1. Plant 'pathogenesis-related' proteins and their role in defense against pathogens
A Stintzi, T Heitz, V Prasad, S Wiedemann-Merdinoglu, S Kauffmann, P Geoffroy, M Legrand, B Fritig Biochimie. 1993;75(8):687-706. doi: 10.1016/0300-9084(93)90100-7.
The hypersensitive reaction to a pathogen is one of the most efficient defense mechanisms in nature and leads to the induction of numerous plant genes encoding defense proteins. These proteins include: 1) structural proteins that are incorporated into the extracellular matrix and participate in the confinement of the pathogen; 2) enzymes of secondary metabolism, for instance those of the biosynthesis of plant antibiotics; 3) pathogenesis-related (PR) proteins which represent major quantitative changes in soluble protein during the defense response. The PRs have typical physicochemical properties that enable them to resist to acidic pH and proteolytic cleavage and thus survive in the harsh environments where they occur: vacuolar compartment or cell wall or intercellular spaces. Since the discovery of the first PRs in tobacco many other similar proteins have been isolated from tobacco but also from other plant species, including dicots and monocots, the widest range being characterized from hypersensitively reacting tobacco. Based first on serological properties and later on sequence data, the tobacco PRs have been classified in five major groups. Group PR-1 contains the first discovered PRs of 15-17 kDa molecular mass, whose biological activity is still unknown, but some members have been shown recently to have antifungal activity. Group PR-2 contains three structurally distinct classes of 1,3-beta-glucanases, with acidic and basic counterparts, with dramatically different specific activity towards linear 1,3-beta-glucans and with different substrate specificity. Group PR-3 consists of various chitinases-lysozymes that belong to three distinct classes, are vacuolar or extracellular, and exhibit differential chitinase and lysozyme activities. Some of them, either alone or in combination with 1,3-beta-glucanases, have been shown to be antifungal in vitro and in vivo (transgenic plants), probably by hydrolysing their substrates as structural components in the fungal cell wall. Group PR-4 is the less studied, and in tobacco contains four members of 13-14.5 kDa of unknown activity and function. Group PR-5 contains acidic-neutral and very basic members with extracellular and vacuolar localization, respectively, and all members show sequence similarity to the sweet-tasting protein thaumatin. Several members of the PR-5 group from tobacco and other plant species were shown to display significant in vitro activity of inhibiting hyphal growth or spore germination of various fungi probably by a membrane permeabilizing mechanism.(ABSTRACT TRUNCATED AT 400 WORDS)
2. Sucrose increases pathogenesis-related PR-2 gene expression in Arabidopsis thaliana through an SA-dependent but NPR1-independent signaling pathway
Marie-Christine Thibaud, Sandrine Gineste, Laurent Nussaume, Christophe Robaglia Plant Physiol Biochem. 2004 Jan;42(1):81-8. doi: 10.1016/j.plaphy.2003.10.012.
Pathogenesis-related (PR) protein-coding gene expression was studied in Arabidopsis thaliana grown in liquid medium in the presence of sugars (sucrose or glucose). PR protein transcripts accumulated in the presence of sugar in the medium. A potential effect linked to osmolarity changes induced by sugar addition in the medium was ruled out using osmotica (NaCl or polyethylene glycol). Two major proteins were purified from the culture medium and found to be homologous to A. thaliana PR-2 (acidic form of beta-1, 3-glucanase) and PR-5 (thaumatin-like PR-protein). The expression of the corresponding genes was increased in the presence of sucrose and was detected exclusively in the green parts of the plant. The use of mutants and transgenic plants of A. thaliana indicated that salicylic acid (SA) was involved in the sugar-dependent activation of these PR protein-coding genes. Activation of the PR-2-coding gene was demonstrated not to be hexokinase-dependent and to be linked to a sugar metabolite acting as an internal signal as shown with non-metabolizable sugars, which were inefficient for the induction of the PR-2-coding gene. Moreover, the activation of this gene occurred in the npr1 mutant suggesting that the sugar signal acts either downstream or independently of NPR1.
3. Antifungal mechanism of a novel antifungal protein from pumpkin rinds against various fungal pathogens
Seong-Cheol Park, Jin-Young Kim, Jong-Kook Lee, Indeok Hwang, Hyeonsook Cheong, Jae-Woon Nah, Kyung-Soo Hahm, Yoonkyung Park J Agric Food Chem. 2009 Oct 14;57(19):9299-304. doi: 10.1021/jf902005g.
A novel antifungal protein (Pr-2) was identified from pumpkin rinds using water-soluble extraction, ultrafiltration, cation exchange chromatography, and reverse-phase high-performance liquid chromatography. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry indicated that the protein had a molecular mass of 14865.57 Da. Automated Edman degradation showed that the N-terminal sequence of Pr-2 was QGIGVGDNDGKRGKR-. The Pr-2 protein strongly inhibited in vitro growth of Botrytis cinerea, Colletotrichum coccodes, Fusarium solani, Fusarium oxysporum, and Trichoderma harzianum at 10-20 microM. The results of confocal laser scanning microscopy and SYTOX Green uptake demonstrated that its effective region was the membrane of the fungal cell surface. In addition, this protein was found to be noncytotoxic and heat-stable. Taken together, the results of this study indicate that Pr-2 is a good candidate for use as a natural antifungal agent.
