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pmid:27490537

Drought-Induced Leaf Proteome Changes in Switchgrass Seedlings.

Zhujia Ye · Sasikiran Sangireddy · Ikenna Okekeogbu · Suping Zhou · Chih-Li Yu · Dafeng Hui · Kevin J Howe · Tara Fish · Theodore W Thannhauser

International journal of molecular sciences · 2016 doi:10.3390/ijms17081251

Paper

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Experiment

complete

Data

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Analysis

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Drought-Induced Leaf Proteome Changes in Switchgrass Seedlings.
doi
10.3390/ijms17081251
pmid
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Zhujia Ye
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Sasikiran Sangireddy
[2]
Ikenna Okekeogbu
[3]
Suping Zhou
[4]
Chih-Li Yu
[5]
Dafeng Hui
[6]
Kevin J Howe
[7]
Tara Fish
[8]
Theodore W Thannhauser
journal
International journal of molecular sciences
year
2016
abstract
Switchgrass (Panicum virgatum) is a perennial crop producing deep roots and thus highly tolerant to soil water deficit conditions. However, seedling establishment in the field is very susceptible to prolonged and periodic drought stress. In this study, a "sandwich" system simulating a gradual water deletion process was developed. Switchgrass seedlings were subjected to a 20-day gradual drought treatment process when soil water tension was increased to 0.05 MPa (moderate drought stress) and leaf physiological properties had expressed significant alteration. Drought-induced changes in leaf proteomes were identified using the isobaric tags for relative and absolute quantitation (iTRAQ) labeling method followed by nano-scale liquid chromatography mass spectrometry (nano-LC-MS/MS) analysis. Additionally, total leaf proteins were processed using a combinatorial library of peptide ligands to enrich for lower abundance proteins. Both total proteins and those enriched samples were analyzed to increase the coverage of the quantitative proteomics analysis. A total of 7006 leaf proteins were identified, and 257 (4% of the leaf proteome) expressed a significant difference (p < 0.05, fold change <0.6 or >1.7) from the non-treated control to drought-treated conditions. These proteins are involved in the regulation of transcription and translation, cell division, cell wall modification, phyto-hormone metabolism and signaling transduction pathways, and metabolic pathways of carbohydrates, amino acids, and fatty acids. A scheme of abscisic acid (ABA)-biosynthesis and ABA responsive signal transduction pathway was reconstructed using these drought-induced significant proteins, showing systemic regulation at protein level to deploy the respective mechanism. Results from this study, in addition to revealing molecular responses to drought stress, provide a large number of proteins (candidate genes) that can be employed to improve switchgrass seedling growth and establishment under soil drought conditions (Data are available via ProteomeXchange with identifier PXD004675).
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Then, log2 fold values from all constituent peptides were subjected to t-test (general linear model procedure) followed by false discovery rate (FDR) corrections to test the statistical significance of the difference in normalized abundance of each protein between the drought-treated and control sample groups
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sections[18] "4.5. Isobaric Tags for Relative and Absolute Quantification (iTRAQ) Labeling and Mass Spectrometry Analysis" paragraph 4
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The mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE [92] partner repository with the dataset identifier PXD004675 and 10.6019/PXD004675.
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followed by nano-scale liquid chromatography mass spectrometry (nano-LC-MS/MS) analysis. Additionally, tota
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However, seedling establishment in the field is very susceptible to prolonged and periodic drought stress. In this study, a \"sandwich\" system simulating a gradual water deletion process was developed. Switchgrass seedlings were subjected to a 20-day gradual drought treatment process when soil water tension was increased to 0.05 MPa (moderate drought stress) and leaf physiological properties had expressed significant alteration. Drought-induced changes in leaf proteomes were identified using the isobaric tags for relative and absolute quantitation (iTRAQ) labeling method followed by nano-scale liquid chromatography mass spectrometry (nano-LC-MS/MS) analysis. Additionally, total leaf proteins were processed using a combinatorial library of peptide ligands to enrich for lower abundance proteins. Both total proteins and those enriched samples were analyzed to increase the coverage of the quantitative proteomics analysis. 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