Abstract
Potato late blight is one the most important crop diseases worldwide. Even though potato has been studied for many years, the potato disease late blight still has a vast negative effect on the potato production [1], [2], [3]. Late blight is caused by the pathogen Phytophthora infestans (P. infestans), which initiates infection through leaves. However, the biological activities during different stages of infection are poorly described, and could enable novel or improved ways of defeating late blight infection [4]. Therefore, we investigated the interactions between P. infestans (mixed strain culture) and potato (Solanum tuberosum). Three commercially available field potato cultivars of different resistance to late blight infection; Kuras (moderate), Sarpo Mira (highly resistant) and Bintje (very susceptable) were grown under controlled green house conditions and inoculated with a diversity of P. infestans populations.We used label-free quantitative proteomics to investigate the infection with P. infestans in a time-course study over 258 h. Several key issues limits proteome analysis of potato leaf tissue [5], [6], [7]. Firstly, the immense complexity of the plant proteome, which is further complicated by the presence of highly abundant proteins, such as ribulose bisphosphate carboxylase/oxygenase (RuBisCO). Secondly, plant leaf and potato, in particular, contain abundant levels amounts of phenols and polyphenols, which hinder or completely prevent a successful protein extraction. Hitherto, protein profiling of potato leaf tissues have been limited to few proteome studies and only 1484 proteins have been extracted and comprehensively described [5], [8], [9]. We here present the detailed methods and raw data by optimized gel-enhanced label free quantitative approach. The methodology enabled us to detect and quantify between 3248 and 3529 unique proteins from each cultivar, and up to 758 P. infestans derived proteins. The complete dataset is available via ProteomeXchange, with the identifier PXD002767.
Highlights
txt para- Detailed description of all settings used for the MaxQuant search
time-course P. infestans infected samples were searched as independent replicates linked by option
This enables statistical analysis based on the search output files
Summary
Raw files and text/excel files Mass Spectrometry Liquid Chromatography High-resolution/high-accuracy mass spectrometer system was used: Q Exactive (Thermo Scientific) Raw and analyzed data. Proteome analysis of three potato cultivars following time-course infection with late blight (Phytophthora infestans) Three potato cultivars were infected with Phytophthora infestans in a time-course over 258 h. Leaf proteins were extracted and analyzed by GelMS using 298 runs of 2.5 h UPLC-OrbitrapMS prior to label-free proteome analysis using MaxQuant. The MS proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD002767. Optimized protein extraction method for potato leaf significantly reducing effects of polyphenols and Rubisco. Most extensive proteomic analysis of potato leaf so far ( 44000 unique proteins). Time-course infection of three cultivars of wide resistance towards late blight enabling study of biological response to infection
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