic relationships and promoter structure. Sequence analysis of the coding regions of all clones resulted in a single protein product which revealed more than 80% similarity to most plant PRFs and a typical organization with an actin-binding and a polybasic phospholipid binding motif at the carboxy terminus. DNA blot hybridization suggested that PRF gene is present with more than one copy in the allotetraploid species G. hirsutum. Expression analysis performed in various organs of cultivated cotton revealed that the PRF gene was preferentially expressed in cotton fibers. Very low levels of expression were observed in whole flowers, while PRF transcripts were not detected in other organs examined. Furthermore, higher levels of expression were observed at the early stages of cotton fiber development (at 10 days post anthesis), indicative that this gene may play a major role in the early stages of cotton fiber development. Quantitation of the expression by real-time PCR revealed higher expression levels in a G. hirsutum variety with higher fiber percentage compared to a variety with lower percentage. In addition, higher levels of expression were found in cultivated allotetraploid G. barbadense cotton species with higher fiber length in comparison to cultivated allotetraploid G. hirsutum.

Keywords

Actin binding Cotton fiber Gossypium hirsutum Profilin 

Supplementary material

11033_2011_1125_MOESM1_ESM.pdf (51 kb)
Fig. S1 Alignment of cotton PRF profilin genomic sequences. Nucleotide sequence alignment was performed using the Clustal method. Boxed areas represent Exons, boxed grey and light grey areas show the nucleotide differences between the four sequences. Start and stop codons, are in bold. (PDF 50 kb)
11033_2011_1125_MOESM2_ESM.pdf (54 kb)
Fig. S2 Aminoacid sequence comparison among plant profilin sequences. Identical amino acids in all proteins are in white letters highlighted by black background, while identical amino acids in more than 75% of the sequences are highlighted by grey background. Dashes indicate gaps to maximize alignment. Underlined are the cotton GhPRF1_A, GhPRF1_D, GaPRF1 and GrPRF1 protein sequences. The conserved polybasic and actin-binding aminoacid stretches are boxed. The alignments were generated using ClustalW [30]. The cotton GhPRF1_A, GhPRF1_D, GaPRF1 and GrPRF1 protein sequences was deduced from the corresponding genomic clones. (PDF 53 kb)
11033_2011_1125_MOESM3_ESM.pdf (54 kb)
Fig. S3 Phylogenetic relation of plant PRFs. The tree was generated by the Neighbor-Joining method using the p-distance correction. Numbers next to the nodes are bootstrap values from 1000 replications. Bootstrap values are shown in the nodes of the tree. The values indicate the number of times the group consisted of proteins at the same side of each branch, out of a hundred trees generated. Underlined are the cotton GhPRF1_A, GhPRF1_D, GaPRF1 and GrPRF1 protein sequences. (PDF 54 kb)

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Copyright information

© Springer Science+Business Media B.V. 2011

Authors and Affiliations

  • Anagnostis Argiriou
    • 1
  • Apostolos Kalivas
    • 1
    • 4
  • Georgios Michailidis
    • 1
    • 3
  • Athanasios Tsaftaris
    • 1
    • 2
  1. 1.Institute of AgrobiotechnologyCERTHThermiGreece
  2. 2.Department of Genetics and Plant BreedingAUThThessalonikiGreece
  3. 3.Department of Animal ReproductionAUThThessalonikiGreece
  4. 4.Cotton and Industrial Plants InstituteNational Agricultural Research FoundationSindosGreece

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