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Table of Content

    01 September 2012, Volume 45 Issue 17
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Detection of QTL for Kernel Weight, Grain Size, and Grain Hardness in Wheat Using DH and Immortalized F2 Population
    LI Wen-Fu, LIU , BIN , PENG , TAO , YUAN Qian-Qian, HAN Shu-Xiao, TIAN Ji-Chun
    Scientia Agricultura Sinica. 2012, 45(17):  3453-3462.  doi:10.3864/j.issn.0578-1752.2012.17.001
    Abstract ( 591 )   PDF (565KB) ( 659 )   Save
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    【Objective】 Grain weight (GW), grain size (GS), and grain hardness (HD) are important complex traits in wheat, which are determined by quantitative trait loci (QTLs). In this study, to attain more genetic information of GW, GS, and HD in wheat, QTLs with additive effects and dominant effects for these traits were detected. 【Method】 Based on a genetic linkage map contains 368 sites, QTLs for GW, GS, and GS were evaluated in five different environments by using a doubled haploid (DH) population lines derived from a cross between two elite Chinese wheat cultivars Huapei 3/Yumai 57 (Triticum aestivum L.) and in two different environments by using an immortalized F2 population generated by randomly permutated intermating of these DHs. 【Result】 A total of 32 additive QTLs and 18 pairs of epistatic QTLs were detected, including 8 additive QTLs and 5 pairs of epistatic QTLs for 1000-kernel weight, 10 additive QTLs and 6 pairs of epistatic QTLs for grain length , 10 additive QTLs and 6 pairs of epistatic QTLs for grain diameter, and 7 additive QTLs and 1 pair of epistatic QTLs for grain hardness. The QTL Qtkw6A for GW was persistently expressed in both of DH and IF2 population, and the contributions of additive effect in the two population were 9.39% and 11.75%, respectively, and 1.37% for that of dominance effect. The QTL Qgd6A for grain diameter was persistently expressed in both of DH and IF2 population, and the contributions of additive effect in DH and IF2 population were, respectively, 15.02% and 15.03%, It was the same as the QTL Qgl6A, the contributions of additive effect in DH and IF2 population were, respectively, 14.96% and 15.10%. 【Conclusion】The results showed that both additive effects and epistatic effects were important genetic bases of GW and GS. Grain hardness is mainly controlled by a major gene located on the short arm of chromosome 5D, and is also influenced by other minor genes and epistatic effects. The QTL detected in this study could be used in MAS. The QTL with dominance effect and those with interactions between dominance and additive, additive and dominance, dominance and dominance detected by IF2 population would provide assistance for the study on heterosis.
    Silencing of Cytochrome P450 in Sitobion avenae and Myzus persicae Through RNA Interference
    WANG Hui, ZHANG Min, ZHANG Xiao-Hong, XIA Lan-Qin
    Scientia Agricultura Sinica. 2012, 45(17):  3463-3472.  doi:10.3864/j.issn.0578-1752.2012.17.002
    Abstract ( 690 )   PDF (1041KB) ( 1086 )   Save
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    【Objective】Silencing of cytochrome P450 in S. avenae and M. persicae through RNA interference by feeding on artificial diet containing double-stranded RNA (dsRNA).【Method】 Fragments of cytochrome P450 in S. avenae and M. persicae were obtained by PCR. The dsRNA of the conserved sequence of cytochrome P450 from S. avenae and M. persicae were synthesized by dsRNA synthesis kit. dsRNA was added to the final concentration of 0, 3, 5, and 7.5 ng•μL-1 in aphid artificial diet, respectively. Three replicates were set up for each experimental group. The number of surviving aphids was counted after 2, 4, 6, and 8 days of feeding. The expression level of target genes were analyzed by real-time fluorescence quantitative PCR.【Result】Cytochrome P450 fragments were cloned from the cDNA of S. avenae and M. persicae, the homology of the two genes was 90.1%. The mortality of aphids increased along with the increased dsRNA concentration as time went on. Total RNA were extracted from aphids fed by dsRNA at concentration of 7.5 ng•μL-1 and collected from different time periods. The results of real-time fluorescence quantitative PCR showed that the target genes were silenced significantly. 【Conclusion】The expression level of cytochrome P450 in S. avenae and M. persicae which fed by the dsRNA of the conserved sequences of cytochrome P450, were suppressed, and thus led to the death of aphids.
    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    The Temporal-Spatial Dynamic Analysis of China Rice Production
    CHENG Yong-Xiang, WANG Xiu-Zhen, GUO Jian-Ping, ZHAO Yan-Xia, HUANG Jing-Feng
    Scientia Agricultura Sinica. 2012, 45(17):  3473-3485.  doi:10.3864/j.issn.0578-1752.2012.17.003
    Abstract ( 646 )   PDF (1171KB) ( 1390 )   Save
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    【Objective】The trajectories of center of gravity rice acreage and production in China’s north and south regions and different rice systems were drawn respectively, the causes are analyzed, and the main factors that affect the total rice production was found at present stage.【Method】Using gravity fitting model, the temporal-spatial dynamic of rice production in China was analyzed. The main factors affecting China’s total rice production at present were found with sensitivity analysis.【Result】 In addition to the 1960s and 1970s center of gravity of rice sown acreage and production in China had skewed toward the southeast and eastern, it general moved toward the northeast direction since the founding of China. In 1979-2009, the center of gravity of rice in north moved quickly to the Songliao Basin of northeast of China. Center of gravity of middle-season rice and single cropping later rice moved firstly toward northeast then toward southeast, and its direction of movement was consistent with the order of the change of southern rice system, i.e. “double cropping rice changes to single cropping rice” from north to south. Center of gravity of early-season rice and double-season later rice both moved toward southwest. The percentage of single cropping rice was increased significantly in south China; the phenomenon “change double rice to single one” was obvious, which was the main reason leading to the decline of China’s rice acreage. Combined with sensitivity analysis, we can see that the sown acreage becomes the key factor most possibly affecting rice production stability.【Conclusion】 Regulation of market mechanism and technological progress were the two main factors currently affecting the temporal-spatial dynamic changes of rice in China. Under the precondition of keeping on the guidance of the market, from the perspective of food security, according to national conditions, especially to regulate early-season rice and double-season late rice that have low costs profit margins and downward trend of acreage, at same time taking into account total rice acreage, to prevent it from large fluctuation, to improve the yield of rice constantly, is the key to ensure steady increase of China rice output at this stage.
    Retrieving LAI of Winter Wheat Based on Sensitive Vegetation ndex by the Segmentation Method
    LI Xin-Chuan, XU Xin-Gang, BAO Yan-Song, HUANG Wen-Jiang, LUO Ju-Hua, DONG Ying-Ying, SONG Xiao-Yu, WANG Ji-Hua
    Scientia Agricultura Sinica. 2012, 45(17):  3486-3496.  doi:10.3864/j.issn.0578-1752.2012.17.004
    Abstract ( 650 )   PDF (410KB) ( 1061 )   Save
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    【Objective】The method of inversion leaf area index (LAI) using a single vegetation index (VI) is influenced by different degrees of saturability and soil background. This paper proposed a method choosing sensitive vegetation index by the segmentation method to form optimal VI combination, and to improve the accuracy of LAI inversion.【Method】In this study the ACRM radiation transmission model was used to simulate data, and the ground measured spectrum data were obtained. The study analyzed soil sensitivity and saturability about the common vegetation index to determine the segment point of LAI, and chose the best vegetation index based on segment point of LAI to form a combination VI for achieving the final inversion of the LAI. This method was also used in the regional winter wheat LAI inversion application with the Landsat5 TM data. 【Result】The analysis showed that, LAI = 3 was the more appropriate segment point, and the use of vegetation index segment combination OSAVI (LAI ≤3) + TGDVI (LAI>3) partly overcame soil factors and the saturation problems. The joint inversion results were significantly superior to the single vegetation index retrieval accuracy.【Conclusion】LAI was effectively inversed with the higher accuracy by choosing the best vegetation index through the segmentation method.
    Effects of Shading on Stalks Morphology, Structure and Lodging of Summer Maize in Field
    CUI Hai-Yan, JIN Li-Bin, LI Bo, ZHANG Ji-Wang, ZHAO Bin, DONG Shu-Ting, LIU Peng
    Scientia Agricultura Sinica. 2012, 45(17):  3497-3505.  doi:10.3864/j.issn.0578-1752.2012.17.005
    Abstract ( 672 )   PDF (532KB) ( 1696 )   Save
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    【Objective】 With the decreasing of the solar radiation in China, effects of shading (low light stress) on stalk morphology and microstructure of summer maize were studied to provide a theoretical basis for summer maize to adapt the current climatic condition of insufficient illumination. 【Method】 Summer maize was grown under the shading condition with a shading degree of 60%. Three shading treatments were designed in growing season ranged from tasseling to maturity (S1), from jointing to tasseling stage (S2) and whole growing period (S3), respectively. Two summer maize cultivars Denghai605 (DH605) and Zhengdan958 (ZD958) were used as experimental materials. Effects of shading in different growing seasons on stem characters and lodging property of summer maize were investigated. 【Result】 Results showed that plant height, ear height, leaf area index, the third internodes length, cross sectional area of stem, and stalk rind penetration strength reduced significantly after shading, and the response to different shading stages showed: S1<S2<S3. Compared to CK, stalk rind penetration strength of DH605 and ZD958 reduced by 22.23% and 24.41%, respectively. Stem rind thickness, vascular bundle number, the area of the central vascular bundle, xylem and phloem reduced significantly. Lodging property in fields increased significantly, and the lodging rate of DH605 and ZD958 in S3 were 36.5% and 24.7%, respectively. Furthermore, grain yield, kernel number per ear and harvest ear number reduced significantly. Barrenness increased and grain yield of S3 reduced. 【Conclusion】Shading had significant effects on stem form. Stem lodging resistance was also reduced by decreased basal internode length, diameter, puncture strength, hard skin tissue thickness and vascular bundle number, thus influencing yield formation. Shading before anthesis had a great influence on the lodging rate.
    PLANT PROTECTION
    Root and Leaf Infection as Revealed by Autofluorescent Reporter Protein GFP Labeled Bipolaris sorokiniana in Wheat
    YAN Li-Min, WANG Xiao-Ming, XU Rong-Qi, DONG Fang-Yang, LI Hong-Jie
    Scientia Agricultura Sinica. 2012, 45(17):  3506-3514.  doi:10.3864/j.issn.0578-1752.2012.17.006
    Abstract ( 619 )   PDF (578KB) ( 749 )   Save
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    【Objective】The objective of this study is to label Bipolaris sorokiniana, which is the causal agent of common root rot and leaf spot blotch of wheat (Triticum aestivum L.), with green fluorescent protein (GFP) for studying infection of B. sorokiniana on wheat roots and leaves, and to establish a direct and non-destructive method for in vivo observing interaction between pathogen and host plant. 【Method】 Gene gfp was transferred into B. sorokiniana strain Bs-1 via Agrobacterium tumefaciens-mediated transformation (ATMT). Based on analysis of fluorescence expression, PCR verification, genetic stability, growth parameters, and metabolism of extra cellular enzymes, a transformed strain Bs-GFP that resembled its wild-type strain, was chosen to investigate infection of B. sorokiniana on roots and leaves of wheat cultivar Aikang 58. 【Result】Bright green fluorescence was observed in hyphae and conidia of the transformant Bs-GFP. The amplification of the diagnostic bands with the gene-specific markers indicated that gfp was integrated into the genome of B. sorokiniana. Analysis of genetic stability and growth parameters demonstrated that gfp was inherited normally, and the growth rate and metabolism of extracellular enzymes were comparable to the wild type strain. The GFP-labeled strain Bs-GFP was able to incite symptoms on both underground and upperground parts of wheat plants, and the quantities of colonization (in terms of colony forming unit) in roots and basal stem tissues of wheat plants by Bs-GFP was similar as those by Bs-1. 【Conclusion】The B. sorokiniana strain expressing GFP obtained via ATMT can be used to directly monitor infection and colonization of the fungus on wheat. The results from this study will facilitate better understanding of the interactions between B. sorokiniana and wheat, as well other cereal crops.
    Identification and rDNA-ITS Sequence Analysis of the Pathogen of Kenaf Anthracnose
    LIU Xiao-Qian, QI Jian-Min, CHEN Yu-Sen, CHEN Mian-Cai, CHEN Mei-Xia, LIU Wei, FANG Ping-Ping, LIN Li-Hui, TAO Ai-Fen
    Scientia Agricultura Sinica. 2012, 45(17):  3515-3521.  doi:10.3864/j.issn.0578-1752.2012.17.007
    Abstract ( 545 )   PDF (630KB) ( 704 )   Save
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    【Objective】The objective of this study is to determine the possible pathogens of kenaf anthracnose of China.【Method】The samples of diseased kenaf were collected and isolated from Anhui, Henan, Zhejiang, Fujian Provinces of China. AH, HN, ZJ, FJ strains were selected from sixteen pathogens and their pathogenicities were determined. Identification was carried out by using traditional morphological methods and sequencing of rDNA-ITS. 【Result】 The differences among AH, HN, ZJ and FJ strains in morphological characteristics and pathogenicities were relatively abundant. The rDNA-ITS length of AH, HN, ZJ, and FJ strains was 541, 545, 541, and 535 bp, respectively. The homology between FJ strain and other strains was 90%-91%, and the homology among AH, HN, ZJ strains was 96%-98%.【Conclusion】The pathogens of kenaf anthracnose were identified by analysis at the molecular level. It is confirmed that kenaf anthracnose in China is caused by C. dematium and C. gioeosporioides, and the pathogenicity of C. dematium is greater than C. gioeosporioides.
    Antenna Ultrastructure of Microplitis mediator
    LI Ke-Ming, ZHANG Yong-Jun, WU Kong-Ming, GUO Yu-Yuan
    Scientia Agricultura Sinica. 2012, 45(17):  3522-3530.  doi:10.3864/j.issn.0578-1752.2012.17.008
    Abstract ( 610 )   PDF (3075KB) ( 619 )   Save
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     【Objective】 The objective of this study is to investigate the types, distribution, morphology and internal structure of antennal sensilla of M. mediator.【Method】 Scanning electron microscope (SEM) and transmission electron microscope (TEM) were used for the observation.【Result】 The results of SEM show that antennae of M. mediator were made up of scape, pedicel and flagellum, which comprises 16 flagellomeres. Both the antennae of male and female contain six kinds of antennal sensilla, namely, sensillum trichodium, sensillum chaeticum, sensillum placodeum, sensillum basiconicumⅠ, sensillum basiconicumⅡ and sensillum coeloconicum. There was no significant difference in length, distribution and amount of each kind of sensillum between male and female M. mediator. The results of TEM show that there were porous thick cuticular wall and dendrites in lumen in sensillum trichodium and sensillum coeloconicum, aporous thick cuticular wall and dendrites in lumen in sensillum chaeticum and sensillum basiconicumⅠ, porous thin cuticular wall and dendrites in lumen in sensillum placodeum and aporous thick cuticular wall and no dendrites in lumen in sensillum basiconicumⅡ. 【Conclusion】 In this work, the internal and external ultrastructure of M. mediator antennal sensilla is obtained, and the results could provide a theoretical basis for olfactory perception behavior of parasitoids.
    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Statistical Analysis of Methane Emission from Rice Fields in China and the Driving Factors
    WEI Hai-苹, SUN Wen-Juan, HUANG Yao
    Scientia Agricultura Sinica. 2012, 45(17):  3531-3540.  doi:10.3864/j.issn.0578-1752.2012.17.009
    Abstract ( 616 )   PDF (515KB) ( 966 )   Save
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    【Objective】 The objectives of this study are to address the spatial characteristics of methane (CH4) emission from rice fields in China and to quantify the relationship between CH4 emission and driving factors associated with environment and field management in different rice cropping systems based on the data extracted from published papers.【Method】By searching literature databases, 111 articles published since 1987 were obtained that related to the field measurements of CH4 emission in different regions. A total of 495 datasets of the seasonal amount of CH4 emission or the mean flux of CH4 emission over a growing season measured at 67 sites were extracted from these articles. ANOVA, correlation analysis and stepwise regression were used to investigate the spatial characteristics of CH4 emission and driving factors. 【Result】The mean and standard error of CH4 emission from single rice, early-rice and late-rice cropping system were (383.5±31.1), (234.3±16.80) and (361.8±25.0) kg•hm-2, respectively. Methane emission from the single rice in southwest China was the highest, approximately being 1.3, 1.2 and 3.3 times higher than those from north, central and northeast China. In double rice cropping system, CH4 emission from the early- and the late-rice in central China was approximately 160% and 90% higher than those in south China (Guangxi and Guangdong Provinces). There is a significant positive correlation between CH4 emission and the input of organic carbon, while CH4 emission was found to be negatively correlated with the rate of synthetic nitrogen application and soil pH. CH4 emission from the single rice system declined with increasing latitude, longitude and soil total nitrogen. By contrast, CH4 emission increased from south to north in the double rice cropping system, and increased with soil total nitrogen in the late-rice system. The spatial variability in seasonal amount of CH4 emission for the single rice can be well quantitatively described by a linear combination of six variables (R2=0.72, P=0.000, n=241), including latitude, soil parameters of C﹕N ratio and sand fraction, irrigation regime, rates of organic carbon input and synthetic nitrogen application. For the early-rice, the spatial variability in CH4 emission was quantitatively determined by a linear combination of five variables (R2=0.35, P=0.000, n=121), including soil parameters of C﹕N ratio and sand fraction, mean temperature from rice transplanting to heading, irrigation regime and rate of organic carbon input. A linear combination of six variables, including latitude, soil parameters of C﹕N ratio, clay fraction and pH, mean temperature over the rice growing season and rate of organic carbon input, determined the spatial variability in CH4 emission for the late-rice system (R2=0.47, P=0.000, n=133).【Conclusion】The seasonal amount of CH4 emission per unit area from rice field ranks in a descent order of single rice, late-rice and early-rice in China, but no significant difference existed between the single and late-rice. CH4 emission in southwest China is significantly higher than those in other single rice regions. For the double rice system, CH4 emission in central China is significantly higher than those in south China. Linear multi-regression models taking into account environment and field management could be used to explain the spatial variability in seasonal amount of CH4 emission for all rice cropping systems, and the explanatory capability of model for the single rice is much better than that for the double rice system.
    Application of the Carbon Isotope in the Partitioning of Soil Respiration in Grassland
    GENG Yuan-Bo, SHI Jing-Jing
    Scientia Agricultura Sinica. 2012, 45(17):  3541-3550.  doi:10.3864/j.issn.0578-1752.2012.17.010
    Abstract ( 560 )   PDF (393KB) ( 1393 )   Save
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    Grassland ecosystem plays an important role in regional climate changes and global carbon cycle. The partitioning of soil respiration in grassland ecosystem is the foundation of quantitative evaluation of grassland vegetation and soil carbon balance, which could help to improve understanding and prediction of the responses of grassland ecosystem to climate changes. In this paper, several methods for separating soil respiration in grassland ecosystem by the application of stable carbon isotope 13C and radioactive carbon isotope 14C were reviewed. Methods for the separation of SOM-derived respiration and root-derived respiration mainly include: 13C natural abundance method, FACE experiment, 13C pulse labeling. Also, the methods for the separation of root and rhizomicrobial respiration mainly include: isotope dilution method, model rhizodeposition method, modeling of 14CO2 efflux dynamics method, and exudates elution method. Carbon isotope methods are novel methods that involve less disturbances to soil-plant system, which ultimately enhance the accuracy of estimation. Overall, carbon isotope methods have a great potential in the study of partitioning of soil respiration in grassland ecosystem.
    HORTICULTURE·STORAGE·FRESH-KEEPING·PROCESSING
    Development and Mapping of Molecular Markers Closely Linked to CRb Gene Resistance to Clubroot Disease in Chinese Cabbage
    CHEN Hui-Hui, ZHANG Teng, LIANG Shan, CHEN Bing, ZHANG Chun-Yu, PU Zhong-Yun
    Scientia Agricultura Sinica. 2012, 45(17):  3551-3557.  doi:10.3864/j.issn.0578-1752.2012.17.011
    Abstract ( 946 )   PDF (590KB) ( 838 )   Save
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    【Objective】The objective of this study is to develop molecular markers closely linked to clubroot resistance (CR) gene CRb in Chinese cabbage, and to evaluate their utility in germplasms of Brasisca rapa.【Method】A F2 mapping population was constructed with a cross between ‘CR Shinkii DH’ line, a Chinese cabbage doubled haploid line containing CRb gene, and a clubroot susceptible Chinese cabbage inbred line ‘07Q69’. Based on the target sequences showing homologous to the flanking markers TCR01, TCR05 and TCR09 linked to CRb on the chromosome A3 of Brassica rapa, primer pairs were designed and markers closely linked to CRb were developed. Forty-two germplasms belonging to different subspecies of B. rapa were employed to evaluate the transferability of markers linked to CRb.【Result】 Four co dominant and one dominant markers closely linked to CRb were developed and mapped. The markers TCR25 and TCR74 were located on one side of CRb, while TCR13, TCR42 and TCR34 were on the other side along with the previous markers TCR01 and TCR05. The CRb gene was flanked by two nearest markers TCR74 and TCR13 with the same genetic distance of 0.09 cM. The transferability evaluation of TCR74 and TCR13 showed that they possessed high polymorphism between clubroot resistant and susceptible sources.【Conclusion】The CRb gene was mapped between two codominant flanking markers with genetic distance of 0.18 cM. The two nearest flanking markers showed high polymorphism and good versatility between resistant and susceptible sources of B. rapa.  
    Simultaneous Determination of 18 Flavonoids in Citrus Fruits y High-Performance Liquid Chromatography
    ZHANG Yuan-Mei, ZHOU Zhi-Qin, SUN Yu-Jing, SHEN Yan, ZHONG Lie-Zhou, QIAO Li-Ping, YE Xing-Qian
    Scientia Agricultura Sinica. 2012, 45(17):  3558-3565.  doi:10.3864/j.issn.0578-1752.2012.17.012
    Abstract ( 712 )   PDF (566KB) ( 1849 )   Save
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    【Objective】This study aimed to investigate a high-performance liquid chromatograph(HPLC)method for the determination of different kinds of flavonoids, namely, eriocitrin, taxifolin, narirutin, naringin, hesperidin, neohesperidin, rhoifolin, quercitrin, eridictyol, didymin, poncirin, naringenin, luteolin, kaempferol, diosmetin, sinensetin, nobiletin and tangeretin in citrus fruits. 【Method】The chromatographic separation was performed on a ZORABX SB-C18 column kept at 25℃ using methanol /formic acid/water mixture as the mobile phase at a flow rate of 0.7 mL•min-1 through gradient elution and the detection wavelength was set at 283 nm for flavanones, 330 nm for flavones and polymethoxylated flavones and 367 nm for flavonol. 【Result】 Results showed that the thorough separation of 18 flavonoids was achieved within 42 min. The method exhibited a linear range of 62.4-960 mg•L-1 (r=0.9998-1.0000) and limits of quantification (LOQ) ranging from 0.05-0.133 μg•mL-1. The average recovery rates of this method were 88.70%-104.76%, with a relative standard derivation range of 0.33%-3.05%. 【Conclusion】This method is simple, rapid, accurate , reliability, and can be used for the determination of flavonoids in citrus fruits.
    ANIMAL SCIENCE·RESOURCE INSECT
    Molecular Cloning and Application in Cell Reprogramming rom Porcine L-Myc Gene
    LI Zhen-Zhen, CUI Yi, GAO Yi, CHENG De, LIU Ya-Jun, WANG Hua-Yan
    Scientia Agricultura Sinica. 2012, 45(17):  3566-3575.  doi:10.3864/j.issn.0578-1752.2012.17.013
    Abstract ( 531 )   PDF (695KB) ( 686 )   Save
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    【Objective】Cloning porcine L-Myc gene and exploring its function in cell reprogramming at the protein level could be helpful to provide a foundation for substituting L-Myc gene for c-Myc gene in the induction of induced pluripotent stem (iPS) cells in porcine. 【Method】Through NCBI sequence alignment, porcine L-Myc gene cDNA was obtained by RT-PCR. The homology of L-Myc gene among these three species of porcine, human and mouse was analyzed by bioinformatics. The fusion expression vector pEGFP/L-Myc-C1 was constructed, and the vector-transfected and western blotting were used to detect the expression of the cloned porcine L-Myc gene cDNA at the protein level. Then the L-Myc gene was inserted into the retroviral vector, and different transcription factors, respectively, induced porcine embryo fibroblast (PEF) cells. Through observing the variation of the induced cells and alkaline phosphatase (AP) staining, the important role porcine L-Myc gene plays in cell reprogramming was verified. 【Result】 A 1 113 bp porcine L-Myc gene cDNA sequence was obtained by RT-PCR, encoding 364 amino acids and the theoretical molecular weight is 40 kD. Bioinformatics analysis showed that a high degree of homology in L-Myc existed among porcine, human and mouse. The western blotting analysis proved that the cloned porcine L-Myc gene cDNA could be expressed at the protein level. After induction, the morphology of induced cells are varied, and the quantity of positive clones induced with the transcription factors Oct4, Sox2, Klf4, and L-Myc (OSKL) was far more than that with Oct4, Sox2, and Klf4 (OSK). 【Conclusion】 The porcine L-Myc, a transcription factor, was cloned, and the results of analysis showed that porcine L-Myc gene plays an important role in cell reprogramming process.
    Construction and Integration Efficiency of PiggyBac Transposon Inducible Cell Immortalization Transgenic Vector
    ZHANG Ting-Ting, XIN Ying, ZHANG Zhi-Qiang, REN Chong-Hua, YANG Han-Jiang, WANG Ling, ZHANG Zhi-Ying
    Scientia Agricultura Sinica. 2012, 45(17):  3576-3583.  doi:10.3864/j.issn.0578-1752.2012.17.014
    Abstract ( 919 )   PDF (405KB) ( 966 )   Save
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    【Objective】 The objective of the study is to construct a Piggybac transposon-based transgenic vector for multiple gene expression and selection of removable marker gene.【Method】 To construct positive and negative selection marker gene cassette based on pXL-BacII, the two marker genes, neo (with SV40 promoter) and tk, were PCR amplified from the corresponding vectors and connected with IRES sequence by an overlap PCR approach. Then LoxP sequence was added into the expression cassette ends and resulted in PB-NIT. The transgenes, SV40 large T and mouse p 53 linked with T2A sequence, were firstly cloned into an expression vector with an inducible promoters Tet-CMVP and SV40 Poly A. The transgene expression cassette was then inserted into the PB-NIT vector and resulted in PB-NIT (LoxP-SV40P neo-IRES-tk-PloyA-LoxP)-STP (Tet-CMV-SV40 T-T2A-p 53-PloyA). Finally, the rtTA gene expression cassette was cloned into the PB-NIT-STP vector and a final transgene vector, PB-rtTA-NIT-STP, was obtained. 【Result】 The final transgene vector was verified by both enzyme digestion and sequencing. The transgene vector transposon activity was examined by co-transfecting the transgene vector and PiggyBac transposase expression vector PB-transposase into mouse ES cells. The results demonstrated that the number of the positive colonies in ES cell transfected with PB-rtTA-NIT-STP and PB-transposase was 20 times more than that transfected with PB-rtTA-NIT-STP alone. 【Conclusion】 The PiggyBac transposon-based transgene vector constructed in this study could be used for inducible multiple gene expression and for selection of marker free transgene generation.
    Effect of Different Levels of Vitamin A on the Colony Development and Larvae Antioxidation of Apis mellifera ligustica During he Period of Spring Multiplication
    FENG Qian-Qian, YANG Wei-Ren, XU Bao-Hua, LI Cheng-Cheng
    Scientia Agricultura Sinica. 2012, 45(17):  3584-3591.  doi:10.3864/j.issn.0578-1752.2012.17.015
    Abstract ( 692 )   PDF (314KB) ( 543 )   Save
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    【Objective】 The objective of this study is to study the effect of different levels of vitamin A on the population and larvae antioxidation of A. mellifera ligustica during the period of spring multiplication. 【Method】 Twenty-five colonies of honeybees (no pollen in colonies) from a local apiary were used in the study. The honeybee colonies were randomly allocated into 5 groups with 5 replicaties for each. The 5 groups of honeybees were fed with basal diet with supplementation of vitamin A at the levels of 0 (group 1), 5 000 (group 2), 10 000 (group 3), 15 000 (group 4) and 20000 IU•kg-1 (group 5), respectively. The colony development and antioxidant activities of larvae of the 5 groups were evaluated. 【Result】 During the period of spring multiplication, there were significant differences in colony development and feeding amounts of the groups fed with different levels of vitamin A. The colony development of group 3 was significantly faster than the control group and other experimental groups (P<0.05) at the end of trial. The antioxidant activities (T-SOD, CAT and T-AOC) of 5-day-old larvae of group 3 and group 4 were significantly higher than the control and other experimental groups (P<0.05). The content of MDA was significantly lower than the control and other experimental groups (P<0.05). Except for group 4, the workers’ emergent weight of group 3 was significantly higher than the control and other experimental groups (P<0.05).【Conclusion】Optimal diet vitamin A supplementation (10 000-15 000 IU•kg-1) will increase the population of bee colony and antioxidant activities of larvae of A. mellifera ligustica during the period of spring multiplication.
    Cloning and Sequence Analysis of JAK/STAT Pathway Related Major Genes of Silkworm (Bombyx mori)
    PENG Jing-Kun, CAO Guang-Li, QIAN Ying, ZHANG Xiao-Li, MA Huan-Yan, HE Lei, XUE Ren-Yu, GONG Cheng-Liang
    Scientia Agricultura Sinica. 2012, 45(17):  3592-3601.  doi:10.3864/j.issn.0578-1752.2012.17.016
    Abstract ( 665 )   PDF (528KB) ( 751 )   Save
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    【Objective】The objective of this study is to identify the elements of JAK/STAT pathway of silkworm, Bombyx mori and determine the function of JAK/STAT pathway related major genes in silkworm. 【Method】JAK/STAT pathway related major genes in silkworm were obtained through electronic cloning and conventional RT-PCR. Domain analysis on the related protein were conducted by using bio-softwares online, and the expression profile of these genes in diverse tissues were analysed through microarray database or qPCR.【Result】The silkworm JAK/STAT pathway related major genes including BmSTAT, BmHOP, BmSOCS2, BmSOCS5A, BmSOCS5B, BmSOCS6, BmDRK, Bmken, BmPIAS1, BmPIAS2 and BmPI3K were cloned, respectively, but the homologues of Drosophila Upd1, Upd2 and Upd3 were not cloned. Sequence analysis showed that BmHOP contained CRD_FZ, Kringle domains, transmembrane region and Pkc superfamily protein kinase catalysis domain TyrKc. BmSTAT had two isoforms, which were consisted of SH2, STAT_bind, STAT_int and STAT_alpha domains. Both BmSOCS2A and BmSOCS6A had a conserved SH2 domain and a SOCS box in cloned three genes of BmSOCS family. BmDRK was consisted of SH3 and SH2 domains that connected in series to form SH3-SH2-SH3. BmKen contained BTB and zinc finger domains. BmPIAS1 and BmPIAS2 contained LCR domains. BmPI3K had two conserved SH2 domains.【Conclusion】The JAK/STAT pathway related major genes were cloned successfully in silkworm. The analysis of conserved structural domains suggests that BmHOP is one of the JAK kinases, which phosphorylates the BmSTAT transcription factor and leads to activation of target genes. BmSOCS members, Bmken, BmPIAS1 and BmPIAS2 are negative feedback regulators in the JAK/STAT pathway.
    VETERINARY SCIENCE
    Antimicrobial Resistance Analysis and Detection of Methicillin- Resistant Staphylococcus aureus (MRSA) Among Staphylococcus aureus Strains Isolated from Bovine Mastitis
    SU Yang-., PU Wan-Xia, CHEN Zhi-Hua, DENG Hai-Ping
    Scientia Agricultura Sinica. 2012, 45(17):  3602-3607.  doi:10.3864/j.issn.0578-1752.2012.17.017
    Abstract ( 632 )   PDF (405KB) ( 574 )   Save
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    【Objective】The aim of the study is to investigate the antimicrobial resistance of S. aureus (Science) and to detect the presence of methicillin-resistant S.aureus (MRSA) among S.aureus strains isolated from bovine mastitis in Inner Mongolia, and to provide credible theory evidence for prevention and treatment of bovine mastitis. 【Method】 K-B disk diffusion method was used to test drug sensitivity of 38 total strains of S.aureus to 17 commonly used antibiotics. Meanwhile, agar screen was performed to identify the minimum inhibitory concentration of oxacillin and vancomycin to all strains. Cefoxitin, oxacillin disk diffusion and oxacillin agar screen were used to detect the phenotype of MRSA, and PCR assay was generated the genotype of MRSA as well. 【Result】 The isolates had different degrees of antimicrobial resistance to each antibiotic, the frequency of resistance to ampicillin, cefradine, penicillin, cotrimoxazole, novobiocin and streptomycin was more than 45%, yet keeping sensitive to ofloxacin, vancomycin, amikacin, ciprofloxacin, gentamicin and cefazolin was over 90%. Two of the strains with vancomycin MIC were ≥16 μg•mL-1. The MICs of oxacillin for eight of total strains were ≥ 8 μg•mL-1, others with oxacillin MICs were ≤2 μg•mL-1. The multidrug resistance was severe, 84.21% of the strains were resistant to at least 3 kinds of antimicrobial agent, four of the total strains can survive in the presence of night various antibiotics. 16(42.11%) S.aureus strains carried mecA gene detected by PCR assay. However, there were only seven of them have the minimum inhibitory concentration over 4 μg•mL-1. When cefoxitin, oxacillin disk diffusion and oxacillin agar screen methods were generated the phenotype of MRSA, only 7, 10 and 7 strains of each were identified. 【Conclusion】 The antimicrobial resistance and multidrug resistance of S aureus were serious. High incidence of MRSA and OS-MRSA has been associated with bovine mastitis in the surveyed region.
    Antibiotic Resistance and Coagulase Typing of Staphylococcus aureus Isolates from Pigs and Cows in Part of China
    LIU Yang, LIANG Yao-Feng, JIAO Xin-An, SONG Li, ZHANG Chun-Ping, NING Yi-Bao, XU Shi-Xin
    Scientia Agricultura Sinica. 2012, 45(17):  3608-3616.  doi:10.3864/j.issn.0578-1752.2012.17.018
    Abstract ( 691 )   PDF (469KB) ( 663 )   Save
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    【Objective】The objective of the experiment is to study the status of antibiotics resistance and coagulase typing distribution of S.aureus in China, and provide a scientific basis for effective control of transmission and prevalence of S.aureus from animals.【Method】Antibiotics resistance was tested by broth dilution method, coagulase serotypes were identified by coagulase grouping serum, coa genes were amplified by PCR method and then sequenced for further analysis by mega software. 【Result】 A total of 124 S.aureus isolates were identified, and the result of resistance detection revealed that the resistant rate of ampicillin and penicillin was near 100%, erythromycin reached 75%, and other 10 antibiotics was below 50%. The positive rate of MRSA was 27%. No vancomycin resistant isolates was found. Six kinds of serotypes were observed and the rate of coagulase serotyping reached 91%: serotypes VII,VI and combined type were prevalent, whereas serotypes Ⅲ and Ⅴwere not found. The phylogenetic tree of coagulase gene was constructed. 【Conclusion】 The status of antibiotics resistance in China was relatively severe, swine isolates were severer than cattle ones. Coagulase serotypes were abundant and the distribution was comparative intensive, and possessed unique prevalence feature. The evolutional characteristic of coa gene had no direct relationship with coagulase serotypes, and this study has provided a theoretical basis for further veterinary research about molecular epidemiology and evolutionary tendency.
    AGRICULTURAL ECONOMY & MANAGMENT
    Patent Quality in China’s Agricultural Sector: A Comparison Across Different Filing Periods, Applicants and Technology Fields
    LIU Li-Jun, SONG Min
    Scientia Agricultura Sinica. 2012, 45(17):  3617-3623.  doi:10.3864/j.issn.0578-1752.2012.17.019
    Abstract ( 475 )   PDF (249KB) ( 961 )   Save
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    【Objective】The study aims to investigate the change in patent quality across different filing periods, applicants and technology fields in China’s agricultural sector when the volume of patent filings rose dramatically. 【Method】Using the total agricultural patents filed in China as a sample, the paper employs statistical and econometric models to examine invention patent  ratios, invention patent grant ratios, grant lapse ratios and patent maintenance period. 【Result】There is no significant difference in grant ratio between domestic applications and foreign applications, but foreign grants are maintained longer than domestic counterparts. At the 0.01 significance level, for domestic applicants, the invention grant ratio of domestic public sector organizations (e.g. research institutes, universities) is a bit higher than that of companies and individuals’ ones; however the probability of renewing patent is significantly smaller than that of companies. The coefficients of time dummies show that both the invention grant ratio and renewal probability of domestic filings are on the rise in recent years. 【Conclusion】It is found that, the quality of Chinese domestic filings has been improved significantly in recent years although it still falls behind that of foreign filings. Particularly the progress in agro-biotechnology is the most striking. Public sectors organizations (universities and public agricultural research institutions) dominate the patenting activities in the agricultural sector and they have a better patent quality than private applicants (companies and individuals), however, most of university patents lapse rapidly in the first three years. The main policy implication is that government should guide and regulate the filing motives of public sector organizations and promote the transfer of agricultural patents from public sector to private sector.
    RESEARCH NOTES
    Functional Analysis of Cyclophilin (GhCYP1) in Gossypium hirsutum
    WU Li, LI Yan-Jun, ZHANG Xin-Yu, WANG Ya-Qin, SUN Jie
    Scientia Agricultura Sinica. 2012, 45(17):  3624-3631.  doi:10.3864/j.issn.0578-1752.2012.17.020
    Abstract ( 487 )   PDF (612KB) ( 568 )   Save
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    【Objective】The objective of this study is to investigate the effects of cotton GhCYP1 gene on drought tolerance of tobacco.【Method】The plant expression vector of GhCYP1 gene was constructed by using gateway cloning technology and transformed into tobacco based on Agrobacterium tumerficiens-mediated transformation approach. Transgenic tobacco plants fused the target genes were generated. Wild type (WT), transgenic lines L1 and L2 of tobacco were used to measure the relative content of chlorophyll in leaf discs, relative water content, malonyldialdehyde content and relative electrical conductivity in plants.【Result】Compared with WT, transgenic tobacco plants form a stronger root system. Under the conditions of water stress, the leaf chlorophyll was less affected in transgenic plants which had obviously higher leaf chlorophyll content in leaf discs. The 3rd day after water stress,relative water content, the malonyldialdehyde content and relative electrical conductivity in L1 and L2 transgenic lines showed no difference with that in WT. After 13 d under water stress conditions, relative water content in transgenic lines was significantly higher than in WT, but the malonyldialdehyde content and relative electrical conductivity were significantly lower than in WT. 【Conclusion】These results suggested that WT tobacco plants were greatly affected under water stress conditions, and the transgenic plants were less affected. Overexpression of GhCYP1 improved the drought tolerance of transgenic tobacco plants.  
    Differential Protein Analysis of Kenaf Leaves Under Drought Stress
    QI Jian-Min, JIANG Hai-Qing, CHEN Mei-Xia, XU Jian-Tang, MA Hong-Bo, FANG Ping-Ping, LIN Li-Hui, TAO Ai-Fen, CHEN Wei
    Scientia Agricultura Sinica. 2012, 45(17):  3632-3638.  doi:10.3864/j.issn.0578-1752.2012.17.021
    Abstract ( 542 )   PDF (501KB) ( 901 )   Save
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    【Objective】 The analysis of stress-responsiveness in kenaf is an important route to the discovery of genes conferring stress tolerance. Proteomic analysis provides a broad view of plant responses to stress at the level of proteins.【Method】A drought-resistant kenaf variety GA42 has been identified as materials by setting the normal water supply and water control comparison test at seeding stage, stress-induced proteome changes were analyzed by two-dimensional gel electrophoresis.【Result】In total, 9 up-regulated protein spots reproducibly presented differential expression patterns on 2-DE maps. Six spots were identified and confirmed by MALDI-TOF-TOF MS, including 2 ribulose-1, 5-bisphosphate carboxylase or large chain (a key enzyme in the first major step of carbon fixation), 1 rubisco activase (a enzyme could regulate the activity of rubisco existing widely in plants), 1 dimethylmenaquinone methyltransferase (a coenzyme participates in methyl transfer reactions), 1 putative plastidic glutamine synthetase (a key enzyme participates in ammonia assimilation in higher plants), and 1 ATP synthase subunit β (an important enzyme that provides energy for the cell to use through the synthesis of ATP, which is the most commonly used “energy currency” of the cells from most organisms). 【Conclusion】 This study preliminarily showed these six protein spots are up-regulated which are associated with GA42 resistance under drought stress.
    Effects of Different Soil Interfaces on Apple Root Architecture and Soil Properties
    HAN Tian-Tian, HU Yan-Li, MAO Zhi-Quan, SHEN Xiang, SHU Huai-Rui
    Scientia Agricultura Sinica. 2012, 45(17):  3639-3645.  doi:10.3864/j.issn.0578-1752.2012.17.022
    Abstract ( 707 )   PDF (377KB) ( 831 )   Save
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    【Objective】The study was to explore apple seedlings root characteristics, physical and chemical properties of the deep soil in three different soil interfaces. 【Method】 This study was conducted with biennial Fuji apple young seedlings in a pot culture experiment, to analyze different effects of glass interface, wood interface and brick interface.【Result】Three interfaces influenced extremely significant difference on the root system, including the root system length, surface area and volume. Brick was the highest in the above indexes, while wood took the second place. Wood was the largest and was the brick second on root weight and average diameter. Glass was the worst on all the indexes. Different interfaces showed different root concentration distribution layers, brick and wood on the second layer (10-20 cm), while glass centralized on the fourth layer (30-40 cm). The discretion of the root activities to root respiration did not show a positive correlation because of the ratio of the absorbed roots to the growth roots. Brick interface had stability on temperature in different layers, glass was in favour of water infiltration, and wood had a strong adsorption effect on salt.【Conclusion】Wood and brick interfaces obviously improved the apple root characteristics and the physical and chemical properties of the deep soil, with few different ways. Brick promoted the root length and increased the root number, and wood obviously promoted the secondary structure of root development role. Interface regulation altered the apple root concentration distribution layer.
    Differential Protein Analysis of Chinese Gooseberry Ovary After Fertilization by a 2D-DIGE Approach
    QI Xiu-Juan, FANG Jin-Bao, CHEN Jin-Yong, GU Hong, ZHANG Shao-Ling
    Scientia Agricultura Sinica. 2012, 45(17):  3646-3652.  doi:10.3864/j.issn.0578-1752.2012.17.023
    Abstract ( 523 )   PDF (581KB) ( 550 )   Save
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    【Objective】The changes of Chinese gooseberry (Actinidia arguta) ovary proteins and morphology before and 120 h after pollination were analyzed to provide valuable evidence for further elucidating the molecular mechanism of double fertilization.【Method】Fluorescence paraffin technique,differential in-gel electrophoresis (DIGE), Matrix-assisted laser-desorption/ionization Time of ?ight/Time of ?ight (MALDI-TOF/TOF) and bioinformatic technology were used in the experiment.【Result】The embryo sac in the ovary completed its double fertilization 120 h after pollination. About 1 500 protein points were detected by DeCyder 6.5 (Amersham Bioscience), and 55 proteins were differentially expressed at ovary before and 120 h after pollination. Thirteen protein spots (differential ratio>2.0) were identified with MALDI-TOF/TOF, of these proteins,8 belong to 6 kinds of proteins. Five of the six proteins are up-regulated while one is down-regulated.【Conclusion】According to the expression differences of the proteins at ovary before and after fertilization, several proteins identified might be related to the process of the double fertilization of Chinese gooseberry.
    Application of Grey Correlation Analysis and AHP Method in Selection of Potted Chrysanthemum 
    WANG Qing, DAI Si-Lan, HE Jing, JI Yu-Shan, WANG Shuo
    Scientia Agricultura Sinica. 2012, 45(17):  3653-3660.  doi:10.3864/j.issn.0578-1752.2012.17.024
    Abstract ( 633 )   PDF (353KB) ( 821 )   Save
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    【Objective】The present study was to establish a synthetic system for assessing the quality of potted chrysanthemum, and to offer a theoretical support for selecting and breeding of spray potted chrysanthemum. 【Method】A total of 430 potted chrysanthemum plants were used as materials and 15 characters were chosen as evaluation factors. The synthetic evaluation system was established by combining the grey correlation analysis and AHP method together. 【Result】AHP method was used to determine the weight value of various characters. The results showed that plant shape, flower dense, plant height and flower color represent a significant share. The weight of each affecting factor was 0.2016, 0.1560, 0.1190 and 0.0944. According to the grey correlation degree of 430 strains, they were divided into 3 grades. And eventually 43 strains with the good comprehensive characters (the good shape, bright and pure color, dense flowers, etc.) were picked out. 【Conclusion】The results show that the combination of the grey correlation analysis and AHP method can be used to evaluate the potted chrysanthemum strains effectively and select the objective ones quickly, it provides a scientific and precise method for selection breeding.
    Effects of Preharvest Bagging Treatment on the Micro-Structure and Chemical Composition of Cuticular Wax in Pingguoli Pear Fruit
    LI Yong-Cai, YIN Yan, CHEN Song-Jiang, BI Yang, GE Yong-Hong
    Scientia Agricultura Sinica. 2012, 45(17):  3661-3668.  doi:10.3864/j.issn.0578-1752.2012.17.025
    Abstract ( 617 )   PDF (729KB) ( 905 )   Save
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    【Objective】 The objective of this study is to further explore the effect of preharvest bagging on micro-structure and chemical composition of cuticular wax in Pingguoli pear fruit (Pyrus bretschneideri Rehd. cv. Pingguoli). 【Method】 Pear fruits were bagged with two-layer paper at 60 days after petal fall, ultrastructure and chemical composition of cuticular wax in Pingguoli pear were measured by scanning electron microscope and gas chromatography-mass spectrometry (GC-MS) respectively. 【Result】No significant differences in cuticular wax content per surface area between bagging and non-bagging fruits were observed. Ultrastructure of cuticular wax in bagged fruit was rougher, uneven, more cracks and no wax crystals formation compared with that in control fruits. Different numbers and kinds of chemical compounds in cuticular wax of bagged and non-bagged fruits were detected. The relative content of alkanes in cuticular wax of bagged pear was higher than the control, on the contrary, the content of alkanoic acids was lower, and the relative content of alkanes, alhydes and triterpenoids had no difference between bagged and the control fruits. Chain-length distribution of saturated n-alkanoic acids made a difference, alkanoic acids with chain lengths of C26 and C28 were present in higher amounts in cuticular wax of bagged and non-bagged fruits, but C30 alkanoic acids were not detected in the wax of bagged fruits.【Conclusion】These results suggested that preharvest bagging significantly influenced micro-structure, and kinds and their chain-length distribution of chemical composition of cuticular wax in Pingguoli pear, but the mechanism of action needs to be further studied.
    Relaxant Effect of Sanguinarine on Ex vivo Intestine Smooth Muscle Movement in Rabbits
    WANG Hui, YU Chun-Hong, XU Jian-Ping, WANG Yue-Shang, LIU Zhao-Ying, CHEN Xiao-Jun, SUN Zhi-Liang
    Scientia Agricultura Sinica. 2012, 45(17):  3669-3676.  doi:10.3864/j.issn.0578-1752.2012.17.026
    Abstract ( 763 )   PDF (478KB) ( 703 )   Save
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    【Objective】 The objective of this study was to investigate the relaxant effect of sanguinarine on intestinal smooth muscle contractions and the mechanisms involved. 【Method】 The experiments were performed in isolated intestinal smooth muscle of rabbits. Inhibitory effects of sanguinarine on contraction of normal intestinal smooth muscle and on that of the intestinal muscle treated with acetylcholine (ACh), phentolamine and Barium Chloride (BaCl2) were compared by analyzing the frequency and amplitude of contractions. 【Result】 Results showed that sanguinarine significantly inhibited the frequency and amplitude of spontaneous contraction of isolated intestinal smooth muscle in a dose-dependent manner. The intestinal smooth muscle contractions caused by Ach and Bacl2 was abolished by sanguinarine (0.4 mg•mL-1), and the frequency and amplitude of the intestinal smooth muscle contraction induced by phentolamine (α-receptor antagonist) decreased after incubation with sanguinarine. 【Conclusion】 These results suggest that the mechanism of relaxant effect of sanguinarine on intestinal smooth muscle contractions may be related to the blockage of M receptor by competing with ACh, to the excitement of α-receptor through competing with phentolamine and to the direct inhibit on of the motility of intestinal smooth muscle.