Effects of a Novel Pathogenesis-Related Class 10 (PR-10) Protein from Crotalaria pallida Roots with Papain Inhibitory Activity against Root-Knot Nematode Meloidogyne incognita

Abstract

A novel pathogenesis-related class 10 (PR-10) protein with papain inhibitory activity, named CpPRI, was purified from Crotalaria pallida roots by ammonium sulfate precipitation followed by three reverse-phase high-performance liquid chromatographies (HPLCs). CpPRI is made up of a single polypeptide chain with a M(r) of 15 kDa, as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). This protein exhibited a K(i) value of 1.8 x 10(-9) M and operates via a noncompetitive inhibition mechanism. The alignment of the N-terminal amino acid sequence of CpPRI with other proteins revealed its identity with PR-10 proteins. CpPRI acts against digestive proteinase from root-knot nematode Meloidogyne incognita and demonstrated nematostatic and nematicide effects on this parasite in bioassays. In a localization study, fluorescein-5-isothiocyanate (FITC)-CpPRI was observed to internalize and diffuse over the entire J2 body after 6 h of incubation. This fact could explain the natural tolerance of this plant species to nematodes.

Overexpression of a Pathogenesis-Related Protein 10 Enhances Biotic and Abiotic Stress Tolerance in Rice

“…Previous studies have shown that PR10 family protein has ribonuclease (Bantignies et al, 2000; Zhou et al, 2002) and cysteine protease inhibition activities (Andrade et al, 2010). Rice JIOsPR10 was also reported to contain RNase activity, suggesting that biochemical activity might be essential for the role of JIOsPR10 (Kim et al, 2008b).…”

“…Those structure elements enclose a large hydrophobic cavity which is most likely related with their functional relevance (Fernandes et al, 2013). Several studies have shown that PR-10 proteins contain RNase activities (Chen et al, 2007; Kim et al, 2008b; Krishnaswamy et al, 2011; Xie et al, 2010), and cysteine protease inhibitor activity (Andrade et al, 2010). …”

“…Previous studies have revealed that PR-10 family proteins are involved in anti-biotic stresses, such as antifungus (Chen et al, 2007; Xie et al, 2010), anti-bacteria (Flores et al, 2002; Xie et al, 2010), anti-virus (Park et al, 2004), and anti-nematode (Andrade et al, 2010). To date, four different PR10 family genes have been characterized in rice: PR10a (McGee et al, 2001), PR10b (McGee et al, 2001), jasmonic acid inducible pathogenesis-related class 10 ( JIOsPR10 ) (Jwa et al, 2001), and RSOsPR10 (Hashimoto et al, 2004).…”

Characterization of resistance to pine wood nematode infection in Pinus thunbergiiusing suppression subtractive hybridization

“…We observed synchronized expression of PR10 with peroxidase in resistant trees, which indicates this gene may be induced by ROS such as H 2 O 2 . However, PR10 (CpPRI) acts against a digestive proteinase from the root knot nematode Meloidogyne incognita [43]. Therefore, PR10 may act as a proteinase against cellulases, beta-1,3-glucanase, and pectate lyases secreted from PWN [44-46].…”

Proteome of Soybean Seed Exudates Contains Plant Defense-Related Proteins Active against the Root-Knot NematodeMeloidogyne incognita

“…The nematostatic and nematicidal effects of the exudates on M. incognita J2s were also verified using a proteinaceous papain inhibitor purified from Crotalaria pallida roots. 7 These effects were found to be lethal, with 95% mortality, when the parasite was exposed to a 50 μg dose of this inhibitor for 48 h. 7 Although the repellent effect of the soybean exudates toward the nematodes was transient (Figure 2), this delay affected their infectivity. This assumption is supported by our experimental findings in which tobacco plants that were inoculated with soybean seed exudatetreated J2s showed around 90 and 94% reductions in the number of galls and egg masses, respectively.…”