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Table of Content

    15 July 2011, Volume 44 Issue 14
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    The Profiling of Protein Expression at Different Developmental Stages of Elite Rice Varieties
    WANG Xian-Yun, LIU Zhao, CAO Ying-Hao, BAI Hui, LIU Li-Juan, LI Li-Yun, LIU Guo-Zhen
    Scientia Agricultura Sinica. 2011, 44(14):  2849-2856 .  doi:10.3864/j.issn.0578-1752.2011.14.001
    Abstract ( 911 )   PDF (1006KB) ( 1156 )   Save
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    【Objective】 The characterization of protein expression at different developmental stages may help to understand the forming mechanisms of elite varieties and then promote their applications in breeding.【Method】In this study, Western blotting were used to investigate the expression of nine proteins, in the categories of photosynthesis, reactive oxygen scavenging, and defense/stress responses, among six rice elite varieties including 9311, PA64s, Teqing, Zhenshan 97B, Guangluai 4 and Aijiaonante, at different developmental stages/tissues including the shoot and root at seedling stage, the flag leaf and panicle at flowering stage, the flag leaf and panicle at filling stage.【Result】The results indicated that polymorphisms of protein expression presented among different rice varieties, the expression patterns in root and panicle varied to a large extent, while the expression in leaves was similar among different rice varieties. Additionally, the polymorphisms also presented among different developmental stages/tissues, indicating the temporal and spatials expressions of these proteins.【Conclusion】This study has established a strategy to investigate the expression of rice proteins by using antibodies.
    QTL Analysis on Grain Yield per Plant and Plant Height in Wheat
    DING An-Ming, CUI Fa, LI Jun, ZHAO Chun-Hua, WANG Xiu-Qin, WANG Hong-Gang
    Scientia Agricultura Sinica. 2011, 44(14):  2857-2867 .  doi:10.3864/j.issn.0578-1752.2011.14.002
    Abstract ( 1129 )   PDF (685KB) ( 1816 )   Save
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    【Objective】 The objective of this study is to reveal the genetic relationship between grain yield (GY) and plant height (PH) and the influence of PH on GY at the QTL level, and provide references for the selection of PH in wheat high yield breeding programs.【Method】Two high density genetic linkage maps were constructed using two associated recombinant inbred lines (RIL) derived from crosses of Weimai8/Yannong19 (hereafter WY) and Weimai8/Jimai20 (hereafter WJ), consisting of 229 and 485 lines, respectively. Both GY and PH of the three parents and the RIL populations were measured in 2009 and 2010 in Tai’an and Zaozhuang. Unconditional and conditional QTL analyse were conducted to detect the genetic relationship between GY and PH, and the QTL effect without the influence of PH. In addition, the effect of population size on the accuracy of QTL detection was discussed. 【Result】In the WY population, 5 and 15 QTLs were detected for GY and PH, respectively, of which totally 8 QTLs explained >10% phenotypic variation (PVE), 3 QTLs were pleiotropy which determined both GY and PH. For GY conditioned on PH, 3 QTLs affecting GY were detected independent of PH, two were totally or partial contributed to PH, and one completely suppressed by PH. In WJ, 7 and 11 QTLs were detected for GY and PH, respectively. One QTL was a major gene affecting PH with additive effect of 8.82 cm, explaining 20.68% of PVE. Conditioned GY on PH, 5 QTLs were identified independent of PH and two were totally due to PH. The QTL effect of the large population WJ was smaller than that of WY, while the LOD scores were higher.【Conclusion】The genetic relationship between GY and PH is contributed and coordinated by various factors, including closely linked or pleiotropy, the contribution or suppression of PH to GY, environmental factors and the interactions between traits, etc. The selection of PH in high yield breeding programs should refer to different genetic backgrounds and ecological environments. A population of large size can detect QTLs more accurate than small ones in QTL mapping.
    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    Climate Suitability of Wheat-Rice Double Cropping System in Huaihe Watershed
    LAI Chun-Jia, QIAN Huai-Sui, DUAN Hai-Lai, SONG Qiu-Hong, YU Fen, ZHANG Yi-Xiu, ZHANG Jing-Fen
    Scientia Agricultura Sinica. 2011, 44(14):  2868-2875 .  doi:10.3864/j.issn.0578-1752.2011.14.003
    Abstract ( 713 )   PDF (514KB) ( 811 )   Save
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    【Objective】 A quantitative research on the response status of wheat - rice double cropping system to the climate change in Huaihe watershed was made in order to make scientific evaluation for improving the breed and cropping system, supply a basis for formulating a reliable and scientific planting system according to the local conditions. 【Method】 The suitability of temperature, precipitation, sunshine and climate for wheat - rice double cropping system in Huaihe watershed was studied by applying the ecological theory of niche-fitness and fuzzy mathematics methods, then the climate suitability function for the crops was established combined with the method of spatial interpolation. 【Result】 The climate suitability of wheat - rice double cropping system showed a downward trend, temperature suitability showed a rising trend, precipitation and sunshine suitability of fitness showed a decreasing trend. The space distribution of climate suitability in Huaihe watershed wheat - rice double cropping system is decreasing from the southeast coast areas and plains to the western mountain areas. In the future climate change scenarios, the Huaihe watershed wheat - rice double cropping system, climate suitability will increase, but little change in spatial pattern. 【Conclusion】 The climate suitability of the wheat-rice double cropping system in Huaihe Watershed had apparent response to the global warming in recent decades.
    The Possible Effects of Global Warming on Cropping Systems in China Ⅶ. The Possible Effects of Climate Warming on Geographical Shift in Northern Limit of Citrus Planting Areas and the Risk Analysis of Freezing Injury in China
    LI Yong, YANG Xiao-Guang, ZHANG Hai-Lin, CHEN Fu
    Scientia Agricultura Sinica. 2011, 44(14):  2876-2885 .  doi:10.3864/j.issn.0578-1752.2011.14.004
    Abstract ( 859 )   PDF (626KB) ( 615 )   Save
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    【Objective】 The period from 1950s to 2007 was divided into two stages, i.e., 1950s-1980 and 1981-2007, then the geographical shift of northern limit for citrus planting areas and the possible risk of freezing injury after the shift were analyzed and compared. 【Method】 Using the standard of China farming system regional planning and some agro-meteorological indicators, the geographical shift of safe northern limit for citrus planting areas were compared and analyzed and the change chart was draw by ArcGIS. Using the risk assessment standard of freezing injury, the venture of freezing injury of citrus planting areas after geographical shift were evaluated. 【Result】 Compared with the results during the period from 1950s to 1980, the average geographical shift of northern boundary for optimum planting areas, suitable planting areas, sub-suitable planting areas, possible suitable planting areas is northward 0.75°, the region of maximum geographical shift is sub-suitable planting areas, the next is suitable planting areas, optimum planting areas, possible suitable planting areas, respectively. The extent of northward shift to the east section of northern limit is obviously larger than west section. The average geographical shift of southern boundary for optimum planting areas and suitable planting areas is northward 0.56° and 0.42°, respectively. The largest increment of areas occurs at optimum planting areas, the next is suitable planting areas. Unable planting areas decreased mostly, the next is possible suitable planting areas and sub-suitable planting areas, respectively. The light and middle freezing injury to the sensitive region of optimum planting areas and suitable planting areas show obviously increase than that of their nonsensitive region, and the occurrence frequency of severe and extra severe freezing injury in sensitive region and nonsensitive region is low. The occurrence frequency of light freezing injury in sub-suitable planting areas and possible suitable planting areas for sensitive region is lower than that of their nonsensitive region, but the risk of freezing injury for severe and extra severe freezing injury of sensitive region is obviously increased than that of their nonsensitive region. 【Conclusion】 The northern limit has shown an obvious northward moving under global warming. The risk of freezing injury of sensitive region for citrus planting areas after the northward shift is obviously higher than that of nonsensitive region.
    Progress in Research on Digital Image Processing Technology for Automatic Insect Identification and Counting
    YAO Qing, 吕Jun , YANG Bao-Jun, XUE Jie, ZHENG Hong-Hai, TANG Jian
    Scientia Agricultura Sinica. 2011, 44(14):  2886-2899 .  doi:10.3864/j.issn.0578-1752.2011.14.005
    Abstract ( 1057 )   PDF (389KB) ( 1784 )   Save
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    As the rapid development of information technology, digitization, precision and intelligence are important characteristics for modern agriculture and automatic identification and counting of agricultural insects has become a hot research topic. Main methods and applications for automatic insect identification and counting by image processing technology were reviewed. The advantages and disadvantages of these methods were compared and the relevant problems and prospect were also discussed.
    PLANT PROTECTION
    Evaluation of Wheat Leaf Rust Resistance of 10 New Wheat Cultivars (Lines)
    SHI Li-Hong, ZHANG Na, HU Ya-Ya, YANG Wen-Xiang, LIU Da-Qun
    Scientia Agricultura Sinica. 2011, 44(14):  2900-2908 .  doi:10.3864/j.issn.0578-1752.2011.14.006
    Abstract ( 888 )   PDF (432KB) ( 1016 )   Save
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    【Objective】 The objective of this study was to investigate the leaf rust resistance of 10 new wheat cultivars (lines) including Henong 5290, Henong 58-3, Henong 825, Henong 826, Henong 827, Henong 6049, Henong 6251, Henong 6425, Henong 7106 and Henong 9206. 【Method】Sixteen Puccinia triticina strains with different virulence patterns were used for seedling genes postulation and 6 strains of them for the test at adult stage. A series of STS and SCAR markers linked to 15 wheat leaf rust resistance genes were used for marker assisted selection of these tested cultivars (lines).【Result】Lr1 was found in cultivars Henong 6425 and Henong 9206. Lr26 was detected in 7 cultivars including Henong 825, Henong 826, Henong 827, Henong 6049, Henong 6251, Henong 6425 and Henong 9206. Henong 5290, Henong 58-3, Henong 825 and Henong 6251 were postulated containning other or unknown leaf rust resistance genes, and they were resistant to most of the P. triticina strains tested. 【Conclusion】 Henong 58-3, Henong 5290, and Henong 6251 showed potential use in wheat breeding for leaf rust resistance.
    Identification of an RNA Silencing Suppressor Encoded by Southern rice black-streaked dwarf virus S6
    LU Yan-Hong, ZHANG Jin-Feng, XIONG Ru-Yi, XU Qiu-Fang, ZHOU Yi-Jun
    Scientia Agricultura Sinica. 2011, 44(14):  2909-2917 .  doi:10.3864/j.issn.0578-1752.2011.14.007
    Abstract ( 853 )   PDF (559KB) ( 896 )   Save
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    【Objective】The objective of this study is to analyze the RNA silencing suppressor activity of SP6 protein encoded by SRBSDV segment 6 (S6) and to clear whether SRBSDV encodes RNA silencing suppressor to interfere with RNA silencing pathway. 【Method】To investigate the suppress activity of SP6 on local and systemic silencing induced by sense RNA, SP6 was coinfiltrated with the reporter green fluorescent protein (GFP) in transgenic Nicotiana benthamiana line 16c carrying GFP. SP6 was also coinfiltrated with GFP and dsGFP to determine whether it interfered with local and systemic silencing induced by dsRNA. GFP and SP6 were infiltrated in different parts of leaves in a same plant, respectively, to characterize whether SP6 can block the spread of RNA silencing signal. SP6 was over-expressed in the PVX heterogenous system to find out whether it was a pathogenicity determinant. 【Result】The local and systemic silencing can be delayed, but not inhibited, in N. benthamiana leaves agroinfiltrated with SP6 and GFP. SP6 can block the spread of the silencing signal and reverse the GFP silencing, but can not inhibit the silence induced by dsRNA. Expression of SP6 enhanced Potato virus X pathogenicity in N. benthamiana. 【Conclusion】SP6 is an RNA silencing suppressor encoded by Southern rice black-streaked dwarf virus. It may target the initiate step and prevent the spread of RNA silencing signal in the RNA silencing pathway.
    Transformed mBt886Cry3Aa into Chrysanthemum Mediated by Agrobacterium tumefaciens and Its Resistance to Phytoecia rufiventris Gautier
    WANG Pei, ZHOU Zhou, SU Li-Juan, YIN Xin-Ming
    Scientia Agricultura Sinica. 2011, 44(14):  2918-2925 .  doi:10.3864/j.issn.0578-1752.2011.14.008
    Abstract ( 656 )   PDF (585KB) ( 696 )   Save
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    【Objective】 The objective of this study is to transform mBtCryAa into chrysanthemum; which has resistance to long-horned beetles; and determine the toxic effect of transgenic plants to Phytoecia rufiventris Gautier. The results of the study will provide a new way to control Coleoptera pests. 【Method】mBtCryAa was transformed into chrysanthemum mediated by Agrobacterium tumefaciens according to the best culture media and the best suitable kanamycin and cefotaxime concentration confirmed for chrysanthemum differentiation and rooting; then artificial feed and transgenic plants were mixed to raise nd instar larvae. 【Result】 Seven transgenic plants including mBtCryAa were obtained by kanamycin riddling and PCR; and four strong transgenic plants mixed with artificial feed were used to raise nd instar larvae. The results showed that the mortality rates between transgenic plants and control were significantly different. 【Conclusion】 Bioassay indicated that the transgenic plants had better toxic effects on nd instar larvae.
    Odorant Binding Characteristics of Chemosensory Protein Alin-CSP6 in Lucerne Plant Bug, Adelphocoris lineolatus (Goeze)
    MA Shuang, WANG Song-Ying, GU Shao-Hua, HAO Zai-Bin, ZHANG Yong-Jun, GUO Yu-Yuan
    Scientia Agricultura Sinica. 2011, 44(14):  2926-2934 .  doi:10.3864/j.issn.0578-1752.2011.14.009
    Abstract ( 922 )   PDF (537KB) ( 630 )   Save
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    【Objective】The objective of this study is to use the mechanism of insect olfactory recognition, making CSPs as potential targets, and interfering with the insect host location and mating behavior for integrate pest management (IPM). 【Method】 In the present study, a novel chemosensory protein Alin-CSP6 in lucerne plant bug Adelphocoris lineolatus (Goeze) was cloned and the recombinant protein was obtained. The binding characteristics of Alin-CSP6 with 113 volatile odors was explored by a fluorescent competitive binding experiment. 【Result】 Alin-CSP6 has a wide binding spectrum with odorant ligands, which can bind to alcohols, aldehydes, ketones, easters, aromatic and so on. The β-citronellol, trans -2 - hexene aldehyde, 6-methyl-5-heptene-2- ketone, 2-undecanone, butyl levulinate, myrcene, (1s)-(-)-β-pinene, nonane, 3,4 - dimethyl benzaldehyde, ethyl ether, ocimene, acetophenone and benzaldehyde have high binding affinities with Alin-CSP6. In order to verify the behavioral influence of the odors on A. lineolatus, Y-tube olfactometer tests were conducted to observe the response of the adult to odorants which exhibited high binding abilities to Alin-CSP6. It was found that trans-2-hexene aldehyde and acetophenone showed strong attraction to lucerne plant bug, whereas 6-methyl-5-heptene-2–ketone, (1s)-(-)-β-pinene and 4-ethylbenzaldehyde displaied strong repellent action. By homologous modeling analysis the 3D structure was predicted, and it was found that Alin-CSP6 is consists of six α-helices located between residues 13-18(α1), 20-32(α2), 39-52(α3), 59-76(α4), 78-88(α5) and 94-102(α6), two pairs of disulfide bridges Cys29-Cys3 and Cys55-Cys58 play an important role in holding the structural stability of the protein. It is speculated that Ile7 and Phe81 may participate in binding to oleic acid amide analogues.【Conclusion】Alin-CSP6 has a wide binding spectrum with odorant ligands which play an important role in olfaction of A. lineolatus.
    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Effect on Net Greenhouse Gases Emission Under Different Conservation Tillages in Jilin Province
    HUANG Jian-Xiong, CHEN Yuan-Quan, LIU Wu-Ren, ZHENG Hong-Bing, SUI Peng, LI Yuan-Yuan, SHI Xue-Peng, NIE Sheng-Wei, GAO Wang-Sheng
    Scientia Agricultura Sinica. 2011, 44(14):  2935-2942 .  doi:10.3864/j.issn.0578-1752.2011.14.010
    Abstract ( 723 )   PDF (311KB) ( 712 )   Save
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    【Objective】 Reveal net greenhouse gases(GHGs)under conservation tillage in farmland in Jilin province. 【Method】 Experimental studies on conservation tillage with 4 treatments (technology of maize planted in ridge side with high stubble; technology of maize planted in wide line and narrow line alternately; technique of seeding straightly with stubble left in the field; technology of maize planted directly on ridge before smashing stubble plowing combined with fertilization) in main producing areas of maize in Jilin province were conducted to figure up the carbon accumulation, GHGs emission from field and input, thus the net emission can be calculated systematically.【Result】Four conservation tillage patterns increased soil organic carbon(SOC) content in arable layer. The highest potential of carbon sequestration was Technique of maize planted in wide line and narrow line alternately(CT2) with a rate of 1 955.07 kg C•hm-2•a-1, while the lowest was showed under technique of seeding straightly with stubble left in the field (CT3) with a rate of 1 492.26 kg C•hm-2•a-1. On the contrary, conventional tillage (CK) decreased SOC content with 173.70 kg C•hm-2•a-1. Average amount of CO2 equivalent (CO2-eq) emission under 4 conservation tillage patterns was 5 259.25 kg•hm-2•a-1, and 5 367.96 kg•hm-2•a-1 emitted from CK. GHGs from input contributed mostly to total emission and production of nitrogen accounted a large proportion for it. The calculated result showed that there was a diversity of GHGs mitigation among 4 conservation tillage patterns. Most CO2-eq mitigated was 1897.56 kg•hm-2•a-1 by CT2 and least by CT3 with 225.75 kg•hm-2•a-1, while CK was -6 004.87 kg•hm-2•a-1. 【Conclusion】 Four conservation tillage patterns in Jilin Province increased the SOC content and offset the GHGs emission form soil and input. They were sinks of GHGs while CK was a source because it decreased the SOC content without mitigation of GHGs.
    Research on the Values of CO2 Fixation and O2 Release by Landuse Ecosystem in Loess Plateau in Northern Shaanxi Province
    LI Jing, REN Zhi-Yuan
    Scientia Agricultura Sinica. 2011, 44(14):  2943-2950 .  doi:10.3864/j.issn.0578-1752.2011.14.011
    Abstract ( 788 )   PDF (379KB) ( 765 )   Save
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    【Objective】 It is important to study quantitative evaluation of CO2 fixation and O2 release function in ecological system. This article attempts to measure and evaluate the value of fixing carbon and releasing oxygen in land ecological system using remote sensing, to provide reference for the regional ecological environment. 【Method】 Based on photosynthesis, using remote sensing estimation model, based on the results from the electroosimosis NPP vegetation, the fixed carbon and released oxygen amount was estimated. Based on this result, by using forestation costs method and the industrial oxygen producing method, the values of fixed carbon and released oxygen in Loess Plateau in Northern Shaanxi Province were estimated. The spatial and temporal changes and distribution of the fixed CO2 and released O2 in loess plateau ecological system were discussed. 【Result】CO2 fixation values in Loess Plateau in Northern Shaanxi Province in 1978, 1990, 2000 and 2005, respectively, were141.27 billion yuan, 177.46 billion yuan, 170.09 billion yuan and 247.08 billion yuan, and the average was 187.72 billion yuan. The value of O2 release from the two evaluation methods, averaged, respectively, 150.07 billion yuan, 188.52 billion yuan, 192.97 billion yuan and 281.48 billion yuan, and the annual average was 203.26 billion yuan, more than an average of CO2 fixation 15.54 billion yuan. From the perspective of land use types, forestland>cultivated land>grassland>garden>other types. 【Conclusion】 The value of the fixed CO2 and released O2 in loess plateau ecological system of northern counties is lower than southern counties and in general distribution along the northern south to increasing gradually. HuangLong Mountain in south has the very high ridge and unit area of value, such as the county, Huangling county, Fu county and Ganquan county. The value of fixing carbon and releasing oxygen in northern agriculture and animal region and the central loess endangers region with the serious soil and water loss has lower value, this fragile ecological environment and relevant.
    Advances in Research of Molecular Ecology of Carbon Fixation Microorganism
    YUAN Hong-Chao, QIN Hong-Ling, LIU Shou-Long, NIE San-An, WEI Wen-Xue, WU Jin-Shui
    Scientia Agricultura Sinica. 2011, 44(14):  2951-2958 .  doi:10.3864/j.issn.0578-1752.2011.14.012
    Abstract ( 983 )   PDF (308KB) ( 2485 )   Save
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    Alleviating the global climate change is one of the most important environmental challenges human faced. Autotrophic microbes have a strong adaptability to environmental changes and potential to CO2 fixation, thus it is important for researchers to study on molecular mechanisms of CO2 fixation by autotrophic microbes. Most of the autotrophic microbes fix CO2 via Calvin cycle, although there are another four alternative pathways. The crucial reaction of this pathway is catalyzed by the enzyme ribulose-1, 5-bisphos-phatecarboxylase/oxygenase (RubisCO). Therefore, the RubisCO and the encode gene can be used as functional gene markers to investigate the community and diversity of the CO2 fixation microorganisms in different ecological systems. The previous molecular analysis of microbial carbon fixation has emphasized on aquatic systems, revealing the characteristics of carbon fixation microbial communities and the response to the different habitats. Recent studies on the terrestrial systems have found out that CO2 fixation microorganism also numerous and widespread in soils whereas the functional implication of those autotrophies is not clear, and need to be future identified. Domestic researches on the molecular ecology of soil CO2 fixation microorganism as yet have not been reported. In the present paper, the species of carbon fixation microorganism, the dynamics of CO2 fixation and the current status concerning the carbon fixation by microorganism were reviewed, the main problems and further researches of the molecular ecology of carbon fixation in agricultural ecosystems were also discussed. The aim of this study is to provide useful information for Chinese molecular ecology research of CO2 fixation microorganism.
    HORTICULTURE
    Molecular Distinction of Two Ogura CMS Sources in Brassica oleracea var. capitata L.
    ZHANG Yang-Yong, FANG Zhi-Yuan, WANG Qing-Biao, LIU Yu-Mei, YANG Li-Mei, ZHUANG Mu, SUN Pei-Tian
    Scientia Agricultura Sinica. 2011, 44(14):  2959-2965 .  doi:10.3864/j.issn.0578-1752.2011.14.013
    Abstract ( 720 )   PDF (772KB) ( 574 )   Save
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    【Objective】 Two feasible Ogura cytoplasmic male sterility sources (CMS HY and CMSR3) in cabbage were used to identify its cytoplasmic DNA distinction. 【Method】 With chloroplast SSR (cpSSR) and mitochondrial SSR (mtSSR) primers, the technologies of polyacrylamide gel electrophoresis and DNA sequencing were used to explore the molecular differences between Ogura CMSHY and CMSR3. The markers with restriction enzyme recognition sites were converted into CAPS markers.【Result】 cpSSR amplicons could not differ two CMS sources on polyacrylamide gel, but the sequence of amplicon ACP9 differed them on SSR repeat numbers. In 21 mtSSR primers, only amplicon of mtSSR2 could distinguish two CMS sources on polyacrylamide gel. Moreover, nine polymorphic differences were obtained with sequencing in other 5 mtSSR primers. Two polymorphic differences with MseⅠ restriction enzyme recognition sites were converted into CAPS markers, named as m92-143 MseⅠ, m1-346 MseⅠ.【Conclusion】 Two cabbage Ogura CMS sources were distinguished with cpSSR and mtSSR markers. CAPS markers could be used as a fast, easy and accurate method to distinguish two CMS sources.
    Proteomics on Somatic Embryogenesis During the Middle Stage in Longan
    FANG Zhi-Zhen, LAI Zhong-Xiong, LAI Cheng-Chun, JIANG Shun-Ri
    Scientia Agricultura Sinica. 2011, 44(14):  2966-2979 .  doi:10.3864/j.issn.0578-1752.2011.14.014
    Abstract ( 840 )   PDF (1825KB) ( 728 )   Save
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    【Objective】 The changes of proteome patterns during the middle stage of longan (Dimocarpus longan Lour.) somatic embryogenesis were analyzed to provide valuable evidence for further elucidating the mechanism of somatic embryogenesis.【Method】Synchronization of longan somatic embryogenesis during the middle stage was carried out, and changes of proteomic patterns during this period were analyzed by 2-DE and MS.【Result】During this period, diversity of expressed proteins declined with the development of somatic embryos of longan, but it showed a increase at torpedo stage. Newly expressed proteins at torpedo stage included those with a MW less than 66.0 kD and a pI of these proteins range between 5.00 and 7.00. Thirty-five differentially expressed proteins were identified, and many of them were related to somatic embryogenesis.【Conclusion】With the development of somatic embryo, somatic cells differentiated progressively and the amount of expressed proteins decreased. Proteins associated with oxidative stress, RAN2 and GTPase ObgE might be related to regulation of somatic embryogenesis in longan.
    Effects of Applying Different Sesame Organic Materials as Nest-Fertilizers on Chemical Components and Aroma Quality of Flue-Cured Tobacco
    HAN Fu-Gen, SHI Jin-Zhong, WANG Xiao-Hui, SONG Peng-Fei, SHEN Zheng, LI Yong-Liang
    Scientia Agricultura Sinica. 2011, 44(14):  2980-2989 .  doi:10.3864/j.issn.0578-1752.2011.14.015
    Abstract ( 812 )   PDF (400KB) ( 670 )   Save
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    【Objective】 The purpose of the experiment is to study the effects of applying different sesame organic materials as nest-fertilizers on chemical components and aroma quality of flue-cured tobacco. 【Method】 Different from the traditional cake fertilizer applied as base fertilizer in row replacement, nest-fertilizers of different sesame organic materials were applied at transplanting period in the field experiment. 【Result】 The quality of three parts of tobacco leaf could be improved when sesame cake fertilize was used as the nest-fertilizer. The quality of middle and lower leaves could be improved when fried sesame was used as the nest-fertilize. The quality of upper leaves could only be improved when the sesame oil was used nest-fertilizer. The ratio of sugar/ nicotine and total N/nicotine were appropriate in three parts of tobacco leaves (with the treatment of cake fertilizer ) and upper & middle leaves (with the treatment of fried sesame), but the quality of three parts leaves with the treatment of sesame oil were no good, and the chemical components contents were less harmonious. The increase of total aroma components contents of lower and middle leaves was from neophytadiene, cembranoids and carotenoids, the increase of total aroma component contents of upper leaf was from phenylalanine products, brown chemical products and cembranoids (with the treatment of sesame cake fertilizer). The increase of total aroma component contents of middle and upper leaves was from neophytadiene, phenylalanine products and cembranoids (with the treatment of fried sesame). The increase of total aroma component contents of upper leaves was from neophytadiene and cembranoids (with the treatment of sesame oil). Sensory smoking results showed that three parts of tobacco leaf (with the treatment of sesame cake fertilizer) were the best in all of the treatments. The characteristics of them were pure aroma, adequate aroma and comfortable aftertaste. 【Conclusion】 The results showed that, when the same amount of sesame cake fertilizer was applied, a part of them applied as base fertilizer and nest-fertilizer was better than as base fertilizer, especially, the contents of neophytadiene (except that in upper leaves compared with CK) and the other aroma components. The other features of this treatment were comfortable sugar/ nicotine and total N/ nicotine ratios, lower contents of protein and Cl, higher K, more harmonious chemical and aroma components. It is better to improve aroma quality of tobacco leaves by combining sesame organic materials applied as basal fertilizer and nest-fertilizer.
    STORAGE·FRESH-KEEPING·PROCESSING
    Effects of Wheat Bran Addition and Particle Size on Mixing Properties of Dough
    CHEN Jian-Sheng, CUI Jin-Long, DENG Zhi-Ying, TIAN Ji-Chun, ZHANG Yong-Xiang, WU Guo-Song, LI Yan
    Scientia Agricultura Sinica. 2011, 44(14):  2990-2998 .  doi:10.3864/j.issn.0578-1752.2011.14.016
    Abstract ( 726 )   PDF (273KB) ( 766 )   Save
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    【Objective】 Mechanism and effect of bran addition and particle size on mixing properties of dough were analyzed to provide a scientific basis for application of wheat bran in food production. 【Method】 Aiming to study the effects of wheat bran addition level and particle size on the mixing property of wheat flour, wheat bran produced from Shannong 2 were added into flour, with bran addition of 5%, 10%, 15%, 20% and with bran particle size of 0.16-0.43 mm, 0.43-1.0 mm, 1.5-2.0 mm and 2.0-2.5 mm. The mixing properties of the mixed flour samples were determined. 【Result】 With the increasing addition level of wheat bran the midline peak value, midline peak width, 8 min value and integral, curve tail value and integral decreased significantly, while the 8 min width, curve width and slope of peak right increased remarkably. There was no remarkable difference in effects of bran addition levels on midline peak time, peak time integral and slope of peak left. With the increasing particle size the midline peak time, 8 min width, curve tail width and slope of peak right increased significantly, but no remarkable different effects were detected in other mixing parameters. 【Conclusion】 Wheat bran decreased significantly the midline peak value,8 min midline value, peak time integral and 8 min midline integral, while increased the midline peak time and 8 min width and slope of right of peak. With the increasing particle size the midline peak time, 8 min width, curve tail width and slope of peak right increased significantly.
    Separation and Detection of Mono-Cellooligosaccharide Purified by Preparative Silica Column Chromatography
    LIU Cheng-Cheng, SHI Bo, YAO Xi-Mei, LIANG Ping, LI Jing-Mei
    Scientia Agricultura Sinica. 2011, 44(14):  2999-3006 .  doi:10.3864/j.issn.0578-1752.2011.14.017
    Abstract ( 1006 )   PDF (543KB) ( 741 )   Save
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    【Objective】The aim of this paper was to explore a simple method to get sizeable amount of mono- cellooligosaccharide with degrees of polymerization(DP)two to DP five. 【Method】 The preparative silica column chromatography was selected to separate and purify the cello-oligosaccharides prepared from absorbent cotton. The separation process was monitored by TLC. FACE was used to test the separated single-component oligosaccharide and ESI-TOF-MS was used to detect their molecular weights. 【Result】 The experimental result showed that the preparative silica column chromatography had a better resolving ability for separating the cello-oligosaccharides with DP between two to five. From the ESI-TOF-MS spectrum, the molecular weights included one sodium ion of the four single-component cello-oligosaccharide were 365 for cellobiose, 527 for cellotriose, 689 for cellotetraose, and 851 for cellopentaose, respectively. Separated 300 mg cello-oligosaccharides once time, could get cellobiose 28 mg, cellotriose 43 mg, cellotetraose 59 mg, and cellopentaose 52 mg, respectively. FACE method was very sensitive to the qualitative analysis of cello-oligosaccharides. 【Conclusion】 The preparative silica column chromatography is a better separation and purification method for preparing mono-cellooligosaccharide with DP two to five.
    ANIMAL SCIENCE·RESOURCE INSECT
    Research on Carbon Balance of Different Grazing Systems in Stipa breviflora Desert Steppe
    HOU Xiang-Yang, XU Hai-Hong
    Scientia Agricultura Sinica. 2011, 44(14):  3007-3015 .  doi:10.3864/j.issn.0578-1752.2011.14.018
    Abstract ( 681 )   PDF (479KB) ( 636 )   Save
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    【Objective】Taking Stipa breviflora desert steppe in Inner Mongolia as the research object, carbon input, carbon output and carbon balance features in plots that affected by fencing, rotational grazing and continuous grazing for ten years (1999-2009) were determined.【Method】Using field investigation and enclosed chamber method (IRGA), the dynamics of biomass and soil respiration rate were determined in Stipa breviflora desert steppe. The effects of grazing systems on carbon input, carbon output and carbon balance was studied in this paper. 【Result】 Compared with continuous grazing, the litter fall, livestock feed intake and below-ground biomass were increased by rotational grazing, while no significant difference the community above-ground biomass was found in between continuous grazing and rotational grazing. Litter fall, above ground and below ground biomass were increased by non-grazing. Compared with continuous grazing, rotational grazing and non-grazing increased the soil respiration rate in dry months. Stipa breviflora desert steppe emerged to be carbon sinks under different grazing systems, and the net ecosystem production was, respectively, 413.78, 401.45, and 416.80 gC•m-2•a-1.【Conclusion】There was little difference between different grazing systems, and carbon sinks decreased in turn from non-grazing, rotational grazing to continuous grazing.
    Effects of Different Selenium Sources on Production Performance, Slaughter Performance, Meat Quality, Immune and Antioxidant in the Early Goose
    WANG Bao-Wei, WANG Na, GE Wen-Hua, YUE Bin, ZHANG Ming-Ai, SHI Xue-Ping
    Scientia Agricultura Sinica. 2011, 44(14):  3016-3026 .  doi:10.3864/j.issn.0578-1752.2011.14.019
    Abstract ( 1085 )   PDF (372KB) ( 774 )   Save
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    【Objective】 The objective of the experiment is to study the effect of different selenium sources on production performance, slaughter performance, meat quality, muscles conventional nutrients,immune function and anti-oxidation function in goose.【Method】Ninety-six one-day-old geese with similar initial body weight (P>0.05) were divided into 4 groups with three replications per group according to a completely random design. The proportion of selenium in the same basal diets was 0.30 mg•kg-1, respectively. The control group was fed with basal diet, the other three groups was fed with basal diet added with sodium selenite (SS), Se-enriched yeast (SY) and Nano-Se, respectively.【Result】Except that breast muscle percentage and leg muscle rate index of SY group were much higher than the control group (P<0.05), the body weight gain, feed conversion ratio and slaughter performance index of 4 and 9-week-old goose in other groups were no obvious difference (P>0.05), and there was no obvious difference in different selenium sources (P>0.05). The water loss rate, hardness, water content and crude fat content index of 9-week-old goodse in other groups were lower than the control group (P<0.05), and there was no obvious difference in different selenium sources (P>0.05). The drip loss of other groups were lower than the control group (P<0.05), the drip loss of SY was lower than SS (P<0.05). The mypglobin content of other groups were higher than the control group (P<0.05), the mypglobin content of SY was higher than SS (P<0.05). The crude protein content of SY and Nano-Se were higher than the control group (P<0.05), and there was no obvious difference in different selenium sources (P>0.05). Various selenium sources increased significantly in immune function of goose, except that spleen index of Nano-Se group compared with the control group was no obvious difference, the immune organ index of 4 and 9-week-old of other groups were much higher than the control group (P<0.05), but there was no obvious difference in different selenium sources.Tlymphocyte transformation rate of Nano-Se group and 4, 9 week-old of SS group were all much higher than the control group (P<0.05), SY group were much higher than SS group (P<0.05) and very higher than the control group (P<0.01). The activity of GSH-PX, T-SOD and T-AOC among various selenium sources both in 4, 9w serum and liver increased significantly (P<0.05). The content of MDA and H2O2 reduced significantly both in 4, 9w serum and liver (P<0.05). There was no difference in the activity of GSH-PX among various selenium sources in 4w serum of geese (P>0.05). Compared with those geese fed by SS, the activity of GSH-PX in serum (9w) and liver (4, 9w) fed by SY increased significantly (P<0.05). There was no difference in the activity of T-SOD among various selenium sources both in 4, 9w serum and liver of geese. The activity of T-AOC in 4, 9w serum and 4w liver fed by SY significantly increased (P<0.05); the activity of T-AOC in 9w liver fed by Nano-Se very significantly increased (P<0.05). The content of MDA in 4, 9w serum and liver fed by SY very significantly reduced (P<0.05). There was no difference in the content of H2O2 among various selenium sources both in 4, 9w serum and liver (P>0.05). The cGPx gene expression of the SS and Nano-Se group were significantly higher than the control group (P<0.05), the SY group was very significantly higher than the control group (P<0.01), the SY group was higher than the SS group and Nano-Se group (P<0.05).【Conclusion】Adding different selenium sources to the diet of goose has no significant effects on production performance and slaughter performance, but can improve the quality of goose and content of muscles conventional nutrients, and promote the immune, antioxidant capacity. Compared with SS and Nano-Se, SY is a better source of selenium for goose.
    SNPs Detection in Chinese Holstein Cattle Transferrin Gene and Its Associations with Milk Performance
    JU Zhi-Hua, LI Qiu-Ling, HUANG Jin-Ming, WANG Ji, LI Rong-Ling, LI Jian-Bin, ZHONG Ji-Feng, WANG Chang-Fa
    Scientia Agricultura Sinica. 2011, 44(14):  3027-3035 .  doi:10.3864/j.issn.0578-1752.2011.14.020
    Abstract ( 723 )   PDF (465KB) ( 522 )   Save
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    【Objective】The polymorphisms of transferrin gene and their correlation to milk performance and healthy traits in Chinese Holstein cattle were investigated, which may help to breed high milk yield and healthy cows.【Method】DNA sequencing, PCR-RFLP and CRS-PCR were applied to analyze the polymorphisms of transferrin gene in 576 Chinese Holstein cattle and its associations with production performance were investigated by SAS software.【Result】Three novel SNPs (g.-1748G>A, g.13942T>C and g.14037A>G) in the 5′flanking region, exon 8 and intron 8 of Tf gene were found, respectively. The locus g.-1748G>A was significantly associated with 305 d milk yield (P<0.05). The locus g.13942T>C was significantly associated with somatic cell score. The cows with genotype CC was significantly higher than TT and TC genotype (P<0.05). At locus g.14037A>G , the cows with genotype AA showed higher 305 d milk yield than those with genotypes AG (P<0.05), with genotype AG showed higher protein rate than those with genotypes GG (P<0.05). Eight haplotype and 19 haplotype combinations were found in 576 Chinese Holstein cattle. The cows with haplotype combinations H4H4 had the highest milk fat and milk protein percentage. The cows with haplotype combinations H2H5 had the highest 305 d milk yield and the cows with haplotype combinations H3H4 had the lowest somatic cell score.【Conclusion】The correlation was detected between different genotypes and haplotype combinations of Tf gene and milk yield, protein rate and somatic cell score, but there was no correlation between the polymorphisms and milk fat rate.
    VETERINARY SCIENCE
    Expression of Outer Membrane Protein P5 Gene of Haemophilus Parasuis and Establishment of an Indirect ELISA Based on the OMP5 Protein
    CHEN Shan-Zhen, LI Chun-Ling, JIA Ai-Qing, WANG Gui-Ping
    Scientia Agricultura Sinica. 2011, 44(14):  3036-3044 .  doi:10.3864/j.issn.0578-1752.2011.14.021
    Abstract ( 745 )   PDF (555KB) ( 788 )   Save
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    【Objective】 In the present study,an indirect ELISA was developed using the purified OMP5 protein to detect the antibody of HPS.【Method】Using gene recombination technology,the OMP5 gene was cloned and inserted into prokaryotic expression vector pET-32a (+) and the recombinant expression vector was identified with PCR,restriction enzyme digestion and sequence analysis.The positive recombinant plasmid was then transformed into E.coli BL21 (DE3) and induced by IPTG.The expressed protein was purified by Ni-NTA affinity chromatography and analyzed by SDS-PAGE.The purified protein was refolded by urea gradient dialysis,and its specificity was tested by Western blot.An indirect ELISA was developed with the purified protein and supersonic bacterial antigen at different concentrations and the optimal antigen concentration and serum dilution were determined by phalanx titration.The other assay conditions were also optimized.【Result】The optimal coating concentration of OMP5 and serum dilution were 1﹕400 and 1﹕160,respectively. Three-hundred and seventeen swine sera collected from healthy pigs which were never immunized with HPS were assayed by the ELISA with urea treated antigen. Seventy-two samples were positive,and the positive rate was 22.71%,which was closed to the isolating rate of 24% from HPS naturally infected pigs. Seventy-eight serum samples were assayed using the ELISA coated with the supersonic HPS antigen. Twenty-seven samples were detected as positive with the ELISA coated with the purified OMP5 protein and the detection rate was 34.62%,and 31 positive was detected with the supersonic bacterial antigen coated ELISA and the detection rate was 39.74%.Coincidence of the two methods was 71.79%.【Conclusion】The developed ELISA had good specificity and reproducibility.It could provide a reliable method for the clinical detection of HPS,epidemiological investigations,and immunization surveillance.
    Co-administration Intranasally the FMDV DNA Vaccine and the Constructed Expressing IL-10 as the Molecular Adjuvant in Enhancement of Mucosal Immune Responses in Murine Model
    WANG Xiao, WANG Zhi-Gang, DU Rui-Ping
    Scientia Agricultura Sinica. 2011, 44(14):  3045-3052 .  doi:10.3864/j.issn.0578-1752.2011.14.022
    Abstract ( 991 )   PDF (590KB) ( 585 )   Save
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    【Objective】 An experiment was carried out to investigate whether co-administration intranasally of the FMDV DNA vaccine, pcD-VP1 and a construct expressing IL-10 as the molecular adjuvant can enhance mucosal immune responses in murine model. 【Method】 proVAX-IL-10 recombinant plasmid was constructed for eukarytic expression, then the mice were randomly divided into 5 groups and they were immunized i.n. with pcD-VP1 alone or co-immunized with the provax-IL-10 molecular adjuvant on days 0, 14 and 28. After immunization, mucosal sIgA ,T cell proliferation and the expession of cytokines were deteced by ELISA, CFSE staining and intracellular cytokine staining method, respectively. 【Result】 Compared to the group intranasally immunized with pcD-VP1 alone, the group immunized with the molecular adjuvant was induced higher level of mucosal sIgA, and intranasal delivery of the IL-10 con struct with pcD-VP1 significantly enhanced the higher level of antigen specific T cell proliferation and higher expession IL-4 in CD4+ cells inform mucosal site compared to the pcD-VP1 alone. 【Conclusion】 The results demonstrated that intranasal delivery of IL-10 as a mucosal adjuvant can enhance the antigen specific mucosal immune responses in a murine model, which may provide a protection against the FMDV initial infection.
    RESEARCH NOTES
    Development of A SCAR Marker for Fusarium Wilt Resistance in Cabbage
    JIANG Ming; ZHAO Yue; XIE Jian-ming; TIAN Ren-peng;CHEN Yan-yang; KANG Jun-gen
    Scientia Agricultura Sinica. 2011, 44(14):  3053-3059 .  doi:10.3864/j.issn.0578-1752.2011.14.023
    Abstract ( 988 )   PDF (510KB) ( 695 )   Save
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    【Objective】The objective of the experiment is to develop a SCAR(sequence characterized amplified region) marker of Fusarium wilt resistant gene FOC-1 in cabbage.【Method】Bulked segregation analysis (BSA) DNA pools were constructed by F2 individuals derived from the cross between resistant ‘8024’× susceptible ‘6A’ cross. An AFLP marker genetically linked to the Fusarium wilt resistant gene was identified and it was converted into a more valuable and simple SCAR marker S46M48199 consequently. Genetic linkage between S46M48199 and FOC-1 was analyzed by using 142 F2 progenies. The validity and practicality of S46M48199 was verified in two different cabbage F2 populations F66 and C1.【Result】One stable SCAR marker S46M48199 linked in repulsion to the dominant resistant allele at 2.78 cM from the FOC-1 locus was developed successfully. The validity and practicality of S46M48199 in two other populations revealed significant consistency with phenotype assay (81% and 83%, respectively). 【Conclusion】The SCAR marker developed in this study could be used for MAS of Fusarium wilt resistance in cabbage.

    Construction of Agrobacterium-mediated Cucumber mosaic virus Infectious cDNA Clones and 2b Deletion Viral Vector

    YAO Min; ZHANG Tian-qi; TIAN Zhi-chao; WANG Yuan-chao; TAO Xiao-rong
    Scientia Agricultura Sinica. 2011, 44(14):  3060-3068 .  doi:10.3864/j.issn.0578-1752.2011.14.024
    Abstract ( 1224 )   PDF (418KB) ( 1353 )   Save
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    【Objective】 The objective of this study is to generate the Agrobacterium-mediated infectious cDNA clones of Cucumber mosaic virus and investigate the phenotype of 2b deletion mutant in Nicotiana benthamiana. 【Method】1x35S-MCS-HDVRZ-NOS cassette was transferred from pHST40 into pBluescrpt SK II and 2x35S enhancer from pRTL2 was used to replace 1x35S on pBluescrpt SK II, then the whole 2x35S-MCS-HDVRZ-NOS cassette was transferred into mini binary vector pCB301 to generate pCB301-2x35S-MCS-HDVRZ-NOS. The full length cDNAs of CMV Fny were inserted between 2x35S promoter and HDV Rybozyme on pCB301-2x35S-MCS-HDVRZ-NOS, then they were introduced into agrobacterium and inoculated N. benthamiana by agro-infiltration. Based on those infectious clones, 2b deletion mutant was constructed to investigate its phenotype in N. benthamiana. 【Result】 A mini binary vector pCB301-2x35S-MCS-HDVRZ-NOS was constructed for making infectious clones of plant RNA virus. The full lengths of three genomic cDNAs of CMV Fny were inserted into this binary vector, then they were introduced into agrobacterium, respectively, and infiltrated into N. benthamiana. The inoculated plant could produce severe symptoms including leaf curl, mosaic and dwarf etc. Further deletion of 2b demonstrate that 2b deletion mutant could induce mild leaf curl symptoms on N. benthamiana in earlier time, but the symptoms disappear in later stage, RT-PCR shows that 2b deletion mutant still could infect the N. benthamiana systemically. 【Conclusion】 The infectious clones of CMV were constructed based on a modified mini binary vector pCB301-2x35S-MCS-HDVRZ-NOS. The Agrobacterium-mediated infection of N. benthamiana by those infectious cDNA clones of CMV was successful and efficient. It is a simple, fast and low cost approach to make viral infectious clones. The phenotype of CMV Fny 2b mutant on N. benthamiana suggest that although 2b deletion mutant induce mild leaf curl symptoms at earlier time, 2b is a key determinant for symptom development for CMV in N. benthamiana. 2b is not required for viral replication and systemic infection in this plant.