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      Comparative Proteomic Analysis of Walnut ( Juglans regia L.) Pellicle Tissues Reveals the Regulation of Nut Quality Attributes

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          Abstract

          Walnuts ( Juglans regia L.) are a valuable dietary source of polyphenols and lipids, with increasing worldwide consumption. California is a major producer, with ’Chandler’ and ’Tulare’ among the cultivars more widely grown. ’Chandler’ produces kernels with extra light color at a higher frequency than other cultivars, gaining preference by growers and consumers. Here we performed a deep comparative proteome analysis of kernel pellicle tissue from these two valued genotypes at three harvest maturities, detecting a total of 4937 J. regia proteins. Late and early maturity stages were compared for each cultivar, revealing many developmental responses common or specific for each cultivar. Top protein biomarkers for each developmental stage were also selected based on larger fold-change differences and lower variance among replicates, including proteins for biosynthesis of lipids and phenols, defense-related proteins and desiccation stress-related proteins. Comparison between the genotypes also revealed the common and specific protein repertoires, totaling 321 pellicle proteins with differential abundance at harvest stage. The proteomics data provides clues on antioxidant, secondary, and hormonal metabolism that could be involved in the loss of quality in the pellicles during processing for commercialization.

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          MetaboAnalyst 4.0: towards more transparent and integrative metabolomics analysis

          Abstract We present a new update to MetaboAnalyst (version 4.0) for comprehensive metabolomic data analysis, interpretation, and integration with other omics data. Since the last major update in 2015, MetaboAnalyst has continued to evolve based on user feedback and technological advancements in the field. For this year's update, four new key features have been added to MetaboAnalyst 4.0, including: (1) real-time R command tracking and display coupled with the release of a companion MetaboAnalystR package; (2) a MS Peaks to Pathways module for prediction of pathway activity from untargeted mass spectral data using the mummichog algorithm; (3) a Biomarker Meta-analysis module for robust biomarker identification through the combination of multiple metabolomic datasets and (4) a Network Explorer module for integrative analysis of metabolomics, metagenomics, and/or transcriptomics data. The user interface of MetaboAnalyst 4.0 has been reengineered to provide a more modern look and feel, as well as to give more space and flexibility to introduce new functions. The underlying knowledgebases (compound libraries, metabolite sets, and metabolic pathways) have also been updated based on the latest data from the Human Metabolome Database (HMDB). A Docker image of MetaboAnalyst is also available to facilitate download and local installation of MetaboAnalyst. MetaboAnalyst 4.0 is freely available at http://metaboanalyst.ca.
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            A Cross-platform Toolkit for Mass Spectrometry and Proteomics

            Mass-spectrometry-based proteomics has become an important component of biological research. Numerous proteomics methods have been developed to identify and quantify the proteins in biological and clinical samples 1 , identify pathways affected by endogenous and exogenous perturbations 2 , and characterize protein complexes 3 . Despite successes, the interpretation of vast proteomics datasets remains a challenge. There have been several calls for improvements and standardization of proteomics data analysis frameworks, as well as for an application-programming interface for proteomics data access 4,5 . In response, we have developed the ProteoWizard Toolkit, a robust set of open-source, software libraries and applications designed to facilitate proteomics research. The libraries implement the first-ever, non-commercial, unified data access interface for proteomics, bridging field-standard open formats and all common vendor formats. In addition, diverse software classes enable rapid development of vendor-agnostic proteomics software. Additionally, ProteoWizard projects and applications, building upon the core libraries, are becoming standard tools for enabling significant proteomics inquiries.
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              Target-decoy search strategy for increased confidence in large-scale protein identifications by mass spectrometry.

              Liquid chromatography and tandem mass spectrometry (LC-MS/MS) has become the preferred method for conducting large-scale surveys of proteomes. Automated interpretation of tandem mass spectrometry (MS/MS) spectra can be problematic, however, for a variety of reasons. As most sequence search engines return results even for 'unmatchable' spectra, proteome researchers must devise ways to distinguish correct from incorrect peptide identifications. The target-decoy search strategy represents a straightforward and effective way to manage this effort. Despite the apparent simplicity of this method, some controversy surrounds its successful application. Here we clarify our preferred methodology by addressing four issues based on observed decoy hit frequencies: (i) the major assumptions made with this database search strategy are reasonable; (ii) concatenated target-decoy database searches are preferable to separate target and decoy database searches; (iii) the theoretical error associated with target-decoy false positive (FP) rate measurements can be estimated; and (iv) alternate methods for constructing decoy databases are similarly effective once certain considerations are taken into account.
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                Author and article information

                Journal
                Life (Basel)
                Life (Basel)
                life
                Life
                MDPI
                2075-1729
                27 November 2020
                December 2020
                : 10
                : 12
                : 314
                Affiliations
                [1 ]Department of Plant Sciences, University of California, Davis, CA 95616, USA; pazaini@ 123456ucdavis.edu (P.A.Z.); ngfeinberg@ 123456ucdavis.edu (N.G.F.); hsaxe@ 123456ucdavis.edu (H.J.S.); chcrisosto@ 123456ucdavis.edu (C.H.C.)
                [2 ]Department of Food Sciences and Technology, University of California, Davis, CA 95616, USA; fsgrilo@ 123456ucdavis.edu
                [3 ]Proteomics Core Facility, University of California, Davis, CA 95616, USA; msalemi@ 123456ucdavis.edu (M.R.S.); bsphinney@ 123456ucdavis.edu (B.S.P.)
                Author notes
                Author information
                https://orcid.org/0000-0003-3535-4776
                https://orcid.org/0000-0002-0431-4037
                https://orcid.org/0000-0003-3870-3302
                https://orcid.org/0000-0001-7925-4086
                Article
                life-10-00314
                10.3390/life10120314
                7760677
                33261033
                0798db9f-0e8b-4054-9ec5-8cde1869096f
                © 2020 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 02 November 2020
                : 25 November 2020
                Categories
                Article

                walnut kernel,pellicle color,lc-ms/ms proteomics,kernel quality,plant genome-wide proteome,small heat shock proteins,fatty acid metabolism,seed coat pigmentation

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