Dual roles for the variable domain in protein trafficking and host-specific recognition of Heterodera glycines CLE effector proteins

Authors: WANG, JIANJING - University Of Missouri; LEE, CHRIS - University Of Missouri; REPLOGLE, AMY - University Of Missouri; JOSHI, SNEHA - University Of Missouri; KORKIN, DMITRY - University Of Missouri; HUSSEY, RICHARD - University Of Georgia; BAUM, THOMAS - Iowa State University; DAVIS, ERIC - North Carolina State University; Wang, Xiaohong; MITCHUM, MELISSA - University Of Missouri

Submitted to: New Phytologist
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/12/2010
Publication Date: 9/1/2010
Citation: Wang, J., Lee, C., Replogle, A., Joshi, S., Korkin, D., Hussey, R., Baum, T., Davis, E., Wang, X., Mitchum, M.G. 2010. Dual roles for the variable domain in protein trafficking and host-specific recognition of Heterodera glycines CLE effector proteins. New Phytologist. 187:1003-1017.

Interpretive Summary: The soybean cyst nematode (SCN, Heterodera glycines) is a serious threat to the U.S. soybean production. In order to successfully parasitize host roots, the nematode secretes effector proteins originated from its esophageal gland cells to transform host plant cells into a specialized site for the nematode to feed. Our previous studies showed that SCN secretes CLE proteins that function as endogenous plant CLE peptides to manipulate plant developmental pathways for the formation of nematode feeding sites. However, the site of action and mechanism of delivery of nematode CLE proteins to host plant cells has not been elucidated. In this paper, we used several approaches to reveal the localization and site of action of SCN secreted CLE proteins in infected roots. Our study revealed for the first time that the variable domain of nematode CLE proteins has an important role in host-specific recognition and in plant trafficking of nematode CLEs. A further study of this nematode CLE-mediated parasitism will likely lead to the development of novel nematode control strategies.

Technical Abstract: Soybean cyst nematodes (Heterodera glycines) produce secreted effector proteins that function as peptide mimics of plant CLAVATA3 / ESR (CLE)-like peptides probably involved in the developmental reprogramming of root cells to form specialized feeding cells called syncytia. The site of action and mechanism of delivery of CLE effectors to host plant cells by the nematode, however, have not been established. In this study, immunologic, genetic and biochemical approaches were used to reveal the localization and site of action of H. glycines-secreted CLE proteins in planta. We present evidence indicating that the nematode CLE propeptides are delivered to the cytoplasm of syncytial cells, but ultimately function in the apoplast, consistent with their proposed role as ligand mimics of plant CLE peptides. We determined that the nematode 12-amino-acid CLE motif peptide is not sufficient for biological activity in vivo, pointing to an important role for sequences upstream of the CLE motif in function. Genetic and biochemical analysis confirmed the requirement of the variable domain in planta for host-specific recognition and revealed a novel role in trafficking cytoplasmically delivered CLEs to the apoplast in order to function as ligand mimics.