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基于iTRAQ的蛋白质组学分析揭示了生防菌Sneb183菌株在增强大豆对大豆胞囊线虫抗性中的作用。

iTRAQ-Based Proteomic Analysis Reveals the Role of the Biological Control Agent, Strain Sneb183, in Enhancing Soybean Resistance Against the Soybean Cyst Nematode.

作者信息

Wang Yuanyuan, Yang Ruowei, Feng Yaxing, Sikandar Aatika, Zhu Xiaofeng, Fan Haiyan, Liu Xiaoyu, Chen Lijie, Duan Yuxi

机构信息

College of Bioscience and Biotechnology, Shenyang Agricultural University, Shenyang, China.

Nematology Institute of Northern China, Shenyang Agricultural University, Shenyang, China.

出版信息

Front Plant Sci. 2020 Dec 11;11:597819. doi: 10.3389/fpls.2020.597819. eCollection 2020.

Abstract

The soybean cyst nematode (SCN), Ichinohe, poses a serious threat to soybean production worldwide. Biological control agents have become eco-friendly candidates to control pathogens. Our previous study indicated that the biocontrol agent, strain Sneb183, may induce soybean resistance to SCN. To study the mechanisms underlying induced disease resistance in the plant by Sneb183, an iTRAQ (isobaric tag for relative and absolute quantitation)-based proteomics approach was used to identify proteomic changes in SCN-infected soybean roots derived from seeds coated with the Sneb183 fermentation broth or water. Among a total of 456 identified differentially expressed proteins, 212 and 244 proteins were upregulated and downregulated, respectively, in Sneb183 treated samples in comparison to control samples. Some identified differentially expressed proteins are likely to be involved in the biosynthesis of phenylpropanoid, flavone, flavanol, and isoflavonoid and have a role in disease resistance and adaptation to environmental stresses. We used quantitative real-time PCR (qRT-PCR) to analyze key genes, including (phenylalanine ammonia-lyase), (chalcone reductase), (chalcone synthase), and (isoflavone synthase), that are involved in isoflavonoid biosynthesis in Sneb183-treated and control samples. The results showed that these targeted genes have higher expression levels in Sneb183-treated than in control samples. High performance liquid chromatography (HPLC) analysis further showed that the contents of daidzein in Sneb183-treated samples were 7.24 times higher than those in control samples. These results suggested that the strain Sneb183 may have a role in inducing isoflavonoid biosynthesis, thereby resulting in enhanced resistance to SCN infection in soybean.

摘要

大豆胞囊线虫(SCN),即北条胞囊线虫,对全球大豆生产构成严重威胁。生物防治剂已成为控制病原体的环保型候选物。我们之前的研究表明,生物防治剂菌株Sneb183可能诱导大豆对SCN产生抗性。为了研究Sneb183诱导植物抗病性的潜在机制,采用了基于iTRAQ(相对和绝对定量等压标签)的蛋白质组学方法,以鉴定用Sneb183发酵液或水处理的种子所衍生的受SCN感染的大豆根中的蛋白质组变化。在总共鉴定出的456种差异表达蛋白中,与对照样品相比,Sneb183处理样品中有212种蛋白上调,244种蛋白下调。一些鉴定出的差异表达蛋白可能参与苯丙烷类、黄酮、黄烷醇和异黄酮的生物合成,并在抗病性和适应环境胁迫中发挥作用。我们使用定量实时PCR(qRT-PCR)分析了参与Sneb183处理和对照样品中异黄酮生物合成的关键基因,包括苯丙氨酸解氨酶、查尔酮还原酶、查尔酮合酶和异黄酮合酶。结果表明,这些靶向基因在Sneb183处理样品中的表达水平高于对照样品。高效液相色谱(HPLC)分析进一步表明,Sneb183处理样品中大豆苷元的含量比对照样品高7.24倍。这些结果表明,菌株Sneb183可能在诱导异黄酮生物合成中发挥作用,从而增强大豆对SCN感染的抗性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ca7f/7759536/56bc09406f1e/fpls-11-597819-g001.jpg

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