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Wheat kernel morphology is a very important trait for wheat yield improvement. This is the first report of association analysis of kernel morphology traits in wheat breeding lines. In Qinghai, China, the research described here involved genome-wide association analysis in breeding lines derived from synthetic hexaploid wheat with a mixed linear model to identify the quantitative trait loci (QTLs) related to kernel morphology. The 8033 effective Diversity Array Technology (DArT) markers produced a genetic map of 5901.84 cM with an average density of 1.36 markers/cM. Population structure analysis classified 507 breeding lines into three groups by Bayesian structure analysis using unlinked markers. Linkage disequilibrium decay was observed with a map coverage of 2.78 cM. Marker-trait association analysis showed that 15 DArT markers for kernel morphology were detected, located on nine chromosomes, and explained 2.6%–4.0% of the phenotypic variation of kernel area (KA), kernel width (KW), kernel length (KL) and thousand-kernel weight (TKW). The marker 1139297 was related to both the KL and KA traits. Only six DArT markers were close to known QTLs. The parent SHW-L1 carried eight favored alleles, while other seven favored alleles were derived from elite common wheat cultivars. These QTLs, identified in elite breeding lines, should help us understand the kernel morphology trait better, and to provide germplasm for breeding new wheat cultivars for Qinghai Province or other regions.

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A novel HMW-GS of Dx5** with slightly faster migration rate than that of Dx5, was found in a Tibet bread wheat landrace using SDS-PAGE. Moreover, Dx5** is the subunit with the fastest migration rate in Glu-Dx locus. The gene for this subunit was isolated and its sequence was obtained in the present study. This gene was very similar to Dx5 both in nucleotide and deduced amino acid sequence. At the nucleotide sequence level, Dx5** different from Dx5 by the deletion of a 27 bp fragment and two nucleotide replacements at position 353(G/C) and 692(C/G), respectively. At the amino acid sequence level, Dx5** different from Dx5 by the deletion of a hexaploid (LGQGQQ) and a tripeptide (GQQ) repetitive motif and two amino acid replacements at position 118(C/S) and 231(A/G), respectively. These results suggested that the Dx5** was a derivation of Dx5 and was formed by replication slippage. Moreover, the specific cysteine (C) located at the beginning of the repetitive domain of Dx5, which proved to be critical for the end-use quality of wheat flours, was replaced by serine (S) in Dx5**.

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Thioacetamide (TAA) is a potent hepatotoxicant in acute and chronic hepatic injury. The study examined the protective effect of sesame oil against TAA-induced hepatic injury in rats. Hepatic injury was induced by intraperitoneal injection of 100 mg/kg of TAA for 24 h. Triple doses of sesame oil (1, 2, or 4 mL/kg) was given orally 0, 6, and 12 h after TAA treatment. TAA significantly increased serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels. Sesame oil decreased serum AST and ALT levels and significantly inhibited hepatic lipid peroxidation and nitric oxide levels compared with TAA-alone group. Further, sesame oil significantly inhibited TAA-induced hepatic neutrophil activation marker myeloperoxidase activity. However, sesame oil did not affect hepatic tumor necrosis factor, IL-1β and IL-10 generation in TAA-treated group. In conclusion, sesame oil protects against TAA-induced hepatic injury and oxidative stress via the inhibition of neutrophil activation. However, inflammatory cytokines may not be involved in sesame-oil-associated hepatic protection against TAA in rats.

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Grains of 12 accessions of Triticum timopheevii (Zhuk.) Zhuk. ssp. timopheevii (AAGG, 2n = 4x = 28) and one bread wheat cultivar Chinese Spring (CS) and one durum wheat cultivar Langdon (LDN) grown across two years were analyzed for grain iron (Fe) and zinc (Zn) concentrations. All the 12 tested T. timopheevii ssp. timopheevii genotypes showed significantly higher concentration of grain Fe and Zn than CS and LDN. Aboundant genetic variability of both the Fe and Zn concentrations was observed among the T. timopheevii ssp. timopheevii accessions, averagely varied from 47.06 to 90.26 mg kg−1 and from 30.05 to 65.91 mg kg−1, respectively. Their grain Fe and Zn concentrations between years exhibited a significantly positive correlation with the correlation coefficients r = 0.895 and r = 0.891, respectively, indicating the highly genetic stability. Flag leaf possessed twice or three times higher concentrations for both Fe and Zn than grain, and a significantly high positive correlation appeared between the two organs with r = 0.648 for Fe and r = 0.957 for Zn concentrations, respectively, suggesting flag leaves might be indirectly used for evaluating grain Zn and Fe contents. Significant correlations occurred between grain Fe and Zn concentrations, and between grain Zn concentration and the two agronomic traits of plant height and number of spikelets per spike. Both the concentrations were not related to seed size or weight as well as NAM-G1 gene, implying the higher grain Fe and Zn concentrations of T. timopheevii ssp. timopheevii species are not ascribed to concentration effects of seed and the genetic control of NAM-G1 gene. There might be some other biological factors impacting the grain’s Zn and Fe concentrations. These results indicated T. timopheevii ssp. timopheevii species might be a promising genetic resource with high Fe and Zn concentrations for the biofortification of current wheat cultivars.

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In this study, the cDNA of homocysteine S-methyltransferase was isolated from Aegilops tauschii Coss., with the gene accordingly designated as AetHMT1. Similar to other methyltransferases, AetHMT1 contains a GGCCR consensus sequence for a possible zinc-binding motif near the C-terminal and a conserved cysteine residue upstream of the zinc-binding motif. Analysis of AetHMT1 uncovered no obvious chloroplast or mitochondrial targeting sequences. We functionally expressed AetHMT1 in Escherichia coli and confirmed its biological activity, as evidenced by a positive HMT enzyme activity of 164.516 ± 17.378 nmol min−1 mg−1 protein when catalyzing the transformation of L-homocysteine. Compared with the bacterium containing the empty vector, E. coli harboring the recombinant AetHMT1 plasmid showed much higher tolerance to selenate and selenite. AetHMT1 transcript amounts in different organs were increased by Na2SeO4 treatment, with roots accumulating higher amounts than stems, old leaves and new leaves. We have therefore successfully isolated HMT1 from Ae. tauschii and characterized the biochemical and physiological functions of the corresponding protein.

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Seed germination is a new beginning for the crop life cycle, which is closely related to seed sprouting and subsequent plant growth and development, and ultimately affects grain yield and quality. Salt stress is one of the most important abiotic stress factors that restrict crop production. Therefore, it is highly important to improve crop salt tolerance and sufficient utilization of saline-alkali land. In this study, we identified the phosphorylated proteins involved in salt stress response by combining SEM, 2-DE, Pro-Q Diamond staining and tandem mass spectrometry. The results showed that salt stress significantly inhibited seed germination and starch degradation. In total, 14 phosphorylated protein spots (11 unique proteins) in the embryo and 6 phosphorylated protein spots (4 unique proteins) in the endosperm were identified, which mainly involved in stress/defense, protein metabolism and energy metabolism. The phosphorylation of some proteins such as cold regulated proteins, 27K protein, EF-1β and superoxide dismutase could play important roles in salt stress tolerance.

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Cereal Research Communications
Authors: Z. L. Li, D. D. Wu, H. Y. Li, G. Chen, W. G. Cao, S. Z. Ning, D. C. Liu, and L. Q. Zhang

Gliadin is a main component of gluten proteins that affect functional properties of bread making and contributes to the viscous nature of doughs. In this study, thirteen novel ω-gliadin genes were identified in several Triticum species, which encode the ARH-, ATDand ATN-type proteins. Two novel types of ω-gliadins: ATD- and ATN- have not yet been reported. The lengths of 13 sequences were ranged from 927 to 1269 bp and the deduced mature proteins were varied from 309 to 414 residues. All 13 genes were pseudogenes because of the presence of internal stop codons. The primary structure of these ω-gliadin genes included a signal peptide, a conserved N-terminal domain, a repetitive domain and a conserved C-terminus. In this paper, we first characterize ω-gliadin genes from T. timopheevi ssp. timopheevi and T. timopheevi ssp. araraticum. The ω-gliadin gene variation and the evolutionary relationship of ω-gliadin family genes were also discussed.

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As one of the world’s earliest domesticated crops, barley is a model species for the study of evolution and domestication. Domestication is an evolutionary process whereby a population adapts, through selection; to new environments created by human cultivation. We describe the genome-scanning of molecular diversity to assess the evolution of barley in the Tibetan Plateau. We used 667 Diversity Arrays Technology (DArT) markers to genotype 185 barley landraces and wild barley accessions from the Tibetan Plateau. Genetic diversity in wild barley was greater than in landraces at both genome and chromosome levels, except for chromosome 3H. Landraces and wild barley accessions were clearly differentiated genetically, but a limited degree of introgression was still evident. Significant differences in diversity between barley subspecies at the chromosome level were observed for genes known to be related to physiological and phenotypical traits, disease resistance, abiotic stress tolerance, malting quality and agronomic traits. Selection on the genome of six-rowed naked barley has shown clear multiple targets related to both its specific end-use and the extreme environment in Tibet. Our data provide a platform to identify the genes and genetic mechanisms that underlie phenotypic changes, and provide lists of candidate domestication genes for modified breeding strategies.

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Cereal Research Communications
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-A1 m allelic variants of the Glu-A1 m x genes in Triticum monococcum ssp. monococcum, designated as 1Ax2.1 a, 1Ax2.1 b, 1Ax2.1 c, 1Ax2.1 d, 1Ax2.1 e, 1Ax2.1 f, and 1Ax2.1 g were characterized. Their authenticity was confirmed by successful expression of the coding regions in E. coli, and except for the 1Ax2.1 a 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-A1 m x alleles. Since variation in the Glu-A1x locus in common wheat is rare, these novel genes at the Glu-A1 m x can be used as candidate genes for further wheat quality improvement.

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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’.

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