Project description:Purpose: this study provided a comprehensive sequence for a systemic view of the transcriptome between mango leaf and fruit, as well as fruit allergens, which will be useful for further genomic research studies and breeding of lower allergenic mango cultivars. Methods:Some allergens have previously been identified in mango (Mangifera indica Linn), including profilins, Bet v 1-like proteins and chitinase. In this paper, 66 potential allergen genes were identified and their relative expressions evaluated in mango fruit and leaf using Illumina RNA-Seq technology. Results:A total of 17.63Gb Clean Data was obtained.The number of %≥Q30 was above 94.58%.RNA-Seq generated 11,751,123 contigs that were assembled into 99,328 unigenes with 16,848 unigenes of >1000 bp. A total of 230,242 unigenes were annotated using public protein databases, with a cut-off E-value above 10−5, of which 27,295, 46,030, 24,227 and 14,023 unigenes were assigned to gene ontology terms, Nr, Swiss-Prot and clusters of orthologous groups, respectively. Allergens mainly belonged to pollen allergen, pathogenesis-related protein Bet v I family and NADPH-dependent FMN reductase.
Project description:Significant progress has been observed in recent years in proteomics tools. However, minimal application of the newest proteomic tools has been massively applied in plant recalcitrant tissues in non-model fruits. This constant occurrence constrains the progress in understanding essential molecular processes during postharvest shelf life (PSL) of fleshy fruits. In fact, comparative proteomic studies in fruit like mango, guava, papaya, and avocado are minimal. Therefore, we set up a label free quantitation (LFQ) and TMT-SPS-MS3 pipeline for comparative study of mango peels during PSL. Our protocol included different protein extraction and peptide fractionation before mass spectrometry analysis. In doing so we were able to confidently identify 4,400 proteins with high confidence, among which 1087 were differentially accumulated. The combination of phenolic protein extraction, high-pH reverse-phase peptide fractionation and LFQ exhibited the best approach for relative quantification, but other protein extraction, peptide fractionation and data acquisition showed significant complementation. LFQ and Byonic also allowed the determination of intact N-glyco peptides on proteins such as the desiccation-related protein, which molecular dynamics analysis support the occurrence of N-glycans providing stability of the protein. Our study represents the first massive proteomics comparison of mango peels during PSL.
