Antimicrobial Peptides from Plants: A cDNA-Library Based Isolation, Purification, Characterization Approach and Elucidating Their Modes of Action

Abstract

Even in a natural ecosystem, plants are continuously threatened by various microbial diseases. To save themselves from these diverse infections, plants build a robust, multilayered immune system through their natural chemical compounds. Among the several crucial bioactive compounds possessed by plants' immune systems, antimicrobial peptides (AMPs) rank in the first tier. These AMPs are environmentally friendly, anti-pathogenic, and do not bring harm to humans. Antimicrobial peptides can be isolated in several ways, but recombinant protein production has become increasingly popular in recent years, with the Escherichia coli expression system being the most widely used. However, the efficacy of this expression system is compromised due to the difficulty of removing endotoxin from its system. Therefore, this review suggests a high-throughput cDNA library-based plant-derived AMP isolation technique using the Bacillus subtilis expression system. This method can be performed for large-scale screening of plant sources to classify unique or homologous AMPs for the agronomic and applied field of plant studies. Furthermore, this review also focuses on the efficacy of plant AMPs, which are dependent on their numerous modes of action and exceptional structural stability to function against a wide range of invaders. To conclude, the findings from this study will be useful in investigating how novel AMPs are distributed among plants and provide detailed guidelines for an effective screening strategy of AMPs.

Isolation of Potato Endophytes and Screening of Chaetomium globosum Antimicrobial Genes

“…However, the accumulation of exogenous gene expression and secretion products to a certain extent may have toxic effects on the host cells, leading to the rupture of the host cells, and finally the phenotype of the recombinant strains will change significantly, resulting in autolysis [55,56]. Using the B. subtilis expression system, we have developed a novel, high-throughput technique for the isolation of novel AMPs [20][21][22][23]. In this study, we screened 10 candidate genes from 4280 clones of C. globosum cDNA library, and finally we selected two antimicrobial peptide genes CgR2150 and CgR3101 with strong antifungal and antibacterial activities for further study.…”

“…Bacillus subtilis has been one of the most extensively utilized prokaryotic expression systems in recent years due to its rapid growth, absence of endotoxin, and minimal damage to organism [18]. Moreover, B. subtilis possesses a robust secretion and expression capability, which enables it to directly secrete recombinant proteins into the extracellular space, thereby contributing significantly to the large-scale development of antibacterial genes [19][20][21]. Using B. subtilis as a bioengineering strain to generate functional protein products has the potential to reduce costs and add economic values [22,23].…”

Nanoparticles in association with antimicrobial peptides (NanoAMPs) as a promising combination for agriculture development

“…AMPs present several beneficial characteristics, including activity against several phytopathogens (fungi, bacteria, virus) and insects ( Mulinari et al, 2007 ; Pinto et al, 2012 ); the capacity to generate direct and durable plant resistance; and small gene nature that facilitates stacking the coding sequence of multiple AMPs on single expression vectors ( Islam et al, 2021 ). Besides, AMPs natural or synthetic also present ease of manipulation and optimization by computational approaches ( in silico design).…”

Isolation and Optimization of a Broad-Spectrum Synthetic Antimicrobial Peptide, Ap920-WI, from Arthrobacter sp. H5 for the Biological Control of Plant Diseases

“…NAMPs are small-molecule polypeptides encoded by genes that are present in organisms [ 7 , 8 ] with broad-spectrum antimicrobial activity [ 9 ]. However, directly isolating NAMPs from natural sources can be expensive, time-consuming, and environmentally unfavorable, especially when peptides are discovered in rare animals with small populations [ 10 ]. Alternatively, SAMPs are produced by cloning foreign genes onto specialized vectors and expressing them in host cells using recombinant DNA technologies, such as the bacterial expression system, which is the most efficient method in recent times [ 10 , 11 ].…”