The Histological Analysis To The Roots Of Setcreasea Purpurea Under Copper Stress And The Screening With Verification Of Related DEPs

Setcreasea purpurea Boom（Setcreasea purpurea Boom）is a copper hyperaccumulator.Setcreasea purpurea would express its protein specifically under copper stress culture.Therefore,TMT proteomics and transcriptome sequencing analysis were performed on the root tissues of Setcreasea purpurea under different copper concentrations.The differentially expressed proteins（DEPs）induced by copper at the protein level and transcription level were screened;bioinformatics methods were used to screen DEPs with consistent expression trends from the combined analysis of transcriptome and proteome,and PRM technical verification on DEPs was performed.The main results are as follows:1.Proteomic sequencing obtained 5718 proteins with quantitative information,of which 4163 differentially expressed proteins were identified.The 300μmol/L Cu²⁺stress group and 1000μmol/L Cu²⁺stress group were compared with the 0.25μmol/L Cu²⁺stress group（control group）,respectively,and 3603 and 3525 differentially expressed proteins were screened.In the analysis of GO function enrichment of differentially expressed proteins and KEGG metabolic pathway enrichment analysis,it is found that the functions of differential proteins were mainly enriched in oxidative stress,lipid metabolism,energy metabolism,transport,catalytic activity,and photosynthesis.2.Transcriptome sequencing obtained a total of 82471 high-quality unigenes.The 300μmol/L Cu²⁺stress group and the 1000μmol/L Cu²⁺stress group were compared with the0.25μmol/L Cu²⁺stress group（control group）,and a total of 5028 were screened.And 9982 differentially expressed genes（DEGs）,using q RT-PCR technology to perform fluorescence quantitative PCR verification on any 10 DEGs,indicating that the RNA-seq data is true and reliable.The transcriptome data and the proteome data were combined and analyzed,and 750 DEPs/DEGs with consistent expression trends were obtained.By bioinformatics analysis and screening,18 key proteins related to copper stress were obtained.3.A parallel reaction monitoring（PRM）experiment was performed on the 18differentially expressed proteins that were screened and compared with TMT quantitative proteomics.It was found that the overall consistency between TMT and PRM of the 18 differentially expressed proteins was higher,indicating these 18 differences The expressed protein is indeed the key protein of Setcreasea purpurea in response to copper stress.