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Transcriptional Profiling During Infection of Potato NLRs and Phytophthora Infestans Effectors Using Cdna Enrichment Sequencing

The Crop Journal(2024)

The James Hutton Institute | Crop Research Centre | University of Dundee

Cited 0|Views17
Abstract
An accurate assessment of host and pathogen gene expression during infection is critical for understanding the molecular aspects of host-pathogen interactions. Often, pathogen-derived transcripts are difficult to ascertain at early infection stages owing to the unfavourable transcript representation compared to the host genes. In this study, we compare two sequencing techniques, RNAseq and enrichment sequencing (RenSeq and PenSeq) of cDNA, to investigate gene expression patterns in the doubled monoploid potato (DM) infected with the late blight pathogen Phytophthora infestans. Our results reveal distinct advantages of cDNA RenSeq and PenSeq over traditional RNAseq in terms of target gene representation and transcriptional quantification at early infection stages. Throughout the infection time course, cDNA enrichment sequencing enables transcriptomic analyses for more targeted host and pathogen genes. For highly expressed genes that were sampled in parallel by both cDNA enrichment and RNAseq, a high level of concordance in expression profiles is observed, indicative of at least semi-quantitative gene expression representation following enrichment.
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Key words
RxLR effector,NLRs,Late blight,Potato,cDNA sequencing,RenSeq,PenSeq,RNAseq
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要点】:该论文通过对比RNA测序和cDNA富集测序技术,研究了马铃薯感染晚疫病菌Phytophthora infestans时的基因表达模式,发现cDNA富集测序在早期感染阶段对目标基因的代表性和转录定量具有明显优势。

方法】:研究使用了两种测序技术:RNA测序和cDNA富集测序(RenSeq和PenSeq),在二倍体马铃薯(DM)上进行实验。

实验】:实验在感染进程的不同时间点进行,使用cDNA富集测序技术对目标和高度表达的基因进行转录组分析。结果显示,对于同时通过cDNA富集和RNA测序平行检测的高度表达基因,表达谱具有较高的一致性,表明富集后至少有半定量的基因表达代表。