Abstract

BackgroundAlthough many genetic manipulations of crops providing biofortified or safer food have been done, the acceptance of biotechnology crops still remains limited. We report on a transgenic barley expressing the multi-functional protein osmotin that improves plant defense under stress conditions.MethodsAn Agrobacterium–mediated technique was used to transform immature embryos of the spring barley cultivar Golden Promise. Transgenic barley plants of the T0 and T1 generation were evaluated by molecular methods. Transgenic barley tolerance to stress was determined by chlorophyll, total protein, malondialdehyde and ascorbate peroxidase content. Methanol extracts of i) Fusarium oxysporum infected or ii) salt-stressed plants, were characterized by their acute toxicity effect on human dermal fibroblasts (HDF), genotoxicity and affection of biodiversity interactions, which was tested through monitoring barley natural virus pathogen–host interactions–the BYDV and WDV viruses transmitted to the plants by aphids and leafhoppers.ResultsTransgenic plants maintained the same level of chlorophyll and protein, which significantly declined in wild-type barley under the same stressful conditions. Salt stress evoked higher ascorbate peroxidase level and correspondingly less malondialdehyde. Osmotin expressing barley extracts exhibited a lower cytotoxicity effect of statistical significance than that of wild-type plants under both types of stress tested on human dermal fibroblasts. Extract of Fusarium oxysporum infected transgenic barley was not able to damage DNA in the Comet assay, which is in opposite to control plants. Moreover, this particular barley did not affect the local biodiversity.ConclusionOur findings provide a new perspective that could help to evaluate the safety of products from genetically modified crops.

Highlights

  • Osmotin is a 26-kDa protein belonging to the 5th class of pathogenesis-related proteins (PR 5) together with thaumatin, zeamatin and others

  • The possibility of tobacco osmotin gene expression under the constitutive promoter of the cauliflower mosaic virus has been already by our group demonstrated in barley previously [29]

  • Transgenic barley plants expressing the osmotin gene were prepared via Agrobacterium-mediated transformation

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Summary

Introduction

Osmotin is a 26-kDa protein belonging to the 5th class of pathogenesis-related proteins (PR 5) together with thaumatin, zeamatin and others. Osmotin was first characterized in Nicotiana tabacum plants adapted to saline conditions [1, 2]. Its homologs were found in both monocots and dicots across the whole plant kingdom [3]. Osmotin is a multifunctional protein that plays an important role in the plant immune system during stress. Abiotic originators of stress such as drought, salt and cold induce osmotin expression. Osmotin plays a role as an osmoprotectant, providing enzyme protection and protein chaperone functions. Biotic stress resistance is related to the ability of osmotin to activate the fungal receptor that induces the programmed cell death of fungi. It has served as a tool for biotechnology engineering for many decades [1]

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