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    20 May 2011, Volume 31 Issue 3
    Phylogenetic Relationship of Thoreales Based on psaA and psbA Genes
    WANG Ya-Nan;FENG Jia;XIE Shu-Lian*
    2011, 31(3):  257-260.  doi:10.7525/j.issn.1673-5102.2011.03.001
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    Two genes, psaA and psbA, were amplified and sequenced for Thorea hispida(Thoreales) and blasted with other groups, then the phylogenetic trees based on two genes were generated by the methods of Neighbor-Joining, Maximum Parsimony and Bayesian. The results showed the sequences of psaA and psbA were 825 bp and 920 bp in length, respectively. A, T, C and G contents of psaA were 29.9%, 35.8%, 16.5% and 17.8% and those of psbA were 27.5%, 35.3%, 16.8% and 20.4%. There was higher content of A+T than that of G+C for two genes. It indicated that these two genes had favor in nucleotide. The phylogenetic trees generated from three methods displayed the similar topologic structures and the species of Thoreales were aggregated in one branch, separated from other groups. This result supported that Thoreales was an independent order.
    Anatomical Study on the Primary Vascular System of Seedling of Catalpa ovata
    YAN Gui-Hua;WANG Li-Jun*;NIE Xiao-Lan
    2011, 31(3):  261-264.  doi:10.7525/j.issn.1673-5102.2011.03.002
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    Based on the cotyledon node zone (CNZ) theory, the anatomical study on primary vascular system at seedling stage of Catalpa ovata. The results indicate that the CNZ of C.ovata is longer, and belongs to basipetal shifting type. From top to bottem, it has pith in the whole CNZ. There is mesarch tetrarch siphonostele in the lower portion of the CNZ, which divides into two parts from protoxylem in the middle portion of the CNZ and gradually form stele structure with eight collateral bundles. In the upper portion of the CNZ, two vascular bundles which formed shorter axile divide into three parts respectively and shape the model of eustele, then connect with endarch eustele of epicotyl. There are three-trace cotyledonary node in the seedling.
    Megasporogenesis and Development of Female Gametophyte in Cynanchum otophyllum Schneid
    WANG Ding-Kang;SUN Gui-Fang;ZHAI Shu-Hua;GUO Li-Hong;WANG Bin
    2011, 31(3):  265-270.  doi:10.7525/j.issn.1673-5102.2011.03.003
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    Using traditional paraffin wax section, fluorescence microscopy, cytochemistry and TEM techniques, the megasporogenesis and development of female gametophyte of Cynanchum otophyllum Schneid were observed in detail. The results showed C.otophyllum was marginal placenta; the ovules are anatropous, tenuinucellate and unitegmic; some endoplasm, starch grains and mitochondria exist in nucellus; the archesporial cell is hypodermal in origin and directly gives rise to the megaspore mother cell; the chalazal megaspore functions to give rise to an 8-nucleate embryo sac, the embryo sac is of the ‘Polygonum type’ and shows the presence of starch grains in the mature condition; fertilization is porogamous; the endosperm passes through an early free nuclear stage before becoming cellular about the stage of 16-32 nucleate; as usual, the endosperm nucleus divides earlier than the zygote; the mature endosperm cells are uninucleate and irregular in outline; the presence of endosperm haustoria was not observed; development of embryo passes through the stages of proembryo, embryo of globular stage and embryo of heart stage and is of the ‘Soland type’; Seeds are comose and the coma develops from the epidermal cells of the integumentary tissue; a lot of fat pellets exist in mature endosperm.
    Pollen Germination and Growth of Platycodon grandiflorum(Jacq.)A.DC
    LIU Zi-Gang;HU Tian-Ming;YANG Ya-Li;SHE Xiao-Ping;WANG Yu-Lin;XUE Fen-Li;LIU Feng-Xia
    2011, 31(3):  271-276.  doi:10.7525/j.issn.1673-5102.2011.03.004
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    Two-year-old Platycodon grandiflorum(Jacq.)A.DC trees grown in the experiment field of Xiangju Pharmacy Limited Company were used as the materials. The effects of culture medium, sucrose, PEG, pH value, culture time and temperature on the pollen germination and tube growth of P.grandiflorum were investigated using the method of liquid culture. The results showed that: (1)the concentration of 100-150 g·L-1 PEG could accelerate pollen germination and the growth of pollen tube, the range of 200-250 g·L-1 PEG could remarkably accelerate pollen germination, however, its function on the growth of pollen tube was not distinct. (2)100 g·L-1 sucrose was helpful for the pollens to germinate and grow, however, high concentration of sucrose can would strongly inhibit pollen germination and the growth of pollen tubes. (3)the optimum culture medium for pollen germination and growth of Platycodon were ME3+BK+10% sucrose+150 g·L-1 PEG (pH 5.8). (4)pollen germination was better at 25-40℃, and it was best at 30℃ for 1.5 h.
    Comparative Investigation of Vessel Perforation Plates of Four Genera in Lardizabalaceae
    ZHANG Xiao-Hui
    2011, 31(3):  277-283.  doi:10.7525/j.issn.1673-5102.2011.03.005
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    The vessel elements and their perforation plates in the secondary xylem of stem of the four genera in Lardizabalaceae were observed under scanning electron microscope (SEM). The results showed that, (1)simple perforations were found on the end wall of the vessel element in four species; (2)Sinofranchetia was distinguished from the others by perforation types on the end wall: reticulate, scalariform, simple, reticulate-scalariform, reticulate-simple, scalariform-simple and reticulate-scalariform-simple perforations; the membrane remnants in perforations were various from web, flake and thread; Sargentodoxa and Holboellia shared some characteristics of scalariform, scalariform-simple and simple perforations plates on the end wall of the vessel element; only simple perforations plate was found in Stauntonia; (3)scalariform and reticulate-scalariform perforations appeared on the lateral wall (except Stauntonia); (4)spirally thicken presented on the lateral wall of the vessel element in Stauntonia. The new characteristics of vessel element of genera can provide important basis for phylogeny analysis in Lardizabalaceae.
    Dynamic Relationships between Structural Characteristic and Saponins Accumulation in Rhizome of Panax japonicus var. major
    WANG Gui-Qin;Abdusalerk N
    2011, 31(3):  284-288.  doi:10.7525/j.issn.1673-5102.2011.03.006
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    Saponin is an important secondary substance stored in the rhizome of Panax japonicus var. major. According to the chemical properties of saponins, the relationships between the anatomical structure and the content of saponins in the rhizome nodes of P.japonicus var. major with different growth years were studied by means of histochemical localization, light microscope and quantitative analysis of saponins. The results showed that the cross-section of rhizome node which has two or more vascular cylinders is much complicated in structure, belongs to abnormal structural; The continual thickening growth of rhizome’s node principally resulted from the differentiation and development of the secondary and tertiary structure. The secretory cells around the cavities, secondary phloem and tertiary phloem are the main stored place of saponins in rhizome node. The numbers of secretory cavities and abnormal vascular cylinders, and saponins content increase as it grows with age. The number of secretory cavities, quantity of the abnormal vascular cylinders and density of nodes are the main structural and morphological indicators in variety breeding of P.japonicus var. major.
    Chlorophyll Fluorescence Analysis of Hermaphrodite and Male Gametophytes from Ceratopteris thalictroides(L.) Brongn. on Different Media
    DAI Xi-Ling;LI Xin-Guo;ZHANG Ying;WANG Quan-Xi*
    2011, 31(3):  289-292.  doi:10.7525/j.issn.1673-5102.2011.03.007
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    The differences of fluorescence parameters of hermaphrodite gametophyte and male gametophyte from Ceratopteris thalictroides cultured in MS medium and modified knop’s medium were investigated. The results showed that the ФPSⅡ and ETRPSⅡ of hermaphrodite gametophyte in MS medium were higher than those of the male gametophyte in MS medium and gametophytes in modified knop’s medium, the differences were significant. The values of Fv/Fm had no significant difference in the gametophytes of different groups. The external nutrient had no obvious effect on the qP and qN in the gametophytes of different teams. In conclusion, external sucrose enhanced photosynthetic activity of the gametophytes in MS medium, which was beneficial for hermaphrodite gametophytes to grow and develop rapidly.
    Effects of Different Aluminum Stress on the Growth of Rice Roots
    LIU Yu-Guo;FENG Han-Qing;SUN Kun*
    2011, 31(3):  293-299.  doi:10.7525/j.issn.1673-5102.2011.03.008
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    Aluminum is one of the primary environmental factors that limit crops production in acid soil. In this study, rice roots were respectively exposed to 0, 50, 100 and 150 mmol·L-1 aluminum stress. The effects of aluminum on the growth of rice root were stuclied. The present results showed that roots growth (root length and fresh weight) of rice decreased with the increase of the concentration of aluminum. The level of cell death of rice root tip and the content of H2O2 in rice root increased. Treatment with dimethylthiourea (DMTU, scavenger of H2O2) decreased the content of H2O2 in rice root. Moreover, DMTU treatments partially alleviated the root cell death and growth inhibition induced by aluminum stress. These results suggested that the effect of aluminum stress on rice root growth could be partly due to the accumulation of H2O2 in roots of rice.
    Protoplast Culture and Plant Regeneration of Celastrus orbiculatus Thunb.
    FAN Xiao-Feng;LI Dong-Bo;LIU Ling-Xia;LIU Xiu-Li;YANG Ying-Li*
    2011, 31(3):  300-305.  doi:10.7525/j.issn.1673-5102.2011.03.009
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    Experimental raw material of protoplast was obtained from embryo callus induction of Celastrus orbiculatus Thunb. under the condition of lower temperature at 4℃ and dark treatment for 24 h. The created culture system of protoplast originated from seedlings differentiation of callus regeneration after the culture of settling the shallow-layer solution and solid-liquid double layer culture as well as agarose-embedding based on MS medium. The results showed that: the lower temperature and dark treatment was beneficial to obtain protoplast with high yield and top quality; the optimum combination for the enzyme activity would be: 0.5% Cellulase+0.5% Pectinase+5 mg·L-1 MES; the optimum time of enzymatic hydrolysis was 12 h; the optimum mannitol concentration was 13%; moreover, the optimum mode of enzymatic hydrolysis would be: standing for 12 h and oscillating for 0.5 h; meanwhile, better protoplast culture efficiency was based on the culture of settling the shallow-layer solution; the optimum differentiation medium of callus was MS+6-BA 2.0 mg·L-1+IBA 0.1 mg·L-1 and rooting medium could be 1/2 MS+NAA 0.1 mg·L-1.
    High Frequency Somatic Embryogenesis and Histological Observation on Linum usitatissimum L. ‘Fany’
    GUO Ling-Ling;MIAO Shu-Kui;JI Qiao-Ling*;JIANG Li
    2011, 31(3):  306-312.  doi:10.7525/j.issn.1673-5102.2011.03.010
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    The cotyledon and hypocotyl of Linum usitatissimum L. ‘Fany’ as explants were respectively inoculated on MB medium (mineral salt of MS with vitamin of B5) with different combinations of plant growth regulators. Callus was induced from all the media. However, embryogenic callus were only obtained on the media with 0.5-4.0 mg·L-1 2,4-D. The 2,4-D concentration needed for embryogenic callus and embryogenesis derived from hypocotyl were both lower than that from cotyledon. The effect of supplementing with 2,4-D only in medium was obviously better than the combinations of any other plant growth regulators in the induction of embryos. The induction number of somatic embryos derived from cotyledon and hypocotyl was 224/g and 265/g, respectively. The histology observation revealed that there were a large number of normal globular embryos, heart-stage embryos, torpedo-stage embryos, a few cotyledon-stage embryos as well as a few abnormal embryos. These results will be a foundation for somatic hybridization between ‘Fany’ and other flax, screening mutant clone of ‘Fany’ as well as relative research on plant gene and cell engineering.
    Construction of Plant Transformation Vector of AtAAPT1 Gene for RNAi
    ZHENG Gui-Ling;LI Peng
    2011, 31(3):  313-317.  doi:10.7525/j.issn.1673-5102.2011.03.011
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    Aminoalcoholphosphotransferase gene AAPT1 from Arabidopsis thaliana was used as target sequence for RNA interference in this paper. The target sequence was linked to pBS-T cloning vector after obtained by RT-PCR reaction. A plant transformation vector pART27-AAPT1(1,2) was constructed from pBS-T, including AAPT1 gene controlled by CaMV35s promoter in this experiment. Then the recombination plasmid was transfected into Agrobacterium C58 by electroporation-mediated way.
    Vector Construction of Shoot-specific Promoter Driven Prosystemin and the Acquisition of Transgenic Plants
    ZHAO Xue-Qin;LIU Wei-Zhong;JIANG Hong-Ling;ZHANG Hai-Yan*
    2011, 31(3):  318-322.  doi:10.7525/j.issn.1673-5102.2011.03.012
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    The objective of this study was to clone a shoot-specific promoter, it is the promoter of cab2(chlorophyll a/b binding protein 2, cab2) gene. The promoter can drive the expression of prosystemin (PS) and GFP fusion vector, then get the transgenic plants. The plasmids were transformed into Arabidopsis by using Agrobacterium tumefaciens-mediated method. The semi-quantitative RT-PCR method and laser scanning confocal microscope (LSCM) were used to observe the expression and subcellular localization of promoter-driven PS-GFP. Using Arabidopsis genomic DNA as a template, the sequence of promoter cab2 was amplified by high fidelity PCR enzyme, then connected it with the PS and GFP fusion vector (SlPS). The LSCM observation showed that the gene driven by this promoter was normally expressed and located in the cytoplasm. The cab2 promoter was cloned, which can drive the expression and localization of the PS and GFP fusion protein.
    Molecular Cloning and Functional Analysis of a Gene Encoding Farnesyl Diphosphate Synthase from Cyclocarya paliurus
    MIAO Qian;CAO Xiao-Ying;LI Chang-Gen;YIN Ting;LI Xiao-Chu;JIANG Ji-Hong*
    2011, 31(3):  323-329.  doi:10.7525/j.issn.1673-5102.2011.03.013
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    Cyclocarya paliurus is a medicinal, timber and ornamental tree. Farnesyl diphosphate synthase (FPS; EC2.5.1.1/EC2.5.1.10) catalyses 1′-4 condensation of the 5-carbon isoprenoid compounds isopentenyl diphosphate (IPP) and 10-carbon geranyl diphosphate (GPP) to form the 15-carbon product farnesyl diphosphate (FPP). FPP is the common precursor of sesquiterpenes, tritepenes, steroids, etc. cDNA cloning and characterization of a novel FPS from C.paliurus were described in this present study. The full-length cDNA named CpFPS(Genbank Accession Number GU121224) contained 1 420 bp with an open reading frame of 1 029 bp encoding a polypeptide of 342 amino acids with a calculated molecular mass of 39.60 kDa. Through sequence analysis by BlASTP online, the resulting protein showed high homology to the FPS of Gossypium arboretum(CAA72793) and Hevea brasiliensis(BAF98301). Deduced amino acid sequence was also analyzed by biological software and the data presented the existence of conserved and functional domains of FPS. The functional analysis of CpFPS was performed by using the sterol-auxotrophic yeast strain CC25. The result indicated that the cloned cDNA of CpFPS encoding a functional FPS in C.paliurus.
    Flowering Biology of Lycium barbarum L. NQ-2
    WANG Ya-Li;WANG Jin-Xiu;CHANG Hong-Yu;NI Xi-Lu;TIAN Ying;QIN Bin-Bin
    2011, 31(3):  330-335.  doi:10.7525/j.issn.1673-5102.2011.03.014
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    To explore the factors that cause the low fruit setting rate of pure line planting of Lycium barbarum NQ-2, the flowering characteristic, pollen viability, stigma receptivity and breeding system of L.barbarum NQ-2 were studied by means of observation on pollen viability, stigma receptivity, pollen-ovule ration, outcrossing index, emasculation, bagging and artificial pollination using Ningqi 1 as control. The results showed that: (1)The florescence of individual last 3-4 days, the pollen viability was high during the 1st-3rd day, then began to decline, the life span of pollen was more than 9 days; (2)The stigma has receptivity one day before flowering, the receptivity was strongest on the 1st day, and on the 3rd day stigma lost its receptivity; (3)The pollen-ovule ratio (p/o) is 8615, out crossing index is 4; Based on the result of emasculation, bagging and artificial pollination studies, the breeding system of NQ-2 should belongs to obligate xenogamy. The self-incompatibility is the main reason for the low fruit setting rate of pure line planting of NQ-2.
    Producing IGF-1 in Transgenic Tobacco
    LI Yuan;LI Hai-Quan;XIAO Gang;LI Jian-Yue*
    2011, 31(3):  336-340.  doi:10.7525/j.issn.1673-5102.2011.03.015
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    In this research, a new expression vector of pCAMBIA1301-35S promoter-igf-1-nos, which included the gene of hIGF-1, was constructed. The gene is promoted by CaMV 35S promoter which can be expressed in tobacco in high performance. The expression vector of IGF-1 cDNA introduced by 35S promoter was transformed into LBA4404 (Agrobacterium tumefacients), and then infected tobacco through leaf-disc infection. 17 transgenic tobacco plants were gained after hygromycin selection, GUS test and PCR test of DNA. It shows IGF-1 gene can be transcribed into normal mRNA in transgenic tobacco by RT-PCR test. In this experiment, the expression of IGF-1 in tobacco lays the foundation for producing IGF-1 in tobacco and other dicotyledon bio-reactors as well.
    Fruit and Seed Variations of Natural Population of Sarcandra glabra in Mt. Jinyun,Chongqing
    QIAO Yuan-Bao;HE Ping*;SHI Zhang-Tian;ZHANG Min
    2011, 31(3):  341-346.  doi:10.7525/j.issn.1673-5102.2011.03.016
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    In order to evaluate the phenotypic variation of eight natural populations of Sarcandra glabra in Mt. Jinyun, Chongqing, 20 characteristics of fruit and seed were investigated. High levels of variations were revealed in these characteristics among/within populations. Fruit and seed production, number of infructescences and infructescence branches varied to a high degree, while the traits of fruit and seed to a low degree. Compared with fruit traits, the traits of seed seemed more stable. The average variation coefficient of different characteristics ranged from 1.477 (percentage of infructescence with four branches) to 0.041 (seed aspect ratio), while the average variation coefficient of populations ranged from 0.564 (population 6) to 0.292 (population 4). There was no significant difference in the sum of infructescence branches among populations, significant differences in the number of infructescences as well as the percentage of di-branch infructescences, and very significant differences in all the other characteristics. Variations within populations (60.7%) were the main source of phenotypic variations, and variations among populations (39.3%) influentially contributed as well. Populations of S.glabra in Mt. Jinyun had differentiated into two groups, which were mainly determined by coenotype and disturbance degree. Cluster of phenotypic characteristics based on Pearson correlation showed that similar characteristics highly correlated in differentiation progress and evolution speeds approximated.
    Soil Characteristics of the Different Habitats of the Chinese Endemic Species Calligonum ebinuricum
    ZHANG Yong-Zhi;ZHANG Qiang;KANG Xiao-Shan;PAN Bo-Rong*;DUAN Shi-Min
    2011, 31(3):  347-353.  doi:10.7525/j.issn.1673-5102.2011.03.017
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    As an endemic plant of genus Calligonum in China, C.ebinuricum is only distributed in the vicinity area of Ebinur Lake in Xinjiang. The soil characteristics of C.ebinuricum were studied through a large number of field investigations in this study. The results showed that soil particle sizes in the habitats were concentrated at 0.25-0.5 mm and 0.5-1.0 mm, and the content of clay (<0.002 mm) was nearly zero. Such soil can be considered as coarse sandy soil under the Chinese soil texture classification criterion. The pH value changed from 7.98 to 10.24 with the averaged value of 8.9, leading to alkaline characteristics of soil in the habitats. The electric conductivity was 0.09-1.41 ms·cm-1; the content of total salt was 0.525-8.400 g·kg-1. The content of salt was low in the whole soil, while some areas showed saline characteristics. The majority of cations and anions were Ca2+ and SO2-4, respectively; the content of soil organic matters was low as 0.829-1.190 g·kg-1, and that of soil organic carbon showed the same situation. By using SPSS Cluster Analysis and Principal Component Analysis, three types of soil habitats were defined for C.ebinuricum in this study, i.e., light salinization sand habitat with sulphate, alkaline sand soil habitat and ordinary sand soil habitat, of which the last one was the main habitat for C.ebinuricum. The population of C.ebinuricum could be renewed naturally in these three types of habitats, indicating that C.ebinuricum adapted to these three habitats. C.ebinuricum had the ecological characteristics, such as saline tolerance, alkaline resistance, infertile soil endurance and fitting to sand environment, which can be concluded from the characteristics of its habitats. Therefore, C.ebinuricum can be used not only as a sand-fixing plant in desert areas with light saline and alkaline, but also as a good plant for windbreak irrigated by relatively high salinity groundwater. The results in this study would give guidance to the application of C.ebinuricum as well as provide accumulative basis data for the further study on the relationship between soil and C.ebinuricum.
    Observation on the Process of Flower Bud Differentiation in Medicinal Plant Tussilago farfara L.
    CUI Gui-Mei;SUN Hai-Feng;HE Run-Li;SUN Yi*;QIN Xue-Mei;LI Zhen-Yu
    2011, 31(3):  354-357.  doi:10.7525/j.issn.1673-5102.2011.03.018
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    Bud primordia of Tussilago farfara L. inflorescence at various growth stages were used as experiment materials in this study. By means of paraffin sections, the morphological process of bud differentiation was observed by optical microscope. The results showed that inflorescence differentiation started in the early July and completed in the early October. The whole differentiation process could be divided into ten stages (phases): pre-differentiation, disc formation, inflorescence primordium differentiation, petal primordium differentiation of central flowers (tubiform florets), stamen primordium differentiation of central flowers, pistil primordium differentiation of central flowers, petal primordium differentiation of marginal flowers (ligulate flowers), pistil primordium differentiation of marginal flowers, pollen differentiation of central flowers, ovary and ovule differentiation. The relationship between bud differentiation and the growth period of T.farfara L. was also discussed.
    Comparison of the Content of Major Flavonoids in the Introduced and Cultivated Radix Astragali
    WANG Xue-Jie;SUN Hai-Feng;XIE Dao-Sheng;WU Bin;QIN Xue-Mei*
    2011, 31(3):  358-362.  doi:10.7525/j.issn.1673-5102.2011.03.019
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    Screening and evaluating the germplasm resources is important for the standardization of cultivation and planting of Radix Astragali (RA). In this study, UV-Vis spectrophotometry and HPLC were used to determine the contents of total flavonoids, calycosin and formononetin in various RAs collected from different sources introduced to Shanxi. The results showed that the contents of the above components were not only related to their culture styles and origins, but also closely related to the germplasms. Astragalus germplasm from Shanxi and Shaanxi were suitable to be widely planted in the local regions, while those introduced from Gansu, Inner Mongolia, and Shanxi to Hunyuan, Shanxi had a higher degree of flavonoids than those from their corresponding original regions, hence were suitable for large introduction. The correlation analysis further showed that the level of calycosin was significantly correlated to the level of formononetin. The study will be informative to the standardization planting, excellent breed screening, and optimization of quality evaluation system of the medical plant.
    Comparisons of the Content and Antioxidant Activity of Polysaccharide from Different Parts of Elaeagnus mollis Diels
    GAO Xiang;WU Jia-Ye;ZHANG Zhi-Feng;YAN Gui-Qin*
    2011, 31(3):  363-366.  doi:10.7525/j.issn.1673-5102.2011.03.020
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    The content of polysaccharide from leaves, branches and seeds of endangered plants Elaeagnus mollis Diels was determined by the phenol-sulfuric acid method under 490 nm. The oxygen anion radical and hydroxyl radical scavenging activity of polysaccharide from three parts of E.mollis was studied by the pyrogallol self-oxidation method and spectrophotometry with safranine O—Mn2+—H2O2 system. The results showed that the average levels of polysaccharide from leaves, branches and seeds of E.mollis were 69.42, 23.15 and 56.47 mg·g-1, respectively. The polysaccharide of three parts showed antioxidant activities in removing superoxide free radicals and hydroxyl radical. The scavenging activity on superoxide free radicals was Vc>seeds>branches>leaves, removing hydroxyl radical activity was leaves>seeds>Vc>branches. The research results showed that the leaves and seeds of E.mollis could be the raw material as natural antioxidant.
    The Extraction Process of Biochanin A from Dalbergia odorifera T. Chen Leaves
    WANG Fang;ZHANG Dong-Yang;MA Fei-Yue;GAI Qing-Yan;JIAO Jiao;FU Yu-Jie;*
    2011, 31(3):  367-370.  doi:10.7525/j.issn.1673-5102.2011.03.021
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    The preliminary experiments and a three-variable, three-level Box-Behnken design were performed to study the microwave-assisted enzymatic extraction of biochanin A from Dalbergia odorifera T. Chen leaves. The optimum parameters were: extraction time 15 min, irradiation power 300 W, irradiation temperature 34℃, pH 5.2, amount of cellulase 3.5 mg·g-1. Under the optimal conditions, the average extraction yield of biochanin A was 1.579 mg·g-1.
    Optimization of Enzyme-assisted Extraction of Total Flavonoids from Elaeagnus mollis Diels Leaves by Using Response Surface Methodology
    GUO Cai-Zhen;XUE Dan;HOU Xue-Min;ZHANG Zhi-Feng;YAN Gui-Qin*
    2011, 31(3):  371-376.  doi:10.7525/j.issn.1673-5102.2011.03.022
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    The process of enzyme-assisted extraction of total flavonoids from the leaves of Elaeagnus mollis Diels was optimized. Based on the single-factor tests, the effects of enzyme amount, time of enzymatic hydrolysis, pH value and temperature of enzymatic hydrolysis on the extraction yields of total flavonoids from E.mollis Diels leaves were studied by response surface methodology (RSM). The results indicated that cellulase showed better ability of enzymatic hydrolysis. The optimum enzyme-assisted extraction process was: enzyme amount 6.5 mg·g-1, time 1.5 h, pH value 4.7 and temperature 46℃. The highest yield of total flavonoids was 7.1%.
    Simultaneous Determination of Five Catechins in Green Tea using High Performance Liquid Chromatography
    SUN Xiao-Li;LIU Shuai-Hua;WANG Lei;LI Wen-Gang;ZHAO Chun-Jian*;ZU Yuan-Gang
    2011, 31(3):  377-380.  doi:10.7525/j.issn.1673-5102.2011.03.023
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    A high performance liquid chromatography (HPLC) method was developed for the simultaneous determination of epicatechin (EC), catechin (C), epigallocatechin gallate (EGCG), epicatechin gallate (ECG) and epigallocatechin (EGC) in Wuzhishan green tea. The separation was performed on a HIQ SIL C18 V column (250 mm×4.6 mm, 5 μm) using gradient elution with acetonitrile-1.5% acetic acid as the mobile phase. The sample was detected by an UV detector at 280 nm, flow rate was 1.0 mL·min-1, the sample volume was 10 μL and column temperature was 35℃. Under the optimized conditions, there was good linearity between the logarithm values and the peak areas of EC, C, EGCG, ECG and EGC in the ranges of 15-300, 10-200, 10-200, 20-400 and 10-200 μg·mL-1, respectively. Recoveries of the five components were between 96.20% and 99.64%, the RSDs were between 1.12% and 2.24%. This method is simple, rapid, accurate and suitable for the quality control of green tea.
    Preparation and Optimization Experiment of Docetaxel-loaded Bovine Serum Albumin Nanoparticles
    ZHANG Zhi-Zhou;JIANG Shou-Gang;ZU Yuan-Gang*;ZHAO Dong-Mei;ZHAO Xiu-Hua;JIN Xiao-Hui
    2011, 31(3):  381-384.  doi:10.7525/j.issn.1673-5102.2011.03.024
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    Docetaxel (DT), the only derivative of paclitaxel for clinical application in tumor, is mainly used for the treatment of breast cancer, nonsmall-cell lung cancer, oophoroma, etc. Because of its low solubility, organic solvents and solution adjuvants are always added to the preparation. However, organic solvents and solution adjuvants are pungent. Solubility of docetaxel can be increased by the preparation of docetaxel-loaded bovine serum albumin nanoparticles (DT-BSA-NPs) through desolvation-chemical crosslinking method. In this study, the important factors during preparation were tested, and the optimal conditions were obtained according to the optimized data by Design-expert. These data were as follows: the concentration of albumin was 35 mg·mL-1, the concentration of DT was 1.03 mg·mL-1, the ratio of ethanol to water was 3∶1, the dripping speed of ethanol was 0.73 mL·min-1, the stirring time was 12 h, cross-link ratio of glutaraldehyde and BSA was 2∶1. The particle size of the product is 185 nm, and the drug-loading rate is 14.4%. The developed method successfully helped to improve the water-solubility of docetaxel and therefore provided a good foundation for the animal experiments and clinical applications in days to come.