PXD009708

PXD009708 is an original dataset announced via ProteomeXchange.

Title	Using RT-qPCR, proteomics and microscopy to unravel the spatio-temporal expression and subcellular localization of hordoindolines across development in barley endosperm
Description	Hordeum vulgare (barley) hordoindolines (HINs), HINa, HINb1 and HINb2, are orthologous proteins of wheat puroindolines (PINs) that are small, basic, cysteine-rich seed-specific proteins and responsible for grain hardness. Grain hardness, is, next to its protein content, a major quality trait. In barley, HINb is most highly expressed in the mid-stage developed endosperm and is associated with both major endosperm texture and grain hardness. However, data required tounderstand the spatio-temporal dynamics of HIN transcripts and HIN protein regulation during grain filling processes are missing. Using reverse transcription quantitative PCR (RT-qPCR) and proteomics we analyzed HIN transcript and HIN protein abundance from whole seeds (WSs) at four ((6 days after pollination (dap), 10 dap, 12 dap and ≥ 20 dap)) as well as from aleurone, subaleurone and starchy endosperm at two (12 dap and ≥ 20 dap) developmental stages. At the WS level, results from RT-qPCR, proteomics and western blot showed a continuous increase of HIN transcript and HIN protein abundance across these four developmental stages. Miroscopic studies revealed HIN localization mainly at the vacuolar membrane in the aleurone, at protein bodies (PBs) in subaleurone and at the periphery of starch granules in the starchy endosperm. Laser microdissetion (LMD) proteomic analyses identified HINb2 as the most prominent HIN protein in starchy endosperm at ≥ 20 dap. Additionally, our quantification data revealed a poor correlation between transcript and protein levels of HINs in subaleurone during development. Here, we correlated data achieved by RT-qPCR, proteomics and microscopy that reveal different expression and localization pattern of HINs in each layer during barley endosperm development. This indicats a contribution of each tissue to the regulation of HINs during grain filling. The effect of the high protein abundance of HINs in the starchy endosperm and their localization at the periphery of starch granules at late development stages at the high end-product quality is discussed. Understanding the spatio-temporal regulated HINs is essential to improve barley quality traits for high end-product quality, as hard texture of the barley grain is regulated by the ratio between HINb/HINa.
HostingRepository	PRIDE
AnnounceDate	2018-07-02
AnnouncementXML	Submission_2018-10-01_08:33:49.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Valentin Roustan
SpeciesList	scientific name: Hordeum vulgare (Barley); NCBI TaxID: 4513;
ModificationList	monohydroxylated residue; acetylated residue; iodoacetamide derivatized residue
Instrument	LTQ Orbitrap Elite

Revision	Datetime	Status	ChangeLog Entry
0	2018-05-08 02:21:53	ID requested
1	2018-07-02 01:12:09	announced
⏵ 2	2018-10-01 08:33:51	announced	Updated publication reference for PubMed record(s): 29951075, 30250475.

Shabrangy A, Roustan V, Reipert S, Weidinger M, Roustan PJ, Stoger E, Weckwerth W, Ibl V, Using RT-qPCR, Proteomics, and Microscopy to Unravel the Spatio-Temporal Expression and Subcellular Localization of Hordoindolines Across Development in Barley Endosperm. Front Plant Sci, 9():775(2018) [pubmed]

Roustan V, Roustan PJ, Weidinger M, Reipert S, Kapusi E, Shabrangy A, Stoger E, Weckwerth W, Ibl V, Microscopic and Proteomic Analysis of Dissected Developing Barley Endosperm Layers Reveals the Starchy Endosperm as Prominent Storage Tissue for ER-Derived Hordeins Alongside the Accumulation of Barley Protein Disulfide Isomerase (HvPDIL1-1). Front Plant Sci, 9():1248(2018) [pubmed]

curator keyword: Biological

submitter keyword: Barley caryopsis

Hordoindoline, spatio-temporal

laser microdissection

endosperm

label-free shotgun proteomics

RT-qPCR

reference genes

end-product quality, Golden Promise

protein bodies

grain hardness

Verena Ibl
contact affiliation	Department of Ecogenomics and Systems Biology, University of Vienna, Althanstrasse 14, 1090 Vienna, Austria
contact email	verena.ibl@univie.ac.at
lab head
Valentin Roustan
contact affiliation	University of Vienna
contact email	valentin.roustan@univie.ac.at
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2018/07/PXD009708

PRIDE project URI

• PRIDE
  1. PXD009708
     1. Label: PRIDE project
     2. Name: Using RT-qPCR, proteomics and microscopy to unravel the spatio-temporal expression and subcellular localization of hordoindolines across development in barley endosperm