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. 2020 Jan 14;10:3004. doi: 10.3389/fmicb.2019.03004

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Copyright © 2020 Santos, Cho, Wu, Chang, Kuo and Lai.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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The four VgrG proteins of 1D1609 are functionally redundant for Hcp secretion. Western blot analysis of cellular and secreted (S) proteins from single vgrG deletion mutants (A), multiple vgrG deletion mutants (B), overexpression of VgrG in 1D1609 quadruple vgrG mutant (Δ4) (C), and in C58 double vgrG deletion mutant (ΔG1,2) (D), and triple vgrG deletion mutants (E). Various A. tumefaciens strains were cultured in LB (A,B,E) or AK (C,E) broth at 25°C and cellular and secreted (S) fractions were subjected to western blotting using anti-Hcp, anti-VgrG, and RpoA as indicated. The T6SS inactive mutant ΔtssL is a negative control and RpoA serves as a non-secreted protein control. Protein markers (in kDa) are indicated on the left. Similar results were obtained from 2–4 independent experiments.