Authors:W.M. Du, F.H. Wang, H.Y. Zhang, B.Z. Jiang, X.Y. Chen, W. Zhang, Y. Xie, and Z.T. Sheng
Present research on prebiotics focuses on either polysaccharides or polyphenols. This study compared the individual and combined impact of polysaccharide, quercetin, and gallic acid (GA) treatment on three human faecal strains. In vitro pure culturing and correlation analysis confirmed that the growth of both beneficial microbe B. longum subsp. longum (0.695, 0.205: R2, slope, respectively) and pathogenic C. perfringens (0.712, 0.085: R2, slope, respectively) increased due to polysaccharide treatment, and only GA treatment would inhibit C. perfringens (0.789, –0.165: R2, slope, respectively) growth. In vivo studies also revealed that genome copies of Bifidobacterium increased and C. perfringens decreased in the faeces, when a blend of the three nutrients rather than single polysaccharide or polyphenols were fed to rats. These data suggested that combined prebiotic treatment improved human faecal strain composition better than single treatment.
Authors:Z.L. Li, H.Y. Li, G. Chen, X.J. Liu, C.L. Kou, S.Z. Ning, Z.W. Yuan, M. Hao, D.C. Liu, and L.Q. Zhang
Seven Glu-A1m allelic variants of the Glu-A1mx genes in Triticum monococcum ssp. monococcum, designated as 1Ax2.1a, 1Ax2.1b, 1Ax2.1c, 1Ax2.1d, 1Ax2.1e, 1Ax2.1f, and 1Ax2.1g were characterized. Their authenticity was confirmed by successful expression of the coding regions in E. coli, and except for the 1Ax2.1a with the presence of internal stop codons at position of 313 aa, all correspond to the subunit in seeds. However, all the active six genes had a same DNA size although their encoding subunits showed different molecular weight. Our study indicated that amino acid residue substitutions rather than previously frequently reported insertions/deletions played an important role on the subunit evolution of these Glu-A1mx alleles. Since variation in the Glu-A1x locus in common wheat is rare, these novel genes at the Glu-A1mx can be used as candidate genes for further wheat quality improvement.
Authors:L. Wei, S.G. Bai, X.J. Hou, J.M. Li, B. Zhang, W.J. Chen, D.C. Liu, B.L. Liu, and H.G. Zhang
Among 20 awnless Tibetan wheat cultivars analyzed by SDS-PAGE, the migration rate of an HMW-GS in XM001584 and XM001593, named 1BX23*. was shown to be slightly faster than 1Bx6. and slower than Bx7. Its nucleotide sequence was isolated based on homology clones. In a phylogenetic tree of 1Bx genes, 1Bx23* was apparently clustered with 1Bx23. Compared with 1Bx23. eight single nucleotide replacements caused four single amino acid replacements in 1Bx23*. The deletion of “G” at base pair 1463 and insertion of “A” at 1509 bps induced a 42-nucleotide frame shift. “GQRQQAGQWQRPGQ” was replaced by “DKGNRQDNGNDRDK”. The new segment cannot be found in other HMW-GSs, and it is very similar to a segment found in collagen. Moreover, an 18-nucleotide deletion made 1Bx23* six amino acids shorter than 1Bx23. The cultivar XM001593 had 28 chromosomes, which signifies that it was tetraploid wheat, and that the new HMW-GS 1Bx23* cannot be used directly for breeding in common wheat.
Authors:G.-F. Shi, Z.-J. Wang, G.-Y. Wang, R.-X. Yao, F.-W. Chen, and Z.-R. Shi
High-performance liquid chromatography-mass spectrometry (HPLC-MS) method coupled with radical reaction for screening active ingredients from perennial fujimoto bean whole herb was established. The active ingredients, present in perennial fujimoto bean whole herb, possess scavenging effects towards 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical, superoxide, peroxy radical, and hydroxyl radical. The radical scavenging abilities of these active ingredients were evaluated based on the relative peak areas in the HPLC chromatogram. The results indicate that potent antioxidants are present in the anhydrous methanol extract of perennial fujimoto bean whole herb. Based on HPLC-MS analysis, it was found that the scavenging ability can be mostly attributed to the presence of three compounds: cyanidin-3-o-β-d-glucopyranoside, troxerutin, and rutin. The structures were identified based on the MS and nuclear magnetic resonance (NMR) data. Free radical scavenging activity decreased in the following order: troxerutin > rutin > cyanidin-3-o-β-d-glucopyranoside.
Authors:Y.Q. Wang, X.J. Hou, B. Zhang, W.J. Chen, D.C. Liu, B.L. Liu, and H.G. Zhang
Red coleoptile is an easily observed agronomic trait of wheat and has been extensively studied. However, the molecular mechanism of this trait has not yet been revealed. In this study, the MYB gene TaMYB-D1 was isolated from the wheat cultivar ‘Gy115’, which possesses red coleoptiles. This gene resided at the short arm of the homoelogous group 7 chromosomes. TaMYB-D1 was the only gene expressed in the coleoptiles of ‘Gy115’ and was not expressed in ‘Opata’ and ‘CS’, which have uncoloured coleoptiles. Phylogenetic analysis placed TaMYB-D1 very close to ZmC1 and other MYB proteins regulating anthocyanin biosynthesis. The encoded protein of TaMYB-D1 had an integrated DNA binding domain of 102 amino acids and a transcription domain with 42 amino acids, similar to the structure of ZmC1. Transient expression analysis in onion epidermal cells showed that TaMYB-D1 was located at the plant nucleus, which suggested its role as a transcription factor. The expression of TaMYB-D1 was accompanied with the expression of TaDFR and anthocyanin biosynthesis in the development of the coleoptile of ‘Gy115’. Transient expression analysis showed that only TaMYB-D1 induced a few ‘Opata’ coleoptile cells to synthesize anthocyanin in light, and the gene also induced a colour change to red in many cells with the help of ZmR. All of these results suggested TaMYB-D1 as the candidate gene for the red coleoptile trait of ‘Gy115’.
Authors:S.F. Dai, D.Y. Xu, Z.J. Wen, Z.P. Song, H.X. Chen, H.Y Li, J.R. Li, L.Z. Kang, and Z.H. Yan
A novel 4.0-kb Fy was sequenced and bacterially expressed. This gene, the largest y-type HMW-GS currently reported, is 4,032-bp long and encodes a mature protein with 1,321 amino acid (AA) residues. The 4.0-kb Fy shows novel modifications in all domains. In the N-terminal, it contains only 67 AA residues, as three short peptides are absent. In the repetitive domain, the undecapeptide RYYPSVTSPQQ is completely lost and the dodecapeptide GSYYPGQTSPQQ is partially absent. A novel motif unit, PGQQ, is present in addition to the two standard motif units PGQGQQ and GYYPTSPQQ. Besides, an extra cysteine residue also occurs in the middle of this domain. The large molecular mass of the 4.0-kb Fy is mainly due to the presence of an extra-long repetitive domain with 1,279 AA residues. The novel 4.0-kb Fy gene is of interest in HMW-GS gene evolution as well as to wheat quality improvement with regard to its longest repetitive domain length and extra cysteines residues.
Premature termination codons (PTCs) are an important reason for the silence of highmolecular- weight glutenin subunits in Triticum species. Although the Glu-A1y gene is generally silent in common wheat, we here isolated an expressed Glu-A1y gene containing a PTC, named 1Ay8.3, from Triticum monococcum ssp. monococcum (AmAm, 2n = 2x = 14). Despite the presence of a PTC (TAG) at base pair positions 1879–1881 in the C-terminal coding region, this did not obviously affect 1Ay8.3 expression in seeds. This was demonstrated by the fact that when the PTC TAG of 1Ay8.3 was mutated to the CAG codon, the mutant in Escherichia coli bacterial cells expressed the same subunit as in the seeds. However, in E. coli, 1Ay8.3 containing the PTC expressed a truncated protein with faster electrophoretic mobility than that in seeds, suggesting that PTC translation termination suppression probably occurs in vivo (seeds) but not in vitro (E. coli). This may represent one of only a few reports on the PTC termination suppression phenomenon in genes.