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. 2018 Mar 13;9(3):162. doi: 10.3390/genes9030162

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© 2018 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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VdPLP gene disruption in V. dahliae and complementation of the ΔVdPLP strains (ΔVdPLP-1 and ΔVdPLP-4). (A) Disruption scheme used for VdPLP in the wild type (wt) strain (Vd-wt); gene disruption was confirmed by PCR. Genomic DNA (gDNA) from the Vd-wt, ΔVdPLP and ΔVdPLP-C strains were used as templates for amplification with primer pairs VdPLP-J-F/VdPLP-J-R, neo-J-F/neo-J-R and hyg-F/hyg-R; (B) A 1.3-kb fragment of VdPLP was amplified from Vd-wt with primer pair VdPLP-J-F/VdPLP-J-R; (C) A 2.6 kb fragment of neo was amplified with primer pair neo-J-F/neo-J-R from the ΔVdPLP mutant strains; (D) Gene complementation of the ΔVdPLP strains (ΔVdPLP-1-C and ΔVdPLP-4-C) was confirmed by amplification of a 1.6 kb fragment of hygromycin B resistance gene (hph) in PCR with primer pair hyg-F/hyg-R.