title: Cloning and characterization of a wheat neutral ceramidase gene Ta-CDase.
abstract: Ceramidases are key enzymes in the regulation of the cellular levels of ceramide, sphingosine and sphingosine-1-phosphate. This study first reports on the molecular cloning, sequencing and expression profile of the gene encoding the wheat neutral ceramidase designated as Ta-CDase. A full length wheat Ta-CDase gene is obtained by rapid amplification of cDNA ends (RACE) based on the sequence of the WSRC36 fragment from an incompatible suppression subtractive hybridization (SSH) cDNA library of wheat leaves infected by Puccinia striiformis f. sp. tritici. The open reading frame (ORF) of 2,839 nucleotides encodes a polypeptide of 785 amino acids with a predicted isoelectric point (pI) of 6.398. The protein conserved domain search indicates that the polypeptide contains the signature of ceramidase, signal peptide sequence and transmembrane region. A phylogenetic analysis reveals that a high degree of relatedness exists among wheat Ta-CDase and ceramidases from other plant species at the amino acid level, while its relationship to that of animals and pathogens is more distant. The expression profile of the Ta-CDase shows a very strong expression of transcripts only at 48 h post inoculation (hpi), while expression level is low at other time points. Southern blot analyses showed that Ta-CDase is a multi-copy gene and located on wheat chromosome 4D and 5A.
authors.affiliation: College of Plant Protection and Shaanxi Key Laboratory of Molecular Biology for Agriculture, Northwest A&F University, Yangling, Shaanxi 712100, People's Republic of China.
authors.forename: Xiumei
authors.initials: X
authors.lastname: Yu
authors.forename: Xiaojie
authors.initials: X
authors.lastname: Wang
authors.forename: Xueling
authors.initials: X
authors.lastname: Huang
authors.forename: Heinrich
authors.initials: H
authors.lastname: Buchenauer
authors.forename: Qingmei
authors.initials: Q
authors.lastname: Han
authors.forename: Jun
authors.initials: J
authors.lastname: Guo
authors.forename: Jie
authors.initials: J
authors.lastname: Zhao
authors.forename: Zhipeng
authors.initials: Z
authors.lastname: Qu
authors.forename: Lili
authors.initials: L
authors.lastname: Huang
authors.forename: Zhensheng
authors.initials: Z
authors.lastname: Kang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001487:Basidiomycota / Q000472:pathogenicity; D055572:Ceramidases / Q000378:metabolism; D032461:Chromosomes, Plant; D003001:Cloning, Molecular; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D007113:Immunity, Innate / Q000235:genetics; D008969:Molecular Sequence Data; D055576:Neutral Ceramidase / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D010802:Phylogeny; D010935:Plant Diseases / Q000382:microbiology; D016415:Sequence Alignment; D013110:Sphingosine / Q000378:metabolism; D014908:Triticum / Q000201:enzymology* / Q000235:genetics* / Q000382:microbiology
chemical_list: D055572:Ceramidases; D055576:Neutral Ceramidase; D013110:Sphingosine