Purification of a Zn-Binding Phloem Protein with Sequence Identity to Chitin-Binding Proteins

Abstract

In citrus blight, a decline disorder of unknown etiology, the tree canopy exhibits symptoms of Zn deficiency while Zn accumulates in the trunk phloem. We have purified a Zn-binding protein (ZBP) from phloem tissue of healthy and blight-affected citrus (Citrus sinensis [L.] Osbeck on Citrus jambhiri [L.]). The molecular weight of the ZBP was estimated to be 5000 by size-exclusion chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Ion-exchange chromatography at pH 8.0 demonstrated the 5-kD ZBP to be anionic. A partial N-terminal amino acid sequence revealed a cysteine-, glycine-rich domain with 45 to 80% identity with the chitin-binding domain of hevein, wheat germ agglutinin, and several class I chitinases. That the abundance of this protein increased 2.5-fold in association with Zn accumulation in the phloem is characteristic of citrus blight. Tissue mass changes of the phloem suggests that altered tissue structure accompanies blight. Phloem accumulation of the 5-kD ZBP may be in response to wounding or other stress of blight-affected citrus.