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Table of Content

    15 October 2010, Volume 43 Issue 20
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Identification and Fine Mapping of a Glume-Like Lodicule Mutant in Rice (Oryza sativa L.)
    YU Xin,WANG Lin-you,ZHANG Li-xia,FAN Hong-huan,ZHU Chao-na,WANG Xi,JIN Qing-sheng,WANG Jian-jun
    Scientia Agricultura Sinica. 2010, 43(20):  4123-4129 .  doi:10.3864/j.issn.0578-1752.2010.20.001
    Abstract ( 781 )   PDF (412KB) ( 752 )   Save
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    【Objective】 The identification and cloning of novel mutant genes of floral organ in rice would play an important role in understanding the molecular genetic mechanisms and molecular signal pathways regulating floral organ development. 【Method】Field identification, comparison between mutant and wild-type floral organs, statistics of segregation of different phenotypes as well as gene mapping based on map-based cloning were used to describe the characteristics of floral organ, to determinate the inheritance of gll mutant and high-resolution mapping of GLL gene. 【Result】 It was shown that a wide variety of spikelet developmental defects exhibited from outer to inner floral organs in gll mutant, and the main mutant phenotypes were glume-like lodicule and the extra glume. The Chi-square test of F2 and F3 individuals derived from four crosses indicated that the segregation ratio of phenotypes was consistent with the inheritance of single recessive nuclear gene. The plants with gll phenotype were obtained from F2 or F3 generations from Nipponbare×gll for mapping GLL gene. One hundred and fifty-six polymorphic molecular markers, which evenly distributed on 12 chromosomes in rice, were used to test 408 individules with mutant phenotype. According to statistical analysis, the GLL gene was located on rice chromosome 1 between RM1068 and RM3482 with a genetic distance of 4.6 and 2.3 cM to gll locus, respectively. Then, four SSR markers were developed within this region, gll locus was finally mapped on a region of 108 kb physical distance. 【Conclusion】 The gll mutant is controlled by a recessive nuclear gene, which is located on the long arm of chromosome1, between RM6097 and RM6827 with 108 kb physical distance.

    Construction of Genetic Linkage Map and QTL Analysis of Agronomic Important Traits Based on a RIL Population Derived from Common Wheat Variety Yanzhan 1 and Zaosui 30
    YAO Qin,ZHOU Rong-hua,PAN Yu-ming,FU Ti-hua,JIA Ji-zeng
    Scientia Agricultura Sinica. 2010, 43(20):  4130-4139 .  doi:10.3864/j.issn.0578-1752.2010.20.002
    Abstract ( 948 )   PDF (698KB) ( 844 )   Save
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    【Objective】 The present study was aimed to develop a RIL (recombinant inbred line) population, construct its genetic map and analyze quantitative trait loci (QTL) associated with important agronomic traits in wheat. This work will accelerate gene discovery and marker-assistant breeding of wheat. 【Method】 A RIL population was derived from the cross of Yanzhan1 and Zaosui 30 by single seed descent approach. SSR (simple sequence repeat), DarT (diversity arrays technology), ISBP (insertion site-based polymorphism) and functional makers associated with days to heading (DH) and plant height (PH) were used to construct genetic map. QTLs related to DH, PH, thousand grain weight (TGW), grain number per spike (GNS), spike length (SL) and spikelet numbers (SPI) in different environments were detected by using the composite interval mapping (CIM). 【Result】 A set of 219 F7 RILs was derived. A genetic linkage map with 481 molecular markers covering the whole wheat genome was constructed. A total of 26 QTLs for the related traits located on 12 chromosomes were detected, among which 9 QTLs were detected in at least two environments. Three QTL clusters were revealed including one cluster for PH and TGW located on dwarf gene Rht2 region on 4DS, one cluster for DH, GNS and SPI located in region Vrn-D1-WMS212 of chromosome 5D and the other cluster for DH, SPI, PH, and SL located in the interval of wPt4230 and wPt4814 on chromosome 7B. 【Conclusion】 The genetic map is favorable to analyze important agronomic traits. Dwarfing gene Rht2 and vernalization gene Vrn-D1, two developmental related genes, are related to some important agronomic traits. There might be a novel gene concerning plant development on chromosome 7B.

    Genetic Analysis of Major Locus Group Constitutions of Growth Stages in Upland Cotton

    AI Ni-jiang,ZHU Xin-xia,GUAN Rong-zhan,ZHAO Jian-jun,ZHANG Tian-zhen
    Scientia Agricultura Sinica. 2010, 43(20):  4140-4148 .  doi:10.3864/j.issn.0578-1752.2010.20.003
    Abstract ( 718 )   PDF (227KB) ( 504 )   Save
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    【Objective】 Early maturity is fundamental to produce high and stable yield in upland cotton. It is of important practical value to dissect the genetic components and analyze the genotypes of beneficial and unbeneficial loci in crosses using early varieties as parents. 【Method】 Six varieties of different origins were used in an incomplete diallel cross design to evaluate the genetic components in parents and the major locus groups related to earliness in F1s by QTL testing system. 【Result】 The genotypes of major locus groups for growth stages were mainly ++ in the early parents Xinluzao 8 and 10, which have the genetic background of 611-b and King’s cotton, respectively. Each growth stage was significantly shorted in these genotypes. The inheritance of earliness related traits was mainly additive. The growth period had the largest additive value, followed by bloom, seedling and square stages. The additive effects of ++ and -- locus group were equally important in the mid-maturity parent Jing 8891. However, almost all ++ and – locus group were unbeneficial in the late parents Ningzao 1, Sumian 3 and TM-1, with each growth stage delayed. Among all major locus groups in all growth stages, the J=16 locus group had the largest additive values, indicating it can significantly reduce the growth period. The performance of earliness related traits in F1s was affected by both heterozygous and homozygous locus groups, but the growth period was primarily determined by the heterogeneity of the major locus groups. 【Conclusion】 Additive effects of major locus groups for earliness related traits were invariably higher than dominance. The additive effects of major locus groups for square and bloom stages played a predominant role in earliness.

    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    Effects of Nitrogen Fertilizer Top-Dressing Based on Canopy Reflectance Spectra in Rice
    CHEN Qing-chun,TIAN Yong-chao,YAO Xia,ZHU Yan,CAO Wei-xing
    Scientia Agricultura Sinica. 2010, 43(20):  4149-4157 .  doi:10.3864/j.issn.0578-1752.2010.20.004
    Abstract ( 764 )   PDF (335KB) ( 667 )   Save
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    【Objective】 The objective of the study is to estimate nitrogen (N) status of rice plant and recommend top-dressing N amount based on canopy reflectance spectra, so as to achieve higher yield, better grain quality and higher economic efficiency. 【Method】 The amounts of nitrogen fertilizer recommended for top-dressing of different treatments with varied basal N rates were calculated with the newly developed N top-dressing model based on plant N accumulation, which was estimated from differential vegetation index (DVI). Then the N top-dressing regulation effect was studied. 【Result】 The results showed that plants of different treatments with varied basal N rates had different N accumulations at the elongation stage. The treatments with high, medium and low basal N rates needed low, relative higher and high N fertilizer supplies, respectively. After N top-dressing, the differences of plant N concentration (PNC) and DVI between the regulation treatments and their CKs were induced gradually. And leaf photosynthetic rates (Pn) of the regulation treatments were significantly increased compared with their CKs. And higher economic benefits were also achieved in regulation treatments than its CKs. Compared with the normal high yield N treatment, the regulation treatments obtained higher Pn, dry matter accumulation, plant N accumulation, grain yield and nitrogen agronomic efficiency.【Conclusion】 Compared with the normal methods, the real-time top-dressing regulation technique based on reflectance spectra can quantify varied N top-dressing amounts induced by different plant and soil N status, so it is recommended for N top-dressing in rice.

    The Harvest Index Model of Winter Wheat in China Based on Meteorological Data
    JI Xing-jie,YU Yong-qiang,ZHANG Wen,YU Wei-dong
    Scientia Agricultura Sinica. 2010, 43(20):  4158-4168 .  doi:10.3864/j.issn.0578-1752.2010.20.005
    Abstract ( 802 )   PDF (355KB) ( 739 )   Save
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    【Objective】 The harvest index model of winter wheat in China based on meteorological data was established for supporting crop yield model. 【Method】 Based on observed data of winter wheat and meteorological data during recent twenty years from thirty agrometeorological stations in Henan, Hebei and Shandong provinces, the trend and meteorological harvest index were extracted by using time-series analysis method. The trend harvest index was separated from the measured harvest index by linear fitting; the meteorological harvest index was then calculated by subtracting the trend harvest index from the measured one, and the relationship between the meteorological harvest index and the meteorological factors was studied by stepwise regression analysis which simulated the influence of the changed meteorological factors on winter harvest index. 【Result】Correlation analysis suggested that the meteorological harvest index of winter wheat significantly correlated with the meteorological factors during the growth period. However, the influencing factors were different in different scales. Both statistical models of single station scale and regional scale were validated, respectively, using other data (n=209). The model validation showed that, the simulated linear regression as against measured harvest index yielded a correlation coefficient (r) of 0.65 with a slope of 0.42, the root mean squared error of 12.2%, the mean bias error of -2.4%, the index of agreement of 75.8% and the model efficiency of 42.3% on the single station scale, and the model almost explained the total variance. The simulated regression against measured harvest index yielded a correlation coefficient (r) of 0.56 with a slope of 0.33, the root mean squared error of 13.3%, the mean bias error of -1.3%, the index of agreement of 69.0% and the model efficiency of 31.7% on regional scale. 【Conclusion】 The established models can well simulate the change of winter wheat harvest index under different meteorological conditions. The winter wheat harvest index model coupled with crop net primary production model can be used to do research on simulating winter wheat yield on the regional scale.

    Relationship Between Fiber Macroelement Content and Fiber Quality in Colored Cotton
    YUAN Shu-na,HUA Shui-jin,NI Mi,LI Yue-you,WEN Guo-ji,SHAO Ming-yan,ZHANG Hai-ping,ZHU Shui-jin,WANG Xue-de
    Scientia Agricultura Sinica. 2010, 43(20):  4169-4175 .  doi:10.3864/j.issn.0578-1752.2010.20.006
    Abstract ( 793 )   PDF (310KB) ( 665 )   Save
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    【Objective】 Colored fiber cotton (Gossypium hirsutum L.) is eco-friendly for the textile industry because the dying process can be omitted and thus reducing the harmful effluent liquor. This experiment was carried out to give more information on the poor fiber quality of colored fiber cotton. 【Method】 Three cotton varieties, brown fiber cotton (X008), green fiber cotton (S029), and white fiber cotton (Xuzhou 142), were used to study the content of macroelement, cellulose, and flavonoid during the development of cotton fiber. 【Result】 Result showed that the cellulose content was 15% lower and flavonoid content was 4-10 times higher in colored cotton fiber than in white fiber cotton; macroelement in colored cotton fiber was 1-2 times higher than in white fiber cotton. Fiber quality properties had a significant positive correlation with fiber cellulose content, but had a significant negative correlation with flavonoid content. N, P and Ca contents had a significant negative correlation with cellulose content and a positive association with flavonoid content, respectively. 【Conclusion】 The results suggested that the higher content of N, P and Ca elements in colored cotton fiber might be necessary for the formation of flavonoid in it, and the higher flavonoid accumulation and lower cellulose content in fiber might be responsible for the inferior fiber quality of colored cotton when compared with the white fiber cotton.

    PLANT PROTECTION
    Involvement of Phospholipase C in Induction of Cell Death and Defense Responses in ParA1-Elicited Tobacco Suspension Cells
    WEI Fang-fang,WANG Lei,WANG Lian-lian,LIU Jin-wei,LIANG Yuan-cun,WANG Yu-jun
    Scientia Agricultura Sinica. 2010, 43(20):  4176-4182 .  doi:10.3864/j.issn.0578-1752.2010.20.007
    Abstract ( 718 )   PDF (449KB) ( 649 )   Save
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    【Objective】 Phospholipase C (PLC) was investigated in response to ParA1, a protein elicitor from Phytophthora parasitica, and could trigger cell death and defense responses in tobacco suspension cells. 【Method】 Using a pharmacological approach, tobacco suspension cells pretreated with PLC inhibitors (neomycin and U73122) in ParA1-induced cells were used to compare cell death and a series of defense responses. 【Result】 Tobacco suspension cells treated with ParA1 resulted in defense responses, such as hypersensitive cell death, an oxidative burst, alkalization of the extracellular medium, expression of 5 defense-related genes and accumulation of scopoletin, defense-related genes hin1, hsr203J, PR-1a, PR-1b and PAL were expressed in ParA1-induced cells. Cell death induced by ParA1 was entirely inhibited by the addition of U73122, a PLC specific inhibitor. Oxidative burst, alkalization of the extracellular medium, expression of 5 defense-related genes and scopoletin accumulation was significantly suppressed by U73122, respectively. 【Conclusion】 PLC is required for ParA1-mediated cell death and defense response in tobacco suspension cells which may involve in signal transduction.

    Potential of Prochloraz-Manganese Chloride in Controlling Sclerotinia Stem Rot of Oilseed Rape
    REN Li,CHEN Kun-rong,WANG Cheng-yu,LUO Li-xia,JIA Jian-guang,WANG Jing,FANG Xiao-ping
    Scientia Agricultura Sinica. 2010, 43(20):  4183-4191 .  doi:10.3864/j.issn.0578-1752.2010.20.008
    Abstract ( 1091 )   PDF (323KB) ( 773 )   Save
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    【Objective】 Sclerotinia stem rot (SSR) caused by Sclerotinia sclerotiorum is a damaging disease of oilseed rape. The sensitivity of S. sclerotiorum to prochloraz-manganese chloride (PMC) and the other three fungicides, carbendazim, dimetachlone-mancozeb and carbendazim-triadimefon was tested. The aim of this study was to assess the potential of PMC in controlling SSR and to screen novel fungicides against Sclerotinia disease. 【Method】 The effects of fungicides on S. sclerotiorum hyphal growth and sclerotial germination, and their protective and therapeutic effects were determined in laboratory. Potential of fungicides in controlling SSR was tested in field. 【Result】 The pathogen was highly sensitive to PMC with the median effective concentration (EC50) to S. sclerotiorum hyphal growth at 0.02 mg?L-1, but less sensitive to carbendazim, carbendazim-triadimefon and dimetachlone-mancozeb. The results showed that myceliogenic germination of S. sclerotiorum sclerotia was delayed by 20 mg?L-1 PMC, and carpogenic germination was inhibited by PMC at 0.5 g?L-1 with the inhibition percentage at 82.9%. PMC had both protective and therapeutic effects on Sclerotinia disease. Field efficacy of PMC on S. sclerotiorum was as high as 88.1%. 【Conclusion】Prochloraz-manganese chloride is a promising fungicide in controlling Sclerotinia disease of oilseed rape.

    Effect of Imazethapyr on Nodule Nitrogenase and Nitrate Reductase Activity in Soybean
    DING Wei,CAO Li-juan,CHENG Zhuo
    Scientia Agricultura Sinica. 2010, 43(20):  4192-4197 .  doi:10.3864/j.issn.0578-1752.2010.20.009
    Abstract ( 822 )   PDF (266KB) ( 596 )   Save
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    【Objective】 To determine the effects of imazethapyr on nitrogenase and nitrate reductase activity in soybean and give evidence to assess the influence on nitrogen metabolism correctly after herbicide application. 【Method】 A field experiment was conducted to study the effects of imazethapyr on nitrogenase and nitrate reductase activity (NRA) after imazethapyr preemergence and postemergence applied. 【Result】 Nitrogenase was significantly reduced by imazethapyr in both treatments of preemergence and postemergence application. Although the reduction of nitrogenase activity was increased with time prolonging after imazethapyr preemergence, the nitrogenase activity was still reduced by 0.2186, 0.2198 and 0.2471 mmolC2H4?g-1?h-1 when imazethapyr was applied at 62.5, 125 and 250 ga.i.?hm-2 75 days after treatment. The reduction of nitrogenase activity was gradually decreased as time goes on after application of imazethapyr postemergence and 38 days after treatment, the nitrogenase activity was reduced by 0.1778 and 0.2177 mmolC2H4?g-1?h-1 at the application rates of 62.5 and 125 ga.i.?hm-2. Root NRA was significantly decreased after application of imazethapyr preemergence and postemergence and the reduced NRA gradually increased over time regardless imazethapyr application rate and application methods. Leaf NRA was strongly reduced within 30 days after application of imazethapyr postemergence. Leaf NRA was significantly reduced only at the application rate of 250 ga.i.?hm-2 75 days after preemergence application. 【Conclusion】 Nodule nitrogenase, root and leaf NRA were significantly reduced after application of imazethapyr preemergence and they have significant effect on inhibition of nitrogen metabolism of soybean.

    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Effect of Long-Term Fertilization on Soil Biological Characteristics in a Paddy Soil Derived from Yellow-Brown Earth
    PEI Xue-xia,ZHOU Wei,LIANG Guo-qing,SUN Jing-wen,WANG Xiu-bin,AI Chao,LI Shuang-lai
    Scientia Agricultura Sinica. 2010, 43(20):  4198-4206 .  doi:10.3864/j.issn.0578-1752.2010.20.010
    Abstract ( 795 )   PDF (434KB) ( 819 )   Save
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    【Objective】 Soil biological fertility reflects soil quality directly. Understanding of soil biological characteristics and microbial community as affected by long-term fertilization could provide a theoretical basis for improvement of soil quality and development of the sustainable agriculture. 【Method】 In this paper, the effects of long-term application of N, NP, NPK, pig manure (M), and MNPK (pig manure plus NPK) on total phospholipids fatty acid (PLFA), soil micro-biomass-C, N and P, and selected hydrolase activities were investigated in a paddy soil derived from yellow-brown earth. 【Result】 Compared to CK or application of chemical fertilizer only, the application of pig manure only or pig manure plus NPK significantly increased SMB-C, N, P, PLFA, fungi PLFA, fungi/bacterial rate, soil alk-phosphatase, protase, and urease activity, fungal-to-bacterial PLFA ratios, and enhanced the growth of AM fungi. Long-term application of N only inhibited soil alk-phosphatase activity and the growth of most microorganisms. Principal Component Analysis (PCA) of PLFA showed that, microbial community structure was affected by manure and chemical fertilizers application in different ways. The loading values for the individual PLFA indicated that fungal and monounsaturated fatty acids (MONO) were mainly influenced by organic manure application, while those of actinomycete was affected by application of chemical fertilization only. 【Conclusion】 Combined application of organic manure and chemical fertilizer could significantly enhance SMB-C, N, P, PLFA, fungi PLFA, fungi/bacterial rate, and improve soil biological fertility. It will be favorable to soil quality.

    Effects of Short-Term No Tillage on Soil Organic Carbon Mineralization Using Least Limiting Water Range
    SHI Xiu-huan,ZHANG Xiao-ping,LIANG Ai-zhen,YANG Xue-ming,SHEN Yan,FAN Ru-qin,CHEN Xue-wen
    Scientia Agricultura Sinica. 2010, 43(20):  4207-4213 .  doi:10.3864/j.issn.0578-1752.2010.20.011
    Abstract ( 759 )   PDF (311KB) ( 853 )   Save
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    【Objective】 Effects of no-tillage (NT), mouldboard plow (MP) and ridge tillage (RT) on the content of soil organic carbon (SOC) in the plow layer (30 cm) of a black soil in Northeast China were studied. 【Method】 The tillage experiment trial was made in Dehui County, Jilin Province in 2001. The effects of the three tillage managements on SOC mineralization were compared using an indicator, least limiting water range (LLWR) under different tillage treatments. 【Result】 NT significantly increased SOC at the top soil (0-5 cm) compared with MP, but SOC content showed a notable reduction at 5-30 cm depth. SOC stratification ratio under NT was higher than that under MP and RT. The values of LLWR under different treatments throughout the study years ranged from 0.129 to 0.170, which were lower under NT than MP, and they went to decrease with the tillage operation. 【Conclusion】 The residue covered on soil surface every year led to a clear effect on SOC accumulation after 7-year field trial. The stratification ratio of SOC under NT was higher than that under MP and RT. Investigating LLWR was helpful to understand SOC fixation related to NT.

    Effect of Fertilization on Forage Yield and Soil Enzymes Under Sudangrass and Ryegrass Rotation Regime
    LI Wen-xi,LI Xiao-kun,LU Jian-wei,ZHAN Li-ping,LU Jun-ming
    Scientia Agricultura Sinica. 2010, 43(20):  4214-4220 .  doi:10.3864/j.issn.0578-1752.2010.20.012
    Abstract ( 976 )   PDF (300KB) ( 643 )   Save
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    【Objective】 The sudangrass and ryegrass rotation is a new type of cropping system in the south of China, and it develops very fast in recent years. Guidelines for proper fertilization are needed for improvement of soil fertility in this rotation. 【Method】 The effects of NPK fertilizers on forage yield, sucrase, urease and phosphatase in soil under sudangrass and ryegrass rotation regime were studied based on the four-year fixed location field experiments. 【Result】 The results indicated that the NPK combination significantly increased the fresh yield of sudangrass and ryegrass. The total yield in NPK treatment was the highest among all treatments from 2005 to 2009, and was 17.9%, 17.3%, 21.7%, and 23.2% higher than that of NP treatment, 26.9%, 20.2%, 23.3%, and 25.9% higher than that of NK treatment, and 312.9%, 339.8%, 231.7%, and 223.1% higher than that of PK treatment, respectively. In the rotation, the activities of soil sucrase and urease in NPK treatment were higher than that in CK and PK treatments, while were similar to that of NP and NK treatments. For soil phosphatase, alkaline phosphatase activity was higher than acid phosphatase activity in the rotation, and NP, NK and NPK treatments were lower than CK and PK treatments. 【Conclusion】 The combination of N, P and K fertilizers could increase yield of forage, influence the activities of sucrase, urease and phosphatase in soil and improve soil fertility in the sudangrass and ryegrass rotation system.

    Effect of CO2 Enrichment on Ca and Mg Concentration in Soil Solution at Arable Layer in the Rice Season
    WANG Xiao-zhi,YIN Wei-qin,SUN Wei,SHENG Hai-jun,FENG Ke,ZHU Jian-guo
    Scientia Agricultura Sinica. 2010, 43(20):  4221-4228 .  doi:10.3864/j.issn.0578-1752.2010.20.013
    Abstract ( 692 )   PDF (412KB) ( 688 )   Save
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    【Objective】 The objective of this study was to understand the response of soil Ca and Mg geochemical recycle in rice paddy ecosystem to elevated atmospheric CO2. 【Method】 Field experiment was carried out at Experiment Station of FACE (free-air carbon dioxide enrichment) in China. There were two treatments, including ambient (the CO2 concentration was approximately 370 μmol?mol-1) and FACE (the target CO2 concentration was 200 μmol?mol-1 higher than that of ambient). Soil solution samples were collected in situ at different stages during the 2004, 2005 and 2007 rice growing seasons. Concentrations of Ca and Mg in soil solution were determined. 【Result】 The concentrations of Ca in soil solution at 5 cm depth in FACE treatment were 25%, 6% and -28% higher than those in ambient treatment during the 2004, 2005 and 2007 rice growing seasons, respectively. The ratios of 15cm/5cm of Ca and Mg in soil solution from FACE treatment were -29%, 14% and 80% and -26%, 4% and 59% higher than those from ambient treatment during the 2004, 2005 and 2007 rice growing seasons, respectively. The concentrations of Ca and Mg in soil solution at 0-15 cm depth in FACE treatment were less than ambient treatment by 6.8% and 4.6% in the year of 2007, respectively. 【Conclusion】 Elevated atmospheric CO2 influenced the distributions of Ca and Mg in soil solution at different depths in arable layer and increased the ratios of 15 cm/5 cm of Ca and Mg in soil solution. This indicates that elevated atmospheric CO2 has a potential impact of soil Ca, Mg in the biogeochemical cycles, and may affect rice yield and quality.

    HORTICULTURE
    Characteristics of Components and Contents of Soluble Sugars in Pear Fruits from Different Species
    YAO Gai-fang,ZHANG Shao-ling,CAO Yu-fen,LIU Jun ,WU Jun,YUAN Jiang,ZHANG Hu-ping,XIAO Chang-cheng
    Scientia Agricultura Sinica. 2010, 43(20):  4229-4237 .  doi:10.3864/j.issn.0578-1752.2010.20.014
    Abstract ( 941 )   PDF (430KB) ( 1597 )   Save
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    【Objective】 The characteristics of components and contents of soluble sugars in 98 cultivars of pear fruit were determined to reveal the differences and characteristics among different cultivars and species. 【Method】 The high performance liquid chromatography (HPLC) was used to detect the components and content of soluble sugar in mature pear fruits, and the data were analyzed by principal component analysis(PCs) and cluster analysis. 【Result】(1) The major components of soluble sugars in mature pear fruits were fructose, glucose, sucrose and sorbitol, and the fructose content was the highest value, with a mean of 51.17 mg?g-1FW. The contents of fructose and glucose were relatively stable among different cultivars, while the contents of sucrose and sorbitol varied greatly, the value ranged from 1.14-47.75 mg?g-1FW and 4.46-47.29 mg?g-1FW, respectively. (2) Among different species, the content of fructose was also the highest one, which accounted for 42.22%-57.02% of the value of total sugars, while the other three compoents of soluble sugars were highly different. The content of glucose was close to sorbitol in both P. bretschnrideri Rehd. and P. sinkiangensis Yü., and sucrose content was the lowest; the contents of fructose and sucrose were equal in both P. communis Linn. and P. ussuriensis Msxim. , however, the sorbitol content was high in P. communis Linn., and P. ussuriensis Msxim. in reverse; the content of sucrose was equal to sorbitol in P. pyrifolia Burm Nakai., and the glucose content was the lowest. (3) The distributive characteristics of soluble sugars in different species: P. bretschnrideri Rehd. mainly distributed in the high glucose and sorbitol area; P. pyrifolia Burm Nakai. mainly distributed in the high sucrose and sorbitol area; P.communis Linn. mainly distributed in high fructose and sorbitol area; P. ussuriensis Msxim. were mainly distributed in the high glucose and sucrose area; P. sinkiangensis Yü. mainly distributed in the high fructose and glucose area. 【Conclusion】 The major components of soluble sugars in pear fruits were fructose, glucose, sucrose and sorbitol, and the content of fructose was the highest one. The content of fructose and glucose kept stable among difference cultivars, while the content of sucrose and sorbitol showed high variability. Comparing different species, the content of fructose kept the highest value, while the other three components differ each other greatly. Based on the distribution of soluble sugars, P. bretschnrideri Rehd. classified as high glucose and sorbitol type, P. pyrifolia Burm Nakai. classified as high sucrose and sorbitol type, P. communis Linn. classified as high fructose and sorbitol type, P. ussuriensis Msxim. classified as high glucose and sucrose type, P. sinkiangensis Yü classified as high fructose and glucose type.

    Research on Improvement of Seedling Rate in Embryo Rescue of Seedless Grapes

    WANG Ai-ling,WANG Yue-jin,TANG Dong-mei,ZHANG Jian-xia,ZHANG Chao-hong
    Scientia Agricultura Sinica. 2010, 43(20):  4238-4245 .  doi:10.3864/j.issn.0578-1752.2010.20.015
    Abstract ( 800 )   PDF (261KB) ( 755 )   Save
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    【Objective】The objective of the study was to improve the breeding efficiency of seedless grape through embryo rescue in vitro and investigate the effects of various factors on seedlings in vitro. 【Method】 Hybrid ovules of 7 cross combinations were used to investigate the effect of different genotypes, different ovule culture media, supplementation of different aminophenols, low-temperature treatment on the proportion of seedlings of harvested fruits in vitro. 【Result】 The results indicated that among the investigated embryoes of seedless grape, the female parent and male parent had influence on seedlings. In cross combinations with diploid seedless grapes as female parent, diploid seedless grapes as male parent was suitable for seedlings than tetraploid seeded grapes. Among the three different media, MM4 medium was most suitable for embryo development and seedlings in vitro culture of seedless grapes. ER medium supplemented with 4 mmol?L-1 proline was most suitable for embryo development in vitro culture of seedless grapes. The effect of low-temperature treatment of Ruby Seedless×Centennial Seedless and Ruby Seedless×Black Olympia for 3 days was the best in seedling growth. 【Conclusion】 ER medium supplemented with 4 mmol?L-1 proline or MM4 medium are suitable for embryo rescue in vitro culture of seedless grapes. Low-temperature treatment of fruits for 3 days, has a positive effect on improvement of embryo rescue efficiency of diploid seedless grape×diploid seedless grape.

    The Degradation Mechanism of Carotenoids in Flue-Cured Tobacco and the Changes of the Related Aroma Components in the Yellowing Stage During the Bulk Curing Process
    SONG Zhao-peng,WU Sheng-jiang,GAO Yuan,XU Zi-cheng,ZHANG Wei-jian,GONG Chang-rong
    Scientia Agricultura Sinica. 2010, 43(20):  4246-4254 .  doi:10.3864/j.issn.0578-1752.2010.20.016
    Abstract ( 1007 )   PDF (357KB) ( 868 )   Save
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    【Objective】 The changes of the contents of carotenoids components, the related enzymes activities, and the aroma components degraded by which in flue-cured tobacco in the yellowing stage during the bulk-curing process were studied in order to provide a theoretical basis for optimization and perfection of curing technology. 【Method】 The effects of different temperature and humidity conditions on the degradation of carotenoids, the related enzymes activities in the yellowing stage and the contents of aroma components in flue-cured tobacco during the bulk curing process were studied by using the electric-heated flue-curing barn designed and made by Henan Agricultural University. 【Result】 The results showed that the contents of carotenoids (β-carotene, lutein, neoxanthin, violaxthin) decreased gradually in the yellowing stage during the bulk curing process. At the end of yellowing stage the β-carotene contents of low temperature with low humidity (T1) and low temperature with middle humidity (T2) treatments were lower than others and the value of them were 17.14 μg?g-1 FW, 19.00 μg?g-1 FW, respectively. The contents of the lutein, neoxanthin, violaxthin decreased gradually in the yellowing stage following the decrease of temperature and humidity. The proportions of the β-carotene to the xanthophylls (lutein, neoxanthin, violaxthin) of the low temperature treatments at the end of yellowing stage were higher than fresh sample, on the contrary, high temperature treatments were lower. The proportions of the lutein to the xanthophylls of different treatments were higher than fresh sample, on the contrary, neoxanthin and violaxthin were lower. The carotenoids contents of T1, T2 treatments were relatively lower than others’, especially the T2 treatment, and its value was 76.31 μg?g-1 in flue-cured tobacco. The contents of aroma components of T2 treatment was the highest and its value was 87.5634 μg?g-1 in flue-cured tobacco while the content value of T1 treatment was 81.1192 μg?g-1, secondly. Both of them were higher than others obviously. The activities of lipoxygenase (LOX) and peroxidase (POD) could express fully under the T1, T2 treatments in the yellowing stage meanwhile the carotenoids degraded fully. Correlation analysis showed that the activities of peroxidase and the carotenoids components were highly significant in the yellowing stage during the bulk curing process. 【Conclusion】 If the high activities of lipoxygenase and peroxidase were prolonged longer appropriately at 36-48 h under low temperature and relatively low humidity in the yellowing stage and the effect between the cooperative and antagonism reach dynamic balance, it would be benefit to degrade carotenoids sufficiently and improve the aroma quality of tobacco leaves.

    Mutagenic Effects on Tissue Culture Plants of Ornamental Crabapple Induced by 60Co-γ Irradiation
    LIU Li-qiang,LIU Jun-li,ZHANG Jie,GENG Hui,ZHANG Yu-ping,SHEN Hong-xiang,GAO Xia-hong,YAO Yun-cong
    Scientia Agricultura Sinica. 2010, 43(20):  4255-4264 .  doi:10.3864/j.issn.0578-1752.2010.20.017
    Abstract ( 817 )   PDF (1066KB) ( 726 )   Save
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    【Objective】 This paper was aimed to discuss the mutagenic effects of the 60Co-γ irradiation treatment on tissue culture seedlings of ornamental crabapple. 【Method】 Seven 60Co-γ doses were selected to irradiate the tissue culture seedlings of crabapple ‘Indian Magic’ (Malus crabapple ‘Indian Magic’), and then the rooting rate, propagation ratio, death ratio of the tissue culture plants, and the variation of the growth status, leaf morphology and ultrastructure of transplanted seedlings were investigated. 【Result】 With the increment of the 60Co-γ irradiation dose, the rooting rate, average root length, seeding height, proliferation rate of the cultured plants in rooting and subculture medium were decreased significantly, in which there were significant differences in index between the treatments of <30 Gy and >30 Gy irradiation dose. As the irradiation dose were amplified, the death rate of the cultured plants rose significantly after irradiation, as a result of that, the median lethal dose(LD50) was defined as 45.5 Gy (r=0.936) by regression analysis. Under the condition of <LD50 irradiation treatments, the morphology of tissue culture plants appeared typical dwarfing characteristics, in which the leaves became smaller and thicker, the internodes of the shoot and root length became shorter and coarser, the apical growth vigor of the transplanted seedlings decreased, and the leaf color, margin, shape varied significantly compared with seedlings in control (CK). Through observation of the leaf ultrastructure, it was found that, with the increment of the irradiation dose, the length, width, perimeter and area of stoma apparatus in the leaves of transplanted seedlings increased, but the ratio of length/width, length, perimeter, width and area of the stoma decreased, and also the thickness of leaves, palisade tissue, spongy tissue as well as its ratio decreased (except 30 Gy). So it was inferred that the variation of the structure and apparent characteristics of leaves are adapted each other in tissue culture seedlings of ornamental crabapple with 60Co-γ irradiation treatments. 【Conclusion】 After radiation treatment at the dose of 30 Gy, the tissue culture seedlings can keep normal growth, and the variation properties appeare obviously, the mutagenic effects are stable. Thirty Gy is an appropriate irradiation dose.

    STORAGE·FRESH-KEEPING·PROCESSING
    Comparisons of Instruments in Measuring Dough Rheological Properties
    WEI Yi-min,ZHANG Bo,GUAN Er-qi,LI Wei-jin,YAN Jun-hui
    Scientia Agricultura Sinica. 2010, 43(20):  4265-4270 .  doi:10.3864/j.issn.0578-1752.2010.20.018
    Abstract ( 981 )   PDF (242KB) ( 962 )   Save
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    【Objective】 The aim of the paper was to investigate the accuracy and precision of Farinographs and Extensographs in measured dough rheological properties from different laboratories. 【Method】 Seventeen Farinographs and 11 Extensographs were tested. Accuracy and precision of Farinographs and Extensographs in measured dough rheological properties were analyzed using three types of wheat flour such as high gluten wheat flour, medium and high gluten wheat flour, and weak gluten wheat flour. 【Result】 The accuracy of 72% Farinographs and the precision of 73% Farinographs were satisfied. There was no significant difference in accuracy of Farinographs among either Farinograph parameters or three types of wheat flour samples. However, the precision of Farinographs among both Farinograph parameters and three types of wheat flour samples was significantly different. The accuracy of 73% Extensographs and the precision of 95% Extensographs were satisfied. Accuracy and precision of Extensographs among extensograph parameters or accuracy of Dxtensographs among the three types of wheat flour samples were insignificantly different. However, the precision of Extensographs among the three types of wheat flour samples was significantly different. 【Conclusion】 The results of comparison show that more than 25% Farinographs can not meet the needs of high quality wheat detection, monitoring or evaluation in national technical standards.

    Correlation Analysis of CIELab Parameters and Anthocyanidins of Different Red Wines
    TAO Yong-sheng,ZHANG Li
    Scientia Agricultura Sinica. 2010, 43(20):  4271-4277 .  doi:10.3864/j.issn.0578-1752.2010.20.019
    Abstract ( 871 )   PDF (333KB) ( 638 )   Save
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    【Objective】 The correlation analysis of color parameters and anthocyanidins was made to find some important information for distinguishing different red wines. 【Method】 Twelve red wines are different in years, cultivars and ageing method from Changli and Shacheng were sampled. Nine fractions of every wine were collected with C18 column, and then they were lyophilized. The residue of each fraction was resolved with synthetic wine of the same volume with the fraction sample. The transmittance spectra of wines and their fractions were measured and the CIELab parameters were calculated. The correlation analysis of transmittance spectra between wines and fractions showed that four fractions were significantly correlated with wines. The anthocyanidins in these four fractions were identified by HPLC. 【Result】 Principle component analysis (PCA) of the data indicated that aged wines were significantly different with young wines in the PCA plot. Young wines were scattered around the anthocyanidin compounds. Anthocyanidins were the main contributors of the colour of young wines, while they played little role in the color of aged wines. The parameter of CIELab a* was related positively with anthocyanidins, and L* and b* were negatively with them. PCA also showed that the four important fractions had their own typical anthocyanidin compounds. 【Conclusion】 It is suggested that the analysis of anthocyanidins from the color point could distinguish different red wines,such as vintage, variety and ageing method, especially for wines from different vintages.

    ANIMAL SCIENCE·RESOURCE INSECT
    Cloning, Tissue Expression Patterns, and Sequence Characters of Fatty Basic Acid-Binding Protein Gene in the Liver of Duck
    ZHONG Wan-fu,LI Jia-qi,YANG Cheng-zhong,WANG Chong,ZHANG Hao,ZHANG Ai-ling
    Scientia Agricultura Sinica. 2010, 43(20):  4278-4285 .  doi:10.3864/j.issn.0578-1752.2010.20.020
    Abstract ( 778 )   PDF (720KB) ( 846 )   Save
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    【Objective】 The cDNA sequence of duck liver basic fatty acid-binding protein(Lb-FABP) gene was cloned and the gene tissue patterns and protein structure were analyzed. 【Method】 Based on comparative genomics, the cDNA sequence of duck Lb-FABP gene was obtained by RT-PCR and RACE. The mRNA expressions of the gene in sixteen tissues were detected through semi-quantitative PCR. And the protein structure was analyzed by bioinformatics methods. 【Result】 The cDNA sequence of the gene contained the 5’ untranslated region (5’UTR, 97 bp), the 3’ untranslated region (3’UTR, 69 bp) and the CDS (381 bp, including the stop condon). The gene was composed of 4 exons and 3 introns, which were 991 bp, 292 bp, and 713 bp, respectively. The semi-quantitative RT-PCR results revealed that Lb-FABP mRNA expression was higher in the liver than that in the other fifteen tissues. The physical and chemical properties of duck Lb-FABP indicated that Lb-FABP is a basic protein. And the protein has no clear signal peptide and transmembrane segments. The secondary structure of duck Lb-FABP was mainly made of β sheets and few α helixes and loops. One cytosol fatty acid-binding protein domain was found on the amino acid region from 5 to 22. The speculated amino acids sequence of duck Lb-FABP shared 97% and above 80% homologies with chicken and other nonmanmalian vertebrate species. The results showed that Lb-FABP is not in mammalian species. 【Conclusion】 The full length of the cDNA and genomic sequences were obtained. And the Lb-FABP mRNA expression was the highest in duck liver. One cytosol fatty acid-binding protein domain was found on the amino acid region from 5 to 22.

    Study on Oxidative Stress-Activated MAPK Signaling Pathway in Broilers Breast Muscle Satellite Cells
    WANG Cheng,WU Shu-geng,ZHANG Hai-jun,YUE Hong-yuan,ZHAI Yong-gong,QI Guang-hai
    Scientia Agricultura Sinica. 2010, 43(20):  4286-4294 .  doi:10.3864/j.issn.0578-1752.2010.20.021
    Abstract ( 780 )   PDF (561KB) ( 1113 )   Save
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    【Objective】 Mitogen-activated protein kinases (MAPKs) signaling pathway was investigated to screen which one plays a key role in broiler skeletal muscle satellite cells (SCs) under oxidative stress induced by dexamethasone (DEX) in vitro. 【Method】 The broilers skeletal muscle SCs were assigned into one of the ten treatments: control (I), DEX (Ⅱ), inhibitor p38 MAPK (Ⅲ), DEX plus inhibitor p38 MAPK (Ⅳ), inhibitor JNK (Ⅴ), DEX plus inhibitor JNK (Ⅵ), inhibitor ERK5 (Ⅶ), DEX plus inhibitor ERK5 (Ⅷ), inhibitor ERK1/2 (Ⅸ), DEX plus inhibitor ERK1/2 (Ⅹ). The treatments exclude treatments I and Ⅱ were treated with inhibitors for 30 min before treating with DEX and normal medium for 24 h. The relative production of malonaldehyde (MDA), reactive oxidative species (ROS), the activities of superoxide dismutase (SOD) and glutathione S-transferase (GST) were measured, respectively. Polymerase chain reaction (PCR) was employed to assess SOD and GST gene expression.【Result】 Compared with DEX, the MDA and ROS concentrations were decreased (P<0.05) in all the inhibitors-treated groups. The concentration of MDA and ROS in treatment with inhibitors were higher than that in treatment with DEX (P<0.05) The MDA and ROS concentrations of treatment with DEX plus inhibitor ERK5 and of treatment with DEX plus inhibitor ERK 1/2 were significantly higher than that of treatment with inhibitor ERK5 and treatment with inhibitor ERK1/2 (P<0.05). However, there were no significant differences in MDA and ROS concentrations among treatment with DEX plus inhibitor p38 MAPK, treatment with DEX plus inhibitor JNK and treatment with inhibitor p38 MAPK, treatment with inhibitor JNK (P>0.05). The activities of SOD and GST of all the inhibitors-treated groups were significantly higher than those of DEX group (P<0.01). The activities of SOD and GST of treatment with DEX plus inhibitor ERK5 and treatment with DEX plus inhibitor ERK 1/2 were significantly lower than those of treatment with inhibitor ERK5 and treatment with inhibitor ERK1/2 (P<0.01), while there were no significant differences in treatment with DEX plus inhibitor p38 MAPK, treatment with DEX plus inhibitor JNK and treatment with inhibitor p38 MAPK, and treatment with inhibitor JNK (P>0.05). DEX could inhibit the expression of SOD and GST gene, the inhibition rate was more than 37%. On the contrary to ERK5 and ERK1/2 pathway, DEX treatment had no significant effects on the expression of SOD and GST gene while the p38 MAPK and JNK pathway were inhibited. 【Conclusion】 The p38 MAPK and JNK pathway play a key role in broiler breast muscle SCs under oxidative stress induced by DEX.

    Zinc Requirement of Yellow Broilers from Forty-Three to Sixty-Three Days of Age
    JIANG Zong-yong,LIU Xiao-yan,JIANG Shou-qun,ZHOU Gui-lian,LIN Ying-cai,CHEN Fang,MA Xian-yong
    Scientia Agricultura Sinica. 2010, 43(20):  4295-4302 .  doi:10.3864/j.issn.0578-1752.2010.20.022
    Abstract ( 724 )   PDF (278KB) ( 623 )   Save
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    【Objective】 An experiment was conducted to investigate dietary zinc level on growth performance, antioxidant capacity, immune function and zinc depositionin in Lingnan yellow broilers from 43 to 63 days of age, and to estimate the optimal dietary zinc level for yellow broilers. 【Method】 A total of 1080 43-day-old male broilers were randomly assigned to six dietary treatments with six replicates and thirty birds with each pen. The groups received the same basal diet supplemented with 0(control group), 20, 40, 60, 80 and 120 mg?kg-1 zinc from zinc sulfate (ZnSO4.H2O), respectively. The trial lasted for 21 days. 【Result】 Dietary zinc supplementation had no effect on the growth of broilers (P>0.05). Zinc supplementation increased the activities of GSH-Px, CuZnSOD and AKP in serum (P<0.05), and significantly raised GSH content in serum, zinc concentrations in serum and tibia, MT content in serum and liver (P<0.05), and addition with 60 mg?kg-1 zinc showed the best result for AKP activity in serum and zinc concentrations in tibia. Supplementation with 20 to 40 mg?kg-1 zinc significantly elevated thymus index, and addition of 20 to 80 mg?kg-1 zinc significantly improved the index of Bursa of Fabricius of broilers at 63d (P<0.05). Zinc addition had no effect on spleen index, CuZnSOD activity in liver and tibia ash percentage (P>0.05). 【Conclusion】 Only take consideration of growth performance of broilers, there’s no need to supplement maize-soybean meal diet with zinc. Based on the criteria including AKP activity in serum and zinc concentration in tibia, the optimal supplemental zinc level estimated by variance analysis and multiple comparisons for broiler chicks was 78 mg?kg-1 for 43-63-day-old broilers, but the optimal supplemental zinc levels estimated by NLIN were 81 and 60 mg?kg-1, respectively.

    VETERINARY SCIENCE

    Development of an Indirect ELISA Diagnostic Method for Detecting Bovine Respiratory Syncytial Virus Using Recombinant Nucleocapsid Protein and Its Preliminary Application

    WANG Hong,YU Li-yun,HOU Xi-lin,PIAO Fan-ze,ZHAI Yan-qing
    Scientia Agricultura Sinica. 2010, 43(20):  4303-4309 .  doi:10.3864/j.issn.0578-1752.2010.20.023
    Abstract ( 958 )   PDF (342KB) ( 587 )   Save
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    【Objective】 The ELISA plate was coated with the purified bovine respiratory syncytial virus (BRSV) HJ strain recombinant nucleocapsid (N) antigen and the optimal reaction conditions of indirect ELISA were determined by experiments. The indirect ELISA method for detecting BRSV antibody was established by determining the cut off value. 【Method】 The recombinant E.coli strain BL21 containing the recombinant plasmid pET-30a-N was induced by IPTG. The recombinant N protein was expressed at high level, then it was purified by electroelution method. The recombinant N protein was identified by western blot. Taking the purified recombinant N protein as diagnostic antigen, the indirect ELISA method for detecting BRSV antibody was established. 【Result】 The recombinant N protein was successfully expressed and purified, it was proved that the recombinant protein has good reactogenicity by western blot. Taking the purified recombinant N protein as diagnostic antigen, the indirect ELISA method for detecting BRSV antibody was established. The results showed that there was no cross reaction between the recombinant N protein and the positive serum of Anaplasmosis, IBRV, BPIV. Compared with the serum neutralization testing, the results indicated that the specificity, sensitivity and coincidence of the developed indirect ELISA was 85.0%, 90.9% and 87.7%, respectively. A total of 600 sera of cattle, which obtained from Mudanjiang, Jamusi, Hegang, Daqing in Heilongjiang province, were tested by the developed indirect ELISA method and the positive rate was 27.33%. 【Conclusion】 The developed indirect ELISA has good specificity, sensitivity and reproducibility, it has been proved that BRSV is existed in some areas in Heilongjing province. It is very important that the development of indirect ELISA and its preliminary application of BRSV in cattle for the preparation of ELISA kit, diagnosis and epidemiological investigation of bovine respiratory syncytial disease.

    Genotypes of TEM-Type and CTX-M-Type Extended-Spectrum β-Lactamases Produced by Escherichia coli Isolated from the Fowl
    YUAN Li,LIU Jian-hua,HU Gong-zheng,PAN Yu-shan,MO Juan,WEI Yong-jun,PEI Ya-ling
    Scientia Agricultura Sinica. 2010, 43(20):  4310-4316 .  doi:10.3864/j.issn.0578-1752.2010.20.024
    Abstract ( 846 )   PDF (373KB) ( 841 )   Save
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    【Objective】 The genotypes and subtypes of ESBLs produced by Escherichia coli isolated from the fowl were detected to elucidate their distributions in Henan Province. 【Method】 Twenty-eight ESBL-producing Escherichia coli were collected from different chicken farms of seven districts in Henan Province, and were amplified by PCR using the primers of TEM, SHV and CTX-M, respectively. Their PCR products were cloned and then the cloned fragments were sequenced. The nucleotide sequences were analyzed by DNAStar software and GenBank database. 【Result】 Twenty-one strains were TEM-type, two isolates were CTX-M-type, and two genotypes were detected in five strains, including TEM-type and CTX-M-type. No SHV-type was found. TEM-1 variant was the most common subtype, and the sequences of 26 isolates shared 98%-99% homology with that of AY293072 (TEM-1 β-lactamase). Except a silent mutation of 18T→C of all, the sequences of C3, X2, and F4 were characterized by another nucleotide base variations (783G→A, 21T→A, and 269G→A, respectively), and only the last variation led to the shift of amino acid (92Gly→Asp). These three sequences were submitted to Genbank database and obtained the accession No (FJ405207, FJ405208, and FJ405211, respectively). The CTX-M-type of ESBLs included two subtypes: three isolates (F2, X3, and B5) CTX-M-gene sequences were identical with EF158299 (CTX-M-14), the others (C2, C3, C4, and K1) CTX-M-gene belonged to CTX-M-65 type (FJ405191, FJ405192, FJ405212, and FJ405214, respectively). 【Conclusion】 The predominant genotype of ESBLs is TEM-1 variant in chicken farms of Henan Province, and then are CTX-M-65 and CTX-M-14.

    New Plasmid-Mediated Quinolone Resistance Gene qnrD Found in an Isolate of Escherichia coli from Pet Animals
    ZHU Heng-qian,LIAO Xiao-ping,SUN Jian,LI Liang,ZHANG Mei-jun,LIU Bao-tao,SUN Ying,ZHONG Guo-guang,LIU Ya-hong
    Scientia Agricultura Sinica. 2010, 43(20):  4317-4322 .  doi:10.3864/j.issn.0578-1752.2010.20.025
    Abstract ( 691 )   PDF (373KB) ( 968 )   Save
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    【Objective】 The objective of this study is to detect plasmid-mediated quinolone resistance qnrD gene in clinical isolates of Escherichia coli from pet animals, analyze the sequence of this gene and study the transmission mechanism of qnrD gene. 【Method】 Susceptibility of positive strain to 15 antimicrobial agents was determined by micro-dilution method; PCR method was used for cloning qnrD gene, clone products and positive plasmid were used for transmission tests, and conjugation experiments were made. 【Results】One (GP2009-036) of the 164 specimens was detected containing qnrD gene, GP2009-036 showed high rate of resistance to 14 antimicrobial agents widely used in veterinary clinical medicine and showed multiple-resistance (14 drugs). PCR products connected with PMD19-T vector could be transformed into DH5α competent cells. The positive plasmid were transmitted into Escherichia coli J53 successfully by conjugation experiments and the positive plasmid could be extracted from GP2009-036 and transconjugants. 【Conclusion】 This qnrD positive strain of Escherichia coli showed high rate of resistance to antimicrobial agents widely used in veterinary clinical medicine, and the positive plasmid can transmit among pathogenic microorganism, and the mechanism of transmission indicates that qnrD gene may spread in pet animal veterinary clinic.

    RESEARCH NOTES
    Cloning and Sequence Analysis of the Aquaporins Gene CaAQP in Pepper
    CHEN Ru-gang,ZHU Wen-chao,GONG Zhen-hui,LI Da-wei,YIN Yan-xu,LU Ming-hui
    Scientia Agricultura Sinica. 2010, 43(20):  4323-4329 .  doi:10.3864/j.issn.0578-1752.2010.20.026
    Abstract ( 1064 )   PDF (751KB) ( 778 )   Save
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    Abstract: 【Objective】 To provide an insight into mechanism of pepper’s cold resistance and cultivar development,the sequence characteristics of the aquaporin gene CaAQP in pepper were analyzed and the expression profiling of the chilling tolerant Capsicum annuum cv. P70 was studied after treatment at 4℃ cold stress. 【Method】 The rapid-amplification of cDNA ends (RACE) was used to amplify the full-length CaAQP gene, and the bioinformatics software was employed to analyze the structures and function of the coded protein. At the same time, the chilling tolerant pepper P70 treated at 4℃ was used as material and the real time-PCR was adoped to study the expression profile of gene CaAQP, the peper cv. treated at 25℃ was used as control. 【Result】 A gene encoding the aquaporin cDNA (designated as CaAQP, GenBank Accessin No. GU116569) was cloned from leaf of Capsicum annuum cv. P70 by RACE. The full-length CaAQP gene cDNA was 1 032 bp in length, which included a potential open reading frame of 753 bp, a 5′-untranslated region (5′-UTR) of 69 bp and a 3′-untranslated region (3′-UTR) of 210 bp, encoded a protein of 250 amino acids with a predicted molecular mass of 25.7 kD. Bioinformatics analysis demonstrated that CaAQP exhibited a typical structure with six membrane-spanning domains and an internal symmetry showing two highly conserved Asn-Pro-Ala (NPA) motifs, and possessing the MIP family signal consensus sequence. The CaAQP amino acids showed high identity with other 10 plant species TIP subfamily by NCBI homology comparison analysis. Phylogenetic analysis indicated that CaAQP was clustered with the SLAQP from Solanum lycopersicum. Real time-PCR analysis showed that the expression of CaAQP was down-regulated by low temperature stress. 【Conclusion】 An aquaporin gene CaAQP was firstly cloned by cDNA-AFLP and RACE from leaf of Capsicum annuum cv. P70. The CaAQP gene is a member of the MIP protein family, and has the typical function region. The expression profiling of this gene indicates that the CaAQP gene may play an important regulating role during low temperature stress. This study provides important information for the future research on the gene-expression regulation during cold stress.