title: Psc-AFP from Psoralea corylifolia L. overexpressed in Pichia pastoris increases antimicrobial activity and enhances disease resistance of transgenic tobacco.
abstract: Psc-AFP, isolated from the seeds of Psoralea corylifolia L., is an antimicrobial protein with trypsin inhibitor activity. Its encoding gene was cloned by 3'- rapid amplification of cDNA ends (RACE) combined with Y-shaped adaptor-dependent extension (YADE) method. The gene Psc-AFP encodes a protein of 203 amino acids with a deduced signal peptide of 24 residues. The growth inhibition effect exerted by the heterologously expressed Psc-AFP in Pichia pastoris revealed that the recombinant Psc-AFP inhibited mycelium growth of Aspergillus niger, Rhizoctonia solani, and Alternaria brassicae and conidial germination of Alternaria alternata. The recombinant Psc-AFP also showed protease inhibitor activity manifested by the inhibition of trypsin. The transgenic tobacco bioassays confirmed that overexpressing Psc-AFP significantly enhanced the disease resistance of tobacco and that some of the transgenic lines were almost fully tolerant to Ralstonia solanacearum and A. alternata, whereas no apparent alteration in plant growth and development was observed. Collectively, these results indicate that the recombinant Psc-AFP is an active antimicrobial protein, with protease inhibitor activity that can be successfully produced in the yeast and tobacco and, therefore, maybe a potential antimicrobial candidate for practical use.
authors: The School of Life Science, Southwest University, Chongqing, 400715, China.|The Chongqing Key Laboratory of Molecular Biology of Plant Environmental Adaptations, Chongqing Normal University, Chongqing, 401331, China.|The College of Life Science, Chongqing Normal University, Chongqing, 401331, China.
authors: Xiu-Mei
authors: XM
authors: Luo
authors: The Chongqing Key Laboratory of Molecular Biology of Plant Environmental Adaptations, Chongqing Normal University, Chongqing, 401331, China.|The College of Life Science, Chongqing Normal University, Chongqing, 401331, China.
authors: Cheng-Jian
authors: CJ
authors: Xie
authors: The School of Life Science, Southwest University, Chongqing, 400715, China.
authors: De
authors: Wang
authors: The Chongqing Key Laboratory of Molecular Biology of Plant Environmental Adaptations, Chongqing Normal University, Chongqing, 401331, China.|The College of Life Science, Chongqing Normal University, Chongqing, 401331, China.
authors: Yun-Min
authors: YM
authors: Wei
authors: The Chongqing Key Laboratory of Molecular Biology of Plant Environmental Adaptations, Chongqing Normal University, Chongqing, 401331, China.|The College of Life Science, Chongqing Normal University, Chongqing, 401331, China.
authors: Jie
authors: J
authors: Cai
authors: The Chongqing Key Laboratory of Molecular Biology of Plant Environmental Adaptations, Chongqing Normal University, Chongqing, 401331, China.|The College of Life Science, Chongqing Normal University, Chongqing, 401331, China.
authors: Shan-Shan
authors: SS
authors: Cheng
authors: The Chongqing Key Laboratory of Molecular Biology of Plant Environmental Adaptations, Chongqing Normal University, Chongqing, 401331, China. suianp@swu.edu.cn.|The College of Life Science, Chongqing Normal University, Chongqing, 401331, China. suianp@swu.edu.cn.
authors: Xing -Yong
authors: X-
authors: Yang
authors: The School of Life Science, Southwest University, Chongqing, 400715, China.
authors: An -Ping
authors: A-
authors: Sui
mesh_terms: D000595:Amino Acid Sequence; D000935:Antifungal Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D001234:Aspergillus niger / Q000187:drug effects; D003001:Cloning, Molecular; D019295:Computational Biology; D060467:Disease Resistance; D010843:Pichia / Q000235:genetics*; D010935:Plant Diseases / Q000517:prevention & control*; D010940:Plant Proteins / Q000235:genetics* / Q000494:pharmacology*; D030821:Plants, Genetically Modified / Q000235:genetics / Q000382:microbiology; D029907:Psoralea / Q000033:anatomy & histology / Q000737:chemistry / Q000235:genetics*; D011994:Recombinant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology; D012232:Rhizoctonia / Q000187:drug effects; D020539:Sequence Analysis, Protein; D014026:Nicotiana / Q000235:genetics / Q000254:growth & development / Q000382:microbiology*; D014361:Trypsin Inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology
chemical_list: D000935:Antifungal Agents; D010940:Plant Proteins; D011994:Recombinant Proteins; D014361:Trypsin Inhibitors
keywords: Antimicrobial protein; Heterologous expression; Pichia pastoris; Psoralea corylifolia; Tobacco; Trypsin inhibitor