Targeting microbial pathogens by expression of new recombinant dermaseptin peptides in tobacco.
Alternaria
/ drug effects
Amphibian Proteins
/ genetics
Anti-Infective Agents
/ metabolism
Antimicrobial Cationic Peptides
/ genetics
Bacteria
/ drug effects
Fungal Proteins
/ genetics
Gene Expression
Microbial Sensitivity Tests
Plants, Genetically Modified
/ genetics
Recombinant Fusion Proteins
/ genetics
Nicotiana
/ genetics
Agrobacterium rhizogenes
antimicrobial peptide
chitin-binding domain
expression
hairy roots
Journal
MicrobiologyOpen
ISSN: 2045-8827
Titre abrégé: Microbiologyopen
Pays: England
ID NLM: 101588314
Informations de publication
Date de publication:
11 2019
11 2019
Historique:
received:
26
12
2018
revised:
28
02
2019
accepted:
28
02
2019
pubmed:
27
3
2019
medline:
20
5
2020
entrez:
27
3
2019
Statut:
ppublish
Résumé
Dermaseptin B1 (DrsB1), an antimicrobial cationic 31 amino acid peptide, is produced by Phyllomedusa bicolor. In an attempt to enhance the antimicrobial efficacy of DrsB1, the DrsB1 encoding 93 bp sequence was either fused to the N or C terminus of sequence encoding chitin-binding domain (CBD) of Avr4 gene from Cladosporium fulvum. Tobacco leaf disk explants were inoculated with Agrobacterium rhizogenes harboring pGSA/CBD-DrsB1 and pGSA/DrsB1-CBD expression vectors to produce hairy roots (HRs). Polymerase chain reaction (PCR) was employed to screen putative transgenic tobacco lines. Semi-quantitative RT-PCR and western blotting analysis indicated that the expression of recombinant genes were significantly higher, and recombinant proteins were produced in transgenic HRs. The recombinant proteins were extracted from the tobacco HRs and used against Pectobacterium carotovorum, Agrobacterium tumefaciens, Ralstonia solanacearum, and Xanthomonas campestris pathogenic bacteria and Alternaria alternata and Pythium sp. fungi. Two recombinant proteins had a statistically significant (p < 0.01) inhibitory effect on the growth and development of plant pathogens. The CBD-DrsB1 recombinant protein demonstrated a higher antibacterial effect, whereas the DrsB1-CBD recombinant protein demonstrated greater antifungal activity. Scanning electron microscopy images revealed that the structure of the fungal mycelia appeared segmented, adhered to each other, and crushed following the antimicrobial activity of the recombinant proteins. Due to the high antimicrobial activity of the recombinant proteins against plant pathogens, this strategy can be used to generate stable transgenic crop plants resistant to devastating plant pathogens.
Identifiants
pubmed: 30912302
doi: 10.1002/mbo3.837
pmc: PMC6854847
doi:
Substances chimiques
AVR4 protein, Cladosporium fulvum
0
Amphibian Proteins
0
Anti-Infective Agents
0
Antimicrobial Cationic Peptides
0
Fungal Proteins
0
Recombinant Fusion Proteins
0
dermaseptin b protein, Phyllomedusa
155483-06-0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
e837Informations de copyright
© 2019 The Authors. MicrobiologyOpen published by John Wiley & Sons Ltd.
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