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Method for Label-Free Quantitative Proteomics for Sorghum bicolor L. Moench

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Relation http://oar.icrisat.org/10608/
http://dx.doi.org/10.1007/s12042-018-9202-6
10.1007/s12042-018-9202-6
 
Title Method for Label-Free Quantitative Proteomics for Sorghum bicolor L. Moench
 
Creator Sharan, A A
Nikam, A N
Jaleel, A
Tamhane, V
Rao, P S
 
Subject Abiotic Stress
Biotic Stress
Sorghum
Genetics and Genomics
 
Description Sorghum (Sorghum bicolor L. Moench) is a rapidly emerging high biomass feedstock for bioethanol and lignocellulosic biomass production. The robust varietal germplasm of sorghum and its completed genome sequence provide the necessary genetic and molecular tools to study and engineer the biotic/abiotic stress tolerance. Traditional proteomics approaches for outlining the sorghum proteome have many limitations like, demand for high protein amounts, reproducibility and identification of only few differential proteins. In this study, we report a gel-free, quantitative proteomic method for in-depth coverage of the sorghum proteome. This novel method combining phenol extraction and methanol chloroform precipitation gives high total protein yields for both mature sorghum root and leaf tissues. We demonstrate successful application of this method in comparing proteomes of contrasting cultivars of sorghum, at two different phenological stages. Protein identification and relative quantification analyses were performed by a label-free liquid chromatography tandem mass spectrometry (LC/MS-MS) analyses. Several unique proteins were identified respectively from sorghum tissues, specifically 271 from leaf and 774 from root tissues, with 193 proteins common in both tissues. Using gene ontology analysis, the differential proteins identified were finely corroborated with their leaf/root tissue specific functions. This method of protein extraction and analysis would contribute substantially to generate in-depth differential protein data in sorghum as well as related species. It would also increase the repertoire of methods uniquely suited for gel-free plant proteomics that are increasingly being developed for studying abiotic and biotic stress responses.
 
Publisher Springer
 
Date 2018
 
Type Article
PeerReviewed
 
Format application/pdf
 
Language en
 
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Identifier http://oar.icrisat.org/10608/1/Method%20for%20Label-Free%20Quantitative%20Proteomics%20for%20Sorghu.pdf
Sharan, A A and Nikam, A N and Jaleel, A and Tamhane, V and Rao, P S (2018) Method for Label-Free Quantitative Proteomics for Sorghum bicolor L. Moench. Tropical Plant Biology (TSI). pp. 1-14. ISSN 1935-9756