Departamento de Ciencia y Tecnología Agroforestal y Genética, ETSIA, Universidad de Castilla-La Mancha, Campus Universitario s/n, Albacete, Spain.
Plant Biol (Stuttg). 2011 Mar;13(2):297-303. doi: 10.1111/j.1438-8677.2010.00359.x.
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.
植物已经发展出许多机制来保护自己免受大多数潜在的微生物病原体和疾病的侵害。在这些机制中，与发病相关的蛋白质作为主动防御的一部分被产生，以防止攻击。在这项研究中，在新鲜藏红花柱头（藏红花）中鉴定出编码 CsPR10 蛋白的全长 cDNA。从编码区的核苷酸序列推导的氨基酸序列与 PR10 蛋白具有同源性。克隆以与 GST 标签融合的形式表达，在大肠杆菌中产生 47 kDa 的蛋白质。CsPR10 具有核糖核酸酶活性，具有 II 型核糖核酸酶的特征；其比活性被量化为 68.8 U·mg（-1）蛋白，因此属于大多数表现出 RNase 活性的 PR10 蛋白的范围内。CsPR10 的抗真菌活性针对黄萎病菌、青霉属和尖孢镰刀菌进行了测定。CsPR10 仅抑制 F. oxysporum 的生长，抗真菌效力反映在 IC（50）为 8.3 μm。表达分析显示在花药和花瓣组织中存在高水平的转录物，在柱头和根中存在低水平的转录物，在叶片中未检测到信号。该蛋白似乎通过激活茉莉酸途径参与主动防御反应。
