TY - JOUR T1 - The value of genotype-specific reference for transcriptome analyses in barley JF - Life Science Alliance JO - Life Sci. Alliance DO - 10.26508/lsa.202101255 VL - 5 IS - 8 SP - e202101255 AU - Wenbin Guo AU - Max Coulter AU - Robbie Waugh AU - Runxuan Zhang Y1 - 2022/08/01 UR - https://www.life-science-alliance.org/content/5/8/e202101255.abstract N2 - It is increasingly apparent that although different genotypes within a species share “core” genes, they also contain variable numbers of “specific” genes and different structures of “core” genes that are only present in a subset of individuals. Using a common reference genome may thus lead to a loss of genotype-specific information in the assembled Reference Transcript Dataset (RTD) and the generation of erroneous, incomplete or misleading transcriptomics analysis results. In this study, we assembled genotype-specific RTD (sRTD) and common reference–based RTD (cRTD) from RNA-seq data of cultivated Barke and Morex barley, respectively. Our quantitative evaluation showed that the sRTD has a significantly higher diversity of transcripts and alternative splicing events, whereas the cRTD missed 40% of transcripts present in the sRTD and it only has ∼70% accurate transcript assemblies. We found that the sRTD is more accurate for transcript quantification as well as differential expression analysis. However, gene-level quantification is less affected, which may be a reasonable compromise when a high-quality genotype-specific reference is not available. ER -