Table of Content

    01 April 2012, Volume 45 Issue 7
    Analysis of the Relationship Between Differential Expression of Rice Gene and Heterosis
    ZHANG Xiao-Meng, XIAO Ning, ZHANG Hong-Xi, FENG Yan-Xia, LIU Zhi-Xiao, GAO Yong, DAI Zheng-Yuan, CHEN Jian-Min
    Scientia Agricultura Sinica. 2012, 45(7):  1235-1245.  doi:10.3864/j.issn.0578-1752.2012.07.001
    Abstract ( 759 )   PDF (664KB) ( 716 )   Save
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    【Objective】The strong and weak heterosis groups in rice were analyzed for investigating the relationship between differential expression patterns of gene and heterosis. 【Method】The differential display-PCR (DD-PCR) technology was used to analyze gene differentially expressed patterns for the nine F1 of Yangdao 6, Minghui 63 and Teqing varieties as male parent in tillering leaves, booting panicles and heading flag leaf. 【Result】At the same growth stage, four kinds of gene differentially expressed patterns, UNP1 (genes specifically expressed in female parent), UNP2 (genes specifically expressed in male parent), UNF1(genes specifically expressed in F1), ABF1 (genes specifically silenced in F1), were presented obvious difference between strong heterosis group G1(Yangdao 6 as male parent) and weak heterosis group G2 (Minghui 63 as male parent). Correlation analysis showed that UNP2 of tillering leaves was significantly positive corelated to yield per plant, and the expressed genes from the paternal were benefit to heterosis. The differentially expressed gene patterns existed obvious difference among three growth stages in the same heterosis group, which indicates that gene expression is spatial and temporal characteristic. Total nineteen differentially expressed gene fragments between G1 and G2 groups were acquired and distributed along one to eleven chromosomes except the twelfth chromosome. The function analysis result showed that they were mainly involved in material synthesis and transport, energy, carbohydrate metabolism pathways and so on. 【Conclusion】 The gene differential expression patterns in strong and weak heterosis group exist significant difference at different growth stages. The phenotypic differences could be reflected by gene differential expression patterns.
    Transcriptomic Analysis of Sesame Development
    WEI Li-Bin, MIAO Hong-Mei, ZHANG Hai-Yang
    Scientia Agricultura Sinica. 2012, 45(7):  1246-1256.  doi:10.3864/j.issn.0578-1752.2012.07.002
    Abstract ( 962 )   PDF (727KB) ( 2818 )   Save
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    【Objective】 To enrich sesame transcriptome data, the transcriptome sequencing and bioinforamtics analysis on sesame growth and development and seed formation processes were performed in this study. 【Method】 Six transcriptome sequencing libraries for developing sesame plants and seeds were constructed and sequenced using Illumina RNA sequencing technique, and the global transcriptome information was analyzed subsequently.【Result】8.80 Gb available trancriptome data were acquired as the adaptor sequences, duplication sequences and low-quality reads were removed from the original 12.69 Gb solexa sequencing data. And 26 837 uni-transcripts, longer than 100 bp, were obtained by de novo assembly method (http://www. ncbi.nlm.nih.gov/genbank/TSA.html, GenBank ID: JP631635-JP668414). The total scaffold sequence length reached 18.35 Mb with the 1 006 bp of N50, the average uni-transcript length was 683 bp. Annotation analysis of uni-transcripts indicated that 25 331 transcripts had homolog in public protein database; however, 1 506 sequences were no hit and might be sesame-specific. With COG and GO functional classifications, all uni-transcripts were grouped into 24 and 42 function categories, respectively, in which many functional categories, such as material and energy metabolism, signaling, transcription regulation and defense reactions, etc. were included. Furthermore, compared with plant growth and development transcripts, 1 277 sequences expressed more than 10-fold low in seed formation process, and the transcript level of 990 uni-trancripts could not be measured. In addition, 660 sequences were found with more than 10-fold high expression level during seed development, in which 296 sequences seemed as seed-specific. 【Conclusion】With the transcriptome sequencing of Sesamum radiatum and several cultivars of S. indicum L., this study gave a global insight into the characteristics of sesame development transcriptome and thousands of transcript sequences with important function were acquired for future genes expression and regulation research about sesame growth and development.
    Molecular Cloning and Analysis of LeWRKY2 Gene
    SUN Qing-Peng, LI Na, YU Yong-Kun, ZHAO Fu-Kuan, WAN Shan-Xia, PAN Jin-Bao
    Scientia Agricultura Sinica. 2012, 45(7):  1257-1264.  doi:10.3864/j.issn.0578-1752.2012.07.003
    Abstract ( 607 )   PDF (554KB) ( 671 )   Save
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    【Objective】Cloning of WRKY transcription factor from tomato can provide a basis for molecular mechanism of pathogen reaction in tomato. 【Method】 Using RT-PCR and RACE technology, the LeWRKY2 full length cDNA was cloned (GenBank accession: EU755368.1), and its function was predicted by bioinformatics tools. The real-time PCR technology was used to analyze the expression levels of LeWRKY2 gene when treated with JA, Botrytis cinerea and cycloheximide. 【Result】 The LeWRKY2 full length cDNA, consisted of 1 007 bp, was isolated from tomato. The bioinformatics analysis showed that it included an ORF of 471 bp. LeWRKY2 protein contained a WRKY domain and a C2H2 zinc finger motif, and it might have the function of transcription, transcriptional regulation and signal transduction. The LeWRKY2 gene expression levels were proportional to the JA treatment time in the range of 0-60 min, while in the range of 60-150 min, the LeWRKY2 gene expression levels were inversely proportional to the JA treatment time when treated with 100 µmol•L-1 JA. LeWRKY2 gene expression was induced by Botrytis cinerea, and the LeWRKY2 gene expression reached maximum abundance at 4 h. The transcript of LeWRKY2 gene was not dependent on the protein biosynthesis. 【Conclusion】 LeWRKY2 gene is an immediate early gene involved in tomato defense response.
    Actual Responses and Adaptations of Rice Cropping System to Global Warming in Northeast China
    ZHANG Wei-Jian, CHEN Jin, XU Zhi-Yu, CHEN Chang-Qing, DENG Ai-Xing, QIAN Chun-Rong, DONG Wen-Jun
    Scientia Agricultura Sinica. 2012, 45(7):  1265-1273.  doi:10.3864/j.issn.0578-1752.2012.07.004
    Abstract ( 887 )   PDF (646KB) ( 719 )   Save
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    【Objective】A better understanding of global warming impacts on grain production can reduce the uncertainties of future food security projection. Rice is one of the most important grain crops in the world. Northeast China is the main region of rice cropping and the greatest warming area in China. It is significant and necessary to learn the potential impacts of warming on rice production in Northeast China for ensuring the future food security.【Method】Based on historical data analysis and three-year field warming experiment (Free air temperature increase, FATI), the actual responses and adaptations of rice cropping system to warming were investigated in Northeast China.【Result】The results showed that there were similar increasing trends between air temperature during rice cropping season and rice yield over the past decades. Significant positive relationship was found between nighttime increment and rice yield enhancement, while no significant relationship was found between rice yield and precipitation. According to the relationship between climate anomaly and rice yield anomaly over the last thirty years, 1℃ increment in daily minimum temperature during rice cropping season might increase rice yield by about 6.0%. Meanwhile, field experiments showed that further  1℃ warming at nighttime might enhance rice yield by about 10.0% in future. The entire growth period of new variety approved and released in different times has been prolonged by about 3.0 d?10a-1 during the past fifty years, which is confirmed by the actual prolonging of rice growth period by 5.0 day observed in field during the last twenty years. Rice sown area in Heilongjiang province increased about 24 times in 2010 than 1970, and the cropping centre moved northward about by 110 km during the past forty years. Similar spatial change was found in ≥10℃ accumulated temperature during rice cropping season over the same years in Northeast.【Conclusion】 The above evidences indicate that warming has contributed a great to the historical increment in rice yield, and further warming will still help rice yield increase in Northeast China. Rice cropping system can progressively adapt to global warming through variety breeding, cropping technique innovation and cropping region adjustment. More efforts should be paid on the great adaptation potential of rice cropping system to warming, so as to fully explore the positive effects of warming on rice production at high latitude area and reduce the risk of yield decrease due to the unstable climate.
    Study on Turbulent Heat Flux over Soybean Field in the Sanjiang Plain
    JIA Zhi-Jun, HUANG Yao
    Scientia Agricultura Sinica. 2012, 45(7):  1274-1284.  doi:10.3864/j.issn.0578-1752.2012.07.005
    Abstract ( 604 )   PDF (868KB) ( 624 )   Save
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    【Objective】 The quantification of variation regularity of surface turbulent heat flux was studied in order to provide a basis for clarifying the energy balance in soybean field. 【Method】 Based on eddy covariance data in soybean field in the Sanjiang Plain, the diurnal and seasonal variation of turbulent heat flux, water balance and energy partitioning were analyzed. 【Result】Diurnal variation curves of latent and sensible heat flux were both obviously single peaked in soybean fields. Seasonal variation of latent heat flux was also clear. Latent heat flux generally reached the maximum during the end of June and early July, which was strongly linearly positive dependent on net radiation. However, when the precipitation became small, latent heat flux was also affected by it. The sensible heat flux showed a declining trend, which was also strongly linearly positive dependent on net radiation, but both sensible heat flux and Hs/Rn are strongly negative dependent on precipitation. Total precipitation was obviously greater than total evapotranspiration during the growing season of soybean, while uneven distribution of precipitation made the water budget become deficit in short term. The characteristics of seasonal variation of Bowen ratio was not exactly same from 2005 to 2007, and Bowen ratio had a strong negative relationship with monthly precipitation and leaf area index. 【Conclusion】 Latent and sensible heat flux in soybean field showed a clear single peak at the diurnal scale. Surface water is surplus and latent heat flux, sensible heat flux and energy partition in soybean field is determined by net radiation, precipitation and plant development.
    Development of a RT-LAMP Assay for Rapid Detection of Rice black-streaked dwarf virus
    ZHOU Tong, DU Lin-Lin, FAN Yong-Jian, ZHOU Yi-Jun
    Scientia Agricultura Sinica. 2012, 45(7):  1285-1292.  doi:10.3864/j.issn.0578-1752.2012.07.006
    Abstract ( 692 )   PDF (744KB) ( 813 )   Save
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    【Objective】The objective of this study is to develop a reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid and sensitive detection of Rice black-streaked dwarf virus (RBSDV) from host plants and insect vector.【Method】Four primers matching a total of 6 sequences of the S10 of RBSDV were synthesized for the RT-LAMP assay. The concentration of the primer and MgSO4 were optimized, and the best temperature and reaction time for detecting the virus were found, respectively. Detection sensitivity comparisons were performed between the RT-LAMP and RT-PCR assay using a single extraction of total RNA from RBSDV-infected rice leaves which was serially diluted in ten-fold increments in DEPC-treated water. In order to determine the specificity of the RT-LAMP assay, total RNA of rice leaves infected with RBSDV or Southern rice black-streaked dwarf virus (SRBSDV) was applied separately to the reaction system. RBSDV-infected rice plants collected from rice field were detected.【Result】 This method demonstrated a high degree of specificity for RBSDV, which can distinguish RBSDV from SRBSDV. The method was also proved to be extremely sensitive, which was as much as the RT-PCR for RBSDV detection. The detection of amplified products was easily monitored.【Conclusion】The RT-LAMP assay is suitable for rapid detection of RBSDV in host plant and vectors.
    Advances in Research on Cell Biology of Potyvirus
    CUI Xiao-Yan, WEI Tai-Yun, CHEN Xin
    Scientia Agricultura Sinica. 2012, 45(7):  1293-1302.  doi:10.3864/j.issn.0578-1752.2012.07.007
    Abstract ( 875 )   PDF (342KB) ( 932 )   Save
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    Potyvirus, family Potyviridae, is the largest genus of plant virus, causing significant losses in a broad range of host plants. In recent years, researchers have contributed to the new advances of cell biology, so in this article the molecular biology involved in genome amplification, cell-to-cell and long-distance movement, symptom during potyviruses systemic infection were referred.
    Isolation and Phylogeny of Nitrogen-Fixing Endophytic Bacteria in Wheat, Rice, Maize, Chinese Cabbage and Celery
    SUN Jian-Guang, LUO Qiong, GAO Miao, HU Hai-Yan, XU Jing, ZHOU Yi-Qing
    Scientia Agricultura Sinica. 2012, 45(7):  1303-1317.  doi:10.3864/j.issn.0578-1752.2012.07.008
    Abstract ( 963 )   PDF (548KB) ( 1524 )   Save
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    【Objective】 The objective of this study is to determine the main groups and phylogenetic position of nitrogen-fixing endophytic bacteria in wheat, rice, maize, Chinese cabbage and celery. 【Method】 Surface sterilization and nitrogen-free medium were used to isolate nitrogen-fixing endophytic bacteria. Acetylene reduction assay was used to measure the nitrogenase activity. 16S rDNA was amplified by PCR and 16S rDNA sequence was analysed with MEGA software. 【Result】 Thirty four nitrogen-fixing endophytic bacteria with nitrogenase activity ranging from 0.30 to 30.24 nmol C2H4/h•mg protein were isolated from field wheat. Based on 16S rDNA sequence similarity, these strains belonged to 21 species of 13 genera, including Pseudomonas, Rhizobium, Bacillus, Flavobacterium, etc. Twenty five nitrogen-fixing endophytic bacteria with nitrogenase activity ranging from 3.12 to 254.12 nmol C2H4/h•mg protein were isolated from field rice. They belonged to 16 species of 9 genera, including Bacillus, Burkholderia, Enterobacter, Klebsiella, etc. Moreover, Burkholderia, Enterobacter and Klebsiella were dominent. Nine maize endophytic diazotrophs with nitrogenase activity ranging from 7.27 to 59.58 nmol C2H4/h•mg protein were identified as 6 species of genera Rhizobium, Sphingomonas, Arthrobacter, Brevibacterium and Microbacterium. Fourteen endophytic diazotrophs with nitrogenase activity ranging from 2.33 to 205.21 nmol C2H4/h•mg protein were isolated from pot trial pakchoi and identified as 8 species of 6 genera of Rhizobium, Arthrobacter, Pseudomonas, etc. Ten endophytic diazotrophs with nitrogenase activity ranging from 1.23 to 46.70 nmol C2H4/h•mg protein were isolated from marcket celery and identified as 8 species of 5 genera of Sphingomonas, Pseudomonas, Brevundimonas, etc. 【Conclusion】 Nitrogen-fixing endophytic bacteria colonized widely in wheat, rice, maize and vegetables. Their nitrogenase activity under pure culture ranged from 0.30 to 254.12 nmol C2H4/h•mg protein. Ninety two endophytic diazotrophs belonged to 56 species of 25 genera, including Pseudomonas, Rhizobium, Bacillus, Burkholderia, Sphingomonas, Enterobacter and Klebsiella, etc. These endophytic diazotrophs have a great potential to agriculture.
    Functions and Cost Benefits of Contour Grass Hedges on Sloping Croplands
    XIAO Bo, WANG Hui-Fang, WANG Qing-Hai, WU Ju-Ying, TENG Wen-Jun, DAI Quan-Hou
    Scientia Agricultura Sinica. 2012, 45(7):  1318-1329.  doi:10.3864/j.issn.0578-1752.2012.07.009
    Abstract ( 818 )   PDF (751KB) ( 612 )   Save
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    【Objective】 The functions of contour grass hedges on sloping croplands were studied, and its cost benefits were analyzed in this research. It is hoped that the results could provide solutions for the serious problems of soil and water loss and agricultural non-point source pollution on sloping croplands in Northern China. 【Method】 The plots with Pennisetum alopecuroide (Pennisetum alopecuroides (Linn.) Spreng) grass hedges and no grass hedges were established. Then the relative parameters, including soil physical properties, soil water and nutrient content, loss of soil and water and soil nutrient, and crop yield, were continuously measured for the plots with grass hedges and no hedges during the period of 2006 to 2010. Finally, the effectiveness of the grass hedges was concluded from the comparative results of the treatments with grass hedges and no hedges. In addition, the cost benefit analysis of grass hedges was also conducted through its costs and effects on crop yield. 【Result】 The soil physical properties were significantly improved by the grass hedges. In the research, the soil bulk density was decreased by 7.0%, and the stable infiltration rate, total soil porosity, soil fine particles (<0.01 mm) and surface soil water content were increased by 157.1%, 11.0%, 10.8% and 3.5%, respectively, by the grass hedges after 5 years. The soil nutrient content was also greatly increased by the grass hedges, especially for organic matter (12.7%) and available phosphate (24.5%). The grass hedges averagely decreased runoff by 63.2%, soil loss by 77.7%, total nitrogen by 69.4%, nitrate nitrogen by 72.7%, ammonium nitrogen by 57.3%, total phosphate by 54.2% and available phosphate by 62.3%. The grass hedges increased the yield of the crop near the hedges by 21.5%. However, the total crop yield of the plots with grass hedges was slightly decreased because that the grass hedges took up some land area which was originally available for crops. The cost of the grass hedges in this study was 2813 RMB per hectare, which is much cheaper than the other practices such as terrace. 【Conclusion】 Pennisetum grass hedges was proved to be very effective and efficient in reducing the loss of soil and water and agricultural non-point source pollution on sloping croplands. Due to its low cost and well performance, its application could be reasonably extended to the whole area of Northern China.
    Effects of Rhizosphere Ventilation at Different Growth Stages on Plant Growth and Yield of Greenhouse Tomato
    LU Ze-Hua, CAI Huan-Jie, WANG Jian, LI Zhi-Jun
    Scientia Agricultura Sinica. 2012, 45(7):  1330-1337.  doi:10.3864/j.issn.0578-1752.2012.07.010
    Abstract ( 1032 )   PDF (592KB) ( 770 )   Save
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    【Objective】By the fact that water is continuously emitted, the root zone of tomato in subsurface drip irrigation is purged of water and therefore experiences insufficient oxygen deficiency, an experiment was conducted to observe the effect of rhizosphere ventilation at different stages on plant growth and yield of greenhouse tomato, and assess its significance.【Method】By comparison with the control group, this study was conducted to evaluate the effects of rhizosphere ventilation at different growth stages on the growth, physiological characteristics and yield of tomato grown in greenhouse. 【Result】 Results showed that all of different rhizosphere ventilation treatments could raise tomato’s growth and yield, and the aeration effect was not superimposed at different growth stages. Aeration at seedling stage could advance the first layer flowering time, and the second and third layer flowering time could be also advanced by the aeration at flowering stage, and also could rise the plant height by 14.8%. The whole growing stages ventilation processing improved chlorophyll by 11.1%. The aeration at fruit-setting stage could improve the plant diameter by 5.6%, thus it could promote dry matter accumulation, improve root/shoot ratio and yield.【Conclusion】Tomato at different growth stages with underground drip irrigation shows different responses to the rhizosphere ventilation, and the optimum aeration period of tomato is fruit-setting stage.
    Cloning and Expression Analysis of S-Adenosyl-L-Homocysteine Hydrolase in Cucumber (Cucumis stavius L.)
    JIN Xiao-Xia, QIN Zhi-Wei, ZHOU Xiu-Yan, WU Tao
    Scientia Agricultura Sinica. 2012, 45(7):  1338-1346.  doi:10.3864/j.issn.0578-1752.2012.07.011
    Abstract ( 876 )   PDF (2275KB) ( 879 )   Save
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    【Objective】 The cDNA sequence of cucumber S-adenosyl-L-homocysteine hydrolase(CsSAHH) gene was cloned and the gene expression patterns and protein structure were analyzed by bioinformatics methods to study the function of the gene. 【Method】 Based on selecting SSH of stem tips treated by ethrel, the cDNA sequence of cucumber SAHH gene (CsSAHH, GenBank accession, HQ444960) was obtained by RT-PCR and in silico cloning. The mRNA expressions of the gene in stem tips treated by ethrel and different tissues of male and female flowers were detected through semi-quantitative RT-PCR. And the protein structure was analyzed by bioinformatics methods.【Result】 The cDNA sequence of the gene was 1 545 bp, contained a predicted protein sequence of 485 amino acids, pI=5.66, MW=53.1 kD. The semi-quantitative RT-PCR results revealed that CsSAHH mRNA expression was increased by ethrel in stem tips and was lower in stamen than that in the other flower tissues. The physical and chemical properties of cucumber SAHH indicated that this protein has no clear signal peptid. The secondary structure of cucumber SAHH was mainly made of loops, α helixes and few β sheets. There were two S-adenosyl-L-homocysteine hydrolase active domains between positions 85-99 and 262-279. The speculated amino acids sequence of cucumber SAHH shared 63% homologies with Medicago truncatula and had low homologies with Oryza sativa, Zea mays and Arabidopsis thaliana.【Conclusion】 The full length of the CsSAHH cDNA were obtained. There are two S-adenosyl-L-homocysteine hydrolase active domains between positions 85-99 and 262-279. The CsSAHH mRNA expression was induced by ethrel in stem tips, and lowest in stamen. The expression of CsSAHH was stronger in female flower buds than small fruits and male flower buds at different development stages of untreated cucumber flowers.
    Cloning and Prokaryotic Expression of MdRGL Gene from Spur-Type Apple (Malus domestica Borkh.)
    SONG Yang, ZHANG Yan-Min, LIU Mei-Yan, WANG Chuan-Zeng, LIU Jin, FENG Shou-Qian, WANG Yan-Ling, CHEN Xue-Sen
    Scientia Agricultura Sinica. 2012, 45(7):  1347-1354.  doi:10.3864/j.issn.0578-1752.2012.07.012
    Abstract ( 904 )   PDF (960KB) ( 747 )   Save
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    【Objective】Cloning, sequence analysis of the DELLA genes from Malus domestica Borkh and its expression it in E. coli were conducted to further explore the relationship between DELLA and mutation shoot of spur type bud sport apple.【Method】The MdRGL cDNA fragments amplified from Malus domestica Borkh by reverse transcription PCR (RT-PCR) and then the cloned genes of MdRGL were inserted into vector pGEX-4T-1. The recombinant plasmids pGEX-4T-MdRGL were expressed in a prokaryotic expression system after its transformation into E. coli BL21 (DE3).【Result】 Five DELLA genes, MdRGL1a/b, MdRGL2a/b and MdRGL3b were isolated. Molecular structure analysis revealed that these five genes exhibited the typical structures of the DELLA and VHYNP domain except MdRGL3b. The results of SDS-PAGE demonstrated that the expressed proteins were consistent with the size of expected protein in the prokaryotic expression system.【Conclusion】The five DELLA genes of spur-type apple were cloned and successfully expressed in E. coli. This study will provide a foundation for studying the function of the target protein.
    The Association Analysis of Phenotypic Traits with SRAP Markers in Chrysanthemum
    LI Ren-Wei, WANG Chen, DAI Si-Lan, LUO Xin-Yan, LI Bao-Qin, ZHU Jun, LU Jie, LIU Qian-Qian
    Scientia Agricultura Sinica. 2012, 45(7):  1355-1364.  doi:10.3864/j.issn.0578-1752.2012.07.013
    Abstract ( 1168 )   PDF (587KB) ( 962 )   Save
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    【Objective】 In order to provide a genetic basis for studies on complex quantitive traits and for molecular assisted breeding of chrysanthemum, the SRAP markers associated with important chrysanthemum horticulture traits were screened.【Method】 The genomic regions with selection sweep were detected through scanning 58 representative chrysanthemum cultivars using 19 SRAP markers. Population structure was firstly analyzed, then association analysis between SRAP markers and 18 important phenotypic traits were performed using TASSEL GLM.【Result】 Genetic structure analysis showed that the selected cultivar population was composed of 5 subpopulations, namely flat type subgroup, tube type subgroup, irregular type subgroup, anemone type subgroup and Japanese subgroup. There were 6 SRAP loci associated with 5 quantitative characters (P<0.01), among which 3 flower traits were associated with 5 loci, while 1 stem and 1 leaf traits were associated with 1 locus, respectively. The rate of explanation on the phenotype of related locus ranged from 0.0738 to 0.4791.【Conclusion】It is feasible to estimate and differentiate chrysanthemum population’s structure effectively using SRAP markers, and the markers obtained in this study are promising in molecular assisted breeding.
    Analysis on Composition and Content of Glucosinolate in Three Color Types of Maca(Lepidium meyenii)
    GAN Jin, FENG Ying, ZHANG Hong, HE Zhao, XU Long-Feng, CHEN Xiao-Ming
    Scientia Agricultura Sinica. 2012, 45(7):  1365-1371.  doi:10.3864/j.issn.0578-1752.2012.07.014
    Abstract ( 941 )   PDF (513KB) ( 1010 )   Save
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    【Objective】 The composition and content of glucosinolate in three color types of maca (Lepidium meyenii) cultivated in Yunnan were analyzed in this study.【Method】HPLC and LC-ESI/MS were applied to analyze glucosinolate compositions and content of the methanol extracted from fresh, vacuum drying and air drying maca hypocotyls samples. 【Result】Two aromatic glucosinolates, benzyl glucosinolate and methoxybenzyl glucosinolate, were identified from three color types of maca. The total glucosinolate content in purple, yellow and white fresh samples was 50.14, 46.35, and 84.57 μmol•g-1, respectively. The total content in air drying samples was 13.05, 14.35, and 14.94 μmol•g-1, respectively, and the total content in vacuum drying samples was 0.24, 0.05, and 0.29 μmol•g-1, respectively. 【Conclusion】 Three color types of maca cultivated in Yunnan contain the same compositions of glucosinolate, the composition was consistent with that of sample cultivated in Peru. The difference in content was significant among three color types of fresh hypcotyle (P<0.01), but all of them were much higher than that from Peru. The content of glucosinolate in white sample was the highest. The content of dried samples was much lower than that in fresh samples due to mostly hydrolyzed during the processing of drying.
    Impact of Caspase-3 Activation on the Tenderness of Duck  Skeletal Muscle During Postmortem Conditioning
    HUANG Wei, CAO Jin-Xuan, WANG Dao-Ying, XU Wei-Min, ZHANG Mu-Han
    Scientia Agricultura Sinica. 2012, 45(7):  1372-1379.  doi:10.3864/j.issn.0578-1752.2012.07.015
    Abstract ( 697 )   PDF (685KB) ( 823 )   Save
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    【Objective】The objective of the experiment is to investigate whether the caspase-3 is activated postmortem and involved in the hydrolysis of the cytoskeleton process in duck, provide new experimental evidence to explain the postmortem conditioning of waterfowl by the mechanism of apoptosis.【Method】The cytoplasm of cytochrome-C levels, active caspase-3, -8 and -9 and cleaved PARP fragment expression of duck breast and thigh meat after slaughter were detected by immunochemical analysis, changes of caspase-3,-8 and -9 activity and the changes of shear force value were also evaluated.【Result】Results showed that cytochrome-c levels in cytosol were increased significantly (P<0.05), caspase-3, -8 and -9 were split into 17 kD,18 kD and 23 kD fragments and there were higher activities at postmortem respectively (P<0.05). PARP as a marker protein of caspase enzyme system was split into small fragments. Shear force values of breast and thigh meat were also significantly decreased at 4-24 h after slaughter (P<0.05). Correlation analysis showed that caspase-8 and -9 activities were positively correlated to the reported caspase-3 activities and the shear force of muscles were negatively correlated with the activities of caspase-3 after slaughter in muscles (P<0.01).【Conclusion】The results of procaspase-3 was significantly activated and the activities were increased which indicated that the postmortem activation of caspase-3 appears to be involved in both caspase-8 and caspase-9 pathway. Cleaved PARP and shear force values were reduced which indicated that caspase protease might have direct or indirect effects on maturation process during postmortem conditioning.
    Role of wnt/β-catenin in the Differentiation of Satellite Cells into Muscle Fibers
    SHI Xin-E, LIU Yue-Guang, YANG Qiu-Mei, CHEN Zong-Zheng, YANG Gong-She
    Scientia Agricultura Sinica. 2012, 45(7):  1380-1386.  doi:10.3864/j.issn.0578-1752.2012.07.016
    Abstract ( 892 )   PDF (664KB) ( 1017 )   Save
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    Satellite cells are established as the major stem cells contributing to gowth and development as well as regeneration of skeletal muscle. Differentiation of satellite cell and muscle fiber type formation are closely related to meat quality. wnt/β-catenin is an important pathway regulating these events. Recently, more and more research focused on skeletal muscle growth and regeneration regulated by wnt/β-catenin. And many interesting results are published. The role of wnt/β-catenin in the differentiation of satellite cells into different type of muscle fiber and turn-over was reviewed.
    IL-8 Gene SNP-180A/G Affects Significantly Somatic Cell Score in Chinese Holstein Cow
    JI De-Jun, CHEN Ren-Jin, YANG Zhang-Ping, ZHU Guo-Qiang, MAO Yong-Jiang, CHEN Ying, LI Rui
    Scientia Agricultura Sinica. 2012, 45(7):  1387-1391.  doi:10.3864/j.issn.0578-1752.2012.07.017
    Abstract ( 803 )   PDF (568KB) ( 420 )   Save
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    【Objective】 Bovine interleukin-8 (IL-8) is one of the major factors recruiting neutrophils during cow mastitis and causing rapid increase of somatic cell counts (SCC). Knowledge of IL-8 polymorphisms would be helpful to genetic selection of low SCC. 【Method】 The SNPs in the 5′ upstream of bovine IL-8 gene were investigated in 610 Chinese Holstein cows from 30 bull families in a dairy farm in Shanghai using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) technique, and milking traits and somatic cell score (SCS) were measured and their relationship was analyzed using a mixed animal model. 【Result】 The results showed that one SNP, A(-180)G, was detected, determining three genotypes, GG, GA and AA with frequencies of 0.41, 0.43 and 0.16, respectively. The SNP showed significant effect on SCS, with GG having a significantly lower SCS than GA or AA (P<0.01), while this SNP had no effect on test-day milk yield, test-day milk fat content, test-day milk protein content, and 305 d corrected milk yield. 【Conclusion】The findings demonstrated that the SNP -180A/G exerted significant effect on SCS.
    Effects of Different Dietary NFC/NDF Ratios on the Ruminal Fermentation and the Changes of the Rumen Microbial  Community of Goats
    WEI De-Yong, ZHU Wei-Yun, MAO Sheng-Yong
    Scientia Agricultura Sinica. 2012, 45(7):  1392-1398.  doi:10.3864/j.issn.0578-1752.2012.07.018
    Abstract ( 1069 )   PDF (546KB) ( 963 )   Save
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    【Objective】This study was conducted to investigate the changes of the ruminal fermentation and the ruminal microbial composition as dietary NFC/NDF ratio increased.【Method】 Four local goats were gelded and designed for a self-controlled trial. The trial consisted of three periods corresponding to three diets with different NFC/NDF ratios (0.42,1.04,2.73). 【Result】The results showed that, as NFC/NDF ratio in the diet increased, the pH value decreased (P<0.05), and the lactate, acetate, propionate, butyrate, isobutyric acid, valerate, isovalerate and TVFA concentration were increased (P<0.05), and the ratio of the acetate to propionate decreased (P<0.05). The analysis of DGGE profiles showed the sample with different NFC/NDF ratios diet fell into different groups. The real-time PCR results showed the copies of the 16S rRNA genes of Lactobacillus and total bacteria were affected by the dietary NFC/NDF ratio (P<0.05), whereas no significant changes were found in the number of E.coli and the Streptococcus bovis (P>0.05). 【Conclusion】 The results indicated that the characteristics of the ruminal fermentation were affected by the ratio of NFC/NDF in the diet, and with the increasing of the ratio of NFC/NDF, the ruminal fermentation was shifted from the acetate type to the propionate type, and the SARA (subacute rumen acidosis) occurred with the change of the ruminal microbiota when the ratio of the dietary NFC/NDF is 2.73.
    Production and Characterization of Anti-PrP Polyclonal Antibody by PRNP Gene Knockout Goat
    WANG Xue-Bin, YU Hui-Qing, WU Xiao-Dong, XU Xu-Jun, CHEN Jian-Quan, CHENG Guo-Xiang
    Scientia Agricultura Sinica. 2012, 45(7):  1399-1405.  doi:10.3864/j.issn.0578-1752.2012.07.019
    Abstract ( 680 )   PDF (537KB) ( 541 )   Save
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    【Objective】The study was conducted in order to produce and characterize anti-PrP polyclonal antibodies by prion protein knockout (PRNP-/-) goat.【Method】Using polymerase chain reaction (PCR) method, the DNA sequence encoding the goat mature PrP (Prion Protein) was amplified from goat DNA. It was then cloned into the vector pET-22b to express recombinant PrP protein. After purification, the recombinant PrP was used as antigen to immunize PRNP-/- goat for producing anti-PrP antibodies. ELISA and Western blot were conducted to characterize the titer and specificity of polyclonal antibodies.【Result】A large scale of anti-PrP polyclonal antibodies was successfully produced by PRNP-/- goat. ELISA assay revealed that the titer of the polyclonal antibodies against PrP was as high as 1﹕25 600. Western blot test showed that the antibody was not only able to react with the native prion proteins (PrPC) of mouse, cattle and goat brain, but also could combine with misfolded prion proteins (PrPSc) from mouse brain.【Conclusion】Ablation of the prion protein(PrP) gene in goat facilitates the production of a large scale anti-PrP polyclonal antibodies, the produced antibody can be widely used in detecting PrPC and PrPSc derived from a variety of animals.
    Construction of Subtractive Hybridization cDNA Library to Screen Differentially Expressed Genes from the Spleen of C57BL/6 and A/J Mouse and Their Functional Analysis
    XIE Zheng-Lu, SHEN Xue-Huai, LIU Lin, YIN Fu-Jian, MA Hai-Tian, FAN Hong-Jie
    Scientia Agricultura Sinica. 2012, 45(7):  1406-1417.  doi:10.3864/j.issn.0578-1752.2012.07.020
    Abstract ( 834 )   PDF (837KB) ( 570 )   Save
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    【Objective】 The aim of this study was to discuss different mechanisms of infecting Streptococcus suis type 2 in C57BL/6 and A/J mouse strains by detecting the immunization parameters and constructed of cDNA library to screen different expressed genes from the spleen of C57BL/6 and A/J mouse strains. 【Method】 Immunization parameters was detected by ELISA. The substracted cDNA library was constructed from the 8-week-old age spleen of C57BL/6 and A/J mouse strains by suppression subtraction hybridization (SSH). 【Result】 The result showed that, the content of IgA was significantly higher in A/J mouse than C57BL/6 mouse,  but there was no significantly differences in IgG and IFN between A/J and C57BL/6 mouse stains. A total of 149 positive clones were screened by PCR and sequenced. After dislodging, clustering and splicing the redundant cDNA sequence, total of 56 differentially expressed sequence tags (ESTs) were obtained in the whole subtractive cDNA library. After comparisons with GenBank using online software of the BLAST, total of 26 specific gene fragment and two unknow sequences were found. 【Conclusion】 Many of those ESTs were highly homologous with the important functional genes with related to signal transduction, apoptosis and immunity and so on, and the results would provide useful baseline for screening and cloning specific resistant genes and understanding the molecular mechanism of Streptococcus suis.
    Osmoregulation of MPK3, MPK4 and MPK6 from Arabidopsis thaliana in Yeast hog1? Mutant
    LI Po, GU Shou-Qin, YANG Yang, WU Min, WANG Mei-Juan, ZHANG Chang-Zhi, DONG Jin-Gao
    Scientia Agricultura Sinica. 2012, 45(7):  1418-1424.  doi:10.3864/j.issn.0578-1752.2012.07.021
    Abstract ( 746 )   PDF (809KB) ( 595 )   Save
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    【Objective】The objective of this research is to study MPK3, MPK4 and MPK6 which are the key genes of MAPK signal transduction pathway, and to identify these MAPKs in osmoregulation of Arabidopsis thaliana. 【Method】The yeast expression vectors pVT102U-MPK3/MPK4/MPK6 were constructed, and were transformed into the yeast HOG1 null mutant (hog1?), and the positive transformants were characterized by complementation. 【Result】 The full length cDNA of MPK3, MPK4 and MPK6 gene was amplified, and then transformed into hog1? of Sacharomyces cerevisiae through yeast expression vector. Under salt stress with 1 mol•L-1 KCl, 0.3 mol•L-1 LiCl, 1 mol•L-1 NaCl and 1 mol•L-1 sorbitol, the growth of transformants was very well which was almost in accordance with wild type strains, and these genes rescued hog1? to phenotype of wild type which is insensitive to salt stress. Under salt stress, the cell morphology of hog1? was aberrant and its intracellular glycerol concentration was lower than WT, but the morphology and glycerol content of transformants was a normal phenotype.【Conclusion】With function of osmoregulation, MPK3, MPK4 and MPK6 could rescue hog1? from loss of resistance to salt.
    Application of Wavelet Analysis for Estimation of Soil Available Potassium Content with Hyperspectral Reflectance
    CHEN Hong-Yan, ZHAO Geng-Xing, LI Xi-Can, LU Wen-Li, SUI Long
    Scientia Agricultura Sinica. 2012, 45(7):  1425-1431.  doi:10.3864/j.issn.0578-1752.2012.07.022
    Abstract ( 817 )   PDF (527KB) ( 950 )   Save
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    【Objective】 This study aimed at improving the precision and practicability of the soil available potassium estimation model by removing the noise of soil hyperspectral reflectance. 【Method】 Seventy-six soil samples with similar soil organic matter, nitrogen and phosphorus element contents and different potassium element contents were selected. The first derivative spectrum of the soil sample logarithmic reflectance was decomposed to multiple levels by using four kinds of wavelet function, respectively. The low frequency wavelet coefficients were obtained, and the hyperspectral estimation models of soil available potassium content were built. 【Result】The results showed that the low frequency wavelet coefficients of 1-3 levels could represent the original spectrum. Based on the low frequency coefficients of different wavelet functions at the same level, the precise of soil available potassium estimation model showed a little difference. The model built with the low frequency coefficient of the second decomposition level using the Bior 1.3 function had comparatively high accuracy and was chosen as the best model. With the data reducing to 25% and reflecting 95.6% information of the input spectrum, the model building R2 reached 0.976 and RMSE was 10.66 mg•kg-1, which was validated to have fairly good forecast accuracy. 【Conclusion】Therefore, wavelet analysis for obtaining wavelet coefficients can not only extract the soil hyperspectral information, but also compress data, which is feasible to forecast soil potassium content in combination with partial least squares regression method.
    Resistance Selection Against Clofentezine in Tetranychus urticae (Koch) and Change of Its Detoxification Enzymes Activity
    GAO Xin-Ju, ZHANG Zhi-Gang, DUAN Xin-Le, SHEN Hui-Min
    Scientia Agricultura Sinica. 2012, 45(7):  1432-1438.  doi:10.3864/j.issn.0578-1752.2012.07.023
    Abstract ( 722 )   PDF (563KB) ( 681 )   Save
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    【Objective】The objective of this study is to assess the development trend of resistance and the activity change of detoxification enzyme of Tetranychus urticae (Koch). 【Method】 The experiment was carried out in laboratory by using the methods of biological assay and biochemical analysis.【Result】After selected with clofentezine for 21 generations, the resistance strain (Clo-R21) increased to 122.26-fold. Clo-R21 exhibited 21.80 and 10.66-fold moderate level cross-resistance to pyr-clofentezine and liuyangmycin. However, the strain had low level cross-resistance to abamectin, fenpropathrin, beta-cyfluthrin, cypermethrin, azocyclotin, and celastrusangulatus with 6.85, 5.45, 5.02, 3.88, 3.48, 1.68-fold, respectively, and negative resistance to fenbutatin oxide, fenpyroximate, hydramethylnon, separately with 0.72, 0.65, 0.55-fold. Synergist experiments with different metabolic inhibitors revealed that triphenyl phosphate (TPP), diethyl maleate (DEM), piperonyl butoxide (PBO) and synergistic phosphorus (SV1), resulted in 1.39, 1.70, 6.78 and 1.64-fold synergist ratios, respectively. The major resistant mechanism to clofentezine was the increasing activities of carboxylesterases (CarE), acid phosphatase (ACP), alkaline phosphatase (ALP), glutathione-S-transferase (GSTs) and mixed function oxidase (MFO). Compared to the susceptible strain, the enzyme activities in Clo-R21 developed 2.25, 2.04, 1.91, 1.98 and 26.68 times.【Conclusion】 Activity increasing of MFO may play an important role in the resistance to clofentezine, meanwhile CarE, ACP, ALP, and GSTs may also confere resistance to the Clo-R21 strain.
    Effects of Different Cryoprotectants on Ovine Preadipocytes Cryopreservation
    CAI Yong, A Yi-Mu-Gu-Li, ZANG Rong-Xin, LIU Yi-Zhong, YANG Ju-Tian, QIAO Zi-Lin, CAO Xin, XU Hong-Wei, WU Jian-Ping
    Scientia Agricultura Sinica. 2012, 45(7):  1439-1446.  doi:10.3864/j.issn.0578-1752.2012.07.024
    Abstract ( 756 )   PDF (604KB) ( 567 )   Save
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    【Objective】 The aim of this experiment is to investigate a suitable cryoprotectant and its optimal concentration for ovine preadipocytes from omental. 【Method】Trypan blue exclusion test, MTT, oil red O staining and real-time PCR were used to test the effects of following cryoprotective agents (CPAs) with different concentrations on post-thaw survival, proliferation, differentiation capacity, PPARγ and LPL mRNA expression of ovine preadipocytes from omental, dimethyl sulphoxide (DMSO), ethylene glycol (EG), propylene glycol (PG), glycerol (G) and polyvinylpyrrolidone (PVP). In addition to the CPAs, the basic medium is DMEM/F12 medium plus 20% FBS. Then karyotype was analyzed. 【Result】Trypan blue exclusion test showed that the highest survival rate, no significant difference with the primary cells, was obtained when cryopreserved with 10% DMSO or 5% DMSO plus 5% PVP among all the CPA treatments in this study. The highest survival rate (94.96%) and cell viability were obtained when cryopreserved with 10% PVP, and showed no significant difference compared with primary cells. Oil red O staining showed no significant difference in lipogenesis among all the CPAs groups and primary cells on 6 th day (P>0.05), while on 11th day, cells cryopreserved with 10% DMSO turned up significantly greater lipogenesis than other CPAs (P<0.05), but had no significant difference with primary cells (P>0.05). Activity of the GPDH, mRNA expression of LPL and PPARγ showed no significant difference among all the groups (P>0.05). Karyotype analysis showed that diploid cells were dominant post thaw and showed no significant difference compared with primary cells (P>0.05). 【Conclusion】Results of the present study indicate that ovine preadipocytes could successfully be cryopreserved with DMEM/F12 medium containing 10% DMSO or 10 % PVP, 5% DMSO plus 5% PVP, while 10% DMSO is a suitable CPA for ovine preadipocytes.
    Empirical Study on Cooperation Performance of Corn Supply Chain—Based on Investigation of Farmers and Companies of Corn Processing and Logistics
    YANG Zi-Gang, YU Hai-Peng, GUO Qing-Hai
    Scientia Agricultura Sinica. 2012, 45(7):  1447-1454.  doi:10.3864/j.issn.0578-1752.2012.07.025
    Abstract ( 777 )   PDF (676KB) ( 854 )   Save
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    【Objective】The objective of this article is to find existing problems of the cooperative partners of supply chain and the factors influencing the cooperative performances of the supply chain.【Method】Through the investigation on the leading corn processing enterprises (above county levels), logistics enterprises and famers among the cooperative partners of corn supply chain of Jilin province, the establishment of structural equation model and computing by AMOS17.0, the influences of the cooperative partner relationship of corn supply chain on the cooperative performances of corn supply chain were analyzed.【Result】The cooperative intentions and cooperative capability among members of the supply chain have a great effect on the cooperative performances, and the total influence is up to 0.825. The cooperative intentions have a little effect on the cooperative performances, but it has a direct positive effect.【Conclusion】The uncertainty of internal and external environments of enterprises militate against the establishment of cooperation among members. The cooperative intentions and cooperative capability among members of the supply chain have a positive effect on the cooperative performances of the whole supply chain, and the influence of cooperative capability is much bigger. The strength of cooperative partner relationship of corn supply chain has no direct influence on the cooperative performances of the supply chain.