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一种 15kDa 的分泌蛋白在可可毛色二孢菌发育和致病性中起着重要作用。

A secreted protein of 15 kDa plays an important role in Phytophthora palmivora development and pathogenicity.

机构信息

Department of Biochemistry, Faculty of Science, Prince of Songkla University, Hat-Yai, Songkhla, 90112, Thailand.

Department of Plant and Environmental Protection Sciences, University of Hawaii at Manoa, Honolulu, HI, 96822, USA.

出版信息

Sci Rep. 2020 Feb 11;10(1):2319. doi: 10.1038/s41598-020-59007-1.

Abstract

Phytophthora palmivora is a destructive oomycete plant pathogen with a wide host range. So far, little is known about the factors governing its infection structure development and pathogenicity. From the culture filtrate of a P. palmivora strain isolated from papaya, we identified a secreted glycoprotein of 15 kDa, designated as Ppal15kDa, using liquid chromatography tandem mass spectrometry. Two gene variants, Ppal15kDaA and Ppal15kDaB were amplified from a P. palmivora papaya isolate. Transient expression of both variants in Nicotiana benthamiana by agroinfiltration enhanced P. palmivora infection. Six Ppal15kDa mutants with diverse mutations were generated via CRISPR/Cas9-mediated gene editing. All mutants were compromised in infectivity on N. benthamiana and papaya. Two mutants with all Ppal15kDa copies mutated almost completely lost pathogenicity. The pathogenicity of the other four containing at least one wild-type copy of Ppal15kDa was compromised at varying levels. The mutants were also affected in development as they produced smaller sporangia, shorter germ tubes, and fewer appressoria. The affected levels in development corresponded to the levels of reduction in pathogenicity, suggesting that Ppal15kDa plays an important role in normal development of P. palmivora infection structures. Consistent with its role in infection structure development and pathogenicity, Ppal15kDa was found to be highly induced during appressorium formation. In addition, Ppal15kDa homologs are broadly present in Phytophthora spp., but none were characterized. Altogether, this study identified a novel component involved in development and pathogenicity of P. palmivora and possibly other Phytophthora spp. known to contain a Ppal15kDa homolog.

摘要

可可疫霉是一种具有广泛宿主范围的破坏性卵菌植物病原体。目前,人们对其侵染结构发育和致病性的调控因素知之甚少。我们从从木瓜中分离出的可可疫霉菌株的培养液中,利用液相色谱串联质谱法鉴定出一种 15 kDa 的分泌糖蛋白,命名为 Ppal15kDa。从可可疫霉木瓜分离株中扩增出两个基因变异体,Ppal15kDaA 和 Ppal15kDaB。通过农杆菌瞬时表达这两个变异体增强了可可疫霉对烟草原生质体的侵染。通过 CRISPR/Cas9 介导的基因编辑生成了 6 个具有不同突变的 Ppal15kDa 突变体。所有突变体在侵染烟草原生质体和木瓜时均受到影响。两个含有所有 Ppal15kDa 拷贝突变的突变体几乎完全丧失了致病性。其他四个至少含有一个野生型 Ppal15kDa 拷贝的突变体的致病性也受到不同程度的影响。这些突变体在发育过程中也受到影响,因为它们产生的游动孢子较小、芽管较短、附着胞较少。发育受到的影响与致病性降低的程度相对应,表明 Ppal15kDa 在可可疫霉侵染结构的正常发育中起着重要作用。与它在侵染结构发育和致病性中的作用一致,Ppal15kDa 在附着胞形成过程中被高度诱导。此外,Ppal15kDa 同源物广泛存在于疫霉属中,但没有被鉴定。总之,这项研究鉴定了一个参与可可疫霉发育和致病性的新成分,可能还有其他已知含有 Ppal15kDa 同源物的疫霉属。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4fa9/7012922/a18ec6f66b7b/41598_2020_59007_Fig1_HTML.jpg

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