title: Isolation and characterization of a fatty acid- and retinoid-binding protein from the cereal cyst nematode Heterodera avenae.
abstract: A gene encoding fatty acid- and retinoid-binding protein was isolated from the cereal cyst nematode Heterodera avenae and the biochemical function of the protein that it encodes was analysed. The full-length cDNA of the Ha-far-1 gene is 827 bp long and includes a 22- nucleotide trans-spliced leader sequence (SL1) at its 5-end. The genomic clone of Ha-far-1 consists of eight exons separated by seven introns, which range in size from 48 to 186 bp. The Ha-far-1 cDNA contains an open reading frame encoding a 191 amino acid protein, with a predicted secretory signal peptide. Sequence analysis showed that Ha-FAR-1 has highest similarity to the Gp-FAR-1 protein from the potato cyst nematode, Globodera pallida and that the protein was grouped with all homologues from other plant-parasitic nematodes in a phylogenetic analysis. Fluorescence-based ligand binding analysis confirmed that the recombinant Ha-FAR-1 protein was able to bind fatty acids and retinol. Spatial and temporal expression assays showed that the transcripts of Ha-far-1 accumulated mainly in the hypodermis and that the gene is most highly expressed in third-stage juveniles of H. avenae. Fluorescence immunolocalization showed that the Ha-FAR-1 protein was present on the surface of the infective second-stage juveniles of H. avenae. Nematodes treated with dsRNA corresponding to Ha-far-1 showed significantly reduced reproduction compared to nematodes exposed to dsRNA from a non-endogenous gene, suggesting that Ha-far-1 may be an effective target gene for control of H. avenae using an RNAi strategy.
authors.affiliation: Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, Department of Plant Pathology, Nanjing Agricultural University, Nanjing, China.
authors.forename: Xiuhu
authors.initials: X
authors.lastname: Le
authors.affiliation: Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, Department of Plant Pathology, Nanjing Agricultural University, Nanjing, China. Electronic address: xuanwang@njau.edu.cn.
authors.forename: Xuan
authors.initials: X
authors.lastname: Wang
authors.affiliation: Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, Department of Plant Pathology, Nanjing Agricultural University, Nanjing, China.
authors.forename: Tinglong
authors.initials: T
authors.lastname: Guan
authors.affiliation: Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, Department of Plant Pathology, Nanjing Agricultural University, Nanjing, China.
authors.forename: Yuliang
authors.initials: Y
authors.lastname: Ju
authors.affiliation: Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, Department of Plant Pathology, Nanjing Agricultural University, Nanjing, China. Electronic address: lihm@njau.edu.cn.
authors.forename: Hongmei
authors.initials: H
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry; D018509:DNA, Helminth / Q000302:isolation & purification; D050556:Fatty Acid-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D005260:Female; D018507:Gene Expression Regulation, Developmental; D020868:Gene Silencing; D015801:Helminth Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D017403:In Situ Hybridization; D008024:Ligands; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012177:Retinol-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D016415:Sequence Alignment; D014158:Transcription, Genetic; D014908:Triticum / Q000469:parasitology; D014431:Tylenchoidea / Q000737:chemistry* / Q000235:genetics
chemical_list: D018076:DNA, Complementary; D018509:DNA, Helminth; D050556:Fatty Acid-Binding Proteins; D015801:Helminth Proteins; D008024:Ligands; D011994:Recombinant Proteins; D012177:Retinol-Binding Proteins
keywords: Developmental expression; Heterodera avenae; Hypodermis; Lipid-binding; RNA interference