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    15 April 2010, Volume 43 Issue 8
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    The Progress and Application of Wheat Transformation Technology
    YU Xiu-dao,XU Zhao-shi,CHEN Ming,LI Lian-cheng,MA You-zhi
    Scientia Agricultura Sinica. 2010, 43(8):  1539-1553 .  doi:10.3864/j.issn.0578-1752.2010.08.001
    Abstract ( 1316 )   PDF (486KB) ( 2567 )   Save
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    In recent years, the world witnessed the rapid development of plant transformation technology. Genetically modified cultivars from soybean, maize, oilseed rape, and cotton have been widely grown and provided enormous economic and social benefits. Wheat is one of the major food crops for the world’s population. Since the first report of wheat transgenic plants in 1992, significant progress has been made in wheat transformation technology that provides a solid foundation for molecular breeding in wheat. Up to date, the particle bombardment and the Agrobacterium-mediated transformation remain to be the two major approaches for wheat transformation, representing 68.8% and 15.9%, respectively, among reported cases. Transgenic wheat have been applied to improve disease and insect resistance, abiotic stress tolerance, quality, and grain yield, with disease resistance (39.7%) and quality improvement (25.6%) as two major areas. In this review, the current research progress in wheat transformation technology and its application status were summarized. The existing problems in wheat transformation were discussed and the development direction in this area was proposed.

    Molecular Cloning and Sequence Analysis of Novel RBR Gene from Barley#br#
    WANG Yan,WANG Ji-rui,ZHENG You-liang,WEI Yu-ming
    Scientia Agricultura Sinica. 2010, 43(8):  1554-1562 .  doi:10.3864/j.issn.0578-1752.2010.08.002
    Abstract ( 900 )   PDF (936KB) ( 1143 )   Save
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    【Objective】 The aim of this study is to isolate and clone RBR (retinoblastoma-related) gene from barley (Hordeum vulgare), which is a negative regulator during the cell cycle progress, to characterize HvRBR sequence, ensure taxonomic status among the homologous genes, and give more information about cell proliferation and differential regulation pathway in animals and plants. 【Method】 After large numbers of RBR genes were bioinformatically analyzed, specific primers were designed based on the conservative sequences. The RBR gene sequences were obtained from both genomic DNA and cDNA of seedlings. DNAMAN software was used to perform sequence analysis, multiple sequence alignment, and phylogenetic tree construction was also carried out by MEGA. 【Result】 The 5 547 bp of DNA sequence (GU121481) and 3 179 bp of cDNA sequence (GU121480) of HvRBR were obtained from GSHO1854 (a shrunken endosperm mutational material of cultivated barley), respectively. The coding region of this gene could encode a protein with 975 amino acid residues. This deduced amino acid had a high identity with the known RBR proteins form plants and animals. Although the lower identity was found in the spacer region between pocket A and B domain, a conservative cysteine residue was found in all the RBR protein family members at similar position. It was suggested that the intra- or inter-molecular disulfide bond probably play an important role in affecting the structure and function of RBR protein. Phylogenetic analysis revealed that the highest degree of identity (84.3%) was between HvRBR and OsRBR2 (Oryza sativa), and a lower value (50%) between HvRBR and dicot plants such as MsRBR (Medicago sativa) and AtRBR1 (Arabidopsis thaliana). 【Conclusion】 It was the first time to obtain the barley HvRBR gene, which regulate cell cycle, proliferation and differentiation. HvRBR is a novel member of subgroup C of plant RBR family by sequence analysis, identification and phylogenetic analysis.

    Role of Small GTP-Binding Protein ROP in Medicago truncatula in Symbiosis: 2. Function Analysis of MtROP5 Associated with Development of Root Hair#br#

    LIU Wei,CHEN Ai-min,FENG Li-xing,CAO Lian-pu,SUN Jie,WANG Yan-zhang

    Scientia Agricultura Sinica. 2010, 43(8):  1563-1570 .  doi:10.3864/j.issn.0578-1752.2010.08.003
    Abstract ( 1167 )   PDF (489KB) ( 879 )   Save
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    【Objective】 The function of MtROP5 in Medicago truncatula was studied for establishment of the groundwork to investigate the regulatory mechanism of ROP in symbiotic interaction. 【Method】 The promoter sequence of MtROP5 was obtained using PCR, and the expression vectors containing MtROP5 were constructed. The method of Agrobacterium rhizogenes-mediated root transformation was used to investigate the function of MtROP5. 【Result】 The sequence of MtROP5 promoter that is at about 2.1 kb upstream of the translation origin point was obtained. The results showed that the MtROP5 promoter allowed GUS expression in every part of the root, GUS displayed more abundant expression level in vascular bundle and initials of lateral root than other parts of root. The transcriptional activity of MtROP5 promoter was induced to increase in the transgenic roots after infected by Sinorhizobium meliloti Rm1021. Compared with root hairs of the transgenic root of control, the length of transgenic root hairs of overexpression and constitutively active mutant of MtROP5 increased markedly by 74.6% and 54.8%, respectively, the length of transgenic root hair of RNAi interference of MtROP5 reduced by 47.6%, but the dominantly negative mutant of MtROP5 did not affect the development of root hairs. 【Conclusion】 MtROP5 in M. truncatula regulated the development of root hair, and probably involved in the signal transduction at the early stage of symbiotic interaction between host plant and Rhizobium.

    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY
    A Knowledge Model for Precision Water Management in Rice#br#
    LIU Xiao-jun,CAO Jing,LI Yan-da,ZHANG Yu-ping,CAO Wei-xing,ZHU Yan
    Scientia Agricultura Sinica. 2010, 43(8):  1571-1576 .  doi:10.3864/j.issn.0578-1752.2010.08.004
    Abstract ( 1044 )   PDF (299KB) ( 1106 )   Save
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    【Objective】 Rice is the most important staple crop and needs much water resource in China, so precision water management is necessary for high efficiency rice production. 【Method】Based on analysis and extraction of the research results in rice cultural theories and technologies, further on the theory of soil water balance, with soil water potential as the irrigation index, a knowledge model for precision water management in rice (KMPWM) was developed by quantifying the relationships of water demand and cultivation technologies, variety traits, and eco-environments. Furthermore, based on the system engineering theory, a decision support system for rice precision water management was established on the platform of Microsoft Visual Studio.NET 2005.【Result】KMPWM realized the functions of irrigation management scheme design and real-time regulation. The field comparison experiments at Hangzhou and Nanjing showed that the average grain yields were improved by 6.9% and 9.1%, respectively, and irrigation water use efficiencies were remarkably increased by 53.61% and 42.73%, respectively, in two eco-sites under the model-designed irrigation strategies, as compared to conventional irrigation regimes. 【Conclusion】 It appears that the present model has good applicability and proper guidance for rice precision water management.

    Induction of H2O2 Burst of Wheat Suspension Cells by IWF and Preliminary Study on Its Productive Mechanism#br#
    REN Li-mei,CHEN Yan,WANG Dong-mei
    Scientia Agricultura Sinica. 2010, 43(8):  1577-1584 .  doi:10.3864/j.issn.0578-1752.2010.08.005
    Abstract ( 957 )   PDF (346KB) ( 993 )   Save
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    【Objective】 Intercelluar washing fluid(IWF-260)of leaf rust - infected wheat leaves ( Lovrin 10 ) was used to stimulate the suspension cell of Lovrin 10 in order to study the H2O2 burst and its productive mechanism. 【Method】 The changes of H2O2 were detected by chemiluminescence method and fluorescent probe DCFHDA staining. The activity of NADPH oxidase was studied by HE staining. Pharmacological experiments were carried out to study the relation between Ca2+ and H2O2。【Result】IWF-260 could cause the outbreak of H2O2 in wheat suspension cultured cells. It played an important role in the programmed death of wheat suspension cells induced by IWF-260. NADPH oxidase might be involved in the production of H2O2. The influx of extracellular Ca2 + could regulate the production of H2O2. 【Conclusion】 IWF can induce H2O2 burst of wheat suspension cells and this process is involved with the activation of NADPH oxidase and the influxing of intercelluar Ca2+.

    Effects of NaCl Stress on the Characteristics of Photosynthesis and Chlorophyll Fluorescence at Seedlings Stage in Different Sea Island Cotton Genotypes #br#
    YANG Shu-ping,WEI Chang-zhou,LIANG Yong-chao
    Scientia Agricultura Sinica. 2010, 43(8):  1585-1593 .  doi:10.3864/j.issn.0578-1752.2010.08.006
    Abstract ( 972 )   PDF (512KB) ( 1435 )   Save
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    【Objective】 The effects of NaCl stress on photosynthesis and chlorophyll fluorescence of sea island cotton genotypes were studied . 【Method】 Two sea island cotton varieties, Xinhai 21 (XH21, NaCl sensitive genotype) and Xinhai 28 (XH28, NaCl tolerant genotype) were selected, and germinated in full nutrient solution. NaCl was added into solution at 3-leaf full expanding stage and NaCl concentration levels at 0, 50, 100, 150, 200 and 250 mmol•L-1 respectively. Photosynthetic parameters, chlorophyll fluorescence variation were investigated after 30 d. 【Result】 The results showed that, under low NaCl concentration stress, the growth of the plants, fresh weight, biomass, chlorophyll content were enhanced, ΦPSⅡ, qP, NPQ were increaseed as well, all these increased Pn of leaves in tolerant genotype Xinhai 28. Under high NaCl stress, plant growth restrained significantly in both genotypes, Pn, Gs, Fv/F0, Fv/Fm, ΦPSⅡ and qP were reduced, while NPQ and Ci increased. 【Conclusion】 The decrease of net photosynthetic rate was considered as a result of no-stomatal restriction turning stomatal restriction along with NaCl concentration increase. Photosynthetic parameters and chlorophyll fluorescence were highly correlated with genotypes’s salt tolerance and salt concentration.

    PLANT PROTECTION
    Identification and Characterization of an Ethoprophos-Degrading Bacteria DS-1 and Its Degradation Characteristics#br#
    LI Yan-fang,SONG Xiao-lei,JI Jing,LIU Feng,MU Wei
    Scientia Agricultura Sinica. 2010, 43(8):  1594-1600 .  doi:10.3864/j.issn.0578-1752.2010.08.007
    Abstract ( 952 )   PDF (334KB) ( 755 )   Save
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    【Objective】 Biodegradation is an effective and economic method to treat the soil environment that has been polluted by ethoprophos.The study focused on the isolating and characterizing bacteria with abilities to degrade ethoprophos.【Method】 The enrichment method was used to find the bacteria which can degrade the ethoprophos from the soil samples of long-term application of ethoprophos. The content of ethoprophos was determined by using UV. 【Result】 A bacterial strain DS-1 capable of utilizing ethoprophos as sole carbon source was isolated. It was preliminarily identified as Bacillus cereus according to its physiological-biochemical characteristics and the similarity analysis of its 16S rDNA sequence (GenBank Accession No. GQ149481). The growth and degrading characteristics of this strain were tested. The highest degradation rate of ethoprophos was up to 77.1% when ethoprophos concentration was 10 mg∙L-1, and the relative degradation amount of 50 mg∙L-1 was 26.8 mg∙L-1 which is the highest. The degradation rate of ethoprophos was 51.6% at 30℃, significantly higher than that at 4℃, 25℃ and 40 ℃. 【Conclusion】 The optimal temperature and pH for the ethoprophos degradation under the action of DS-1 were 30℃ and 7.0, respectively. The optimal carbon and nitrogen sources were sucrose, ammonium sulfate, and the optimum molar ratio of carbon to nitrogen was 4:1.

    Influence of α-Terthienyl on the Mitochondrial Membrane Potential and Cell Cycle of Spodoptera litura Cell#br#
    WANG Yu-jian,ZHANG Zhi-xiang,XU Han-hong,LIAO Mei-de,ZHANG Qian-feng
    Scientia Agricultura Sinica. 2010, 43(8):  1601-1607 .  doi:10.3864/j.issn.0578-1752.2010.08.008
    Abstract ( 862 )   PDF (368KB) ( 668 )   Save
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    【Objective】 To obtain the influence of α-T on the mitochondrial membrane potential and cell cycle of insect cell is essential. 【Method】 Rhodamine123 and flow cytometer were employed to acquire the influence of α-T on the mitochondrial membrane potential of Spodoptera litura cell after 24 h and 48 h treatments. The cell cycle phases were determined with propidium iodide (PI) through FCM. 【Result】 After 24 h treatment, the mitochondrial membrane potential of cells treated with 0.0625 μg•mL-1and 0.1250 μg•mL-1 changed little. While the mitochondrial membrane potential of cells treated with 0.5000 μg•mL-1 increased obviously. After 48 h, the mitochondrial membrane potential of the treatment with high dose (1.0000 μg•mL-1) depolarized. Moreover , the cell cycle was arrested in S phase when SL cell was treated with low dose (0.0625 μg•mL-1) after 24 h treatment. However, it was arrested in G2/M phase when treated with higher dose. After 48 h, the cell cycle was arrested in G2/M phase with lower dose (≤0.1250 μg•mL-1) and in S phase with higher dose (≥0.1250 μg•mL-1). 【Conclusion】 After treatment, the cells were in a serial complicated status. When the cells couldn’t resist the oxidation damage caused by ROS, the mitochondrial membrane potential and cell cycle phase changed violently. These changes decided the path of cells damage and death.

    RNAi Effect of Cathepsin B Gene on Reproduction of Radopholus similis#br#
    LI Yu,XIE Hui,XU Chun-ling,LI Dan-lei,ZHANG Chao
    Scientia Agricultura Sinica. 2010, 43(8):  1608-1616 .  doi:10.3864/j.issn.0578-1752.2010.08.009
    Abstract ( 919 )   PDF (472KB) ( 702 )   Save
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    【Objective】 In order to elucidate the function of cathepsin B gene from Radopholus similis and provide a scientific basis for the further research on this cathepsin B gene of plant-parasitic nematodes, and also for developing new control strategies, the RNAi effect of cathepsin B gene on reproduction of R. similis was studied. 【Method】 Cathepsin B gene of R. similis (Rs-cb-1) was cloned from R. similis by designed primers and used as the target gene for RNA silencing. The specific double-stranded RNA (dsRNA) was synthesized by using plasmid DNA containing target gene as template. The nematodes of R. similis were soaked with dsRNA of Rs-cb-1 and RNA interference (RNAi) effects were evaluated by examining the reproduction of R. similis on carrot discs and detecting the gene transcription by RT-PCR. 【Result】 A significant difference in reproduction factors was observed in treatments with different Rs-cb-1 dsRNA soaking time. The average reproduction factor of R. similis on carrot discs was 165,93,54,53 after dsRNA soaked for 12,24,48, 72 h,respectively. The reproduction factors of those treatments without Rs-cb-1dsRNA soaking were greater than 420. RT-PCR revealed that the transcription of target gene was significantly degenerated after 12 h Rs-cb-1 dsRNA soaking. After soaked for 24 h, the transcription was decreased more with only a weak band appeared in electrophoresis gel. No transcription of Rs-cb-1 was observed after 48 h and 72 h Rs-cb-1 dsRNA soaking. 【Conclusion】 The function of Rs-cb-1 is probably related to the reproduction ability of R. similis Rs-cb-1 dsRNA and soaking inhibited the transcription of cathepsin B gene of R. similis, which may further affected the reproduction of R. similis. The RNAi efficiency of Rs-cb-1 was different in Rs-cb-1 dsRNA soaking times, 48 h dsRNA soaking may produce the best silencing effect on target gene.

    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Effects of Glucose Addition on N Transformations in Paddy Soils with a Gradient of Organic C Content in Subtropical China#br#
    JIA Jun-xian,LI Zhong-pei,CHE Yu-ping
    Scientia Agricultura Sinica. 2010, 43(8):  1617-1624 .  doi:10.3864/j.issn.0578-1752.2010.08.010
    Abstract ( 1075 )   PDF (285KB) ( 876 )   Save
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    【Objective】 In order to understand the mechanisms of N transformation and manage N fertilization better, the effect of glucose addition on N transformation in paddy soils with a gradient of organic C content were determined. 【Method】 Changes of mineralization, nitrification and denitrification in paddy soils, as well as their response to glucose addition were measured by incubation experiment. 【Result】 Intensity of mineralization and denitrification were: soils with high fertility > soils with middle fertility > soils with low fertility. During the first week of incubation, net mineralization and denitrification rate in paddy soils with high fertility were 1.9 and 1.1 times of that in soil with middle fertility and 5.3 and 2.9 times of that in soils with low fertility, respectively. Addition of glucose decreased approx. 78.8%, 109.2%, and 177.4% of net mineralization in soils with high, middle, and low fertility, respectively. However, denitrification rate in soils with middle and low fertility increased by 14.4% and 166.2%. The highest nitrate content in all the tested soils was 0.62 mg•kg-1 and the highest nitrification ratio was 0.33%. Addition of glucose had no obvious effects on nitrate content and nitrification ratio. 【Conclusion】 It was suggested that the intensity of mineralization and denitrification were quite different in soils with different fertility. The intensity was increased with the increase of soil organic C content. Addition of glucose decreased mineralization, but increased denitrification. The effects of glucose addition were significantly different in different soils. The effect of glucose addition on soils with low organic C content were greater than that on soils with high organic C content. Neither addition of glucose nor inherent soil organic C had obvious effects on nitrification of test paddy soils.

    Effect of Different Irrigation Patterns on Soil Dissolved Organic Carbon and Microbial Biomass Carbon in Protected Field#br#
    HAN Lin,ZHANG Yu-long,JIN Shuo,WANG Jiao,WEI Yan-yan,CUI Ning,WEI Wei
    Scientia Agricultura Sinica. 2010, 43(8):  1625-1633 .  doi:10.3864/j.issn.0578-1752.2010.08.011
    Abstract ( 1203 )   PDF (297KB) ( 1557 )   Save
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    【Objective】 The objective of this study was to research the distribution characteristics in the profile of dissolved organic carbon (DOC), microbial biomass carbon (MBC) in protected field under three irrigation patterns(subsurface irrigation, drip irrigation and furrow irrigation). 【Method】The modes of irrigation are subsurface irrigation, drip irrigation and furrow irrigation. The same irrigation experiments were conducted for ten years. Soil samples at different depths within 0-100 cm were collected and the contents of soil total organic carbon (TOC), dissolved organic carbon (DOC), microbial biomass carbon (MBC) and their profile distribution were determined. 【Result】 The results showed that the content of soil total organic carbon, dissolved organic carbon, and microbial biomass carbon were all showed the biggest amount in the soil surface and their content increased with layers’ depth, but there were significant differences within irrigation methods. Soil total organic carbon content in 0-10 cm and 80-100 cm soil layers were furrow irrigation>subsurface irrigation>drip irrigation and in 10-80 cm soil layers was subsurface irrigation>furrow irrigation>drip irrigation. In 0-100 cm profile layers, DOC content showed furrow irrigation>drip irrigation>subsurface irrigation. MBC content showed drip irrigation>furrow irrigation>subsurface irrigation. The content of DOC and MBC accounted for 4.98%-12.87% and 1.48%-2.82% of the total organic carbon, respectively. Their ratios were all showed drip irrigation>furrow irrigation>subsurface irrigation. There were significant positive correlation between the content of soil dissolved organic carbon, the microbial biomass carbon and the soil total organic carbon content. 【Conclusion】 Furrow irrigation is conducive to accumulation of SOC and DOC; drip irrigation is conducive to an increase in MBC; subsurface irrigation is the least conducive to the accumulation of soil organic matter comparison, not only the content of TOC is low but also the content of DOC in the ratio of TOC is small.

    HORTICULTURE
    Recent Progress in Research on Anthocyanin and Its Genetics in Potatoes#br#
    CHEN Hai-xia,NIE Xian-zhou,LIU Ming-yue,XIONG Xing-yao
    Scientia Agricultura Sinica. 2010, 43(8):  1634-1642 .  doi:10.3864/j.issn.0578-1752.2010.08.012
    Abstract ( 952 )   PDF (564KB) ( 892 )   Save
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    Recent advances in anthocyanin research in potatoes was reviewed. The types and contents of anthocyanin as well as their antioxidant abilities in potatoes were elucidated. Genes involved in anthocyanin biosynthesis in diploid potatoes were compared to that in tetroploid potatoes. Moreover, the role of R and P genes as well as their cloning and expression was reviewed.
    Advance in Research on Domestication and Taxonomy of Eurasian Grape (Vitis vinifera L.)#br#
    WANG Jun,DUAN Chang-qing
    Scientia Agricultura Sinica. 2010, 43(8):  1643-1654 .  doi:10.3864/j.issn.0578-1752.2010.08.013
    Abstract ( 945 )   PDF (464KB) ( 2175 )   Save
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    The Eurasian grape (Vitis vinifera L.) is the most widely cultivated and economically important fruit crop in the world. Cultivated grapevines (V. vinifera L. ssp. sativa) are considered to have been domesticated from wild population of V. vinifera L. ssp. sylvestris. The domestication process involves the selection of hermaphrodite genotypes producing larger and sweeter berries of attractive colors and the development of techniques for their vegetative propagation. Major questions regarding grapevine domestication concern the time and number of domestication events, and the geographic locations where they took place. In this article, the advances in research on the domestication and taxonomy of Eurasian grape according to eco-geographic origin and distribution are reviewed.

    STORAGE·FRESH-KEEPING·PROCESSING
    Establishment of Rapid Method for Detecting Staphylococcus aureus in Food by Loop-Mediated Isothermal Amplification #br#
    XU Yi-gang,LI Su-long,LI Dan-dan,JIANG Yan-chun,XIE Xiao-feng
    Scientia Agricultura Sinica. 2010, 43(8):  1655-1663 .  doi:10.3864/j.issn.0578-1752.2010.08.014
    Abstract ( 916 )   PDF (583KB) ( 1072 )   Save
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    【Objective】 Staphylococcus aureus is an important pathogen of zoonose, mainly initiates its transmission through food contamination and causes food poisoning, suppurative inflammation of tissues and organs. Thus, the development of a technique that can identify the microbe quickly and accurately is of great importance. 【Method】 Loop-mediated isothermal amplification (LAMP) for DNA is a novel nucleic acid amplification method with rapidity and high sensitivity. In this study, a LAMP method for detecting Staphylococcus aureus was developed. Four primers including two outer primers F3, B3 and two inner primers FIP, BIP were designed according to highly conserved Sa442 gene of Staphylococcus aureus. Genomic DNA extracted from 45 bacteria strains including 20 Staphylococcus aureus strains and 25 other bacterial strains were amplified using the primers with Bst DNA polymerase at 65oC, only Staphylococcus aureus strains were LAMP positive result, which indicated that the LAMP primers designed were highly specific for target bacteria. 【Result】 The detection sensitivity of the LAMP method for Staphylococcus aureus purely cultured was 25 CFU/mL and for Staphylococcus aureus in contaminated food was 42 CFU/g, which could complete the test in 40-60 min. In practice, 115 pieces of samples detected from 958 pieces of meat samples, egg, dairy products and artificial contamination samples were positive using the LAMP method, which accorded with the detection result by GB. 【Conclusion】 The LAMP assay established in this study is a sensitive, rapid and simple tool for detecting Staphylococcus aureus.

    Kinetic Characteristics of Calpain System from Beef of Chinese Yellow Cattle #br#
    HUANG Ming,TANG Xiao-yan,HUANG Feng,XUE Mei,ZHOU Guang-hong
    Scientia Agricultura Sinica. 2010, 43(8):  1664-1669 .  doi:10.3864/j.issn.0578-1752.2010.08.015
    Abstract ( 1079 )   PDF (313KB) ( 753 )   Save
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    【Objective】 The objective of this experiment was to demonstrate the kinetic characteristics of calpain system from beef. 【Method】 The kinetics of purified calpain system including µ-calpain, m-calpain and calpastatin were investigated by changes of some parameters such as storage temperature and time, calcium concentration, reaction time, enzyme amount and substrate concentration. 【Result】 When frozen, calpain system activity decreased with storage time, calpastatin, however, was far more sensitive to freezing than calpains. The calcium concentrations required for half maximal activity of µ-calpain and m-calpain were 50 µmol•L-1 and 320 µmol•L-1, respectively. To assay calpain activity accurately, the reaction time should be within 60 min, and activity less than 0.45. When casein was used as substrate, the Km and Vmax of m-calpain was 3.185 mg•mL-1 and 0.015 U•min-1, µ-calpain was 5.320 mg•mL-1, and 0.017 U•min-1, respectively. 【Conclusion】 Storage temperature and time, calcium concentration, reaction time, enzyme amount and substrate concentration had great influence on the activity of calpain system.

    Analysis of Stable Carbon and Nitrogen Isotope Compositions and Geographical Origins of Sheep Tissues #br#
    SUN Shu-min,GUO Bo-li,WEI Yi-min,FAN Ming-tao
    Scientia Agricultura Sinica. 2010, 43(8):  1670-1676 .  doi:10.3864/j.issn.0578-1752.2010.08.016
    Abstract ( 928 )   PDF (392KB) ( 930 )   Save
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    【Objective】 The carbon and nitrogen isotope compositions of sheep tissues were investigated, and their variations among the regions were discussed, the aim is to provide the scientific basis for using stable isotope analysis to trace the geographical origin of mutton; 【Method】 The δ13C and δ15N values of sheep muscle, hair and their feeds originating from three pastoral areas including Alxa League, Xilin Gol League and Hulunbuir City of Inner Mongolia, and two agricultural areas including Chongqing City and Heze City were determined by Isotope Ratio Mass Spectrometry(IRMS). The differences of carbon and nitrogen compositions in sheep tissues from above regions were compared, and the correlations of isotope compositions between different tissues were analyzed. Moreover, the change of δ13C and δ15N values in sheep tissues under different feeding modes and environmental conditions were also investigated; 【Result】 C and N isotope ratios of sheep tissues from different regions showed significant differences; the δ13C values of tissues were highly correlated with the δ13C value of their feeds, mainly affected by the kinds of forage; the δ15N values were related with feeds and geographical origin. In addition, the δ13C and δ15N values between defatted muscle, crude fat and sheep hair were both significantly correlated. 【Conclusion】 The stable carbon and nitrogen isotopes could be used as reference indexes for tracing geographical origin and feeding practice of sheep. The defatted muscle, crude fat, and sheep hair could be used in mutton geographical origin assessment.

    Ultrasound-Assisted Extraction and Antioxidation Stability of Silkworm Pupa Oil#br#
    WU Xiao-xia,LI Jian-ke,ZHANG Yan-yu
    Scientia Agricultura Sinica. 2010, 43(8):  1677-1687 .  doi:10.3864/j.issn.0578-1752.2010.08.017
    Abstract ( 1028 )   PDF (572KB) ( 1121 )   Save
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    【Objective】 The optimum conditions of Ultrasound-assisted extraction of silkworm pupa oil (SPO), physicochemical property of SPO, factors influencing SPO oxidation, and antioxidant effects of different antioxidants and several polyphenols extracts for SPO were all investigated in this paper. 【Method】 Response surface method (RSM) was used to establish the mathematical model of ultrasound-assisted extraction of SPO to obtain the optimum conditions. Fatty acid compositions and contents of SPO were tested by gas chromatography (GC). Factors influencing SPO oxidation and antioxidation stability of different antioxidants and several polyphenol extracts for SPO were investigated by Schaal oven method. 【Result】 The optimum conditions obtained through RSM analysis were as follows: ultrasonic power was 102 W, ultrasonic time was 20 min and ultrasonic temperature was 40℃. The average yield of SPO was 28.9% and the extraction rate was 96.3%. The relative content of unsaturated fatty acid in SPO was 74.2%, where oleic acid was 38.3%, α-linolenic acid was 30.2%, linoleic acid was 4.3%, palmitoleic acid was 1.0% and arachidonic acid was 0.4%. Light, metallic ions and temperature can significantly speed up SPO oxidation. Among several polyphenol extracts, gallnut tannin had the strongest antioxidant capacity. 【Conclusion】 These results suggest that Ultrasound- assisted extraction of SPO had a higher extraction rate, a less extraction time and a good product quality. SPO should be stored and processed in a lower temperature condition and avoid light and metallic ions. Gallnut tannin could be used as a good natural antioxidant in SPO.

    ANIMAL SCIENCE·RESOURCE INSECT
    Effect of Castration on the Growth Performance and Carcass Quality Traits of Male Pigs#br#
    CAI Zhao-wei,ZHAO Xiao-feng,YAO Yu-chang,ZHANG Li-fan,ZHAO Chun-jiang,XU Ning-ying,WU Chang-xin
    Scientia Agricultura Sinica. 2010, 43(8):  1688-1695 .  doi:10.3864/j.issn.0578-1752.2010.08.018
    Abstract ( 903 )   PDF (347KB) ( 1238 )   Save
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    【Objective】 The present study was performed to investigate the effect of castration on the growth performance,carcass quality traits and muscle fatty acid composition of male pigs. 【Method】 Eighteen pairs of male pigs were selected from six litters according to a matched-pairs design. Each pair was composed of two male full siblings with similar body weight, and one of the pigs from each pair was randomly selected to be castrated at 35 days old. The pigs were weighed at 5, 12, 21 and 30 weeks old and blood samples were collected for subsequent determination of hormones. Nine pairs of pigs were randomly selected to be slaughtered at 21 weeks of age, the other pairs were slaughtered at 30 weeks of age, and the carcass traits were determined then. 【Result】 Boars had significant higher serum testosterone concentrations compared with barrows at 12, 21 and 30 weeks of age (P<0.01). The barrows grew faster than the boars before 21 weeks, but the barrows had a high significant growth difference at 30 weeks. The barrows had a significant higher average back fat thickness, leaf fat percentage and fat percentage, but lower average skin thickness and lean meat percentage than boars at both 21 and 30 weeks (P<0.01), significant higher carcass weight,foot weight and loin eye area were found in boars than barrows at 30 weeks (P<0.05). Compared to boars, higher intramuscular fat content (IMF) was found in barrows at 21 weeks. The content of several saturated fatty acids and total monounsaturated fatty acid (MUFA) were significantly higher (P<0.01), and total polyunsaturated fatty acid (PUFA)was significantly lower in the longissimus dorsi muscle of barrows than in boars. Serum testosterone concentrations was significantly negatively correlated with C16:0, C18:1and MUFA, but positively correlated with C18:0 and PUFA. 【Conclusion】 The growth performance and carcass quality traits of male pigs were significantly influenced by castration.

    Characters of Metabolism, Endocrine and Tissues Genes Expression of Dairy Cows Affected by Fatty Liver#br#
    XIA Cheng,WANG Zhe,ZHANG Hong-you,XU Chuang,ZHANG Cai
    Scientia Agricultura Sinica. 2010, 43(8):  1696-1702 .  doi:10.3864/j.issn.0578-1752.2010.08.019
    Abstract ( 903 )   PDF (421KB) ( 957 )   Save
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    【Objective】 To understand the changes of metabolism, endocrine and mRNA expression of three genes in liver and adipose of fatty liver cows. 【Method】 Ten fatty liver cows and ten health control cows were chosen from the same dairy cow farm were used in the experiment. Fourteen blood parameters and liver fat content were detected in the tested cows. Meanwhile, mRNA expression of liver PEPCK-C was measured by a semi-quantitative RT-PCR, of adipose Lp and HSL by a real time RT-PCR in the tested cows. 【Result】 In fatty liver cows, the concentration of plasma glucose decreased very significantly (P<0.01), plasma NEFA and BHBA increased very markedly (P<0.01), the liver fat content (about 41.88%) and activity of serum AST increased very significantly (P<0.01), the level of serum TBI, γ-GT and CHE increased significantly (P<0.05), compared with the health control cows. In fatty liver cows, the concentration of plasma Ins and ratio of Ins to Gn all increased significantly (P<0.05), mRNA expression level of liver PEPCK-C increased markedly (P<0.05), and of adipose HSL also decreased significantly (P<0.05), compared with the health control cows. mRNA expression level of adipose Lp , the concentration of plasma Lp and NPY all were lower markedly in fatty liver cows than that in health control cows(P<0.05). 【Conclusion】 These results suggested that fatty liver cows have serious energy metabolism disorder, and marked liver disfunction. It might be an important mechanism of energy metabolic disorder for fatty liver cows to have a disturbance between Ins and Gn, Lp and NPY, and a decreased mRNA level of liver PEPCK-C, fat Lp and HSL.

    Proteome Comparison Between Worker Pupal Head of Native Italian Honeybee (Apis mellifera L.) and Higher Royal Jelly Producing Strain#br#
    ZHENG Ai-juan,FANG Yu,FENG Mao,WU Jing,SONG Fei-fei,LI Jian-ke
    Scientia Agricultura Sinica. 2010, 43(8):  1703-1715 .  doi:10.3864/j.issn.0578-1752.2010.08.020
    Abstract ( 912 )   PDF (979KB) ( 730 )   Save
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    【Objective】 To investigation into the differentially expressed proteins between the higher royal jelly producing (HRJB) and the native Italian honeybee (NIB) (Apis mellifera L.), the proteome profilings of the worker pupal head of the two honeybee strains were compared on 3 time-points (day 13, 15, 17) for providing a theoretical principle of clarifing honeybee developmental biology. 【Method】 Two-dimensional gel electrophoresis (2-DE) was employed to establish the proteomic map of the worker pupal head of the two honeybee strains on 13, 15 and 17 days, then the protein expression abundance, molecular weight and pI was obtained. Part of differentially expressed proteins was identified by analysis of matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF/MS) and database research. 【Result】 A total of 279, 296 and 340 proteins were detected on 3 time-points in HRJB, and 268, 277 and 279 were observed in NIB, respectively. A significantly higher protein spot numbers were observed in HRJB than those in NIB. Two hundred and twelve proteins were presented in the worker pupal head of these two bees on day 13, 67 proteins were specific in the worker pupal head of HRJB and 56 proteins were exclusive to NIB. While on day 15, 203 shared proteins and 93, 74 unique proteins were detected in HRJB, NIB, respectively. Until day17, 246 proteins were resolved in the head of both bees strains in the pupal period, 94 and 33 proteins were found unique to the worker pupal head of the HRJB and NIB, respectively. During the determination of PMF, 11 proteins were unambiguously identified. Hierarchical cluster analysis divided the 11 proteins into two categories, i. g. tubulin, actin 88F, ATP synthase, glutathione S transferase S1, arginine kinase, heat shock protein, proteasome 2 subunit, vacuolar ATP synthase catalytic subunit A and eukaryotic translation initiation factor 5A were up-regulated their expression, while only peroxiredoxin 2 540 was down-regulated in the head during the pupal period in both strains. 【Conclusion】 The HRJB and NIB have significant differences in protein expression in the head during the pupal development. The increased numbers of total and specific proteins expressed in the HRJB compared with that in the NIB suggested the significant change in the pupal development in the two bee strains. Furthermore, altered expression of antioxidant, metabolism and cytoskeleton confirmed that the development and metabolism of salivary gland and hypopharyngeal gland are accelerated and the nevous system is restructured during the pupal period. Finally, the data of proteomic analysis have contributed to the well understanding of the function or roles of proteins towards the development of honeybee worker pupal head of each type.

    VETERINARY SCIENCE
    Detection of Plasmid-Mediated Quinolone Resistance of Enterobacteria in Nursing Sow#br#
    LI Shu-juan,YUE Lei,LIAO Xiao-ping,JIANG Zhi-gang,CHEN Xue-ying,YANG Yan-lin,ZHANG Yue,HUANG Chu-yan,LIU Ya-hong
    Scientia Agricultura Sinica. 2010, 43(8):  1716-1722 .  doi:10.3864/j.issn.0578-1752.2010.08.021
    Abstract ( 919 )   PDF (743KB) ( 670 )   Save
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    【Objective】 To investigate the prevalence of the plasmid-mediated quinolone resistance PMQR genes and their molecule epidemiological relationship with class 1 integrons of nursing sow origin Enterobacteria in Guangdong Province. 【Method】 Seventy-four Escherichia coli, 20 Salmonella were screened for the PMQR(qnrA, qnrB, qnrS, aac(6')-Ib-cr, qepA) genes by PCR and then conformed with sequencing. intIⅠ was screened from the PMQR positive strains. Antimicrobial susceptibility tests for the PMQR positive strains were performed by the agar dilution method. 【Result】 PMQR genes were detected in 14 (14.9%) of 94 strains, including 2 strains carried qnrB,11 strains carried qnrS, and 4 strains carried aac(6')-Ib-cr gene. The strain Salmonella 14 harboured 3 PMQR genes (qnrB, qnrS and aac(6')-Ib-cr), and one E. coli 2 harboured qnrB and qnrS. qnrA and qepA were not detected. intI1 was detected in all the 14 PMQR positive strains and they showed high rate of resistance to antimicrobial agents widely used in veterinary medicine. 【Conclusion】 The prevalence of PMQR genes existed from strains of Enterobacteria in nursing sow in Guangdong Province. And had a highest prevalence of qnrS was detected among Enterobacteria strains from the farms. The co-exist of class Ⅰintegron and PMQR genes played an important role in multi antimicrobial resistance in Enterobacteria.

    Study on Pharmacokinetic/Pharmacodynamic Model of Amoxicillin Against Staphylococcus aureus in Serum and Tissue Fluid of Pigs ex vivo#br#
    YANG Yu-hui,LI Xiao-chun,HAN Xin-chou,WANG Xue-mei,WU Ke-bang,HU Ri-cha
    Scientia Agricultura Sinica. 2010, 43(8):  1723-1729 .  doi:10.3864/j.issn.0578-1752.2010.08.022
    Abstract ( 892 )   PDF (380KB) ( 1279 )   Save
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    【Objective】 The research was performed for the rational usage of amoxicillin in treating pigs infected by staphylococcus aureus. 【Method】 The combined method of pharmacokinetics in vivo and pharmacodynamics in vitro was used to survey and evaluate the pharmacokinetics-pharmacodynamics relationship of amoxicillin against staphylococcus aureus. 【Result】The MICs of amoxicillin against Staphylococcus aureus in serum or tissue cage fluid were 0.2µg•mL-1<MIC<0.4µg•mL-1 in vitro. The concentration of amoxicillin that is more than MIC in serum or tissue fluid will be needed to inhibit the bacteria if lots of staphylococcus aureus was added. EC50 were 143.63±54.35 and 29.61±5.07 respectively in serum and tissue fluid of pigs after intramuscular administration at 10 mg•kg-1 body weight. Amoxicillin produced 50% of the maximal antibacterial effect when the concentration of amoxicillin was 2.39 μg•mL-1 and 0.49 μg•mL-1, respectively, in serum and tissue fluid. 【Conclusion】 Dosage regime should be designed according to specific case when amoxicillin was employed to treat pigs infected by staphylococcus aureus. I.M injections should be administered in the amount of 20mg daily per kilo of body weight, subdivided into injections every 12 hours when pigs are infected by low-grade infection of staphylococcus aureus. When the infection become more serious, dosage regime of 30-40mg daily per kilo body weight should be designed and subdivided into injections every 6-8 hours.

    Development Change of Growth Hormone, Insulin and Their Receptors in Mammary Gland of Dairy Goats#br#
    LI Zhen,LI Qing-zhang
    Scientia Agricultura Sinica. 2010, 43(8):  1730-1737 .  doi:10.3864/j.issn.0578-1752.2010.08.023
    Abstract ( 1005 )   PDF (490KB) ( 819 )   Save
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    【Objective】 The aim of the experiment was to investigate the change of growth hormone (GH) and insulin (INS) in development of dairy goat mammary gland, and the expression of growth hormone receptor (GHR) and insulin receptor (INSR) in the same periods. 【Method】The mammary glands of Chinese Guanzhong dairy goats based on the development character were divided into 4 periods, virgin, pregnancy, lactation and involution. The levels of GH and INS in serum were measured by radioimmunoassay (RIA), and the expressions of GHR and INSR in tissue were measured by immunofluorescence (IF).【Result】 No remarkable difference in GH changes (P>0.05) in serum was observed, but in mammary gland the level of GH reached the peak in late pregnancy (2.599±0.459 ng•mL-1, P<0.01). The level of INS in serum had no significant difference in virgin (P>0.05) and in early pregnancy it reached 31.664±6.416 µIU•mL-1 (P<0.01). In mammary gland INS began to increase after pregnancy and maintained to lactation. The expression of GHR in pregnancy was higher than in virgin and in early metaphase of lactation, and reached maximum at the late of lactation. INSR increased slowly from virgin and reached the top after the lactogenesis. 【Conclusion】 In the development of mammary gland, GH not changed obviously in serum and mammary gland. The content of serum INS gradually reduced during the pregnancy period. While in mammary gland it got down gradually in the lactogenesis. GHR expressed higher in pregnancy, the later lactation and the early involution periods. The expression of INSR maintained at a high level during the pregnant and lactation periods.

    AGRICULTURAL ECONOMY & MANAGMENT
    Ecological Footprint Analysis of Food Consumption of Chinese Rural Households in the Latest 30 Years#br#
    CHEN Dong-dong,GAO Wang-sheng
    Scientia Agricultura Sinica. 2010, 43(8):  1738-1747 .  doi:10.3864/j.issn.0578-1752.2010.08.024
    Abstract ( 925 )   PDF (403KB) ( 1929 )   Save
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    【Objective】 Food consumption appropriate not only arable land but grassland and other various agricultural land resources . It is of great use of dynamic analysis of ecological appropriation for food consumption of Chinese rural households from the perspective of total agricultural land resources and then overall evaluation of resource basis to ensure food security with structure upgrading of food consumption. 【Method】 The appropriated land categories and domestic conversion factors of food consumption items were firsty discussed. Accordingly, the ecological footprint (EF) model with refined factors was applied to macro-estimate the ecological pressure of food consumption and the changes of Chinese rural residents in the latest 30 years. 【Result】 The study indicated that ecological appropriation increased from 1.465 national standard mu (nsm) in 1980 to 1.782 nsm in 2006 with the changes of Chinese food consumption structure of rural residents, and especially the direct appropriated arable land has declined dramatically since 2000. Food EF had a strong correlation with expenditure on food over the period, furthermore, it can be evidently proved by an elasticity of per capita meat and aquatic products EF 0.43. 【Conclusion】 The overall ecological pressure of food EF of Chinese rural residents has increased unsteadily, as the grain-consuming animal products and aquatic product contributed to this. The comparison of dynamic analysis of food EF calculated by fixed yields per unit area and variable yields per unit area indicated that higher productivity of arable land has greatly alleviated the pressure on resources demanded by the process of structure upgrading of the food consumption of rural residents in China.

    RESEARCH NOTES
    Effects of Light Spectral Energy Distribution on Growth and Development of Tomato Seedlings#br#
    CHANG Tao-tao,LIU Xiao-ying,XU Zhi-gang,YANG Yang
    Scientia Agricultura Sinica. 2010, 43(8):  1748-1756 .  doi:10.3864/j.issn.0578-1752.2010.08.025
    Abstract ( 1271 )   PDF (562KB) ( 1491 )   Save
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    【Objective】 Effects of different light spectral energy distributions on growth and development of tomato was studied in order to provide theoretic references for the commercial production of seedlings. 【Method】 The dysprosium lamp used as a control light source, six kinds of spectra were obtained by using light-emitting diode(LED). Tomato seedlings were grown in AGRI-LED plants cultivation system where other environmental factors were the same. Morphological parameters such as plant height, stem diameter, leaf area and physiological parameters such as root activity, content of sugar and nitrogen, enzyme activities of leaves were studied at seeding stage. At the same time, the stomata traits on leaf subsurface were observed. 【Result】 Plants under blue light, combinations of light (RB, RBG) treatment grown stronger and had a larger root-shoot ratio and also had smaller specific leaf area, but the red light had the opposite effect. Combinations of light benefited the growth and morphogenesis of seedlings. Monochrome red, yellow and green light induced seedling spindling and blue light inhibited the effects. Under blue light treatment, plants exhibited the highest total soluble sugar content, total amino acid content, and the dry weight was also the largest. The root activity and total starch content of tomato under the combinations of light (RBG) treatment was the highest. The combination of light (RB) made the leaves show highest photosynthetic rate and enzyme activity and accelerated carbon and nitrogen balanced distribution in leaves, crowns and roots. Meanwhile, the density of stomata of tomato leaves was higher and the area of stomata, stomata area per unit leaf area was larger under blue light and the combinations of light (RB, RBG). Though the total sucrose of tomato in yellow light treatment was the highest, but other kinds of indexes were smaller and it had unfavorable effect on growth and development of seedling. 【Conclusion】 It is concluded that blue light or the combinations of light (RB, RBG) benefit the growth and development of tomato seedling.

    Histomorphology of Antheraea pernyi Colleterial Glands at Different Developmental Stages
    ZHU Bao-jian,LIU Chao-liang,WU Hong-zhen
    Scientia Agricultura Sinica. 2010, 43(8):  1757-1760 .  doi:10.3864/j.issn.0578-1752.2010.08.026
    Abstract ( 845 )   PDF (422KB) ( 778 )   Save
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    【Objective】 The experiment on morphological characteristics and microstructure of colleterial glands from Antheraea pernyi female silkmoths at different developmental stages was carried out to investigate the secretion mechanism of glue materials in the colleterial glands. 【Method】 The morphological characteristics and microstructure of colleterial glands were observed by anatomy and tissue paraffin section at different developmental stages and tissues. 【Result】 The colleterial glands became bigger with the development and reached the peak as the silkmoth emerges while the color of its inclusion was changed remarkably from transparent to blue black. Differences were observed in the microstructure between secretion section and reservoir section, the former showed the characteristics of secreting cells while the later did not. 【Conclusion】 Differences in morphological characteristics and microstructure of colleterial glands were observed at various stages and in different tissues.