Functional Analysis of the C-5 Sterol Desaturase PcErg3 in the Sterol Auxotrophic Oomycete Pathogen Phytophthora capsici.
Functional Analysis of the C-5 Sterol Desaturase PcErg3 in the Sterol Auxotrophic Oomycete Pathogen Phytophthora capsici.
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DOI：
10.3389/fmicb.2022.811132
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发表时间：
2022
影响因子：
5.2
通讯作者：
Liu, Xili
中科院分区：
文献类型：
--
作者：
Wang, Weizhen;Cui, Tongshan;Zhang, Fan;Xue, Zhaolin;Zhang, Borui;Liu, Xili
虽然甾醇在大多数真核生物中起着重要作用，但一些卵菌，包括疫霉属，已经失去了固醇合成途径。然而，编码固醇合成途径中的C-5固醇去饱和酶的ERG 3基因仍然存在于疫霉属物种的基因组中。辣椒疫霉是一种危害性很强的病原菌，其寄主范围很广，对农业生产构成严重威胁。本研究以辣椒疫霉中的ERG 3基因（PcERG 3）为研究对象，探讨其在辣椒疫霉中的功能。结果表明，PcERG 3基因在辣椒疫霉各发育阶段均能表达，其中孢子囊和菌丝体表达量较高。利用Erg 3底物处理辣椒疫霉野生型菌株和PcERG 3 Δ菌株，GC-MS分析其甾醇谱，野生型菌株能将星甾醇转化为下游产物，而Δ菌株不能，表明PcERG 3保留了C-5甾醇去饱和酶活性。通过比较不同菌株的生物学特性，发现PcERG 3基因对辣椒疫霉的生长发育并不重要。PcERG 3 Δ转化子和野生型菌株的致病性相当，表明PcERG 3不是辣椒疫霉与寄主互作所必需的。进一步的研究表明，PcERG 3 Δ转化体和野生型菌株对外界逆境如不合适的温度、高渗透压和过度的pH表现出相似的耐受性，这表明PcERG 3不是辣椒疫霉科普这些环境胁迫所必需的。
Although sterols play an important role in most eukaryotes, some oomycetes, including Phytophthora spp., have lost the sterol synthesis pathway. Nevertheless, the ERG3 gene encoding C-5 sterol desaturase in the sterol synthesis pathway is still present in the genomes of Phytophthora spp. Phytophthora capsici, a destructive pathogen with a broad range of plant hosts, poses a significant threat to the production of agriculture. This study focused on the ERG3 gene in P. capsici (PcERG3) and explored its function in this pathogen. It showed that the PcERG3 gene could be expressed in all tested developmental stages of P. capsici, with sporangium and mycelium displaying higher expression levels. A potential substrate of Erg3 (stellasterol) was used to treat the P. capsici wild-type strain and a PcERG3Δ transformant, and their sterol profiles were determined by GC-MS. The wild-type strain could convert stellasterol into the down-stream product while the transformant could not, indicating that PcErg3 retains the C-5 sterol desaturase activity. By comparing the biological characteristics of different strains, it was found that PcERG3 is not important for the development of P. capsici. The pathogenicity of the PcERG3Δ transformants and the wild-type strain was comparable, suggesting that PcERG3 is not necessary for the interaction between P. capsici and its hosts. Further investigations revealed that the PcERG3Δ transformants and the wild-type strain displayed a similar level of tolerance to external adversities such as unsuitable temperatures, high osmotic pressures, and intemperate pH, signifying that PcERG3 is not essential for P. capsici to cope with these environmental stresses.
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