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Identification of a Novel Proline-Rich Antimicrobial Peptide from Brassica napus

Proline-rich antimicrobial peptides (PR-AMPs) are a group of cationic host defense peptides that are characterized by a high content of proline residues. Up to now, they have been reported in some insects, vertebrate and invertebrate animals, but are not found in plants. In this study, we performed...

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Autores principales: Cao, Huihui, Ke, Tao, Liu, Renhu, Yu, Jingyin, Dong, Caihua, Cheng, Mingxing, Huang, Junyan, Liu, Shengyi
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4575134/
https://www.ncbi.nlm.nih.gov/pubmed/26383098
http://dx.doi.org/10.1371/journal.pone.0137414
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author Cao, Huihui
Ke, Tao
Liu, Renhu
Yu, Jingyin
Dong, Caihua
Cheng, Mingxing
Huang, Junyan
Liu, Shengyi
author_facet Cao, Huihui
Ke, Tao
Liu, Renhu
Yu, Jingyin
Dong, Caihua
Cheng, Mingxing
Huang, Junyan
Liu, Shengyi
author_sort Cao, Huihui
collection PubMed
description Proline-rich antimicrobial peptides (PR-AMPs) are a group of cationic host defense peptides that are characterized by a high content of proline residues. Up to now, they have been reported in some insects, vertebrate and invertebrate animals, but are not found in plants. In this study, we performed an in silico screening of antimicrobial peptides, which led to discovery of a Brassica napus gene encoding a novel PR-AMP. This gene encodes a 35-amino acid peptide with 13 proline residues, designated BnPRP1. BnPRP1 has 40.5% identity with a known proline-rich antimicrobial peptide SP-B from the pig. BnPRP1 was artificially synthetized and cloned into the prokaryotic expression vector pET30a/His-EDDIE-GFP. Recombinant BnPRP1 was produced in Escherichia coli and has a predicted molecular mass of 3.8 kDa. Analysis of its activity demonstrated that BnPRP1 exhibited strong antimicrobial activity against Gram-positive bacterium, Gram-negative bacterium, yeast and also had strong antifungal activity against several pathogenic fungi, such as Sclerotinia sclerotiorum, Mucor sp., Magnaporthe oryzae and Botrytis cinerea. Circular dichroism (CD) revealed the main secondary structure of BnPRP1 was the random coil. BnPRP1 gene expression detected by qRT-PCR is responsive to pathogen inoculation. At 48 hours after S. sclerotiorum inoculation, the expression of BnPRP1 increased significantly in the susceptible lines while slight decrease occurred in resistant lines. These suggested that BnPRP1 might play a role in the plant defense response against S. sclerotiorum. BnPRP1 isolated from B. napus was the first PR-AMP member that was characterized in plants, and its homology sequences were found in some other Brassicaceae plants by the genome sequences analysis. Compared with the known PR-AMPs, BnPRP1 has the different primary sequences and antimicrobial activity. Above all, this study gives a chance to cast a new light on further understanding about the AMPs’ mechanism and application.
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spelling pubmed-45751342015-09-25 Identification of a Novel Proline-Rich Antimicrobial Peptide from Brassica napus Cao, Huihui Ke, Tao Liu, Renhu Yu, Jingyin Dong, Caihua Cheng, Mingxing Huang, Junyan Liu, Shengyi PLoS One Research Article Proline-rich antimicrobial peptides (PR-AMPs) are a group of cationic host defense peptides that are characterized by a high content of proline residues. Up to now, they have been reported in some insects, vertebrate and invertebrate animals, but are not found in plants. In this study, we performed an in silico screening of antimicrobial peptides, which led to discovery of a Brassica napus gene encoding a novel PR-AMP. This gene encodes a 35-amino acid peptide with 13 proline residues, designated BnPRP1. BnPRP1 has 40.5% identity with a known proline-rich antimicrobial peptide SP-B from the pig. BnPRP1 was artificially synthetized and cloned into the prokaryotic expression vector pET30a/His-EDDIE-GFP. Recombinant BnPRP1 was produced in Escherichia coli and has a predicted molecular mass of 3.8 kDa. Analysis of its activity demonstrated that BnPRP1 exhibited strong antimicrobial activity against Gram-positive bacterium, Gram-negative bacterium, yeast and also had strong antifungal activity against several pathogenic fungi, such as Sclerotinia sclerotiorum, Mucor sp., Magnaporthe oryzae and Botrytis cinerea. Circular dichroism (CD) revealed the main secondary structure of BnPRP1 was the random coil. BnPRP1 gene expression detected by qRT-PCR is responsive to pathogen inoculation. At 48 hours after S. sclerotiorum inoculation, the expression of BnPRP1 increased significantly in the susceptible lines while slight decrease occurred in resistant lines. These suggested that BnPRP1 might play a role in the plant defense response against S. sclerotiorum. BnPRP1 isolated from B. napus was the first PR-AMP member that was characterized in plants, and its homology sequences were found in some other Brassicaceae plants by the genome sequences analysis. Compared with the known PR-AMPs, BnPRP1 has the different primary sequences and antimicrobial activity. Above all, this study gives a chance to cast a new light on further understanding about the AMPs’ mechanism and application. Public Library of Science 2015-09-18 /pmc/articles/PMC4575134/ /pubmed/26383098 http://dx.doi.org/10.1371/journal.pone.0137414 Text en © 2015 Cao et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.
spellingShingle Research Article
Cao, Huihui
Ke, Tao
Liu, Renhu
Yu, Jingyin
Dong, Caihua
Cheng, Mingxing
Huang, Junyan
Liu, Shengyi
Identification of a Novel Proline-Rich Antimicrobial Peptide from Brassica napus
title Identification of a Novel Proline-Rich Antimicrobial Peptide from Brassica napus
title_full Identification of a Novel Proline-Rich Antimicrobial Peptide from Brassica napus
title_fullStr Identification of a Novel Proline-Rich Antimicrobial Peptide from Brassica napus
title_full_unstemmed Identification of a Novel Proline-Rich Antimicrobial Peptide from Brassica napus
title_short Identification of a Novel Proline-Rich Antimicrobial Peptide from Brassica napus
title_sort identification of a novel proline-rich antimicrobial peptide from brassica napus
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4575134/
https://www.ncbi.nlm.nih.gov/pubmed/26383098
http://dx.doi.org/10.1371/journal.pone.0137414
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