Authors:J. Wu, H. Xing, D. Tang, Y. Gao, X. Yin, Q. Du, X. Jiang, and D. Yang
The method of high-performance liquid chromatography (HPLC) with diode array detector (DAD) was used and validated for the simultaneous determination of nine flavonoids (rutin, myricetin, quercitrin, quercetin, luteolin, genistein, kaempferol, apigenin, and isorhamnetin) in beagle dog plasma. Plasma sample was pre-treated with acetonitrile (containing 0.05% formic acid). Chromatographic separation was performed on a kromasil C18 column (250 × 4.6 mm, 5 µm) maintained at 35 °C. The mobile phase was a mixture of methanol and 0.2% formic acid with a step linear gradient. At 1.0 mL min−1 flow rate, the eluent of other eight flavonoids was detected simultaneously at 360 nm with good separation except genistein (detected at 254 nm). Under optimum conditions, the correlation coefficient between the peak area and the concentrations for each analyte was all above 0.999. The intra-day and inter-day precisions were less than 10% for all analytes. The limit of detection and the limit of quantification for the selected nine flavonoids were 0.006–0.03 and 0.02–0.12 g mL−1, respectively. The extracted recoveries of selected nine flavonoids were 74.02%–99.37%. The assay has been successfully applied to determine concentrations of nine flavonoids in plasma from beagle dog after being intravenously administrated Ginkgo biloba extract.
Authors:P.Y.Z. Jiang, Y.F. Tian, X.Y. Liu, L.L Wei, Y.X. Bai, X.L. Liu, and S.B. Li
Marine organisms have attracted considerable attention in recent years. In this study, peptides with osteogenic activity from Pinctada martensii were isolated and identified. Additionally, the effects of the hydrolysates on MC3T3-E1 cell proliferation and differentiation were evaluated using the MTT and alkaline phosphatase (ALP) assays, respectively. First, trypsin, pancreatin, and neutral protease were used to hydrolyse the intact shellfish. The hydrolysates with the greatest effects on osteoblast proliferation and ALP activity were separated and purified. Second, fraction WP2 was isolated and purified using a Sephadex G-25 column. WP2, which had the highest osteogenic activity, increased cell growth by 48.57 ± 0.05% and ALP activity by 6.27 ± 0.07 mU. Finally, four novel peptides were identified in WP2 (FDNEGKGKLPEEY, IVLDSGDGVTH, IVLDSGDGVSH, and SSENSDLQRQ) by Orbitrap Fusion Lumos Tribrid orbital liquid chromatography-mass spectrometry. Our findings revealed that P. martensii contains peptides with potential osteogenic activity.
Aegilops sharonensis (Sharon goatgrass) is a valuable source of novel high molecular weight glutenin subunits, resistance to wheat rust, powdery mildew, and insect pests. In this study, we successfully hybridized Ae. sharonensis as the pollen parent to common wheat and obtained backcross derivatives. F1 intergeneric hybrids were verified using morphological observation and cytological and molecular analyses. The phenotypes of the hybrid plants were intermediate between Ae. sharonensis and common wheat. Observations of mitosis in root tip cells and meiosis in pollen mother cells revealed that the F1 hybrids possessed 28 chromosomes. Chromosome pairing at metaphase I of the pollen mother cells in the F1 hybrid plants was low, and the meiotic configuration was 25.94 I + 1.03 II (rod). Two pairs of primers were screened out from 150 simple sequence repeat markers, and primer WMC634 was used to identified the presence of the genome of Ae. sharonensis. Sequencing results showed that the F1 hybrids contained the Ssh genome of Ae. sharonensis. The sodium dodecyl sulfate polyacrylamide gel electrophoresis profile showed that the alien high molecular weight glutenin subunits of Ae. sharonensis were transferred into the F1 and backcross derivatives. The new wheat-Ae. sharonensis derivatives that we have produced will be valuable for increasing resistance to various diseases of wheat and for improving the quality of bread wheat.
Authors:W. Li, Z.Y. Chen, Z. Li, X.F. Zhao, Z.E. Pu, G.Y. Chen, Q.T. Jiang, Y.M. Wei, and Y.L. Zheng
To study the development of starch granules in polyploid wheats, we investigated the expression of starch synthetic genes between the synthetic hexaploid wheat SHW-L1, its parents T. turgidum AS2255 and diploid Ae. tauschii AS60. The synthetic hexaploid wheat SHW-L1 showed significantly higher starch content and grain weight than its parents. Scanning electron microscopy (SEM) showed that SHW-L1 rapidly developed starch granules than AS2255 and AS60. The amount of B-type granule in AS60 was less than that in SHW-L1 and AS2255. RT-qPCR result showed that the starch synthetic genes AGPLSU1, AGPLSU2, AGPSSU1, AGPSSU2, GBSSI, SSIII, PHO1 and PHO2 expressed at earlier stages with larger quantity in SHW-L1 than in its parents during wheat grain development. The expression of the above mentioned genes in AS60 was slower than in SHW-L1 and AS2255. The expression pattern of starch synthase genes was also associated with the grain weight and starch content in all three genotypes. The results suggested that the synthetic hexaploid wheat inherited the pattern of starch granule development and starch synthase gene expression from tetraploid parent. The results suggest that tetraploid wheat could plays more important role for starch quality improvement in hexaploid wheat.
Authors:Q. Chen, J. Song, W.P. Du, L.Y. Xu, Y. Jiang, J. Zhang, M. Zhang, and G.R. Yu
Chinese endemic wheat landraces possess unique morphological features and desirable traits, useful for wheat breeding. It is important to clarify the relationship among these landraces. In this study, 21 accessions of the four Chinese endemic wheat landrace species were investigated using single-copy genes encoding plastid Acetyl-CoA carboxylase (Acc-1) and 3-phosphoglycerate kinase (Pgk-1) in order to estimate their phylogenetic relationship. Phylogenetic trees were constructed using maximum parsimony (MP), maximum likelihood (ML) and Bayesian, and TCS network and gene flow values. The A and B genome sequences from the Pgk-1 loci indicated that three accessions of Triticum petropavlovskyi were clustered into the same subclade, and the T. aestivum ssp. tibetanum and the Sichuan white wheat accessions were grouped into a separate subclade. Based on the Acc-1 gene, T. aestivum ssp. tibetanum and T. aestivum ssp. yunnanense were grouped into one subclade in the A genome; the B genome from T. petropavlovskyi and T. aestivum ssp. tibetanum, and the Sichuan white wheat complex and T. aestivum ssp. tibetanum were grouped in the same clades. The D genome of T. aestivum ssp. yunnanense clustered with T. petropavlovskyi. Our findings suggested that (1) T. petropavlovskyi is distantly related to the Sichuan white wheat complex; (2) T. petropavlovskyi, T. aestivum ssp. tibetanum and T. aestivum ssp. yunnanense are closely related; (3) T. aestivum ssp. tibetanum is closely related to T. aestivum ssp. yunnanense and the Sichuan white wheat complex; and (4) T. aestivum ssp. tibetanum may be an ancestor of Chinese endemic wheat landraces.
Authors:Y.-F. Yang, X.-Y. Lai, G.-L. Huang, Y.-H. Chen, X.-P. Du, Z.-D. Jiang, F. Chen, and H. Ni
Bee pollen is a health food with a wide range of nutritional and therapeutic properties. However, the bioactive compounds of bee pollen have not been extensively revealed due to low efficacy in separation. High-speed counter-current chromatography (HSCCC) and solvent extraction were applied to separate tyrosinase inhibitors from camellia pollen in this study. The camellia pollen extracts prepared with petroleum ether, ethyl acetate, and n-BuOH have tyrosinase inhibitory activity. Acidic hydrolysis could promote the tyrosinase inhibitory activity of crude sample. Three fractions with tyrosinase inhibitory activity were separated from the hydrolysate by a one-step HSCCC procedure. Among the fractions, two chemicals were sufficiently purified and identified to be levulinic acid (LA) and 5-hydroxymethylfurfural (5-HMF). The recovery was 0.80 g kg−1 pollen for LA and 1.75 g kg−1 pollen for 5-HMF; and their purity was all over 98%. The study demonstrates that HSCCC method is powerful for preparative separation of tyrosinase inhibitors from camellia pollen.
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:S. Wang, D. Chen, G. Guo, T. Zhang, S. Jiang, X. Shen, D. Perovic, S. Prodanovic, and Y. Yan
In this work, 9 novel LMW-GS genes (6 LMW-m and 3 LMW-i type) from 4 diploid and 1 tetraploid Aegilops species were amplified and cloned by allelic-specific PCR. Analysis of the deduced amino acid sequences showed that 7 and 2 LMW-GS had 9 and 7 cysteines, respectively. Four LMW-m type subunits genes had an extra cysteine at the C-terminal III, which could form intermolecular disulphide bonds to extend the chains, and therefore would facilitate to form larger gluten polymers. This suggested that these genes are expected to be used as candidate genes for wheat quality improvement. The correlation between specific N-terminal sequences and a decapeptide deletion in the C-terminal II in LMW-GS encoded by D genome was found. Particularly, if LMW-GS possessed a METRCIPG-N-terminal beginning sequences and a decapeptide (LGQCSFQQPQ) deletion in the C-terminal II, they could be encoded by D genome.
Authors:M.Y. Jiang, Z.R. Wang, K.W. Chen, J.Q. Kan, K.T. Wang, Zs. Zalán, F. Hegyi, K. Takács, and M.Y. Du
After suffering from mechanical injury and fungal infection, grapes are perishable. Botrytis cinerea, the causal agent of gray mould, is a critical pathogen for grapes. In this study, the inhibitory effect of Pseudomonas fluorescens on the formation of gray mould on grapes during the postharvest storage was investigated on “Kyoho” grape. The results suggest that a living cell suspension of P. fluorescens significantly inhibited spore germination of B. cinerea, and significantly reduced the incidence of grape gray mould. Moreover, compared with the control, the fruit inoculated with P. fluorescens had elevated activities of polyphenol oxidase (PPO), peroxidase (POD), catalase (CAT), phenylalanine ammonia-lyase (PAL), chitinase (CHI), and β-1,3-glucanase (GLU). Increase in enzyme activity correlated with enhanced host resistance. In addition, there was little difference in storage quality between the treated group and control group, indicating no adverse effects of the induced defence response on fruit quality.
Authors:W. Xiong, R.Q. Yan, Y.N. Liu, S.W. Peng, Z.Z. Jiang, X. Chai, A.D. Qi, and Y.F. Wang
Compound danshen preparations (CDPs) are used clinically for the treatment of cardiovascular and cerebrovascular diseases. By using the quantitative analysis of multi-components by single-marker (QAMS) method, sixteen compounds (danshensu, protocatechuic acid, protocatechuicaldehyde, caffeic acid, rosmarinic acid, lithospermic acid, notoginsenoside R1, salvianolic acid B, ginsenoside Rg1, ginsenoside Re, salvianolic acid A, salvianolic acid C, ginsenoside Rb1, ginsenoside Rd, cryptotanshinone, and tanshinone IIA were quantified on an ACQUITY ultraperformance liquid chromatography (UPLC) HSS T3 column (2.1 × 100 mm, 1.8 μm) with the mobile phase consisting of 0.1% formic acid aqueous solution (A) and acetonitrile (B) using a gradient elution at the flow rate of 0.30 mL/min in 30 min at 30°C, which was also validated by UPLC-diode array detection (DAD) and UPLC-electrospray ionization multistage/mass spectrometry (ESI-MS/MS) for assuring the feasibility and accuracy. Tested by robustness experiment under slightly changeable conditions, the stability of relative correction factor (RCF) proved to be stable, with RSDs below 5.69%, except for notoginsenoside R1 with relative standard deviation (RSD) 7.83%. This reliable and convenient QAMS method resolved the problem of standard substance insufficiency and improved the quality assessment of preparations consisting of complex compounds with different chemical structures, such as CDPs.