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Published on April 29, 2009; 10.1104/pp.109.138669

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Received March 13, 2009
Accepted April 20, 2009

Isolation and Characterization of Hydroxyproline-rich Glycopeptide Signals in Black Nightshade Leaves

Gregory Pearce *, Ramcharan Bhattacharya , Yu-Chi Chen , Guido Barona , Yube Yamaguchi , and Clarence A. Ryan

Institute of Biological Chemistry, Washington State University, Pullman, Washington 99164-6340

* Corresponding author; email: pearce{at}wsu.edu.

A gene encoding a preprohydroxyproline-rich systemin, SnpreproHypSys, was identified from the leaves of black nightshade, Solanum nigrum (Sn), which is a member of a small gene family of at least three genes that have orthologs in tobacco (NtpreproHypSys), tomato (SlpreproHypSys), petunia (PhpreproHypSys), potato (PhpreproHypSys), and sweet potato (IbpreproHypSys). SnpreproHypSys was induced by wounding and by treatment with methyl jasmonate (MeJA). The encoded precursor protein contained a signal sequence and was post-translationally modified to produce three hydroxyproline-rich glycopeptide signals (HypSys peptides). The three HypSys peptides isolated from nightshade leaf extracts were called SnHypSys I (19 amino acids with 6 pentoses), SnHypSys II (20 amino acids with 6 pentoses), and SnHypSys III (20 amino acids with either 6 or 9 pentoses) by their sequential appearance in SnpreproHypSys. The three SnHypSys peptides were synthesized and tested for their abilities to alkalinize suspension culture media, with synthetic SnHypSys I demonstrating the highest activity. Synthetic SnHypSys I was capable of inducing alkalinization in other Solanaceae cell types (or species), indicating that structural conformations within the peptides are recognized by the different cells/species to initiate signal transduction pathways, apparently through recognition by homologous receptor(s). To further demonstrate the biological relevance of the SnHypSys peptides, the early defense gene lipoxygenase D was shown to be induced by all 3 synthetic peptides when supplied to excised nightshade plants.






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