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Table of Content

    01 January 2011, Volume 44 Issue 1
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Global Genome Expression Analysis of Photosynthesis-Related Genes Under Low Nitrogen Stress in Rice Flag Leaf
    ZHAO Ming-hui,SUN Jian,WANG Jia-yu,XU Hai,TANG Liang,CHEN Wen-fu
    Scientia Agricultura Sinica. 2011, 44(1):  1-8 . 
    Abstract ( 834 )   PDF (350KB) ( 1092 )   Save
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    【Objective】 The response of photosynthesis-related genes to low nitrogen stress were researched, thus providing a molecular basis for improving the absorption efficiency and utilization efficiency of nitrogen fertilizer in rice. 【Method】 The Agilent rice genome arrays were used to study the expression change of photosynthesis-related genes in two rice cultivars with different chlorophyll contents under low nitrogen fertilizer stress. 【Result】 The results showed that chlorophyll content and photosynthetic rate of the flag leaf were decreased under low nitrogen stress in SN19-6 and Fengjin, but the degree of decrease was significantly different between SN19-6 and Fengjin. A total of 41 photosynthesis-related genes (14 photosynthesis-related genes were down-regulated and 27 photosynthesis-related were up-regulated at the transcription level) in flag leaf of supper-green rice SN19-6 and 29 photosynthesis-related genes (15 photosynthesis-related genes were down-regulated and 14 photosynthesis-related genes were up-regulated at the transcription level) in flag leaves of Fengjin were affected by low nitrogen stress. The photosynthesis-related genes for expressed cultivar special-response to low nitrogen stress, with 29 photosynthesis-related genes for SN19-6 and 17 for Fengjin. There was an overlap of photosynthesis-related genes in response to low nitrogen stress between SN19-6 and Fengjin, with 5 up-regulated and 7 down-regulated at the transcription level, respectively.【Conclusion】 Expression of photosynthesis-related genes were affected by low nitrogen stress in flag leaf, and there are genes of cultivar special-response as well as overlap between SN19-6 and Fengjin.

    Genetic Analysis and SSR Molecular Mapping of New Stripe-Rust Resistance Gene YrWV Derived from Triticum aestivum-Haynaldia villosa Translocation Line V9125-2
    WANG Rui,ZHANG Shu-ying,XU Zhong-qing,CHEN Jie,LI Qiang,HOU Lu,JING Jin-xue
    Scientia Agricultura Sinica. 2011, 44(1):  9-19 . 
    Abstract ( 753 )   PDF (1038KB) ( 672 )   Save
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    【Objective】 The high stripe rust resistance material, translocation line V9125-2, from Triticum aestivum-Haynaldia villosa was studied. Inheritance, location and SSR molecular mapping of stripe rust resistance genes in V9125-2 are important for providing a basis for breeding high stripe rust resistance materials. 【Method】 To identify and tag new resistance genes to stripe rust in the translocation line V9125-2 from Triticum aestivum-Haynaldia villosa, the yellow rust races(Puccinia striiform f.sp. stritici)CYR29, CYR30, CYR31, CYR32, CYR33, Su11-4 and Su11-11 were selected as innoculum of genetic analysis of resistance to stripe rust in the progeny population of V9125-2×Mingxian169 at seedling stage. Using the F2 population inoculated with CYR29 to test SSR analysis, combined with bulked segregation analysis (BSA), 289 pairs of primers were used. The F3 population were used to test the linked markers. And the homologous rate between the stripe rust resistance genes of V9125-2 and the varieties in Huang-Huai wheat grown region was detected. 【Result】 The results showed that one dominant gene conferred resistance to stripe rust race CYR29. One dominant gene and one recessive gene indenpently conferred to CYR30, CYR32, CYR33 and Su11-11. And two pairs of independent dominant genes conferred to CYR31 and Su11-4, respectively. The F2 population inoculated with CYR29 were used for SSR analysis, temporarily designated as YrWV. SSR markers, as well as 289 pair primers, combined with bulked segregation analysis (BSA) revealed that six SSR markers, Xbarc87, Xwmc463, Xwmc405, Xbarc126, Xwmc438 and Xgwm473, located on chromosome 7DS were linked to YrWV, and their genetic distances to YrWV were 9.1, 3.9, 5.1, 12.6, 29.0 and 57.4cM, respectively, and the six sites linked to YrWV were validated by a segregated ratio of F3 lines. These markers would be used for breeding new wheat cultivars with marker-assisted selection. A very low homologous rate was found in 43 varieties in Huang-Huai wheat grown region tested with YrWV tags. 【Conclusion】 The result realed that the six SSR markers, Xbarc87, Xwmc463, Xwmc405, Xbarc126, Xwmc438 and Xgwm473, located on the short arm of chromosome 7D in wheat were linked to YrWV. It may be a new wheat stripe rust resistant gene which comes from Haynaldia villosa.

    Construction of DNA Fingerprinting and Analysis of Genetic Diversity with SSR Markers for Cotton Major Cultivars in China
    KUANG Meng,YANG Wei-hua,XU Hong-xia,WANG Yan-qin,ZHOU Da-yun,FENG Xin-ai
    Scientia Agricultura Sinica. 2011, 44(1):  20-27 . 
    Abstract ( 1024 )   PDF (397KB) ( 1165 )   Save
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    【Objective】The aim of this study was to construct a DNA fingerprinting database of 32 cotton major cultivars from three main cotton regions (including eight main cotton production provinces), and the genetic diversity was analyzed based on simple sequence repeats (SSR).【Method】Thirty six evenly distributed SSR primer pairs with high polymorphisms and good repeatability were successfully screened out from 214 candidates to construct the fingerprinting database.【Result】Among the 32 varieties, 36 primer pairs had 142 polymorphic genotypes, and 3.94 genotypes were detected by each SSR primer pair on an average with the range from 2 to11. Ten cultivars had specific genotypes by 9 primer pairs. Thirty two major cultivars could be identified by 5 primer combinations at least. The genetic diversity analyzed by the software of NTSYS-pc V 2.10 indicated that Yangtze River valley cotton region had the highest polymorphic level, followed by Xinjiang, and the lowest was the varieties from the Yellow River valley. The genetic basis of conventional cultivars was narrower than that of hybrids. 【Conclusion】 Core primer combination method is better than distinct band method for the construction of DNA fingerprinting of cotton major cultivars in China. The genetic relationship of the cotton cultivars derived from the same district is similar to a certain extent.

    In Silico Cloning and Bioinformatic Analysis of TPK1 Gene in Tobacco
    LU Li-ming
    Scientia Agricultura Sinica. 2011, 44(1):  28-35 . 
    Abstract ( 714 )   PDF (454KB) ( 1055 )   Save
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    【Objective】 The aim of this study is to clone K+ transporter gene in tobacco, analyze its genetic relationship, and predict its structure, nature, function and to provide a basis for K+ gene function studies of tobacco. 【Method】 Cloning tobacco K+ transporter gene by means of the combination of laboratory work and e-cloning techniques. Using tobacco K+ transporter gene 3' end sequence, cloned by RACE, as a probe to search the tobacco EST databases, filter out the ESTs sequences with the homology of 95% of the probe, and spliced the two segments using DNAStar software. Then, analyze gene structure and properties of the full cDNA sequence, and predict its function by applying a variety of bioinformatics databases and softwares. 【Result】A new tobacco K+ gene (TPK1) with full cDNA sequence was found. The molecular weight of the protein encoded by this gene is 26.21 kD, and the theoretical isoelectric point pI is 9.72. TPK1 is a hydrophobic membrane protein, containing six distinct transmembrane helix topologies. In secondary structure, a total of 11 α-helix, and 16 β-fold were found in TPK1. At subcellular level, TPK1 protein located in the plastid, vacuole and endoplasmic reticulum. The signal peptide splice site and the serine, threonine and tyrosine kinase phosphorylation sites in TPK1 amino acids sequence were found as well. In the function, TPK1 protein contains a typical conserved K+ uptake transporter function domain, and is highly of homology to the major K+ uptake transporter genes in higher plants. Northern blotting analysis indicated that TPK1 was expressed in root, stem, leaf and flower.【Conclusion】A new K+ transporter TPK1 was successfully cloned in tobacco, and its function maybe involved in K+ transportation in tobacco plants. Therefore, further investigation still needed to understand its role in tobacco K+ uptake and transportation.

    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY

    Relationships of Rice Root Morphology and Physiology with the Formation of Grain Yield and Quality and the Nutrient Absorption and Utilization

    YANG Jian-chang
    Scientia Agricultura Sinica. 2011, 44(1):  36-46 . 
    Abstract ( 1082 )   PDF (446KB) ( 2407 )   Save
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    Roots are an integral part of plant organs and involved in acquisition of nutrients and water, synthesis of plant hormones, organic acids and amino acids, and anchorage of plants. Root morphology and physiology are closely associated with the growth and development of above-ground part of plant. In this review, the relationships of root morpho-physiological traits with the formation of grain yield and the absorbtion of water and nutrients in rice were summerized. Recent advances in research on the role of the root chemical signals, such as hormones and organic acids, in the formation of grain quality of rice and the relation of ultra-structure of root tip cells with the growth and development of above-ground part of plant were intruduced. The existing problems and futher studies on rice roots were discussed.

    Effect and Preliminary Analysis of Chromosomal Control on the Chlorophyll Fluorescence Parameters of Wheat Substitution Lines Between Synthetic Hexaploid Wheat and Chinese Spring Under Drought Stress

    BAI Zhi-ying,LI Cun-dong,ZHAO Jin-feng,WU Tong-yan,ZHENG Jin-feng,BI Chang-rui
    Scientia Agricultura Sinica. 2011, 44(1):  47-57 . 
    Abstract ( 773 )   PDF (265KB) ( 927 )   Save
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    【Objective】 Chromosome substitution lines between Chinese Spring and synthetic hexaploid wheat (CS-Synthetic 6x) were used to study the regulation effect of foreign added chromosome on chlorophyll fluorescence parameters under drought stress. 【Method】 The chlorophyll fluorescence parameters,including Fo, Fm, Fv/Fm and Fv/Fo in flag leaves at different developing stages in different water treatments were measured during 2005-2007. 【Result】 The results showed that Fm and Fo of 5B substitution line were significantly different from Chinese Spring in the control treatment. Drought stress resulted in inhibition of PSⅡ photochemical activity and reduction of Fm, Fv/Fm, Fv/Fo as well as accumulation of Fo; Fo of 3A substitution line and Fm of 4D substitution line were significantly different from Chinese Spring; Fv/Fm and Fv/Fo of 3A, 7A substitution lines were significantly different from Chinese Spring from booting stage to filling stage under drought stress. 【Conclusion】 The genes regulating Fo and Fm may be located on 5B chromosome of Synthetic 6x in the control treatment. In drought treatment the genes regulating Fo and Fm may be located on 3A and 4D chromosome, respectively, and the genes regulating Fv/Fm and Fv/Fo may be located on 3A and 7A chromosomes, the problem is up for further study.

    PLANT PROTECTION
    Development of Ralstonia solanacearum Quantification Method and Its Application in Peanut with Bacterial Wilt Disease
    HUANG Jia-quan,YAN Li-ying,YE Xiao-wen,LEI Yong,LIAO Bo-shou
    Scientia Agricultura Sinica. 2011, 44(1):  58-66 . 
    Abstract ( 835 )   PDF (477KB) ( 975 )   Save
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    【Objective】The aim of this study was to build rapid and accurate detection and quantification of R. solanacearum and study the interaction between bacterial and its host. 【Method】 In this study, real time quantitative PCR (RTQ-PCR) targeting the hrpB was developed for the quantification of bacteria load and the method was used to analyze the bacterial population dynamics in peanut after R. solanacearum inoculation.【Result】The results demonstrated that both the extracted DNA and whole cell of R. solanacearum could be used as template for bacterial quantification. Without enrichment step, the detection limit of the method using whole cell as template was 103 CFU/mL, the liner range was in 103-108 CFU/mL, the established RTQ-PCR method could be used for R. solanacearum quantification but the calculated number was about 1.5-fold compared with the traditional plating method. The application of the method in peanut suggested that 3-5 d post inoculation is of vital importance for peanut and R. solanacearum interaction and the node site in peanut may play a role in restriction of bacterial proliferation.【Conclusion】 RTQ-PCR is a simple, rapid and high throughput method developed for R. solanacearum quantification.

    Purification and Antifungal Mechanism of a Kind of Antifungal Protein from Strain A of Bacillus subtilis
    PENG Bing,ZHANG Shu-bin,JIA Yu,WANG Li-an
    Scientia Agricultura Sinica. 2011, 44(1):  67-74 . 
    Abstract ( 803 )   PDF (690KB) ( 1347 )   Save
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    【Objective】The objective of this study is to separate and purify a kind of antifungal protein from Bacillus subtilis strain A(CGMCC No.1982), and study the antifungal mechanism using Bipolaris maydis as a indicator. 【Method】 The crude protein was subjected to precipitation with ammonium sulfate and then a DE52 anion exchange chromatography column was employed for further purification of the protein with antifungal properties. The active fraction was identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF-MS). The mechanism of antifungal protein on the conidium germination and appressorium formation, on the mycelium morphology and intracellular structure of B. maydis was studied. 【Result】 The analysis of the antifungal protein (FV) by MALDI-TOF-MS suggested that it might be a kind of RNA-binding protein Hfq gi|16078797, with a relative molecular mass of 8 508.5 Da and isoelectric point (pI) of 8.04. The similarity of amino acid sequence between the antifungal protein from the strain A and a kind of RNA-binding protein Hfq was 86.484%. The IC50 for inhibiting spore germination was 138.32 μg?mL-1, while the IC50 for inhibiting appressorium formation was 22.23 μg?mL-1. The mycelia treated by FV with a concentration of 34.58 μg?mL-1 exhibited misshapen and existing intumescentia vesicles in top or middle. Under scanning electron microscope, the surface of the deformed mycelia became rough and the swollen vesicle appearance exhibited sags and crests and had some chips. Under transmission electron microscope, the protoplasm of the treated mycelia cell became vacuolization and in some mycelia existing daughter mycelia. 【Conclusion】 The amino acid sequence between the antifungal protein(FV) and a kind of RNA-binded protein Hfq was of higher homology. This protein could inhibit mycelial growth by changing the morphological structure of mycelial cells and affect conidium germination and appressorium formation.

    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Soil Labile Organic Carbon as Affected by Non-flooded Rice Cultivation with Straw Mulching Under Different Tillages
    WANG Dong,LI Hui-xin,LI Xiao-hong,WANG Jing,HU Feng
    Scientia Agricultura Sinica. 2011, 44(1):  75-83 . 
    Abstract ( 937 )   PDF (327KB) ( 829 )   Save
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    【Objective】 The objective of this study was to investigate the characteristics of soil total organic carbon, labile organic carbon and carbon pool management index under non-flooded rice cultivation with straw mulching, and no-tillage and non-flooded rice cultivation with straw mulching. 【Method】 A field experiment was conducted since 2003 in double-rice cropping system in the Rice Yuanzhong Farm in Yujiang County, Jiangxi Province in southeast China. Soil samples at 0-15 cm depth were collected at the late-season rice harvest period in 2007, and contents of soil total organic carbon and different fractions of labile organic carbon under different rice cultivations were determined. The rice cultivations included conventional flooded rice cultivation (CF), non-flooded rice cultivation with zero straw mulching (ZM), non-flooded rice cultivation with straw mulching (SM), no-tillage and non-flooded rice cultivation with zero straw mulching (N-ZM), and no-tillage and non-flooded rice cultivation with straw mulching (N-SM). 【Result】 Compared with CF, SM and N-SM could significantly increase soil total organic carbon content, particulate organic carbon (53-2 000 μm) content, hot water-extractable organic carbon content, readily oxidizable organic carbon (oxidized by 333 mmol?L-1 KMnO4) content, and the percentage of labile organic carbon to soil total organic carbon, and soil organic carbon pool management index (CPMI). However, ZM could significantly decrease the parameters mentioned above compared to CF.【Conclusion】 As a novel water-saving technique and straw management, SM and N-SM applied to late-season rice cropping system might be an alternative to CF for improving and increasing soil fertility under double-rice cropping system, in the seasonal drought region of southeast China.

    Establishing Fertilization Recommendation Index of Paddy Soil Based on the “3414” Field Experiments in the Middle of Sichuan Hilly Regions
    JI Lin,ZHANG Xi-zhou,LI Ting-xuan
    Scientia Agricultura Sinica. 2011, 44(1):  84-92 . 
    Abstract ( 859 )   PDF (257KB) ( 912 )   Save
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    【Objective】 In order to provide reference for rational application of fertilizer, decreasing resource waste and increasing economic benefit to paddy soil in the middle of Sichuan hilly regions, gradation index system of soil nutrition and fertilization recommendation index were established. 【Method】 Data analysis were based on the “3414” field experiments of paddy soil in 2006. Taking plentiful-lack of soil nutrition as an index, soil available nitrogen (AN), available phosphorus (AP) and available potassium (AK) were classified by relative yield of 75%, 80%, 85%, 90% and 95%. Fertilizer recommended rates were simulated by the models of three-factor, one-factor, liner and platform for each “3414” field experiment. 【Result】 Nitrogen fertilizer rates (N) are 170-220, 140-170, 110-140, 90-110, and 0-90 kg?hm-2 when soil fertilizer is classified by low, lower, medium, higher and the highest (AN<100, 100-140, 140-180, 180-220 and >220 mg?kg-1 ), respectively, phosphorus fertilizer rates (P2O5) are 100-120, 80-100, 60-80, 40-60, 0-40 kg?hm-2 (AP<5, 5-10, 10-20, 20-30 and >30 mg?kg-1), while potassium fertilizer rates (K2O) are 140-170, 120-140, 100-120, 80-100, 0-80 kg?hm-2 (AK<30, 30-50, 50-70, 70-90 and >90 mg?kg-1). 【Conclusion】Compared with the fertilization recommendation rate calculated by original system, the fertilization recommendation rate of this research suggested that nitrogen fertilizer rates must be decreased, but phosphorus and potassium fertilizer rates must be increased, especially of K2O. It objectively reflected the abundance or deficiency of nutrients and the characteristics of fertilizers input in this study area.

    Effects of Agricultural Management Practices on Carbon Emissions in Paddy Fields

    CAO Cou-gui,LI Cheng-fang,ZHAN Ming,WANG Jin-ping
    Scientia Agricultura Sinica. 2011, 44(1):  93-98 . 
    Abstract ( 660 )   PDF (204KB) ( 764 )   Save
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    Paddy soil is one of the most important carbon storages, which is affected by agricultural management practices. The achievements obtained in past researches were reviewed and the effects of agricultural management practices on carbon emissions, including the stocking of duck in paddy field, irrigation, N fertilization and tillage, were discussed. The results indicated that raising ducks in paddy field significantly decreased greenhouse effects, intermittent irrigation reduced CH4 emissions, N fertilization did not affect carbon emissions, and no-tillage had a significant decrease in carbon emissions. However, due to the complexity of soil carbon emission process, the heterogeneity of paddy environment and the human activity, it is inaccurate to assess the effects of agricultural management practices on carbon emissions, and thus it is necessary to further discuss the spatio-temporal changes in carbon emissions under different agricultural management practices for accurate assessment of the effects of carbon emissions on global warming.

    HORTICULTURE
    Analysis of Genetic Diversity and Relationship of Tea Cultivars and Lines Suitable for Making Green and Black Tea Using SSR Markers
    DUAN Yun-shang,JIANG Yan-hua,WANG Li-yuan,CHENG Hao,WANG Yu-hua,LI Xing-hui
    Scientia Agricultura Sinica. 2011, 44(1):  99-109 . 
    Abstract ( 895 )   PDF (478KB) ( 690 )   Save
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    【Objective】 The genetic diversity and relationship of tea cultivars and strains suitable for manufacturing green and black tea were studied in order to learn the genetic effect on the manufacture suitability of tea plant and to provide some molecular information to support the effective tea breeding. 【Method】 The genetic diversity and cluster analysis of 157 cultivars and strains of tea plant were analyzed using 39 pairs of SSR markers. 【Result】 Totally 191 alleles and 440 genotypes were amplified by 39 SSR markers, with the average of 4.90 and 11.28. The average polymorphism information content (PIC) , genetic diversity (H) and genetic distance were 0.57, 0.53 and 0.65, respectively. Genetic diversity of tested tea cultivars and stains from Guangdong was higher than that from other provinces. Evaluated samples were classified by their manufacture suitability and the genetic diversity of black tea cultivars was higher than that of double-edged cultivars for both black tea and green tea and that of green tea cultivars. The level of genetic diversity among cultivars bred by individual selection was higher than that of cultivars bred by other methods. Almost all the tested cultivars and strains were clustered according to their geographic source and breeding source using UPGMA method rather than manufacture suitability. 【Conclusion】 The genetic diversity of 157 tested tea cultivars was relatively abundant and the most abundant genetic diversity was identified among cultivars suitable for making black tea. In addition, the samples were clustered based on geographic and breeding source.

    Changes of Aroma Components in Oncidium Sharry Baby in Different Florescence and Flower Parts
    ZHANG Ying,LI Xin-lei,WANG Yan,TIAN Min,FAN Miao-hua
    Scientia Agricultura Sinica. 2011, 44(1):  110-117 . 
    Abstract ( 824 )   PDF (283KB) ( 1407 )   Save
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    【Objective】 The aim of this experiment was to analyze the changes of aroma components in Oncidium Sharry Baby in different florescence and flower parts, and provide reference for finding out the formation and release mechanism of Oncidium.【Method】 Aroma components in different florescence and flower parts were analyzed by solid phase micro-extraction and GC/MS technology. Volatile changes were also studied. 【Result】 There were 7 components identified at bud stage, 22 at first flowering stage, 24 at flowering stage and 41 at declining period. With the flower blooming and senescence, the contents of alkenes, alcohol and esters raised, and the contents of aldehydes and ketones, and alkanes decreased. There were 97 components identified in gynandrium, 71 in leans petal and 89 in labellum. In gynandrium and leans petal, the relative content of alcohol compounds was the highest, 25.36% and 58.16%, respectively. In labellum, the relative content of alkenes compounds was 49.17%, which was the highest.【Conclusion】 On the basis of above organoleptic evaluation and GC/MS results, alkenes is the most important compounds for Onc. Sharry Baby volatile. 3,7-dimethyl-1,3,6-Octatriene, (E)-2-Butenoic acid-2-(methylenecyclopropyl)prop-2-yl ester, 3,7-dimethyl-1,6- Octadien-3-ol and (Z)-3,7-dimethyl-2,6-Octadien-1-ol are the characteristic constituents of aroma. Labellum is probably the most important part that could influence the volatile releasing.

    STORAGE·FRESH-KEEPING·PROCESSING
    The Role of Lipoxygenase in Banana Fruit Ripening
    HE Quan-guang,KUANG Jian-fei,CHEN Jian-ye,LU Wang-jin
    Scientia Agricultura Sinica. 2011, 44(1):  118-124 . 
    Abstract ( 959 )   PDF (477KB) ( 780 )   Save
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    【Objective】 The role of lipoxygenase (LOX) in banana fruit ripening was studied. 【Method】 Fruit ripened naturally and fruit treated with 1 000 μL?L-1 propylene for 10 h and 48 h after harvest were used as experimental materials. LOX activity and the expression patterns of MaLOX, MaACS, MaACO genes and ethylene production and its relation to fruit ripening were analyzed. 【Result】 The results showed that the peak of LOX activity and transcript accumulation firstly decreased, then increased, moreover, the increase in LOX activity and transcript accumulation appeared earlier than that of mRNA accumulations of MaACS and MaACO gene in the peel of propylene treated fruit at 48 h after harvest. Although propylene could induce the accumulations of MaACS and MaACO gene in the fruit of propylene treated fruit at 10h after harvest, but could not increase LOX activity and transcript accumulation, and thus could not accelerate banana fruit ripening.【Conclusion】These results suggest that LOX, ACS and ACO are likely to play a role in regulating the climacteric ethylene production and banana fruit ripening coordinately.

    Changes of Cell Ultrastructure and Some Physiological Indexes and Physical Properties of Tobacco Leaves During Bulk Flue-Curing
    WU Sheng-jiang,SONG Zhao-peng,HE Fan,SUN Jian-feng,GONG Chang-rong
    Scientia Agricultura Sinica. 2011, 44(1):  125-132 . 
    Abstract ( 742 )   PDF (1223KB) ( 981 )   Save
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    【Objective】 In order to provide a theoretical basis for optimization and perfection of curing technology, changes of cell ultrastructure and some physiological indexes and physical properties of tobacco leaves were studied during bulk-curing.【Method】 Changes of cell ultrastructure, some physiological indexes and physical properties were studied at key temperature points (fresh, 38℃, 42℃, 48℃, 54℃, dry sample) during flue-curing using the electric-heated flue-curing barn designed and made by Henan Agricultural University. 【Result】 The contents of the starch, chlorophyll and carotenoids of tobacco leaves changed greatly in the early yellowing stage. The differences of moisture content, thickness, density of tobacco leaves in the yellowing and leaf-drying stages were highly significant. The contents of the pectin and cellulose changed mainly in the leaf-drying stage, but leaf specific weight had a relatively small change during flue-curing. Cell ultrastructure observation indicated that cell wall, chloroplast and nucleus changed obviously, but mitochondrias were not easy to be found during flue-curing. The cell structure of the fresh leaves was normal, but chromatin decreased and part of the nuclear membrane disappeared at 38℃. Cells showed plasmolysis, chloroplast and other cellular organs freed to the central cell, chloroplast outer membrane and nuclear membrane fractured, most osmiophilic granules presented half-hollow state, grana lamella and most starch grains disappeared at 42℃. The obvious twist distortion of cell wall and nucleus pyknosis appeared at 48℃; Individual shapes of chloroplast were not apt to differentiate and the remains were basically osmiophilic materials of the weak electronic density after 54℃.【Conclusion】The changes of the cell ultrastructure of tobacco leaves were closely related to physical properties and physiological and biochemical reactions and extent of degradation and transformation of inclusion during flue-curing.

    ANIMAL SCIENCE·RESOURCE INSECT
    Effects of Dietary Lysine to Methionine Ratio on Growth Performance, Nutrient Digestibility, and Metabolism in Weaned Calves
    YUN Qiang,DIAO Qi-yu,TU Yan,ZHANG Nai-feng,WANG Jian-hong,ZHOU Meng
    Scientia Agricultura Sinica. 2011, 44(1):  133-142 . 
    Abstract ( 756 )   PDF (249KB) ( 796 )   Save
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    【Objective】 This trial was designed to study the effects of dietary lysine to methionine ratio on growth performance, nutrient digestibility and nitrogen metabolism in weaned calves. 【Method】 Twenty four weaned calves were randomly allocated into four treatments either fed a control diet containing 14.76%DM CP or one of the three trial diets containing 12.02% DM CP and the dietary lysine/ methionine ratios were 2.5﹕1, 3.0﹕1 or 3.5﹕1, respectively, for 8 weeks. The calves were weighed and blood samples were collected on the initial and final day of the experiment. Four calves were selected from each group and metabolism trials were conducted from the third week of the feeding experiment by total feces collection method. 【Result】 The results indicated that the gain of calves in group 3.0﹕1 was higher than that of calves in other groups numerically (P>0.05). Serum concentrations of urea nitrogen in calves fed test diets was lower than that in the control (P<0.01). The concentrations of free amino acid in serum were lower (except methionine) in calves fed test diets than those in control group (P>0.05). No difference was observed in serum concentration of urea nitrogen and free amino acids (except arginine and valine) among test groups. The utilization and apparent biological value of nitrogen in group 3.0﹕1 and group 3.5﹕1 were higher than group 2.5﹕1 and control group numerically (P>0.05). 【Conclusion】 The gain of calves fed diet addition of RPLys and RPMet with a crude protein level of 12.02% was higher than that fed diet with a crude protein level of 14.67%. The ratio of lysine and methionine at 3.0﹕1 in diets could enhance growth performance and utilization of nitrogen in weaned calves.

    Tracing of the Origin and the Spreading Track of Silkworm White Muscardine in Southwestern Anhui
    LI Jia-li,LUAN Feng-gang,LI Zeng-zhi
    Scientia Agricultura Sinica. 2011, 44(1):  143-152 . 
    Abstract ( 703 )   PDF (579KB) ( 811 )   Save
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    【Objective】To understand and define the pathogen origin and spreading track of white muscardine caused by entomopathogenic fungus Beauveria bassiana in southwestern Anhui and to control prevalence of silk disease.【Method】Seventy isolates of B. bassiana from silkworm cadavers, rearing room and soil from surrounding mulberry garden, cropland, and adjacent pine plantations were tested for clustering and 3-d principal coordinate analyses with the ISSR data.【Result】The local B. bassiana population is heterogenous, with obvious dominance. Among the 6 groups, only group II-1 (the primary disease origin) and group IV (the secondary origin) caused enzootic muscardine, while both of them were neither related to the isolate used as fungal insecticide applied against the masson pine caterpillar nor to the indigenous isolates prevailing in pine caterpillar population. A bioassay revealed that LC25, LD25 and LT25 caused by the strain isolated from infected pine caterpillars were 1 327, 1 378, and 1.5 times of LC75, LD75 and LT75 caused by a typical pathogenic strain isolated from silkworm, respectively. 【Conclusion】 Based on obvious host specificity of the isolates from the pine caterpillars, the use of B. bassiana insecticide developed from these isolates against the pine caterpillars is comparatively safe to sericulture. The epizootic caused by B. bassiana in surrounding mantid populations is not associated to the white muscardine of silkworms.

    VETERINARY SCIENCE
    Detection and Analysis of Low Pathogenic Avian Influenza in Poultry in Eastern China from 2002 to 2009
    ZHAO Guo,LIU Xiao-wen,QIAN Zhong-ming,XUE Feng,PENG Yi,PENG Da-xin,LIU Xiu-fan
    Scientia Agricultura Sinica. 2011, 44(1):  153-159 . 
    Abstract ( 930 )   PDF (316KB) ( 627 )   Save
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    【Objective】 This study engaged in epidemiological surveillance of low pathogenic avian influenza A viruses in poultry in Eastern China. 【Method】 From July 2002 to September 2009, 11 645 cloacal swab samples were collected from poultry in the LPMs (live–poultry markets), which were from different provinces. All cloacal swabs were tested for hemagglutinating viruses by inoculating embryonated hens eggs. 【Result】 There were 1 158 strains of AIVs were isolated with an isolation rate of 9.94%. These strains of AIV belonged to eight HA subtypes, the isolation rate of high to low were H3, H6, H11, H1, H4, H9, H10, and H8. 【Conclusion】The isolation rate of AIVs was distributed with significant seasonal variations, higher in winter and spring and lower in summer. The distribution of HA subtypes of AIVs was also uneven in different months. So far, eight HA subtypes (H1, H3, H4, H6, H8, H9, H10 and H11) and seven NA subtypes (N1, N2, N3, N4 , N5, N6 and N8) were isolated from domestic ducks. Strains in HA and NA subtypes combination included the following 17 subtypes. Domestic duck is still the most prone to mixed infections, and since 2007 H6 subtypes have played an important role that are able to mix infect poultry with H3, H11, H4, H9 subtype etc, providing a good vector to generate new subtypes and variation of virulence.

    Biofilm Formation and Analysis of Associated Genes Involved in Staphylococcus Isolates from Bovine Mastitis
    LI Li,YANG Hong-jun,LIU Dai-cheng,HE Hong-bin,WANG Chang-fa,ZHONG Ji-feng,GAO Yun-dong
    Scientia Agricultura Sinica. 2011, 44(1):  160-166 . 
    Abstract ( 728 )   PDF (362KB) ( 837 )   Save
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    【Objective】The objective of the study was to investigate biofilm forming ability, distribution of biofilm-associated genes of the clinically isolated bovine mastitis Staphylococcus and the correlation of them. 【Method】Staphylococcus biofilm formation was conducted using a silicone elastomer slices plate assay in 24-well plates, after rinsing planktonic bacteria away, the biofilm forming abilities of them were determined by silver staining qualitatively and by crystal violet staining quantitatively, and the structure of biofilm was observed by using scanning electron microscopy. bap, icaAD, icaBC, sar, agr, sigB, clfaA, clfaB, fnbpA and fnbpB were amplified by PCR.【Result】 Formation of biofilm could be found macroscopically in 120 out of 137 strains by silver staining, and the biofilm formation rate was 87.6%. It showed that 5 strains didn’t adhere to the surface of silica gel by crystal violet staining, while the rest 132 isolates did. bap was amplified in 57 isolates and icaAD and icaBC in 43 and 54 strains, respectively. sigB, sar and agr were amplified in 73, 49 and 38 isolates, and clfaA and clfaB in 76 and 50 strains, respectively. fnbpA was present in 52 strains and fnbpB in 26 isolates. 【Conclusion】 It reveals that bap, sigB, sar, icaAD and icaBC may be significant biofilm-associated genes, for these genes are present more in biofilm-positive strains than in biofilm-negative strains. The roles of agr, clfaA, clfaB, fnbpA, and fnbpB genes in biofilm development are unassured.
    16SrRNA Methylase Resistant Gene in Enterobacter cloacae from Pig Farms

    CHEN Lin,LIU Jian-hua,ZHANG Jun-feng,CHEN Zhang-liu,ZENG Zhen-ling,WEI Dong-xia,HUANG Dong-zhang

    Scientia Agricultura Sinica. 2011, 44(1):  167-175 . 
    Abstract ( 747 )   PDF (393KB) ( 684 )   Save
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    【Objective】 To analyze the plasmid-mediated 16SrRNA methylase genes and their horizontal transferable mode of Enterobacter cloacae GZL42B which was isolated from pig farm. 【Method】 Polymerase chain reaction (PCR) and sequencing were applied to detect and identify the 16SrRNA methylase resistant gene. Conjugation experiments were used to study the horizontal transferable mode of 16SrRNA methylase in E. cloacae by using the E.coli Rif+488(rifampin-resistant) as the recipient. MICs were evaluated by using the micro-dilution method. Comparison of the neighboring sequences of rmtB between E. cloacae GZL42B and Escherichia coli GZL42B which isolated from the same rectal swab of one pig and the prevalence of 16SrRNA methylases genes in 172 E. coli were investigated by PCR and sequencing. 【Result】 The fragment of rmtB was amplified succeessfully. The restriction fragments were corresponded with the expectant, and the rmtB was confirmed by using sequencing. The accession number of the rmtB sequence submitted to GenBank was EF017943. Transconjugant was obtained successfully by using the E. coli Rif+488 as the recipient. The MICs of the parental E. cloacae GZL42B and its transconjugant to kanamycin, amikacin, tobramycin, sisomicin, netilmicin and gentamycin were ≥256 μg?mL-1. The right-hand end of transposon Tn3 and the orf1 which was part of groEL/intI1 on genomic island SG1 of Salmonella enterica serovar Typhimurium were located on the upstream and downstream of rmtB in Escherichia coli GZL42H, respectively. But they were not amplified in E. cloacae GZL42B. Tweenty-five and one of all 172 isolates (15% and 0.6%) were rmtB and armA positive, respectively. 【Conclusion】 It was the first report that the 16SrRNA methylase rmtB emergenced in E. cloacae from pig farm. High resistance to 4,6- deoxystreptamine was mediated by rmtB and can be transferred to E. coli Rif+488. The rmtB can be transferred between different species of bacteria. The genetic basis for dissemination of rmtB was different between E. coli GZL42H and E. cloacae GZL42B. And the rmtB was highly prevalent in animal E. coli.

    AGRICULTURAL ECONOMY & MANAGMENT
    Study on the Influencing Factors of Grain Production in Hebei Province Based on Gray Correlation Degree Method
    WANG Shu-tao,LI Xin-wang,MEN Ming-xin,XU Hao
    Scientia Agricultura Sinica. 2011, 44(1):  176-184 . 
    Abstract ( 758 )   PDF (378KB) ( 777 )   Save
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    【Objective】 The changes of food production in Hebei Province and the causes of fluctuations in grain yield were studied, in order to guide the production of food security in Hebei Province and provide a scientific and theoretical method. 【Method】 EMD and residual method were used comprehensively to separate food production and its influencing factors of Hebei, and a gray correlation method based on the results compared with paired comparison to determine matrix was used to study the causes of food production fluctuation. 【Result】 The characteristics influencing the fluctuations of grain production in Hebei Province are that natural resources and the amplitude of various index is between -113.43-34.38; the amplitude of various indicators of policy and economy capacity is -34.59-31.79, which has large fluctuation in intensity; the amplitude of investment of fixed assets is between -17.17-10.52, which has less intensity fluctuations. Index of variable input is between -11.39-9.16. Disaster area, the effective irrigated area, crop acreage, support agricultural production and agricultural business costs, precipitation, chemical fertilizer, and agriculture, forestry, animal husbandry and fishery workers are 6 controlling factors. The degrees of impacts were as follows: 19.08%, 17.8%, 12.84%, 11.03%, 10.28% and 7.79%, to represent the main elements of fluctuations in farmland productivity. 【Conclusion】 Results showed that the integrated use of EMD method and gray correlation method based on the results compared with paired comparison to determine matrix method, has overcome the shortcomings of the traditional method which can be used for qualitative analysis. It is a new scientific theory and method of objectively reflecting the real fluctuations of food production in Hebei Province, which can be used to quantitatively study the degree of impacting factors.

    RESEARCH NOTES
    Study on Main Na+ Exclusion Localization and Na+/H+ Antiport Activity of Buckwheat
    MA De-yuan,ZHAN Wei-yan,YANG Hong-bing,YI Yan-jun
    Scientia Agricultura Sinica. 2011, 44(1):  185-191 . 
    Abstract ( 732 )   PDF (248KB) ( 625 )   Save
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    【Objective】The objective of this experiment is to determine the main Na+ exclusion localization of buckwheat (Fagopyrum esculentum Moench) and the correlation between proton pump and Na+/H+ antiport activity with Na+ exclusion property. 【Method】Buckwheat varieties Chuanqiao 1 (salt-tolerant variety) and TQ-0808 (salt-sensitive variety) were used as experimental materials. Na+ exclusion capability of different parts was determined through “pressure chamber” method extruding xylem juice. Proton pump and Na+/H+ antiport activity were also determined by sucrose density gradient centrifugation method separating plasmolemma and tonoplast. 【Result】The main Na+ exclusion localization of salt-sensitive variety was in roots, while that of salt-tolerant one was in roots and stem base, and the whole Na+ exclusion capability of salt-tolerant variety was obviously higher than that of salt-sensitive one. Proton pump and Na+/H+ antiport activity of salt-tolerant variety were obviously higher than that of salt-sensitive one in the same salinity treatment. It indicated that Na+ extrusion capability and Na+ circumscription capability of the main Na+ exclusion localization cell in salt-tolerant variety were obviously higher than that in salt-sensitive one. 【Conclusion】Na+ circumscription of the main Na+ exclusion localization in salt-tolerant variety played an important role in restricting transportation Na+ to shoot, which was the main way to Na+ exclusion of main Na+ exclusion localization in salt-tolerant buckwheat variety.

    Effects of Shading Stress on Qualities of Purple Sweetpotato Storage Roots
    WANG Qing-mei,HOU Fu-yun,WANG Bao-qing,WANG Zhen-lin,DONG Shun-xu,ZHANG Hai-yan, LI Ai-xian,ZHANG Li-ming,XIE Bei-tao
    Scientia Agricultura Sinica. 2011, 44(1):  192-200 . 
    Abstract ( 744 )   PDF (302KB) ( 663 )   Save
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    【Objective】 The study is to elucidate the effects of shading stress on root qualities of different genotypes purple-freshed sweetpotato. 【Method】 Using Jishu18 and Ayamuraski as materials, the plants were shaded with nets which had 40%, 70% transmittance of natural light at the root expanding peak period (50-100 d after transplantion), and the natural light was used as the control. Dry matter, starch content, amylose and amylopection content, starch accumulation ratio, soluble sugar content, protein content, the parameters of viscosity and anthocyanidin content of tuber root were determined. 【Result】 Results indicated that dry matter, total starch content and starch accumulation rate in the root of purple-freshed sweetpotato decreased significantly compared with the control under shading treatments. Shading treatments not only significantly increased amylose content and the ratio between amylose and amylopectin, but also significantly decreased peak viscosity, break down, set back and pasting temperature of Jishu18. However, the above-mentioned starch qualities of Ayamuraski were against to those of Jishu18. Soluble sugar and soluble protein increased significantly accompaned by rising of shading degree in two cultivars. In addition, anthocyanidin content and accumulation rate declined significantly under shading treatments, compared with Ayamuraski, the response to stress of Jishu18 was more sensitive. Researches showed that shading treatments changed root qualities of purple-freshed sweetpotao and displayed difference between varieties. 【Conclusion】 Shading treatments can significantly decrease dry matter, starch content and starch accumulation ratio, anthocyanidin content and anthocyanidin accumulation in tuber root of purple-freshed sweetpotato, while the quality of root starch varied in the different sweetpotato varities under shading treatments.

    Effects of Water Stress on Respiration and Other Physiological Metabolisms of Cerasus sachalinensis Kom. Seedlings
    QIN Si-jun,Lü De-guo,LI Zhi-xia,MA Huai-yu,LIU Ling-zhi,LIU Guo-cheng
    Scientia Agricultura Sinica. 2011, 44(1):  201-209 . 
    Abstract ( 679 )   PDF (441KB) ( 706 )   Save
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    【Objective】 The mechanisms of Cerasus sachalinensis Kom. seedlings responding to water stress were studied in order to provide a academic and practical basis for evaluating the resource characteristics.【Method】 Changes of root respiratory pathways, related enzyme activities, respiratory substrates and chlorophyll fluorescence parameters of leaves were determined using continual drought and waterlogging treatments. Different mechanisms of C. sachalinensis Kom. seedlings responding to water stress in different extents were compared and analyzed. 【Result】 Seedling growth and respiratory metabolism were both badly influenced by continuous drought and waterlogging stresses. Root activities were badly restrained after 1-day drought, but rapidly depressed after 3-day waterlogging. The roots respiratory rates under drought stress were firstly increased then rapidly dropped, and the peak value appeared at 5 d. However, the respiratory rates hardly changed at the first five days of waterlogging and then decreased. The root biochemical respiratory pathways and electron transfer pathways were not changed significantly under 1-day water stress, and then the percentages of TCA and CP dropped sharply, PPP and AP became the leading way. After 7-day water stress, the proportions of each pathway were all decreased greatly. Root respiratory metabolisms were badly checked. The key enzyme activities of TCA pathway such as SDH were reduced whereas the key enzyme activities of PPP pathway such as G-6-PDH were induced under drought and waterlogging stress in some extent. Tolerance to drought and waterlogging of seedlings stretched to the limit after 7 days of treatment. Each key enzyme activity and the regulation were greatly declined. The respiratory substrate contents such as soluble sucrose and starch and the respiratory intermediate products pyruvic acid were firstly increased and then rapidly dropped during the water stress. However, the citric acid contents were continually decreased. Moreover, continual drought and waterlogging stress reduced the Fv/Fm value in leaves and the dry matter accumulations of plants and obviously restrained the photosynthesis.【Conclusion】At the beginning period of water stress, C. sachalinensis Kom. seedlings could adapt ill environments through regulating the key enzyme activities such as SDH and changing the proportions of respiratory pathways, which adjusted the consumptions of respiratory substrates and formations of respiratory intermediate products. Seedlings under waterlogging had a lingering responses than drought. The physiological metabolisms of seedlings such as root respiration and leaf photosynthesis were all greatly influenced by continuous water stress.
    Cloning, Expression and Polymorphism Analysis of Pig ERK2 Gene for Reproduction Traits
    FANG Mei-xia,ZHANG Wei,HU Yong-sheng,OUYANG Hong-jia,JIA Xin-zheng,NIE Qing-hua,ZHANG Xi-quan
    Scientia Agricultura Sinica. 2011, 44(1):  210-217 . 
    Abstract ( 656 )   PDF (414KB) ( 609 )   Save
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    【Objective】 Extracellular signal-regulated kinase-2 (ERK2) gene plays important roles in regulating cell proliferation and differentiation, as well as starting the ovulation of ovary. It is an important candidate gene for porcine reproduction traits. In this study, the pig ERK2 gene was cloned and its genomic organization, tissue expression patterns, and gene polymorphism were investigated.【Method】 In this study, the pig ERK2 gene was cloned by RT-PCR and its mRNA distribution in different tissues was studied by real time PCR, moreover, the ERK2 gene structure and polymorphisms were analyzed. 【Result】 The cloned pig ERK2 cDNA was 1 888 bp long, and contained an open reading frame of 1 080 bp, which encoded a peptide of 359 amino acid residues. The encoded ERK2 was highly similar with predicted pig ERK2 and the human and mouse counterpart. The ERK2 gene was widely expressed in many porcine tissues. The highest expression was found in spleen, however almost no expression in adipose tissue, muscle tissues from front and back legs. The pig ERK2 gene was located at the 14th chromosome, and composed of 9 exons and 8 introns. Variation detection of exon 2-9 and intron-exon boundary sequences indicated that a total of 11 SNPs and 1 indel were identified, most of which were in introns and only one was in 3’ UTR. 【Conclusion】The obtained cDNA of pig ERK2 gene was 1 888 bp long and encoded a 359-amino acid peptide. The ERK2 gene was found to express in many tissues of pig with highest level in spleen. The ERK2 gene comprised 9 exons and 8 introns and showed high conservation. A total of 11 SNP and 1 indel were found, but none of them was in coding regions.

    Derivation and Karyotype Analysis of Porcine Haploid Parthenogenetic Embryos

    ZHANG Man-ling,ZHAO Li-hua,ZHOU Xin,LI Rong-feng
    Scientia Agricultura Sinica. 2011, 44(1):  218-224 . 
    Abstract ( 629 )   PDF (382KB) ( 923 )   Save
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    【Objective】 In order to obtain high efficient method to produce porcine haploid parthenogenetic embyos, different physical and chemical methods were applied to activate MII porcine oocytes. 【Method】 In experiment 1, electrical activation, a combination of electrical activation and cytochalasin B (CB),and ethanol activation at different concentrations and durations were used to activate porcine oocytes. In experiment 2, electrical activation followed by MG-132 or Thimerosal/DTT treatments were investigated for their effects on porcine oocytes activation. 【Result】 The results of experiment 1 indicated that the development rates of blastocysts in CB group(27.34%)were significantly higher than that of electrical activation alone (16.92%) (P<0.05), and both of them were significantly higher than that of all ethanol groups (P<0.05). In all ethanol groups, 9% ethanol treatment for 11 min got the highest blastocyst rate (10.20%) . The results of experiment 2 showed that the rate of blastocysts formation in MG-132 group (24.69%) was not significantly different with that of CB group (27.50%), but they were significantly higher than that of Thimerosal/DTT (14.10%) and electrical activation (17.5%) groups (P<0.05). The ploidy analysis of blastocysts derived from different groups indicated that the percentages of haploid blastocysts in electrical activation, 9% ethanol and Thimerosal /DTT group were 41.7%, 40%, and 35.3% respectively, they were significantly higher than that of CB (0) and MG-132 (6.7%) groups (P<0.05)【Conclusion】 Considering both blastocysts formation rate (out of oocytes) and haploid rate (out of total blastocysts), the final haploid blastocyst percentages (out of oocytes) in electrical activation, 9% ethanol, Thimerosal/DTT and MG-132 groups were 7.3% (17.5%×41.7%), 4.1% (10.2%×40%), 5.0% (14.1%×35.3%), and 1.7% (24.69%×6.7%), respectively. The electrical activation might be the best activation way to obtain haploid embryos.