Abstract

Subtilisin-like proteases (SBTs) constitute a large family of extracellular plant proteases, the function of which is still largely unknown. In tomato plants, the expression of SBT3 was found to be induced in response to wounding and insect attack in injured leaves but not in healthy systemic tissues. The time course of SBT3 induction resembled that of proteinase inhibitor II and other late wound response genes suggesting a role for SBT3 in herbivore defense. Consistent with such a role, larvae of the specialist herbivore Manduca sexta performed better on transgenic plants silenced for SBT3 expression (SBT3-SI). Supporting a contribution of SBT3 to systemic wound signaling, systemic induction of late wound response genes was attenuated in SBT3-SI plants. The partial loss of insect resistance may thus be explained by a reduction in systemic defense gene expression. Alternatively, SBT3 may play a post-ingestive role in plant defense. Similar to other anti-nutritive proteins, SBT3 was found to be stable and active in the insect's digestive system, where it may act on unidentified proteins of insect or plant origin. Finally, a reduction in the level of pectin methylesterification that was observed in transgenic plants with altered levels of SBT3 expression suggested an involvement of SBT3 in the regulation of pectin methylesterases (PMEs). While such a role has been described in other systems, PME activity and the degree of pectin methylesterification did not correlate with the level of insect resistance in SBT3-SI and SBT3 overexpressing plants and are thus unrelated to the observed resistance phenotype.

Highlights

  • Plant subtilases (SBTs) constitute a large family of mostly extracellular proteases of unknown function

  • A reduction in the level of pectin methylesterification that was observed in transgenic plants with altered levels of SBT3 expression suggested an involvement of SBT3 in the regulation of pectin methylesterases (PMEs)

  • While such a role has been described in other systems, PME activity and the degree of pectin methylesterification did not correlate with the level of insect resistance in SBT3 expression (SBT3-SI) and SBT3 overexpressing plants and are unrelated to the observed resistance phenotype

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Summary

Introduction

Plant subtilases (SBTs) constitute a large family of mostly extracellular proteases of unknown function. Structural modeling of representative Arabidopsis subtilases indicated that PA domain-mediated dimerization and enzyme activation are unlikely to be a general property of all plant SBTs, since residues that were found to be important for dimerization and an auto-inhibitory β-hairpin are only partially conserved in the Arabidopsis SBT family (Rose et al, 2010) Consistent with this notion, the monomeric state was found to be the predominant form of cucumisin in solution (Murayama et al, 2012). The defensive role of SBT3 may rely on its post-ingestive activity in the insect gut

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