Enhancing disease resistance against microbial pathogens by expression of Et-AMPs in Arabidopsis plants

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AmranAmmar_2010_10_08.pdf (10.91 MB)

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2010

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http://dx.doi.org/10.22029/jlupub-17029

Abstract

The aim of the study was to detect the potency of the novel insect antimicrobial peptidesEt-AMP1 and Et-AMP2 derived from drone fly Eristalis tenax to engineer diseaseresistance in the model plant Arabidopsis thaliana against the fungal pathogen grey moldBotrytis cinerea and the bacterial pathogen Pseudomonas syringae pv. tomato strainDC3000. For the first antimicrobial peptide (Et-AMP1), a protocol for the production ofrecombinant protein in E. coli expression system was established by inserting the sequencefor mature Et-AMP1 peptide in frame downstream of the pEXp5-CT/Topo expressionvector. The recombinant protein was evaluated in vitro as to its effects on the devastatingphytopathogen Fusarium culmorum by using spore germination inhibition assays. Resultsof this assay exhibited a weak effect with an inhibition percentage about 10.5% against thisplant pathogenic fungus with the maximum concentration of produced recombinant protein(0.9 μM).Both antimicrobial peptides (Et-AMP1 and Et-AMP2) were introduced into Arabidopsisplants via Agrobacterium-mediated transformation. After the production of transgenicArabidopsis plants, in vitro antifungal assays were carried out by using T3 plants forevaluating their resistance against the fungal pathogens B. cinerea and the bacterialpathogen P. syringae. Our results show that Et-AMP1 transgenic plants did not exhibitresistance against B. cinerea. In contrast, these plants were more resistant to P. syringae,especially lines 5, 3, 7 and 2. On the other hand, the Et-AMP2 transgenic plants wereresistant both to B. cinerea and P. syringae. Lines 1, 5 and 4 reduced significantly thenecrotic lesion size caused by B. cinerea pathogen. Lines 1, 2 and 5 were more resistant toP. syringae infection. Subsequently, further research is required to identify the potential ofEt-AMP1 and Et-AMP2 for plant protection approaches.

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Anthology

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