title: Molecular characterisation of two homoeologous elicitor-responsive lipin genes in cotton.
abstract: The identification and molecular characterisation of two lipin-like gene copies (GhLIPN) in cotton, Gossypium hirsutum, an allotetraploid derived from two progenitor diploid Gossypium species, is described. Sequence analyses of the GhLIPN copies, designated GhLIPN-1 and -2, revealed that they contain 11 exons, separated by ten introns. They each have a 2,643 bp open reading frame that encodes 880 aa proteins, and share a 97.7 and 95.5 % sequence similarity at the translated nucleotide and amino acid level, respectively. The GhLIPN genes have a distinct domain architecture consisting of an archetypical N-terminal lipin domain, followed by a haloacid dehalogenase (HAD) domain towards the C-terminus. A Southern blot did not distinguish between the two gene copies, which suggests that they may be homoeologs rather than paralogs. GhLIPN-2 is more similar to a homoeologous sequence from G. raimondii, representing the ancestral D-genome, compared to GhLIPN-1 that matches G. herbaceum and that represents the A-genome. Our data indicate that GhLIPN-1 and GhLIPN-2 are homoeologs that derive from the A- and the D-diploid genomes, respectively. The promoter sequences of GhLIPN-1 and -2 differ by 56 %, as a result of multiple indels. In silico analysis of the promoter regions revealed that both genes contain numerous putative defence-related and elicitor-responsive cis-elements that support a role for GhLIPN in defence responses. Relative quantification real-time PCR confirmed the up-regulation in response to a cell-wall-derived V. dahliae elicitor, which supported the association of GhLIPN with defence signalling. The results add a new dimension to the proposed roles of lipins in plants by suggesting that lipins may have a role in defence signalling.
authors.affiliation: Department of Biochemistry, University of Johannesburg, Kingsway Campus, P.O. Box 524, Auckland Park, 2006, South Africa.
authors.forename: Sonia M
authors.initials: SM
authors.lastname: Phillips
authors.forename: Ian A
authors.initials: IA
authors.lastname: Dubery
authors.forename: Henriette
authors.initials: H
authors.lastname: van Heerden
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D015139:Blotting, Southern; D017931:DNA Primers / Q000235:genetics; D040461:Gene Components; D017343:Genes, Plant / Q000235:genetics*; D003368:Gossypium / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D010711:Phosphatidate Phosphatase / Q000235:genetics*; D011401:Promoter Regions, Genetic / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D013045:Species Specificity
chemical_list: D017931:DNA Primers; D010711:Phosphatidate Phosphatase
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