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Sugar-Binding Activity of Pea Lectin Expressed in White Clover Hairy Roots
C. L. Diaz1, TJJ. Logman, H. C. Stam
1Center for Phytotechnology Rijksuniversiteit Leiden-The Netherlands Organization for Applied Scientific Research TNO (C.L.D., J.W.K.), and Institute of Molecular Plant Sciences (T.J.J.L., H.C.S., J.W.K.), Wassenaarseweg 64, 2333 AL Leiden, The Netherlands.
Plant Physiology
|December 1, 1995
Summary
Transgenic white clover expressing the pea lectin (PSL) gene shows functional PSL on root surfaces, enabling rhizobial infection. This demonstrates correct protein processing and targeting for symbiotic interactions.
Area of Science:
- Plant molecular biology
- Microbial symbiosis
- Legume genetics
Background:
- Pea lectin (PSL) gene introduction into white clover facilitates nodulation by Rhizobium leguminosarum.
- PSL normally facilitates nodulation in pea, not white clover.
Purpose of the Study:
- To confirm functional expression and localization of PSL in transgenic white clover hairy roots.
- To investigate PSL's role in rhizobial infection of white clover.
Main Methods:
- Genetic transformation of white clover with the PSL gene.
- Affinity chromatography for PSL isolation.
- Immunoanalysis to detect PSL subunits.
- Glycan-based localization technique using rat IgE.
Main Results:
- PSL was functionally expressed and isolated from transgenic white clover hairy roots.
- Immunoanalysis confirmed the presence of PSL precursor and beta subunits.
- PSL localized to the external cell surface of epidermal cells and root hair tips, similar to pea roots.
- PSL was absent in normal white clover roots and non-transgenic hairy roots.
Conclusions:
- Transgenic white clover hairy roots correctly process and target PSL to infection-susceptible root cells.
- PSL is surface-bound in white clover roots, with available binding sites for rhizobial ligands, mimicking its function in pea roots.