Lectin-like glycoprotein PsNLEC-1 is not correctly glycosylated and targeted in boron-deficient pea nodules.
Symbiosome development was studied in pea root nodules from plants growing in the absence of boron (B). Rhizobia released into the host cells of nodules from B-deficient plants developed to abnormal endophytic forms with an altered electrophoretic lipopolysaccharide pattern. Immunostaining after sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electroblotting of nodule homogenates with antibodies that recognize glycoprotein components showed that two previously described lectin-like glycoproteins (PsNLEC-1A and PsNLEC-1B) did not harbor the carbohydrate epitope normally recognized by specific monoclonal antibodies. Material derived from B-deficient nodules, however, still contained three antigenic isoforms with similar electrophoretic mobilities to PsNLEC-1 isoforms A, B, and C. These could be detected following immunoblotting and immunostaining with a specific antiserum originating from the purified PsNLEC protein that had been heterologously expressed in Escherichia coli. Immunogold localization of PsNLEC-1 sugar epitopes in B-deficient nodules showed that they were associated mostly with cytoplasmic vesicles rather than normal localization in the symbiosome compartment of mature infected cells. These results suggest that a modification of the glycosyl-moieties of PsNLEC-1 and an alteration of vesicle targeting occur during the development of pea nodules in the absence of B, and that these changes are associated with the development of aberrant nonfunctional symbiosomes.
Bolaños
L
L
Departmento de Biología, Facultad de Ciencias, Universidad Autónoma de Madrid, Spain.
Cebrián
A
A
Redondo-Nieto
M
M
Rivilla
R
R
Bonilla
I
I
D000818:Animals; D000911:Antibodies, Monoclonal; D001895:Boron / Q000172:deficiency*; D006023:Glycoproteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D006031:Glycosylation; D015151:Immunoblotting; D008854:Microscopy, Electron; D018532:Pisum sativum / Q000382:microbiology / Q000502:physiology*; D010940:Plant Proteins / Q000032:analysis / Q000737:chemistry / Q000378:metabolism*; D018517:Plant Roots / Q000382:microbiology / Q000502:physiology / Q000648:ultrastructure; D051381:Rats; D016961:Rhizobium leguminosarum / Q000302:isolation & purification / Q000472:pathogenicity / Q000502:physiology*
D000911:Antibodies, Monoclonal; D006023:Glycoproteins; D010940:Plant Proteins; C099998:PsNlec1 protein, Pisum sativum; D001895:Boron