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利用荧光蛋白和细胞活力标记物观察油棕小叶中的感染情况

Visualization of Infection in Oil Palm Leaflets with Fluorescent Proteins and Cell Viability Markers.

作者信息

Ochoa Juan C, Herrera Mariana, Navia Mónica, Romero Hernán Mauricio

机构信息

Oil Palm Biology and Breeding Research Program, Colombian Oil Palm Research Center (CENIPALMA), Bogotá, Colombia.

Departamento de Biología, Universidad Nacional de Colombia, Bogotá, Colombia.

出版信息

Plant Pathol J. 2019 Feb;35(1):19-31. doi: 10.5423/PPJ.OA.02.2018.0034. Epub 2019 Feb 1.

Abstract

Bud rot (BR) is the most devastating disease affecting oil palm () crops in Colombia. Its causal agent, , initiates the infection in immature oil palm leaflets producing necrotic lesions, followed by colonization of opportunistic necrotrophs, which increases disease damage. To improve the characterization of the disease, we transformed using -mediated transformation (ATMT) to include the fluorescent proteins CFP-SKL (peroxisomal localization), eGFP and mRFP1 (cytoplasmic localization). The stability of some transformants was confirmed by Southern blot analysis and single zoospore cultures; additionally, virulence and in vitro growth were compared to the wild-type isolate to select transformants with the greatest resemblance to the WT isolate. GFP-tagged was useful to identify all of the infective structures that are commonly formed by hemibiotrophic oomycetes, including apoplastic colonization and haustorium formation. Finally, we detected cell death responses associated with immature oil palm tissues that showed reduced susceptibility to infection, indicating that these tissues could exhibit age-related resistance. The aim of this research is to improve the characterization of the initial disease stages and generate cell biology tools that may be useful for developing methodologies for early identification of oil palm materials resistant or susceptible to BR.

摘要

芽腐病(BR)是影响哥伦比亚油棕作物的最具毁灭性的病害。其病原菌,在未成熟的油棕小叶中引发感染,产生坏死病变,随后机会性坏死营养菌定殖,这加剧了病害损害。为了更好地描述该病害,我们使用农杆菌介导的转化(ATMT)对进行转化,使其包含荧光蛋白CFP-SKL(过氧化物酶体定位)、eGFP和mRFP1(细胞质定位)。通过Southern印迹分析和单游动孢子培养证实了一些转化体的稳定性;此外,将毒力和体外生长与野生型分离株进行比较,以选择与野生型分离株最相似的转化体。带有绿色荧光蛋白标记的有助于识别半活体营养型卵菌通常形成的所有感染结构,包括质外体定殖和吸器形成。最后,我们检测了与未成熟油棕组织相关的细胞死亡反应,这些组织对感染的易感性降低,表明这些组织可能表现出与年龄相关的抗性。本研究的目的是更好地描述病害的初始阶段,并生成细胞生物学工具,这些工具可能有助于开发早期鉴定对芽腐病具有抗性或易感性的油棕材料的方法。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dcb1/6385658/3129d6921054/ppj-35-019f1.jpg

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