<br>云杉两个新 PR4 基因的克隆和表征。,International Journal of Molecular Sciences

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云杉两个新 PR4 基因的克隆和表征。
International Journal of Molecular Sciences ( IF 4.9 ) Pub Date : 2022-11-28 , DOI: 10.3390/ijms232314906
Weidong Zhao ₁ , Lijuan Liu ₁ , Chengsong Li ₁ , Chunlin Yang ₁ , Shujiang Li ₁ , Shan Han ₁ , Tiantian Lin ₁ , Yinggao Liu ₁

Affiliation

病程相关 (PR) 蛋白在植物病原抗性中发挥重要作用，由 17 个家族组成，其中包括 PR4 家族，具有抗真菌和抗病原功能。 PR4 蛋白含有 C 端 Barwin 结构域，并根据 N 端几丁质结合结构域 (CBD) 的存在分为 I 类和 II 类。本研究首次从云杉中分离出两个PR4基因PaPR4-a和PaPR4-b，分别编码PaPR4-a和PaPR4-b。序列分析表明它们是 II 类蛋白质，因为存在 N 端信号肽和 C 端 Barwin 结构域，但没有 CBD。使用木瓜类 Barwin 蛋白作为模板的三级结构分析揭示了蛋白质之间的结构相似性，因此也揭示了功能相似性。预测结果揭示了 N 末端跨膜结构域，亚细胞定位研究证实了其在细胞膜和细胞核上的位置。实时定量 PCR (RT-qPCR) 表明，云杉感染后 PaPR4-a 和 PaPR4-b 表达水平上调。此外，PaPR4-a和PaPR4-b在大肠杆菌中被诱导，重组蛋白以包涵体形式存在。复性的纯化蛋白质显示出抗真菌活性。此外，过表达PaPR4-a和PaPR4-b的转基因烟草表现出对真菌感染的抵抗力的提高。该研究可以为进一步了解 PR4 诱导的防御反应的分子机制提供基础。

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Cloning and Characterization of Two Novel PR4 Genes from Picea asperata.

Pathogenesis-related (PR) proteins are important in plant pathogenic resistance and comprise 17 families, including the PR4 family, with antifungal and anti-pathogenic functions. PR4 proteins contain a C-terminal Barwin domain and are divided into Classes I and II based on the presence of an N-terminal chitin-binding domain (CBD). This study is the first to isolate two PR4 genes, PaPR4-a and PaPR4-b, from Picea asperata, encoding PaPR4-a and PaPR4-b, respectively. Sequence analyses suggested that they were Class II proteins, owing to the presence of an N-terminal signal peptide and a C-terminal Barwin domain, but no CBD. Tertiary structure analyses using the Barwin-like protein of papaya as a template revealed structural similarity, and therefore, functional similarity between the proteins. Predictive results revealed an N-terminal transmembrane domain, and subcellular localization studies confirmed its location on cell membrane and nuclei. Real-time quantitative PCR (RT-qPCR) demonstrated that PaPR4-a and PaPR4-b expression levels were upregulated following infection with Lophodermium piceae. Additionally, PaPR4-a and PaPR4-b were induced in Escherichia coli, where the recombinant proteins existed in inclusion bodies. The renatured purified proteins showed antifungal activity. Furthermore, transgenic tobacco overexpressing PaPR4-a and PaPR4-b exhibited improved resistance to fungal infection. The study can provide a basis for further molecular mechanistic insights into PR4-induced defense responses.

更新日期：2022-11-28

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