Conservation analysis of the COL2A1 p.Gly207 amino acid residue.

<p>Conservation analysis of the COL2A1 p.Gly207 amino acid residue.</p

Heterozygous c.620G>A (p.Gly207Glu) mutation in the <i>COL2A1</i> gene.

<p>Heterozygous c.620G>A (p.Gly207Glu) mutation in the <i>COL2A1</i> gene.</p

The schematic structure and the mutations of the human myosin XVa.

<p>The myosin XVa consists of 3530 amino acids, including an N-terminal extension domain and Motor domain, two light chain binding IQ motifs, two myosin-tail homology 4 (MyTH4) domains and band 4.1/ezrin/radixin/moesin (FERM) domains, a Src-homology-3 (SH3) domain and a C-terminal class I PDZ-ligand domain. The novel <i>MYO15A</i> mutation in this study is showed with red box at the bottom of the figure, and previously reported mutations are displayed at the top of the figure. <i>MYO15A</i>, the myosin XVa gene.</p

Identification of Leaf Proteins Differentially Accumulated between Wheat Cultivars Distinct in Their Levels of Drought Tolerance

<div><p>The drought-tolerant ‘Ningchun 47’ (NC47) and drought-sensitive ‘Chinese Spring’ (CS) wheat (<i>Triticum aestivum</i> L.) cultivars were treated with different PEG6000 concentrations at the three-leaf stage. An analysis on the physiological and proteomic changes of wheat seedling in response to drought stress was performed. In total, 146 differentially accumulated protein (DAP) spots were separated and recognised using two-dimensional gel electrophoresis. In total, 101 DAP spots representing 77 unique proteins were identified by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry. These proteins were allocated to 10 groups according to putative functions, which were mainly involved in carbon metabolism (23.4%), photosynthesis/respiration (22.1%) and stress/defence/detoxification (18.2%). Some drought stress-related proteins in NC47, such as enolase, 6-phosphogluconate dehydrogenase, Oxygen-evolving enhancer protein 2, fibrillin-like protein, 2-Cys peroxiredoxin BAS1 and 70-kDa heat shock protein, were more upregulated than those in CS. Multivariate principal components analysis revealed obvious differences between the control and treatments in both NC47 and CS, while cluster analysis showed that the DAPs displayed five and six accumulation patterns in NC47 and CS, respectively. Protein–protein interaction network analysis showed that some key DAPs, such as 2-Cys peroxiredoxin BAS1, RuBisCO large subunit-binding protein, 50S ribosomal protein L1, 6-phosphogluconate dehydrogenase, glyceraldehyde 3-phosphate dehydrogenase isoenzyme and 70-kDa heat shock protein, with upregulated accumulation in NC47, had complex interactions with other proteins related to amino acid metabolism, carbon metabolism, energy pathway, signal transduction, stress/defence/detoxification, protein folding and nucleotide metabolism. These proteins could play important roles in drought-stress tolerance and contribute to the relatively stronger drought tolerance of NC47.</p></div

Pedigree and sequence analysis of an ARNSHL family.

<p>(A) Pedigree of the ARNSHL family. N, normal; M, the <i>MYO15A</i> c.9316dupC variant. (B) The homozygous <i>MYO15A</i> c.9316dupC variant of the affected individual (IV:2). (C) The heterozygous <i>MYO15A</i> c.9316dupC variant of the unaffected individual (III:1). (D) The <i>MYO15A</i> gene sequence of a normal control. ARNSHL, autosomal recessive nonsyndromic hearing loss; <i>MYO15A</i>, the myosin XVa gene.</p

Table_1.xlsx

<p>In this study, we performed the first comparative proteomic analysis of wheat flag leaves and developing grains in response to drought stress. Drought stress caused a significant decrease in several important physiological and biochemical parameters and grain yield traits, particularly those related to photosynthesis and starch biosynthesis. In contrast, some key indicators related to drought stress were significantly increased, including malondialdehyde, soluble sugar, proline, glycine betaine, abscisic acid content, and peroxidase activity. Two-dimensional difference gel electrophoresis (2D-DIGE) identified 87 and 132 differentially accumulated protein (DAP) spots representing 66 and 105 unique proteins following exposure to drought stress in flag leaves and developing grains, respectively. The proteomes of the two organs varied markedly, and most DAPS were related to the oxidative stress response, photosynthesis and energy metabolism, and starch biosynthesis. In particular, DAPs in flag leaves mainly participated in photosynthesis while those in developing grains were primarily involved in carbon metabolism and the drought stress response. Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) further validated some key DAPs such as rubisco large subunit (RBSCL), ADP glucose pyrophosphorylase (AGPase), chaperonin 60 subunit alpha (CPN-60 alpha) and oxalate oxidase 2 (OxO 2). The potential functions of the identified DAPs revealed that a complex network synergistically regulates drought resistance during grain development. Our results from proteome perspective provide new insight into the molecular regulatory mechanisms used by different wheat organs to respond to drought stress.</p

Morphological and physiological changes in seedlings of two wheat (<i>Triticum aestivum</i> L.) cultivars, ‘Chinese Spring’ (CS) and ‘Ningchun 47’ (NC47), during 48 h of PEG-mediated drought stress.

<p>(A) Morphological changes of wheat seedlings in response to different PEG6000 concentration treatments. (B) Leaf relative water content (RWC) analysis. (C) Proline content analysis. (D) Soluble sugar content analysis.</p

Hierarchical cluster analysis of DAP spots in NC47 and CS.

<p>Red colour indicates a positive abundance in protein spots; green colour denotes a negative abundance in protein spots. (A) Cluster analysis of 69 differentially accumulated proteins in NC47. (B) Cluster analysis of 77 differentially accumulated proteins in CS. 0, 15%, 20%, 25% and 30% represent the PEG6000 concentration treatment.</p

Principal components analysis (PCA) of the set of 101 DAP spots in NC47 and CS.

<p>(A) PCA of individual experimental samples in NC47. (B) PCA of 69 differentially accumulated protein spots in NC47. (C) PCA of individual experimental samples in CS. (D) PCA of 77 differentially accumulated protein spots in CS. Red spots indicate an upregulated protein; green spots indicate a downregulated protein. CS-0, CS-15, CS-20, CS-25 and CS-30 represent the 0, 15%, 20%, 25% and 30%, respectively, of the PEG6000 concentration treatment in CS. NC47-0, NC47-15, NC47-20, NC47-25 and NC47-30 represent the 0, 15%, 20%, 25% and 30%, respectively, of the PEG6000 concentration treatment in NC47.</p

Image_1.pdf

<p>In this study, we performed the first comparative proteomic analysis of wheat flag leaves and developing grains in response to drought stress. Drought stress caused a significant decrease in several important physiological and biochemical parameters and grain yield traits, particularly those related to photosynthesis and starch biosynthesis. In contrast, some key indicators related to drought stress were significantly increased, including malondialdehyde, soluble sugar, proline, glycine betaine, abscisic acid content, and peroxidase activity. Two-dimensional difference gel electrophoresis (2D-DIGE) identified 87 and 132 differentially accumulated protein (DAP) spots representing 66 and 105 unique proteins following exposure to drought stress in flag leaves and developing grains, respectively. The proteomes of the two organs varied markedly, and most DAPS were related to the oxidative stress response, photosynthesis and energy metabolism, and starch biosynthesis. In particular, DAPs in flag leaves mainly participated in photosynthesis while those in developing grains were primarily involved in carbon metabolism and the drought stress response. Western blotting and quantitative real-time polymerase chain reaction (qRT-PCR) further validated some key DAPs such as rubisco large subunit (RBSCL), ADP glucose pyrophosphorylase (AGPase), chaperonin 60 subunit alpha (CPN-60 alpha) and oxalate oxidase 2 (OxO 2). The potential functions of the identified DAPs revealed that a complex network synergistically regulates drought resistance during grain development. Our results from proteome perspective provide new insight into the molecular regulatory mechanisms used by different wheat organs to respond to drought stress.</p