Potato Root Diffusate-Induced Secretion of Soluble, Basic Proteins Originating from the Subventral Esophageal Glands of Potato Cyst Nematodes

Self Cite

243

229

Plants lack the seemingly unlimited receptor diversity of a somatic adaptive immune system as found in vertebrates and rely on only a relatively small set of innate immune receptors to resist a myriad of pathogens. Here, we show that disease-resistant tomato plants use an efficient mechanism to leverage the limited nonself recognition capacity of their innate immune system. We found that the extracellular plant immune receptor protein Cf-2 of the red currant tomato (Solanum pimpinellifolium) has acquired dual resistance specificity by sensing perturbations in a common virulence target of two independently evolved effectors of a fungus and a nematode. The Cf-2 protein, originally identified as a monospecific immune receptor for the leaf mold fungus Cladosporium fulvum, also mediates disease resistance to the root parasitic nematode Globodera rostochiensis pathotype Ro1-Mierenbos. The Cf-2-mediated dual resistance is triggered by effector-induced perturbations of the apoplastic Rcr3 pim protein of S. pimpinellifolium. Binding of the venom allergen-like effector protein Gr-VAP1 of G. rostochiensis to Rcr3 pim perturbs the active site of this papain-like cysteine protease. In the absence of the Cf-2 receptor, Rcr3 pim increases the susceptibility of tomato plants to G. rostochiensis, thus showing its role as a virulence target of these nematodes. Furthermore, both nematode infection and transient expression of Gr-VAP1 in tomato plants harboring Cf-2 and Rcr3 pim trigger a defense-related programmed cell death in plant cells. Our data demonstrate that monitoring host proteins targeted by multiple pathogens broadens the spectrum of disease resistances mediated by single plant immune receptors.parasitism | secretions | SCP/TAPS proteins | hypersensitive response

Section: Methodsmentioning

confidence: 99%

Dual disease resistance mediated by the immune receptor Cf-2 in tomato requires a common virulence target of a fungus and a nematode

Lozano‐Torres¹,

Wilbers²,

Gawroński³

et al. 2012

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229

“…12. MGR48 is a mouse monoclonal IgG1 that has been expressed in tobacco plants (41). A homozygous plant expressing this antibody was crossed bidirectionally with selected line xylGalT12.…”

Section: Methodsmentioning

confidence: 99%

An antibody produced in tobacco expressing a hybrid β-1,4-galactosyltransferase is essentially devoid of plant carbohydrate epitopes

Bakker

Rouwendal

Karnoup

et al. 2006

126

N-glycosylation of a mAb may have a major impact on its therapeutic merits. Here, we demonstrate that expression of a hybrid enzyme (called xylGalT), consisting of the N-terminal domain of Arabidopsis thaliana xylosyltransferase and the catalytic domain of human ␤-1,4-galactosyltransferase I (GalT), in tobacco causes a sharp reduction of N-glycans with potentially immunogenic corebound xylose (Xyl) and fucose (Fuc) residues as shown by Western blot and MALDI-TOF MS analysis. A radioallergosorbent test inhibition assay with proteins purified from leaves of WT and these transgenic tobacco plants using sera from allergic patients suggests a significant reduction of potential immunogenicity of xylGalT proteins. A mAb purified from leaves of plants expressing xylGalT displayed an N-glycan profile that featured high levels of galactose, undetectable xylose, and a trace of fucose. Hence, a transgenic plant expressing the hybrid GalT might yield more effective and safer monoclonals for therapeutic purposes than WT plants and even transgenic plants expressing the unchanged GalT.biopharmaceutical ͉ glycosylation ͉ immunogenicity

“…The effect of expression of GalT on a plantibody (Mgr-48) was investigated. Plantibodies were isolated by affinity chromatography on protein G from tobacco plants expressing Mgr-48 and from two selected plants (GalT-8 ϫ Mgr-48-11 and -12) resulting from a crossing between GalT plants and tobacco expressing Mgr-48 (28). Isolated Mgr-48 from hybridoma cells, tobacco, and the crossings of GalT-8 ϫ Mgr-48 were analyzed on blot with sheep-anti-mouse-IgG and RCA (Fig.…”

Section: Effects Of Human Galt On N-linked Glycan Structures In Tobaccomentioning

confidence: 99%

Galactose-extended glycans of antibodies produced by transgenic plants

Bakker

Bardor²,

Molthoff

et al. 2001

303

196

Plant-specific N-glycosylation can represent an important limitation for the use of recombinant glycoproteins of mammalian origin produced by transgenic plants. Comparison of plant and mammalian N-glycan biosynthesis indicates that ␤1,4-galactosyltransferase is the most important enzyme that is missing for conversion of typical plant N-glycans into mammalian-like N-glycans. Here, the stable expression of human ␤1,4-galactosyltransferase in tobacco plants is described. Proteins isolated from transgenic tobacco plants expressing the mammalian enzyme bear N-glycans, of which about 15% exhibit terminal ␤1,4-galactose residues in addition to the specific plant N-glycan epitopes. The results indicate that the human enzyme is fully functional and localizes correctly in the Golgi apparatus. Despite the fact that through the modified glycosylation machinery numerous proteins have acquired unusual N-glycans with terminal ␤1,4-galactose residues, no obvious changes in the physiology of the transgenic plants are observed, and the feature is inheritable. The crossing of a tobacco plant expressing human ␤1,4-galactosyltransferase with a plant expressing the heavy and light chains of a mouse antibody results in the expression of a plantibody that exhibits partially galactosylated N-glycans (30%), which is approximately as abundant as when the same antibody is produced by hybridoma cells. These results are a major step in the in planta engineering of the N-glycosylation of recombinant antibodies.