Molecular cloning and characterisation of a pathogenesis‐related protein CsPR10 from Crocus sativus

Abstract

Plants have developed many mechanisms to protect themselves against most potential microbial pathogens and diseases. Among these mechanisms, pathogenesis-related proteins are produced as part of the active defence to prevent attack. In this study, a full-length cDNA encoding the CsPR10 protein was identified in fresh saffron stigmas (Crocus sativus). The deduced amino acid sequence from the nucleotide sequence of the coding region showed homology with PR10 proteins. The clone expressed as a protein in fusion with a GST tag produced a 47-kDa protein in E. coli. CsPR10 had ribonuclease activity, with features common to class II-type ribonucleases; its specific activity was quantified as 68.8 U·mg(-1) protein, thus falling within the range of most PR10 proteins exhibiting RNase activity. Antifungal activity of CsPR10 was assayed against Verticillium dahliae, Penicillium sp. and Fusarium oxysporum. CsPR10 inhibited only F. oxysporum growth, and antifungal potency was reflected in a IC(50) of 8.3 μm. Expression analysis showed the presence of high transcript levels in anther and tepal tissues, low levels in stigmas and roots, and no signal detected in leaves. This protein seems to be involved in the active defence response through activation of the jasmonic acid pathway.

The pathogenesis-related protein PR-4b from Theobroma cacao presents RNase activity, Ca2+ and Mg2+ dependent-DNase activity and antifungal action on Moniliophthora perniciosa

“…However, several characterized PR proteins showed enzymatic activities related to antimicrobial properties. Among the 17 PR protein families already described, at least 9 present enzymatic activity such glucanases (PR-2; [6]), osmotins and thaumatins (PR-5; [7-10]), protease inhibitors (PR-6; [11]), lysozymes (PR-8; [12]), peroxidase (PR-9; [13,14]), ribonucleases (PR10; [15-17]) and chitinases (PR-3, PR-4, PR-8, PR-11; [18-20]). Focusing on PR-4 proteins, several studies showed the involvement of this family in plant defense responses regulated by signal molecules, such as salicylic acid (SA), abscissic acid (ABA), jasmonate (JA) and ethylene (ET).…”

A novel ribonuclease with antiproliferative activity toward leukemia and lymphoma cells and HIV-1 reverse transcriptase inhibitory activity from the mushroom, Hohenbuehelia serotina

“…Ribonucleases (RNases) have been isolated and characterized from a multitude of organisms, including parasites, bacteria, fungi, plants and a variety of tissues from mammals (9)(10)(11)(12)(13)(14)(15)(16)(17) RNases display different activities, such as antitumor (18)(19)(20)(21)(22)(23)(24)(25), immunosuppressive (26), antifungal (27), and antiviral (28,29) activities. Due to the array of potentially exploitable activities, RNases have drawn the attention of many researchers.…”

Pathogenesis-Related Gene, JcPR-10a from Jatropha curcas Exhibit RNase and Antifungal Activity

“…RNase activity is reported in PR-10 proteins from birch [51], lupine [52], cotton [17], hot pepper [10] and saffron [53]. JcPR-10a protein showed RNase activity at a wide pH range (4-9) and at short time duration, indicating that JcPR-10a protein shows strong RNase activity.…”