8 research outputs found

    Differences in Grain Ultrastructure, Phytochemical and Proteomic Profiles between the Two Contrasting Grain Cd-Accumulation Barley Genotypes

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    <div><p>To reveal grain physio-chemical and proteomic differences between two barley genotypes, Zhenong8 and W6nk2 of high- and low- grain-Cd-accumulation, grain profiles of ultrastructure, amino acid and proteins were compared. Results showed that W6nk2 possesses significantly lower protein content, with hordein depicting the greatest genotypic difference, compared with Zhenong8, and lower amino acid contents with especially lower proportion of Glu, Tyr, Phe and Pro. Both scanning and transmission electron microscopy observation declared that the size of A-type starch molecule in W6nk2 was considerably larger than that of Zhenong8. Grains of Zhenong8 exhibited more protein-rich deposits around starch granules, with some A-type granules having surface pits. Seventeen proteins were identified in grains, using 2-DE coupled with mass spectrometry, with higher expression in Zhenong8 than that in W6nk2; including z-type serpin, serpin-Z7 and alpha-amylase/trypsin inhibitor CM, carbohydrate metabolism, protein synthesis and signal transduction related proteins. Twelve proteins were less expressed in Zhenong8 than that in W6nk2; including barley trypsin inhibitor chloroform/methanol-soluble protein (BTI-CMe2.1, BTI-CMe2.2), trypsin inhibitor, dehydroascorbate reductase (DHAR), pericentrin, dynein heavy chain and some antiviral related proteins. The data extend our understanding of mechanisms underlying Cd accumulation/tolerance and provides possible utilization of elite genetic resources in developing low-grain-Cd barley cultivars.</p></div

    Proteins whose expression were significantly higher expressed (+) in Zhenong8 compared with W6nk2 grains (Zhenong8 <i>vs</i> W6nk2).

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    [1]<p>Protein spot ID refers to numbers in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0079158#pone-0079158-g004" target="_blank">Figure 4</a>.</p>[2]<p>Accession number of top database match from the NCBInr database. Grain proteins induced in Zhenong8 <i>vs</i> W6nk2. Fold increase was calculated as Zhenong8/W6nk2 for higher expressed proteins in Zhenong8 compared with W6nk2 grains (Zhenong8 <i>vs</i> W6nk2). All ratios shown are statistically significant (<i>p</i><0.05).</p

    Representative 2-DE maps comparing two grain proteins of Zhenong8 (a) and W6nk2 (b).

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    <p>Total grain proteins were extracted and separated by 2-DE. In IEF, 90 µg of proteins were loaded onto pH 4–7 IPG strips (24 cm, linear). SDS-PAGE was performed with 12.5% gels. The spots were visualized by silver staining. Differentially accumulated protein spots are indicated by arrowheads. Seventeen higher expressed spots (U1∼U17) and 12 suppressed spots (D1∼D12) are indicated on the map of high-grain-Cd-accumulate genotype Zhenong8 (Zhenong8 <i>vs</i> W6nk2).</p

    Scanning electron microscopy (SEM) image of matured grains in two barley genotypes W6nk2 (left panel) and Zhenong8 (right panel).

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    <p>Photo is representative from six different experiments. <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0079158#pone-0079158-g003" target="_blank">Figure 3</a> (a)–(d), SEM image illustrating the starch–protein interface in Zhenong8 (b, d) compared with that exhibited by W6nk2 (a and c). Note the higher amount of associated protein in a, c, which surrounds the large A-type starch granules (S<sup>a</sup>) and engulfs the smaller B-type starch granules (S<sup>b</sup>). PD, protein deposits; PM, protein matrix among large and small starch granules. (e) and (f), SEM image of the nature fracture surface of aleurone layer, arrow shows aleurone grain; AC, aleurone cell. (g) and (h), SEM image of the epidermis of grain.</p

    Concentrations of Cd (a and c) and microelements (b and d) in grains of Zhenong8 and W6nk2 determined by flame atomic absorption spectrometry (a, b) and by ICP-OES (c, d). Means with the same letters are not significantly different at 0.05 level between the two genotypes.

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    <p>Concentrations of Cd (a and c) and microelements (b and d) in grains of Zhenong8 and W6nk2 determined by flame atomic absorption spectrometry (a, b) and by ICP-OES (c, d). Means with the same letters are not significantly different at 0.05 level between the two genotypes.</p

    The ‘spot view’ of grain proteins higher expressed or suppressed in Zhenong8 <i>vs</i> W6nk2.

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    <p>The areas in the arbitrary polygon of barley grain proteins from high-grain-Cd-accumulate genotype Zhenong8 have been enlarged and placed side by side with the corresponding areas of gel obtained from low-grain-Cd- accumulate genotype W6nk2. Protein spot ID refers to numbers in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0079158#pone-0079158-g005" target="_blank">Figure 5</a>.</p
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