A high-performance liquid chromatography (HPLC) method has been developed for simultaneous determination of six alkaloids, i.e., (−)-(R)-platydesmin, noroxyhydrastinine, berberine, skimmianine, canthin-6-one, and pteleine in the herbal medicine of Phellodendron amurense Rupr. The optimal condition for extraction and separation was achieved with a linear mobile phase gradient prepared from 0.1% phosphoric acid and acetonitrile. The LODs and LOQs for the analytes ranged from 0.06 to 0.22 μg mL−1 and from 0.25 to 0.80 μg mL−1, respectively. The optimized method was applied to the determination of alkaloids in P. amurense Rupr. and was found to be efficient. This method can provide a scientific and technical platform to the manufacturers for setting up a quality control standard as well as to the public for quality and safety assurance of the proprietary traditional Chinese medicines.
Authors:Z. Xiao, D. Liu, C. Wang, Z. Cao, X. Zhan, Z. Yin, Q. Chen, H. Liu, F. Xu, and L. Sun
The effect of mechanical alloying on Zn-Sb alloy system is investigated with X-ray diffraction (XRD), laser grain size analysis
and differential scanning calorimetry (DSC) respectively. The results of laser particle size analysis shows that the particle
size decreases with increasing of the grinding time between 0 and 24 h. XRD and DSC results indicate that longer the grinding
time of Zn-Sb is, the more content of Zn4Sb3 become in the product in this process.
Authors:X.G. Hu, J. Liu, L. Zhang, B.H. Wu, J.L. Hu, D.C. Liu, and Y.L. Zheng
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.
Authors:L.J. Wu, Y. Shang, T. Liu, W.J. Chen, B.L. Liu, L.Q. Zhang, D.C. Liu, B. Zhang, and H.G. Zhang
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.
Authors:W. Zhou, X. Y. Zhang, Y. P. Lv, X. D. Liu, C. Xu, and G. L. Duan
Radix Isatidis has widely useful activities including anti-virus, anti-bacterial. Tryptanthrin, indigo, and indirubin are active ingredients in R. Isatidis. Response surface methodology (RSM)-optimized infrared-assisted extraction (IRAE) was developed and combined with HPLC for simultaneous determination of tryptanthrin, indigo, and indirubin from R. Isatidis. IRAE were investigated through extraction yields of the three components and optimized by RSM. The optimum conditions were as follows: infrared power of 129 W, solid/liquid ratio of 1:40 g/mL, and irradiation time of 22.5 min. IRAE conditions obtained by RSM were not only accurate, but also had practical value reflecting the expected optimization. Subsequently, this novel IRAE method was evaluated by extraction yield of the components of R. Isatidis samples from different regions. Compared with common extraction methods including maceration extraction (ME), reflux extraction (RE), ultrasound-assisted extraction (UAE), and microwave-assisted extraction (MAE), IRAE showed higher yield with advantages of no limitation of solvent selection, low cost, convenience under optimum extraction conditions. These results suggested the potential of RSM-optimized IRAE for extraction and analysis of the water-/fat-soluble compositions of Chinese herbal medicine. A simple chromatographic separation for simultaneous determination of tryptanthrin, indigo, and indirubin from Chinese herbal medicine R. Isatidis was performed on a C18 column (Diamonsil 150 mm × 4.6 mm i.d., 5 μm) with a mobile phase isocratic consisting of methanol and water at a flow-rate of 0.8 mL min−1. The retention times of tryptanthrin, indigo, and indirubin were 15.4, 31.9, and 58.6 min, respectively. The linear equations were obtained as follows: y = −3094.5744 + 21208.792x for tryptanthrin (R = 0.9998, 0.9–18.0 μg mL−1), y = 4730.0448 + 30180.567x for indigo (R = 0.9997, 0.5–10.0 μg mL−1) and y = −6582.9045 + 67069.312x for indirubin (R = 0.9997, 0.4–8.0 μg mL−1). The result showed that RSM-optimized IRAE was a simple, efficient pretreatment method for the analysis of complex matrix.
Authors:X. Luo, C. Han, X. Deng, D. Zhu, Y. Liu, and Y. Yan
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.
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: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.
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:J.Q. Xu, L. Wang, B.L. Liu, T.F. Xia, D.C. Liu, X. Chang, T.W. Zhang, H.G. Zhang, and Y.H. Shen
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.