Authors:W. N. Zhou, J. Ouyang, Z. H. Wang, X. Y. Wang, Y. R. Suo, Z. Zhang and H. L. Wang
Elsholtzia densa Benth. var. densa (Lamiaceae) is a famous medicinal herb which has been widely used for treatment of colds, headaches, pharyngitis, fever, diarrhea, digestion disorder, rheumatic arthritis, nephritises, and nyctalopia in China. In this study, fraction of the ethyl alcohol extract of E. densa (aerial part) by different polarity solvents indicated that the ethyl acetate soluble fraction exhibited a potent 1,1-diphenyl-2-picryhydrazyl (DPPH) radical scavenging activity with the IC50 value of 148.2 μg/mL. Under the target guidance of DPPH experiment, isoquercitrin, trachelogenin, ethyl caffeate, and arctigenin were separated with purities 95.98%, 92.98%, 96.07%, and 88.83%, respectively, by a dual-mode high-speed counter-current chromatography (HSCCC) method using n-hexane–ethyl acetate–methanol–water (4.5:5:3:4, v/v/v/v) as the solvent system. In order to evaluate the scientific basis, antioxidant activity of four isolated compounds was assessed by the radical scavenging effect on DPPH radical; isoquercitrin and ethyl caffeate showed stronger antioxidant activities with IC50 values of 9.4 μg/mL and 9.2 μg/mL, respectively, while trachelogenin and arctigenin showed weak antioxidant activities with IC50 values of >500 μg/mL and 72.8 μg/mL, respectively. Results of the present study indicated that the combinative method using DPPH antioxidant assay and dual-mode HSCCC could be widely applied for rapid screening and isolating of antioxidants from complex traditional Chinese medicine extract.
Authors:Y. Wang, M. Xu, G. Yin, L. Tao, D. Wang and X. Ye
Although significant progress has been made on
-mediated wheat transformation, the current methodologies use immature embryos as recipient tissues, a process which is labor intensive, time consuming and expensive. In this study, we have managed to develop an
-based transformation scheme using explants derived from mature embryos. Based on transient expression of
-glucuronidase (GUS) marker, mature embryo halves prepared from freshly imbibed seeds were generally most susceptible to
-mediated T-DNA transfer. According to the results of callus induction and shoot production, Yumai 66 and Lunxuan 208 showed higher selection and regeneration efficiency than Bobwhite. In line with this finding, fertile T
transgenic plants were most readily obtained for both spring and winter wheat when mature embryo halves were used for co-inoculation by
cells. The presence of the antibiotic selection marker (
, encoding neomycin phosphotransferase II) in the T
plants was revealed by both genomic PCR amplification and the enzyme-linked immunosorbent assay (ELISA). Additional analysis showed that the transgene was stably inherited from the two different generations and segregated normally among the T
progenies. Further development along this line will raise the efficiency of wheat transformation and increase the use of this approach in the molecular breeding of wheat crop.
Authors:C. Wang, L. Li, X. Zhang, Q. Gao, R. Wang and D. An
Molecular markers are important tools that have been used to identify the short arm of rye chromosome 1R (1RS) which contains many useful genes introgressed into wheat background. Wheat expressed sequence tag (EST) sequences are valuable for developing molecular markers since ESTs are derived from gene transcripts and more likely to be conserved between wheat and its relative species. In the present study, 35 sequence-tagged site (STS) primers were designed based on EST sequences distributed on homology group 1 chromosomes of
and used to screen specific markers for chromosome 1RS of
. Two primer pairs different from the early studies, STS
, which amplified a 1680-bp and a 1750-bp fragment, and STS
, which produced a 850-bp fragment from rye genome, were proved to be specific to chromosome 1RS since the corresponding fragments were only amplified from 1R chromosome addition line and wheat-rye lines with chromosome 1RS, but not from wheat-rye 2R-7R chromosome addition lines and the other lines lacking chromosome 1RS. Eleven wheat-rye lines derived from ‘Xiaoyan 6’ and ‘German White’ were used to test the presence of specific markers for 1RS. The specific fragments of 1RS were amplified in 4 wheat-rye lines, but not in the other lines. The testing results using EST-STS markers of 1RS were consistent with those obtained from fluorescence
hybridization (FISH), suggesting that these markers specific to 1RS could be used in marker-assisted selection (MAS) for incorporating 1RS into wheat cultivars in breeding.
Authors:Y. Zhang, X. An, X. Li, S. Chen, L. Gao, K. Wang, S. Wang and Y. Yan
Two new y-type HMW-GSs in
with the mobility order of 1Dy12.2
>1Dy12, were identified by both SDS-PAGE and MALDI-TOF-MS. Molecular cloning and sequencing showed that the genes encoding subunits 1Dy12.1*
had identical nucleotide acid sequences with 1,947 bp encoding a mature protein of 627 residues. Their deduced molecular weights were 67,347.6 Da, satisfactorily corresponding to that of 1Dy12.2
subunit determined by MALDI-TOF-MS (67,015.7 Da), but was significantly smaller than that of the the 1Dy12.1*
subunit (68,577.1 Da). Both subunits showed high similarities to 1Dy10, suggesting that they could have a positive effect on bread-making quality. Interestingly, the expressed protein of the cloned ORF from accessions TD87 and TD130 in
co-migrated with subunit 1Dy12.2
, but moved slightly faster than 1Dy12.1*
on SDS-PAGE. The expressed protein in transgenic tobacco seeds, however, had the same mobility as the 1Dy12.1*
subunit, as confirmed by both SDS-PAGE and Western blotting. Although direct evidence of phosphoprotein could not be obtained by specific staining method, certain types of post-translational modifications (PTMs) of the 1Dy12.1*
subunit could not be excluded. We believe PTMs might be responsible for the molecular weight difference between the subunits 1Dy12.1*
Authors:B. Peng, P. Li, S. Lai, Y. Wang, L. Yang and Y. Wang
High ozone (O3) can cause great damage to plants. However, the effect of high O3 on nitrogen (N) absorption, distribution, and utilization in rice at different growth stages under different planting densities is poorly understood. In the present study, a conventional cultivar (Yangdao 6) and a hybrid cultivar (II You 084) with different planting densities were exposed to an elevated amount of O3 (E-O3; 50% higher than that of the control, C-O3) under a freeair gas concentration enrichment (FACE) system. N absorption, distribution, and utilization of the green leaves, stems, and shoots at tillering, jointing heading, and maturity were investigated. Results showed that E-O3 significantly increased the N content in the shoots of Yangdao 6 by 7.5%, 12.7%, and 19.6%, respectively, at jointing, heading, and maturity. Also, the N content in the shoots of II You 084 increased by 5.4%, 6.5%, and 8.4% at the corresponding growth stage upon E-O3 application. E-O3 significantly decreased N accumulation of II You 084 by 8.3%, 4.9%, 4.7%, and 19.2%, respectively, at tillering, jointing, heading, and maturity. Further, E-O3 had a decreasing effect on the N distribution in green leaves (p ≤ 0.05) of both cultivars, but exerted an increasing effect on that in the stems of both cultivars (p ≤ 0.05). In addition, E-O3 significantly decreased the N use efficiency (NUE) for biomass of the two cultivars in all growth stages. These results revealed that E-O3 could increase the N content in rice plants but decrease the N accumulation and utilization in both cultivars. The effects of E-O3 on N absorption, distribution, and utilization were not affected by planting density.
Authors:B. Zhou, S. Jiang, L. Zou, X. Wang, D. Wang, J. Liu, S. Shang and D. Zhou
A new compound cyclohexyl-t-butyldimethylammonium tetraphenylborate, [C6H11N(CH3)2(C(CH3)3)]BPh4 has been prepared, and its decomposition mechanism was studied by TG. The IR spectra of the products of thermal decomposition were examined at every stage. Kinetic analysis for the first stage of thermal decomposition process was obtained by TG and DTG curves, and kinetic parameters were obtained from the analysis of the TG-DTG curves with integral and differential equations. The most probable kinetic function was suggested by comparison of kinetic parameters.
Authors:Y. Duan, J. Li, X. Yang, X. Cao, L. Hu, Z. Wang, Y. Liu and C. Wang
The thermal decomposition of strontium acetate hemihydrate has been studied by TG-DTA/DSC and TG coupled with Fourier transform
infrared spectroscopy (FTIR) under non-isothermal conditions in nitrogen gas from ambient temperature to 600°C. The TG-DTA/DSC
experiments indicate the decomposition goes mainly through two steps: the dehydration and the subsequent decomposition of
anhydrous strontium acetate into strontium carbonate. TG-FTIR analysis of the evolved products from the non-oxidative thermal
degradation indicates mainly the release of water, acetone and carbon dioxide. The model-free isoconversional methods are
employed to calculate the Ea of both steps at different conversion α from 0.1 to 0.9 with increment of 0.05. The relative constant apparent Ea values during dehydration (0.5<α<0.9) of strontium acetate hemihydrate and decomposition of anhydrous strontium acetate (0.5<α<0.9)
suggest that the simplex reactions involved in the corresponding thermal events. The most probable kinetic models during dehydration
and decomposition have been estimated by means of the master plots method.
Authors:P. Zhu, S. Wang, J. Wang, L. Zhou and P. Shi
In order to assess the contribution of adenosine triphosphate and its metabolites to the cellular metabolism process in Saccharomyces cerevisiae, it is very important to simultaneously determine the relative concentrations of ATP and its metabolites. In this study, a fast, simple reversed-phase high-performance liquid chromatography with high selectivity was developed to simultaneously measure adenosine triphosphate and its metabolites (adenosine diphosphate, adenosine monophosphate, and cyclic adenosine monophosphate) in yeast. The method was performed under the gradient grogram, and the detection was monitored at 254 nm. Analysis was achieved within 25 min. The four components can be detected with linear response over the concentration range from 1 to 100 mg L−1 with excellent correlation coefficients (r2) > 0.999. The recovery of the four analytes was 92.9%, 90.4%, 99.1%, and 105.1%, respectively. To demonstrate the good analysis of yeast samples, changes in the four adenine nucleotides levels caused by caloric restriction in yeast were determined. It is expected that the current method may contribute to further metabolomics and system biology investigations of yeast.
Authors:D. Wang, A. Bai, X. Lin, L. Fang, X. Shu, X. Shi, Q. Sun and X. Wang
Supercritical fluid extraction (SFE) was used to extract shionone from Aster tataricus L. f. The effect of various parameters, i.e., temperature, pressure and sample particle size on yield was investigated with an analytical-scale SFE system to find the optimal conditions. The process was then scaled up by 50 times with a preparative SFE system under the optimized conditions of temperature 40 °C, pressure 30 MPa, and a sample particle size of 40–60 mesh. Then preparative high-speed counter-current chromatography was successfully used for isolation and purification of shionone from the SFE extract with a two-phase solvent system composed of n-hexane-methanol (2:1, volume ratio). The separation produced a total of 75 mg of shionone from 500 mg of the crude extract in one step separation with the purity of 98.7%, respectively, as determined by high-performance liquid chromatography (HPLC) and 92% recovery. The structure of shionone was identified by electrospray ionization-mass spectrometry (ESI-MS), hydrogen-1 nuclear magnetic resonance (1H-NMR), and carbon-13 nuclear magnetic resonance (13C-NMR).
Authors:L. Liu, T. Zhao, X. M. Cheng, C. H. Wang and Z. T. Wang
Harmaline and harmine accounted for more than 70% in composition in extracts of P. harmala. More attention, however, should be paid to the other alkaloids which would be favorable or unfavorable to the efficacy and safety of the products. It was necessary to determine these trace alkaloids in the extracts; thereafter, most of them have been characterized. Diglycoside vasicine, vasicine, vasicinone, harmalol, harmol, tetrahydroharmine, 8-hydroxy-harmine, ruine, harmaline, and harmine were separated and identified with reference substances and characteristic MS spectra in extracts by liquid chromatography-electrospray ionization-mass spectrometry (LC-ESI-MS) and high-performance liquid chromatography (HPLC). Three trace alkaloids, vasicine, harmalol, and harmol were determined using the developed chromatographic separation method subsequently. The average contents of vasicine, harmalol, and harmol in extracts of ten batches were 2.53 ± 0.73, 0.54 ± 0.19, and 0.077 ± 0.03%, respectively. The total content of the three alkaloids was 3.23 ± 0.90% (from 1.81 to 4.48%). For rough estimation of all the relative alkaloids except of harmaline and harmine, the average total areas of all peaks in extracts varied from 4.35 to 26.64% detected at 220, 254, 265, 280, and 380 nm, respectively. The results indicated that area normalization method was powerless for the quality evaluation for traditional herb medicine consisting of numerous compounds with highly differential features. It might be concluded that LC-MS or HPLC could be utilized as a qualitative and quantitative analytical method for quality control of the extracts from seeds of P. harmala L.