Novel expression system for enhanced synthesis of antimicrobial peptide cecropin P1 in plants

Abstract

A gene of antimicrobial peptide cecropin P1 (CP1) was inserted into the vector plasmid pPCV91 under the control of promoter 35S RNA of cauliflower mosaic virus (CaMV 35S) containing four enhancer sequences CaMV 35S and nontranslated leader sequence Ω RNA of tobacco mosaic virus. The recombinant vector obtained was used for agrobacterial transformation of tobacco plants (Nicotiana tabacum L., variety Sam sun) with the polymerase chain reaction (PCR) based method. The presence of gene CP1 in the genome of plants was proven by western blot analysis and testing the antibiotic activity of plant extracts. In different plant lines, the level of cecropin P1 synthesis amounted 0.02-0.2% of total soluble plant leaf protein. The transgenic plants, unlike the control ones, displayed enhanced tolerance to phytopathogenic microorganisms and oxidative stress. It was established that the ability of the transgenic plants to express cecropin P1 is transmitted to progeny.Abbreviations: SOD-superoxide dismutase; Cf-cefatoxime; CaMV 35S-promoter 35S RNA cauliflower mosaic virus (CaMV); CP1-cecropin P1; Hm-hygromycin; hpt-hygro mycin phosphotransferase gene; pnos-promoter of agrobacterial nopalin synthase.

The Brassica napus L. plants expressing antimicrobial peptide cecropin P1 are safe for colonization by beneficial associative microorganisms

“…It was proven by the activity of SOD, along with decreased content of superoxide radical and peroxide in oxidative stress conditions. Increased antioxidant resistance of cec-P1 plants is observed not only in experiments in the effect of hers-P1 plants with herbicide paraquat, but also in the infection with phytopatogenes (Zakharchenko et al 2015(Zakharchenko et al , 2019. Also, a substantial decrease in the chlorophyll content was found in non-transgenic but not in transgenic plant leaves.…”