Figure 4

Subcellular localisations of Globodera pallida effector fusions with the enhanced green fluorescent protein (eGFP) in leaves of Nicotiana benthamiana . (A) Localisation of free eGFP. (B) Localisation of GPLIN_000015300 (ortholog of Heterodera glycines G7E05 sequence) in the cytoplasm and nucleoplasm but excluded from the nucleolus. (C) Localisation of GpA42 _ (ortholog of Globodera rostochiensis A42 sequence) in the nucleoli. (D) Faint expression of GPLIN_000235400 (ortholog of G. rostochiensis 1106 sequence) in the cytoplasm with accumulation in the nucleus, particularly in the nucleoli. (E & F) Exclusion of SPRYSEC effector GPLIN_001465500 from the cytoplasm but accumulation in the nucleoplasm, particularly surrounding the nucleolus. (G-H) Expression of GPLIN_000662500 (ortholog of H. glycines G20E03 sequence) in peroxisomes. (I-J) Expression of GPLIN_000457000 (ortholog of H. glycines hgsec4 sequence) in the peroxisome membrane (arrows). In all images the eGFP-effector fusion is seen in green, with monomeric-red fluorescent protein (mRFP) co-label in magenta, from either transgenic expression of an mRFP-histone fusion (A-F) or co-expression of an mRFP-tagged peroxisome marker (G-J), and autofluorescent chloroplasts displayed in blue. Scale bars represent 50 μm for A, D, 20 μm for C, 10 μm for G, H, and 5 μm for E, F, I and J. Localisation of Agrobacterium-mediated transient expression of eGFP-effector fusions was observed 48 h post inoculation by confocal microscopy.