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首页> 外文期刊>Plant Pathology >The peach [Prunus persica) defensin PpDFNI displays antifungal activity through specific interactions with the membrane lipids
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The peach [Prunus persica) defensin PpDFNI displays antifungal activity through specific interactions with the membrane lipids

机译：桃（Prunus persica）防御素PpDFNI通过与膜脂的特异性相互作用显示出抗真菌活性

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摘要

Ppdfnl is a defensin gene previously identified in peach {Prunus persica). The biological role of Ppdfnl was investigated by analysing its expression profile in leaves, flowers and fruits, either inoculated with the Monilinia laxa fungalpathogen or mock-inoculated. Ppdfnl expression was highest in flowers and, in fruits, did not vary upon M. laxa inoculation. To characterize the PpDFNI antifungal activity, the recombinant mature peptide was expressed in Escherichia coli and purified; recombinant PpDFNI displays antifungal activity against Botrytis cinerea, M. laxa and Penicillium expansutn, with IC_(50) values of 15T, 9·9 and IT μg rnL~(-1), respectively. Treatment of fungal hyphae with FITC-labelled PpDFNI indicated that the peptide is not internalized by fungal hyphae, but localizes on their external cell surface. At this site, PpDFNI is capable of membrane destabilization and permeabilization, as demonstrated by SYTOX Green fluorescence uptake by the treated mycelia. Using artificial lipid monolayers, it was shown that PpDFNI interacts with sphingolipid-containing membranes; however the strongest interaction occurs with monolayers composed of lipids extracted from sensitive fungi, such as P. expansutn. These data suggest that the lipid composition of fungal membranes is of key relevance for defensin specificity.
机译：Ppdfnl是先前在桃子（Prunus persica）中鉴定的防御素基因。通过分析接种了念珠菌疏松真菌病原体或模拟接种的Ppdfnl在叶片，花朵和果实中的表达谱，研究了其生物学作用。 Ppdfnl表达在花中最高，而在水果中，接种Laxa并没有变化。为了表征PpDFNI的抗真菌活性，将重组成熟肽在大肠杆菌中表达并纯化。重组PpDFNI显示出对灰葡萄孢，松弛杆菌和青霉的抗真菌活性，IC_（50）值为15T，9·9和ITμgrnL〜（-1）。用FITC标记的PpDFNI处理真菌菌丝表明该肽未被真菌菌丝内化，而是位于其外部细胞表面。在这个部位，PpDFNI能够使膜不稳定和通透，如处理过的菌丝体对SYTOX Green荧光的吸收所证明。使用人工脂质单层，显示PpDFNI与含鞘脂的膜发生相互作用。然而，最强烈的相互作用发生在由敏感真菌（例如扩张性假单胞菌）提取的脂质组成的单分子层上。这些数据表明，真菌膜的脂质组成与防御素特异性具有关键的关系。

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