Sterol-Sensing Domain (SSD)-Containing Proteins in Sterol Auxotrophic Phytophthora capsici Mediate Sterol Signaling and Play a Role in Asexual Reproduction and Pathogenicity.
Sterol-Sensing Domain (SSD)-Containing Proteins in Sterol Auxotrophic Phytophthora capsici Mediate Sterol Signaling and Play a Role in Asexual Reproduction and Pathogenicity.
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DOI：
10.1128/spectrum.03797-22
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发表时间：
2023-02-14
影响因子：
3.7
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中科院分区：
文献类型：
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疫霉属（Phytophthora）是一种破坏性的丝状植物病原体，属于卵菌纲（Oomycetes），卵菌纲是一组在形态上与真菌相似但在遗传学上不同的微生物。它们是甾醇营养缺陷型的，但仍然利用外源甾醇进行生长和发育。然而，到目前为止，疫霉属中甾醇利用的潜在机制尚不清楚。在这项研究中，我们确定了四个基因在辣椒疫霉编码的蛋白质含有一个甾醇敏感结构域（SSD），蛋白质结构域约180个氨基酸，包括五个跨膜片段，并已知在动物甾醇信号转导的功能。使用改良的CRISPR/Cas9系统，我们成功地敲除了名为PcSCP 1至PcSCP 4的四个基因（用于辣椒疫霉含SSD的蛋白1至4），无论是单独敲除还是顺序敲除，从而产生了单敲除、双敲除、三敲除和四敲除转化体。结果表明，敲除四种PcSCP中的一种不足以阻断甾醇信号传导。然而，与在甾醇处理下产生游动孢子的野生型辣椒疫霉相反，四重“全部四个”PcSCP敲除转化体在无性繁殖中不再响应甾醇处理。这四个PcSCP在辣椒疫霉甾醇信号转导中发挥着重要作用，具有功能冗余性。转录组分析表明，一个子集的基因的表达调控外源甾醇通过PcSCP。进一步的研究表明，甾醇可以通过PcSCP控制肌动蛋白介导的膜运输来刺激游动孢子的分化。此外，PcSCPs基因敲除后的致病性显著降低，许多致病性相关基因表达下调，表明PcSCPs基因也参与了植物与病原菌的互作。疫霉属是卵菌纲的一个重要属，由许多破坏性的植物病原体组成。由于甾醇合成途径的不完全性，疫霉属（Phytophthora spp.）他们没有能力生产类固醇。因此，这些甾醇营养缺陷型卵菌需要从环境（如宿主植物）中募集甾醇以支持生长和发育，这在病原体-植物相互作用中似乎至关重要。然而，潜在的机制甾醇利用疫霉属。但基本上仍不为人所知。在这里，我们表明，一个家庭的甾醇敏感结构域包含蛋白（SCP）的四个成员组成的辣椒疫霉在甾醇信号与功能冗余的关键作用。此外，这些SCP在不同的生物过程中发挥作用，包括无性繁殖和致病性。我们的研究总体上揭示了PcSCP的多种功能，并解决了外源甾醇如何调节异宗配合疫霉菌发育的问题。通过含有SSD的蛋白质。
Phytophthora species are devastating filamentous plant pathogens that belong to oomycetes, a group of microorganisms similar to fungi in morphology but phylogenetically distinct. They are sterol auxotrophic, but nevertheless exploit exogenous sterols for growth and development. However, as for now the mechanisms underlying sterol utilization in Phytophthora are unknown. In this study, we identified four genes in Phytophthora capsici that encode proteins containing a sterol-sensing domain (SSD), a protein domain of around 180 amino acids comprising five transmembrane segments and known to feature in sterol signaling in animals. Using a modified CRISPR/Cas9 system, we successfully knocked out the four genes named PcSCP1 to PcSCP4 (for P. capsici SSD-containing protein 1 to 4), either individually or sequentially, thereby creating single, double, triple, and quadruple knockout transformants. Results showed that knocking out just one of the four PcSCPs was not sufficient to block sterol signaling. However, the quadruple “all-four” PcSCPs knockout transformants no longer responded to sterol treatment in asexual reproduction, in contrast to wild-type P. capsici that produced zoospores under sterol treatment. Apparently, the four PcSCPs play a key role in sterol signaling in P. capsici with functional redundancy. Transcriptome analysis indicated that the expression of a subset of genes is regulated by exogenous sterols via PcSCPs. Further investigations showed that sterols could stimulate zoospore differentiation via PcSCPs by controlling actin-mediated membrane trafficking. Moreover, the pathogenicity of the “all-four” PcSCPs knockout transformants was significantly decreased and many pathogenicity related genes were downregulated, implying that PcSCPs also contribute to plant-pathogen interaction. IMPORTANCE Phytophthora is an important genus of oomycetes that comprises many destructive plant pathogens. Due to the incompleteness of the sterol synthesis pathway, Phytophthora spp. do not possess the ability to produce sterols. Therefore, these sterol auxotrophic oomycetes need to recruit sterols from the environment such as host plants to support growth and development, which seems crucial during pathogen-plant interactions. However, the mechanisms underlying sterol utilization by Phytophthora spp. remain largely unknown. Here, we show that a family of sterol-sensing domain-containing proteins (SCPs) consisting of four members in P. capsici plays a key role in sterol signaling with functional redundancy. Moreover, these SCPs play a role in different biological processes, including asexual reproduction and pathogenicity. Our study overall revealed the multiple functions of PcSCPs and addressed the question of how exogenous sterols regulate the development of heterothallic Phytophthora spp. via SSD-containing proteins.
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影响因子：
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作者：
Jee, HJ;Ko, WH
通讯作者：
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影响因子：
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Waterman MR
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通讯作者：
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