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 ascert...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.展开更多
More than 170 years after causing the potato famine in Ireland,late blight is still considered one of the most devastating crop diseases.Commercial potato breeding efforts depend on natural sources of resistance to pr...More than 170 years after causing the potato famine in Ireland,late blight is still considered one of the most devastating crop diseases.Commercial potato breeding efforts depend on natural sources of resistance to protect crops from the rapidly evolving late blight pathogen,Phytophthora infestans.We have identified and mapped a novel broad-spectrum disease resistance gene effective against P.infestans from the wild,diploid potato species Solanum bulbocastanum.Diagnostic resistance gene enrichment sequencing(dRenSeq)was used to confirm the uniqueness of the identified resistance.RenSeq and GenSeq-based mapping of the resistance,referred to as Rpi-blb4,alongside recombinant screening,positioned the locus responsible for the resistance to potato chromosome 5.The interval spans approximately 2.3 Mb and corresponds to the DM reference genome positions of 11.25 and 13.56 Mb.展开更多
基金supported by the Rural & Environment Science & Analytical Services (RESAS) Division of the Scottish Government through project JHI-B1-1the Biotechnology and Biological Sciences Research Council (BBSRC) through awards BB/ S015663/1+2 种基金BB/X009068/1Research Leaders 2025 fellowship funded by European Union’s Horizon 2020 research and innovation programme under Marie Sklodowska-Curie grant agreement no. 754380the Research/Scientific Computing teams at The James Hutton Institute and NIAB for providing computational resources and technical support for the “UK’s Crop Diversity Bioinformatics HPC” (BBSRC grant BB/ S019669/1)。
文摘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.
基金the Rural&Environment Science&Analytical Services(RESAS)Division of the Scottish Government(JHI-B1-1)the Biotechnology and Biological Sciences Research Council(BBSRC,BB/S015663/1)+3 种基金the Royal Society(NAF\R1\201061)the National Natural Science Foundation of China(32061130211,32372558)AK was supported through a Research Leaders 2025 fellowship funded by European Union’s Horizon 2020 Research and Innovation Programme under Marie Sklodowska-Curie Grant Agreement(754380)The authors acknowledge the Research/Scientific Computing teams at The James Hutton Institute and NIAB for providing computational resources and technical support for the‘‘UK’s Crop Diversity Bioinformatics HPC”(BBSRC Grant BB/S019669/1),use of which has contributed to the results reported within this paper.
文摘More than 170 years after causing the potato famine in Ireland,late blight is still considered one of the most devastating crop diseases.Commercial potato breeding efforts depend on natural sources of resistance to protect crops from the rapidly evolving late blight pathogen,Phytophthora infestans.We have identified and mapped a novel broad-spectrum disease resistance gene effective against P.infestans from the wild,diploid potato species Solanum bulbocastanum.Diagnostic resistance gene enrichment sequencing(dRenSeq)was used to confirm the uniqueness of the identified resistance.RenSeq and GenSeq-based mapping of the resistance,referred to as Rpi-blb4,alongside recombinant screening,positioned the locus responsible for the resistance to potato chromosome 5.The interval spans approximately 2.3 Mb and corresponds to the DM reference genome positions of 11.25 and 13.56 Mb.
