Anil Kumar
Nathalia Fitoussi
Payal Sanadhya
Natalia Sichov
Patricia Bucki
Menachem Borenstein
Eduard Belausov
Sigal Brown
During parasitism, root-knot nematodes Meloidogyne spp. inject molecules termed effectors, having multifunctional roles in construction and maintenance of nematodes' feeding sites. As an outcome of transcriptomic analysis of Meloidogyne javanica, we identified and characterized two differentially expressed genes encoding the predicted proteins MjShKT carrying a Stichodactyla toxin (ShKT) domain, and MjPUT3 carrying a ground-like domain, both expressed during nematode parasitism of tomato plant. Fluorescence in-situ hybridization revealed expression of MjShKT and MjPUT3 in the dorsal esophageal glands, suggesting their injection into host cells. MjShKT expression was upregulated during the parasitic life stages to a maximum at mature female stage, whereas MjPUT3 expression increased in third- to fourth-stage juveniles. Subcellular in-planta localization of MjShKT and MjPUT3 using a fused fluorescence marker indicated MjShKT co-occurrence with endoplasmic reticulum, the perinuclear endoplasmatic reticulum and the Golgi organelles markers, while MjPUT3 localized to some extent within the endoplasmatic reticulum and clearly observed within the nucleoplasm. MjShKT inhibited programmed cell death induced by overexpression of MAPKKKα and Gpa2/RBP-1. Overexpression of MjShKT in tomato hairy roots allowed an increase in nematode reproduction as indicated by the high number of eggs produced on roots overexpressing MjShKT. Roots overexpressing MjPUT3 were characterized by enhanced root growth, with no effect on nematode development on those roots. Investigation of the two candidate effectors suggested that MjShKT is mainly involved in manipulating the plant's effector-triggered immune response toward establishment and maintenance of active feeding sites, whereas MjPUT3 might modulate roots morphology in favor of nematode fitness in the host roots.
Anil Kumar
Nathalia Fitoussi
Payal Sanadhya
Natalia Sichov
Patricia Bucki
Menachem Borenstein
Eduard Belausov
Sigal Brown
During parasitism, root-knot nematodes Meloidogyne spp. inject molecules termed effectors, having multifunctional roles in construction and maintenance of nematodes' feeding sites. As an outcome of transcriptomic analysis of Meloidogyne javanica, we identified and characterized two differentially expressed genes encoding the predicted proteins MjShKT carrying a Stichodactyla toxin (ShKT) domain, and MjPUT3 carrying a ground-like domain, both expressed during nematode parasitism of tomato plant. Fluorescence in-situ hybridization revealed expression of MjShKT and MjPUT3 in the dorsal esophageal glands, suggesting their injection into host cells. MjShKT expression was upregulated during the parasitic life stages to a maximum at mature female stage, whereas MjPUT3 expression increased in third- to fourth-stage juveniles. Subcellular in-planta localization of MjShKT and MjPUT3 using a fused fluorescence marker indicated MjShKT co-occurrence with endoplasmic reticulum, the perinuclear endoplasmatic reticulum and the Golgi organelles markers, while MjPUT3 localized to some extent within the endoplasmatic reticulum and clearly observed within the nucleoplasm. MjShKT inhibited programmed cell death induced by overexpression of MAPKKKα and Gpa2/RBP-1. Overexpression of MjShKT in tomato hairy roots allowed an increase in nematode reproduction as indicated by the high number of eggs produced on roots overexpressing MjShKT. Roots overexpressing MjPUT3 were characterized by enhanced root growth, with no effect on nematode development on those roots. Investigation of the two candidate effectors suggested that MjShKT is mainly involved in manipulating the plant's effector-triggered immune response toward establishment and maintenance of active feeding sites, whereas MjPUT3 might modulate roots morphology in favor of nematode fitness in the host roots.