title: Molecular Cloning of HbPR-1 Gene from Rubber Tree, Expression of HbPR-1 Gene in Nicotiana benthamiana and Its Inhibition of Phytophthora palmivora.
abstract: This is the first report to present a full-length cDNA (designated HbPR-1) encoding a putative basic HbPR-1 protein from rubber tree (Hevea brasiliensis) treated with salicylic acid. It was characterized and also expressed in Nicotiana benthamiana using Agrobacterium-mediated transient gene expression system in order to investigate the role of HbPR-1 gene in rubber tree against its oomycete pathogen Phytopthora palmivora and to produce recombinant HbPR-1 protein for microbial inhibition test. The HbPR-1 cDNA was 647 bp long and contained an open reading frame of 492 nucleotides encoding 163 amino acid residues with a predicted molecular mass of 17,681 Da and an isoelectric point (pI) of 8.56, demonstrating that HbPR-1 protein belongs to the basic PR-1 type. The predicted 3D structure of HbPR-1 was composed of four α-helices, three β-sheets, seven strands, and one junction loop. Expression and purification of recombinant HbPR-1 protein were successful using Agrobacterium-mediated transient expression and one-step of affinity chromatography. Heterologous expression of HbPR-1 in N. benthamiana reduced necrosis areas which were inoculated with P. palmivora zoospores, indicating that the expressed HbPR-1 protein played an important role in plant resistance to pathogens. The purified recombinant HbPR-1 protein was found to inhibit 64% of P. palmivora zoospore germination on a water agar plate compared with control, suggesting that it was an antimicrobial protein against P. palmivora.
authors.affiliation: Department of Biochemistry, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, Thailand.|Department of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Manoa, HI, United States of America.
authors.forename: Uraiwan
authors.initials: U
authors.lastname: Khunjan
authors.affiliation: Department of Biochemistry, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, Thailand.|Department of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Manoa, HI, United States of America.
authors.forename: Kitiya
authors.initials: K
authors.lastname: Ekchaweng
authors.affiliation: Digital Media Program, Prince of Songkla University International College, Prince of Songkla University, Hat Yai, Songkhla, Thailand.|Center for Genomics and Bioinformatics Research, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, Thailand.
authors.forename: Tanate
authors.initials: T
authors.lastname: Panrat
authors.affiliation: Department of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Manoa, HI, United States of America.
authors.forename: Miaoying
authors.initials: M
authors.lastname: Tian
authors.affiliation: Department of Biochemistry, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, Thailand.
authors.forename: Nunta
authors.initials: N
authors.lastname: Churngchow
mesh_terms: D000977:Antiparasitic Agents / Q000494:pharmacology; D001665:Binding Sites; D003001:Cloning, Molecular; D017343:Genes, Plant; D028482:Hevea / Q000235:genetics* / Q000469:parasitology; D015394:Molecular Structure; D010838:Phytophthora / Q000187:drug effects*; D010936:Plant Extracts / Q000494:pharmacology*; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics* / Q000494:pharmacology; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology; D014026:Nicotiana / Q000235:genetics
chemical_list: D000977:Antiparasitic Agents; D010936:Plant Extracts; D010940:Plant Proteins; D011994:Recombinant Proteins