Psc-AFP from Psoralea corylifolia L. overexpressed in Pichia pastoris increases antimicrobial activity and enhances disease resistance of transgenic tobacco

Autor: Jie Cai, Chengjian Xie, De Wang, An Ping Sui, Yun-Min Wei, Shan-Shan Cheng, Xing Yong Yang, Xiu-Mei Luo
Rok vydání: 2016
Předmět:
0106 biological sciences
0301 basic medicine
Antifungal Agents
Psoralea corylifolia
Trypsin inhibitor
Biology
01 natural sciences
Applied Microbiology and Biotechnology
Alternaria alternata
Pichia
Psoralea
Rhizoctonia
Pichia pastoris
Microbiology
03 medical and health sciences
chemistry.chemical_compound
Sequence Analysis
Protein
Tobacco
Amino Acid Sequence
Cloning
Molecular
Disease Resistance
Plant Diseases
Plant Proteins
Ralstonia solanacearum
fungi
digestive
oral
and skin physiology
Computational Biology
food and beverages
General Medicine
Plants
Genetically Modified
biology.organism_classification
Recombinant Proteins
digestive system diseases
Alternaria brassicae
030104 developmental biology
chemistry
Aspergillus niger
Heterologous expression
Growth inhibition
Trypsin Inhibitors
010606 plant biology & botany
Biotechnology
Zdroj: Applied Microbiology and Biotechnology. 101:1073-1084
ISSN: 1432-0614
0175-7598
DOI: 10.1007/s00253-016-7768-1
Popis: 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.
Databáze: OpenAIRE
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