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    10 May 2008, Volume 41 Issue 5
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    In search of Genetic Architecture of Complex Traits
    Scientia Agricultura Sinica. 2008, 41(5):  1255-1266 .  doi:10.3864/j.issn.0578-1752.2008.05.001
    Abstract ( 1385 )   PDF (342KB) ( 1805 )   Save
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    Complex traits are features whose properties are determined by multiple factors ,which can be genetic or environmental, and whose inheritance does not follow the simple rules of Mendelian genetics. Most human complex diseases and most economically important traits in plants and animals are complex traits. The molecular basis of complex traits is theoretical and applied important for genetic improvement in plant and animal breeding. We summarized the conception and research background of complex traits and reviewed the strategies identifying genes underlying complex traits. Based on these strategies, the great progress had been made in dissecting genetic basis of complex traits. At last, we also discussed the challenges and possible developments in future research.
    Screening Differential Profile of Genes Expressed during Compatible Interactions between Puccinia striiformis and Wheat
    Scientia Agricultura Sinica. 2008, 41(5):  1267-1273 .  doi:10.3864/j.issn.0578-1752.2008.05.002
    Abstract ( 1393 )   PDF (437KB) ( 1168 )   Save
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    【Objective】Isolation of differentially expressed genes induced by Puccinia striiformis is the first step to reveal the molecular mechanism of interaction between wheat and yellow rust fungus. 【Method】A SSH-cDNA library was constructed with wheat variety shuiyuan-11 and a Chinese virulent race CY31 of Puccinia striiformis, 250 positive clones were selected for sequencing.【Result】In total 149 nonredundant ESTs were assembled with CAP3, then function annotation and classification were performed. There were 50 unigenes (33.6%) without or with weak similarities to the existent proteins in the nr database from NCBI and 25 unigenes (16.8%) significantly matched with hypothetical protein. Whilst the unigenes assigned to primary metabolism, energy and disease/defence accounted for 8.7%, 6.7%, 4.0%, respectively. Only 2 unigenes, however, had higher similarity with pathogenic protein. Four genes were randomly selected to investigate their expression profiles by reverse transcription PCR (RT-PCR). 【Conclusion】Some genes which had higher similarity with catalase, elongation factor and cell death suppressor protein were supposed to have important roles during compatible interaction between Puccinia striiformis and wheat.
    A Study on Variability of Fat-related Traits in Cultivated and Wild Soybean Germplasm in China
    Scientia Agricultura Sinica. 2008, 41(5):  1283-1290 .  doi:10.3864/j.issn.0578-1752.2008.05.004
    Abstract ( 1342 )   PDF (316KB) ( 936 )   Save
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    Abstract 【Objective】The present paper was aimed to evaluate the variability of content of fat and major fatty acids (hereafter referred as fat-related traits) in cultivated and wild soybean germplasm in China and to screen for accessions superior in fat-related traits so as to provide materials and guidelines in the improvement of fat-related traits in various eco-regions. 【Methods】The cultivated and wild soybean germplasms from various eco-regions[10] were sampled, tested, and analyzed for their fat-related traits. Based on single trait analyses, multivariate index, principal component analysis and cluster analysis were carried out to investigate the comprehensive variability of all fat-related traits in cultivated and wild soybeans from various eco-regions. 【Results】The results were as follows: (1) The average fat content of cultivated soybean in whole country was 17.21%, 6.22% higher than that of wild soybean; that of oleic was 23.25%, 7.75% higher than the wild; that of linoleic acid was 53.53%, 2.57% lower than the wild; and that of linolenic acid was 8.00%, 4.23% less than the wild; however, the variability of the fat-related traits in cultivated soybean was smaller than those in wild soybean. A similar tendency was observed in various eco-regions. As for the saturated fatty acids, there showed no large difference between the two species in whole country as well as in various eco-regions. (2)The multivariate variability in wild soybean was larger than that in cultivated soybean in whole country as well as in various eco-regions. The long term artificial selection reduced the variability, but made a wider direction of variation in cultivated soybean. (3) There showed a significant positive correlation between fat content and source latitude of accessions in cultivated soybean, but not in wild soybean, indicating the correlation in cultivated soybean was mainly due to artificial selection for fat content in the history. (4) Elite accessions with high fat content, high oleic acid content, high linoleic acid, low linolenic acid were screened out, respectively. Among them, N23547 and N23697 were the best with high fat content more than 23%, oleic acid more than 30% and linolenic acid less than 5%.【Conclusions】 The average contents of fat and oleic acid of cultivated soybean were significantly higher than, the average content of linolenic acid was significantly lower than, and the average content of linoleic acid was a little lower than that of the wild soybean, while no large difference in saturated fatty acids between the two species in China. Large variation of fat-related traits existed within all the eco-regions in the two species, even larger than those among eco-regions. The significant correlation between fat content and source latitude was mainly due to the long term artificial selection for fat content in the history.
    Heredity Analysis and Gene Mapping of Bruchid Resistance of A Mungbean Cultivar V2709
    Lei SUN Xu-zhen CHENG
    Scientia Agricultura Sinica. 2008, 41(5):  1291-1296 .  doi:10.3864/j.issn.0578-1752.2008.05.005
    Abstract ( 1362 )   PDF (376KB) ( 906 )   Save
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    By using V2709, a mungbean cultivar with bruchid resistance from India, and VC1973A, a bruchid susceptible variety form AVRDC as parents, The segregation of bruchid resistance in the F2 and BC1F1 and F3 populations showed that the bruchid resistance of V2709 is controlled by one dominant gene. The local linkage map of one dominant gene harbored in V2709 was constructed with F2 population and three kinds of molecular marker techniques. The results showed that the genetic distances from the target gene to two markers, RAPD and STS, were 11.0cM and 5.8cM, respectively.
    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY
    Effect of Non-flooded Cultivation Patterns on the Quality of Rice and Its Relations with the Concentrations of Hormones in Grains
    Kai LIU Zichang ZHANG Guowei XU
    Scientia Agricultura Sinica. 2008, 41(5):  1297-1307 .  doi:10.3864/j.issn.0578-1752.2008.05.006
    Abstract ( 1337 )   PDF (358KB) ( 1093 )   Save
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    【Objective】The purpose of this study was to investigate the quality of rice under different non-flooded cultivations and its relationship with the concentrations of hormones in grains during grain filling. 【Method】Two rice cultivars, Zhendao 88 (japonica) and Yangdao 6 (indica), were used as materials. Four treatments, traditional flooding (TF), bare and non-flooded cultivation (BN), non-flooded plastic film mulching (PM) and non-flooded wheat-straw mulching (SM), were conducted during the whole growth season. 【Result】Compared with TF, both BN and PM significantly reduced grain yield, whereas there was no significant difference in the yield between TF and SM. SM significantly reduced the percentage of chalky kernels, chalkiness and setback value, while markedly increased gel consistency, alkali spreading value and breakdown value. The effects of BN and PM on the above quality traits were opposite to those of SM. No significant differences were observed in amylose content and protein content among the four treatments. The two cultivars behaved the same. Concentrations of indole-3-acetic acid (IAA), gibberellins (GA1 + GA4) and zeatin (Z) + zeatin riboside (ZR) in grains differed little among the treatments at the early grain filing stage, and showed SM > TF > PM > BN at the mid and late filling stages. Ethylene evolution rate of grains exhibited BN > PM > TF > SM. There were no significant differences in abscisic acid (ABA) concentrations in grains among the TF, PM and SM during the grain filling period. BN had the lowest ABA concentration at the early filling stage, whereas had the highest concentration at the mid and late filling stages among the all treatments. Concentrations of IAA and GA1 + GA4 at the mid and late filling stages, ABA concentrations at the early and mid filling stage and Z + ZR concentrations during the whole grain filling period significantly or very significantly and positively correlated with grain weight, brown rice, milled rice and breakdown value (r = 0.71* ~ 0.96**), and negatively correlated with setback value (r = -0.76* ~ -0.91**). Concentrations of IAA and Z + ZR at the late filling stage significantly or very significantly and negatively correlated with the percentage of chalky kernels and chalkiness (r = -073* ~ -0.85**). The ethylene evolution rates at each filling stage significantly or very significantly and negatively correlated with grain weight, brown rice, milled rice, alkali spreading value and breakdown value (r = -0.71* ~ 0-.91**), and positively correlated with the percentage of chalky kernels, chalkiness and setback value (r = 0.73* ~ 0.93**). 【Conclusion】The results demonstrate that SM improve, whereas BN and PM degrade, rice quality, and the formation of good or poor quality is closely associated with the concentration of hormones in grains during grain filling.
    Size distribution of starch granules in wheat endosperm under low light environment
    Scientia Agricultura Sinica. 2008, 41(5):  1308-1316 .  doi:10.3864/j.issn.0578-1752.2008.05.007
    Abstract ( 1296 )   PDF (495KB) ( 1096 )   Save
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    Abstract: 【Objective】The purpose of this research was to reveal the size-distribution characteristics of starch granules in wheat endosperm and to study the effect of different-stage shading on the size distribution of wheat starch granules.【Method】In this study, we used a self-made tent to shade wheat plants under field conditions, selected two strong-gluten wheat cultivars: GC8901 and JN17, and set four shading stages: flagging to flower, 1-10, 11-20 and 21-30 days after anthesis(DAA) to study the changes of number, volume and surface area distribution of starch granules in wheat endosperm.【Result and conclusion】The results showed that wheat endosperm had a unimodal curve in starch granule number distribution, and a bimodal curve in starch guanule volume or surface area distribution. On average, B-type granules (diameter<10µm) made up to 99.89% of the totle number of starch granules in wheat endosperm, and accounted for 84.43% of the total starch surface area, but contributed only about 46.85% of the total starch volume. On the other hand, A-type granules (diameter≥10µm) made up to 0.11% of the totle number of starch granules in wheat endosperm, and accounted for 15.57% of the total starch surface area, but contributed about 53.15% of the total starch volume. Size distribution of starch granules in wheat endosperm were influenced markedly by different-stage low light after flagging stage. Low light in 11-20 DAA could decrease the number of A-type starch granules in wheat endosperm, but the other shading treatment influenced it slightly. Low light influenced the inner size distribution of B-type starch granules significantly , and the influence varied with shading time, genotype and grain position in wheat spike. After shading from flagging stage to flower stage, the number of BS-type granules (diameter<1µm) decreased and BL-type granules (diameter 1~10µm) increased in GC8901 superior grains, but those changes in GC8901 inferior grains were on the contrary. The number distribution of starch granules in JN17 superior grains was influenced slightly by shading from flagging stage to flower stage, but in JN17 inferior grains BS-type granules number decreaed and BL-type granules increased. Shading from 1 DAA to 10 DAA increased the number of BS-type granules in wheat endosperm, but low light in the middle or latter grain filling stage increased BL-type granule number, and the function of middle grain filling stage shading was higher than latter grain filling stage shading. The volume and surface area percents of B-type starch granules in the endosperm of two wheat cultivars were increased after shading from 1 DAA to 10 DAA, but those in A-type granules were decreased. After shading from flagging stage to flower stage and low light from 11 DAA to 30 DAA, the volume and surface area percents of A -type starch granules increaesd but those in B-type granules decreased.
    Effects of Different Copper Levels on Photosynthetic Parameters and Active Oxygen Metabolism of Amaranthus tricolor Seedlings
    Shi-sheng KE
    Scientia Agricultura Sinica. 2008, 41(5):  1317-1325 .  doi:10.3864/j.issn.0578-1752.2008.05.008
    Abstract ( 1298 )   PDF (369KB) ( 819 )   Save
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    【Objective】The objective of this study was to gain better insight into the physiological mechanisms of copper (Cu) on photosynthesis and active oxygen metabolism in three-coloured amaranth plants (Amaranthus tricolor). 【Method】The experiments were performed to investigate the parameters of growth, photosynthesis, mineral elements contents and active oxygen metabolism. Three-coloured amaranth seedlings were subjected to different Cu levels in soil at the whole experimental period. 【Result】2.0-4.0 mmol•kg-1 Cu treatment decreased the whole plant dry mass to 90%-73%. The net photosynthetic rate (Pn) and stomalal conductance (gs) in the third leaves of three-coloured amaranth seedlings treated with 2.0-4.0 mmol•kg-1 Cu were similarly reduced. None of the investigated Cu levels decreased internal CO2 concentration (Ci). The effect of Cu on potential efficiency of photosystem Ⅱ, PSⅡ (Fv/Fm) was negligible. The Cu dependent reduction of PSⅡ quantum efficiency (ΦPSⅡ) after plant adaptation in actinic irradiation was more noticeable. This could imply that reduced demand for ATP and NADPH in a dark phase of photosynthesis caused a down-regulation of PSⅡ photochemistry. Furthermore, in third leaves of Cu-treated plants the decrease in water percentage as well as lower contents of photosynthetic pigments and mineral elements including Fe, K and Mg were observed. Superoxide dismutase (SOD) and guaiacol peroxidase (POD) activities and proline (Pro) content in third leaves of three-coloured amaranth seedlings treated with 2.0-4.0 mmol•kg-1 Cu significantly increased, while catalase (CAT) and ascorbate peroxidase (APX) activities and the contents of carotenoid (Car), glutathione (GSH) and ascorbic acid (AsA) decreased accompanied by the contents of hydrogen peroxide (H2O2), superoxide (O2-•) and malondialdehyde (MDA) and electrolyte leakage increased. 【Conclusion】 Treated with 2.0-4.0 mmol•kg-1 Cu, Pn and ΦPSⅡ decreased, and peroxidization aggravated by unbalance of active oxygen metabolism, so the growth of three-coloured amaranth were significantly inhibited.
    Physiochemical character and ability to promote cotton growth of bacteria strains Rs-5 and Rs-198
    Scientia Agricultura Sinica. 2008, 41(5):  1326-1332 .  doi:10.3864/j.issn.0578-1752.2008.05.009
    Abstract ( 1162 )   PDF (339KB) ( 1033 )   Save
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    Two plant growth-promoting bacteria, strains Rs-5 and Rs-198, isolated from salt-stressed cotton rhizosphere, could improve germination rate of cottonseed under salt stress. The ability of solubilizing tricalcium phosphate and secreting IAA (indo-3-acetic acid) of strains Rs-5 and Rs-198 were tested.Cotton seeds germination was tested by pot experiment. Spectrophotometrically with Mo-Sb-VC and Salkowski methods were used to determine content of solubilizing tricalcium phosphate and IAA.The two bacteria strains were characterized as Rs-5 Klebsiella.sp and Rs-198 Pseudomonas.sp. The germination rates was 26.3 % and 34.4 % more than control. During seven days fementation, the soluble phosphate contents solubilized by the strains Rs-5, Rs-198 in fementation broth reached to 73.38 mg•L-1 after incubating 24 hours and 82.41 mg•L-1 after 48 hour, respectively. The pH value in fermentation broth of the two strain decreased from 7.48 to 5.22 and 4.54, respectively. Contents of IAA secreted by Rs-5 and Rs-198 in cultures were accumulated following the incubation time. The contents of IAA were accumulated to 7.09 mg•L-1mg•L-1 and 10.77 mg•L-1 each at 96 hours fermentation. Phosphate solubilizing and IAA secretion might be the main reason for promoting cotton germination under salt stress.
    PLANT PROTECTION
    Molecular Diversity of 23 Wheat Leaf Rust Resistance Near-isogenic Lines Determined by Sequence-Related Amplified Polymorphism
    Ya-Hui LIU Hong-Fei RAN Wen-Xiang YANG Xing LI Ya-Ning LI Qing-Fang MENG Li-Rong ZHANG Ting ZHANG Da-Qun LIU
    Scientia Agricultura Sinica. 2008, 41(5):  1333-1340 .  doi:10.3864/j.issn.0578-1752.2008.05.010
    Abstract ( 1153 )   PDF (353KB) ( 1127 )   Save
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    【Objective】The objective of the research were analysis the Genetic diversity of Wheat Leaf Rust Resistance Near-isogenic Lines in Thatcher Background, and investigating the feasibility of SRAP marker applied on study of Wheat Leaf rust Resistance Genes marker and clone. 【Method】Total 23 wheat leaf rust resistance near-isogenic lines and Thatcher were carried out using SRAP(sequence-related amplified polymorphism)markers. 【Result】41of 128 SRAP primer combinations identified a total of 537 reproducible fragments. Among them, 49.5% were polymorphic, and 115 of them were specific bands (10.6%). Between 6 and 41 fragments were amplified per primer combination, with an average of 13.1 bands. The number of polymorphic fragments for each primer combination varied from 1 to 13, with an average of 2.8. A cluster analysis was performed using NTSYSpc Version 2.10e. Cluster analysis revealed 23 wheat leaf resistance genes and Thatcher into two clusters of A and B with the similarity coefficient of 0.72 . 96% of them were clustered into cluster B, which was divided into two groups,ⅠandⅡ. 95% of the near isogenic lines in cluster B fell into group Ⅱ. Group Ⅱ included four small groups with 0.785 similarity coefficient. 【Result】It was concluded that the 23 wheat leaf rust resistance near isogenic lines have more differences in molecular level. All these results showed SRAP marker is simple, economic and effective,it could be widely used in study of Wheat Leaf Rust resistance genes.
    Transcriptional profile of plant pathogenic bacteria revealed by DNA microarray analysis
    Shi-Qiangg GAO Chenyang He
    Scientia Agricultura Sinica. 2008, 41(5):  1341-1346 .  doi:10.3864/j.issn.0578-1752.2008.05.011
    Abstract ( 1180 )   PDF (280KB) ( 992 )   Save
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    With the completion of full genomic sequences of many bacterial phytopathogens, the development of microarray profiling technology provides new insights into the study of pathogen-host interactions. Using DNA microarray technology to detect the transcriptional profiles of bacterial pathogens under different conditions (especially during in vivo infection) could identify pathogenicity-related genes and their regulatory networks. These would gain helpful information to understand the gene function and pathogenic mechanisms of pathogens. Microarrays therefore provide new ideas for the prevention of plant pathogenic bacteria. Here we will review the latest application of DNA microarray technology in the transcriptional profile study of plant pathogenic bacteria.
    Competition Effects between Ageratina adenophora (asteraceae) and Setaria sphacelata (gramineae)
    Zhi-Lin JIANG Zheng-Yue LI
    Scientia Agricultura Sinica. 2008, 41(5):  1347-1354 .  doi:10.3864/j.issn.0578-1752.2008.05.012
    Abstract ( 1295 )   PDF (428KB) ( 998 )   Save
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    Abstract:【Objective】 The experiment was conducted to investigate the intra- and inter-specific competition interaction between Ageratina adenophora (Sprengel) R. King & H. Robinson and Setaria sphacelata (Schum) Stapf ex Massesy cv. Narok at different densities and its succession in the field, and to further explore the methods of controlling the invasive weed A. adenophora displaced by S. sphacelata.【Method】 We grew two perennial grasses species (A. adenophora and S. sphacelata) for 2 yr in monoculture and in mixture on an experimental densities gradient. The above- and blow-ground biomass of two species were measured and the growth response and relative competitive ability were discussed.【Result】 The biomass total and relative yield (RY) and competitive balance index (CB) of S. sphacelata were significant higher than those of A. adenophora, the relative yield total (RYT) was significant lower than 1. The relationships mentioned above become more obvious with density decreasing. The total biomass of A. adenophora at the densities of 19, 46 and 174 plants/m2 in mixture decreased by 33.0%、38.5% or 70.1% compared to those in monoculture, respectively. However, the total biomass of S. sphacelata at the same density between in monoculture and in mixture showed no significant difference. The ratios of root mass: shoot mass of S. sphacelata in mixture were significant lower than those in monoculture. However, the ratios of root mass: shoot mass of A. adenophora in the mixture were significant higher than those in monoculture.【Conclusion】There was significant mutual antagonism between A. adenophora and S. sphacelata. Compared to A. adenophora, the relative competitive ability of S. sphacelata was higher which decreased heavily the growth of A. adenophora mainly by shoot competition. It will be a good strategy for controlling of A. adenophora and restoring its invaded habitats by S. sphacelata in the future.
    Sensitivities of Phytophthora infestans to metalaxyl, cymoxanil, and dimethomorph
    ZHU Gui-Ning
    Scientia Agricultura Sinica. 2008, 41(5):  1355-1365 .  doi:10.3864/j.issn.0578-1752.2008.05.013
    Abstract ( 1218 )   PDF (321KB) ( 901 )   Save
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    【Objective】Isolates of Phytophthora infestans on tomato in Guangxi province were determined for the sensitivities to metalaxyl, cymoxanil and dimethomorph in order to give the basic information for integrate management of this disease.【Method】Sensitivities were determined by measuring radial growth on agar medium amended with fungicide, compared with the floating-leaf-disk method. 【Result】 Two hundred and thirty-nine isolates were collected from eight tomato growing areas during 2000 to 2006. The results of determine showed that the frequencies of sensitive, intermediate and resistant isolates to metalaxyl were 42.26%, 35.98% and 23.53%, respectively, while the frequencies of sensitive, intermediate isolates to cymoxanil were 37.24% and 62.76%, respectively, and that to dimethomorph were 54.81% and 45.19%, respectively. However, no isolate resistant either to cymoxanil or to dimethomorph was detected. The baselines of sensitivity to metalaxyl, cymoxanil, and dimethomorph were 0.003599µg/ml, 0.07156µg/ml and 0.0627µg/ml, respectively. Variations in sensitivities among isolates from different areas or different years were low for cymoxanil and dimethomorph, but very high for metalaxyl. All isolates from Tianlin and Wuxuan were sensitive to metalaxyl. But metalaxyl-resistant isolates predominated in Tianyang, with the frequency of 51.35%. The EC50 values of some isolates from Tianyang were higher than 500µg/ml and their resistance levels were over 100000 folds. The comparison with the floating-leaf-disk method indicated both techniques provided equivalent results.【Conclusion】 These studies suggested that metalaxyl could still be used in Tianlin, Wuxuan and Nanning, where the resistant isolates had not been found, but in other areas, the use of metalaxyl should be reduced or stopped, and cymoxanil or dimethomorph was recommended.
    Study of resistance of Spodoptera exigua to tebufenozide and the effect of tebufenozide on its biological characteristics
    Scientia Agricultura Sinica. 2008, 41(5):  1366-1372 .  doi:10.3864/j.issn.0578-1752.2008.05.014
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    【Objective】In this paper the resistance risk of Spodoptera exigua to tebufenozide and the biological characteristics of Spodoptera exigua treated with tebufenozide were studied.【Method】Using dipping method, selections for resistant population of spodoptera exigua to tebufenozide were carried out from laboratory strain by treating alternative generation’s 3rd-instar larvae at LC50 dose and treating every generation’s larvae with LC10 dose. The biological characters of each generation were determined. Dipping exposure of 3rd-instar larvae and egg mass of Spodoptera exigua with tebufenozide under different dose, the effects on biology characters in current generation and followed generation was also studied.【Result】Treated alternative generation’s larvae with LC50 dose, Spodoptera exigua had no loss in susceptibility to tebufenozide from F1, F3, F5, F7 ,F9 to F11. But as the selection procedure, the pupation rate, average pupa weight and eggs per female of corresponding followed generation were decreased. Another selection experiment of successive treating every generation’s larvae with LC10 dose of tebufenozide to F19 only obtained 3.52-fold resistance ratio to the initial laboratory strain of spodoptera exigua. The pupation rate, average pupa weight, eclosion rate, adult life and eggs per female had no significant changes in each generation. Dipping exposure of 3rd-instar larvae to tebufenozide solutions of 5-40µg•ml-1, the survival rate of larvae was decreased after three days, and the larval period time, pupation rate and the mean pupa weights were significantly shorter or lower than those of control larvae as the dose increasing. But the eclosion and female/male rate
    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Evaluation of Nitrogen Application Based on Model Combined with GIS in 2000,Jiangsu Province
    Da-Wei Zhu Zhi-Qing Jin Wen Zhang Jun Jiang
    Scientia Agricultura Sinica. 2008, 41(5):  1373-1382 .  doi:10.3864/j.issn.0578-1752.2008.05.015
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    【Objective】This study was conducted to evaluate the adequacy of nitrogenous fertilizer application and to provide scientific references for nitrogen management.【Method】At the basis of the soil mineralization model, organic manures nitrogen releasing model and nitrogenous fertilizer use efficiency model, the models combined with GIS was applied to analyse the spatial variability of nitrogen nutrition from soil and organic manures and its contributing factors in 2000,Jiangsu province. Then the model simulation cropland requirement nitrogenous fertilize was compared with the actual nitrogenous fertilizer application to evaluate its adequacy.【Result】There was about 71.8% cropland overmuch of nitrogen when calculated by actual crop yield, the total quantity was 760 kiloton, accounted for 41.5% of all nitrogen application. While calculated by actual tiptop yield, there was 64.3% cropland short of nitrogen, the total quantity was about 800 kiloton. Nitrogenous fertilizer was used in very low efficiency in Taihu lake area, Nanjing-Zhenjiang hilly area and Xuzhou area, while the deficient nitrogenous fertilizer region mostly distributed at east beach area.【Conclusion】It was great advantageous for transferring superfluous nitrogenous fertilizer from excessive application area to lower area.
    Resolution on the Liquation Character of Nitrogen from Coated Urea by Reaction Rate Theory
    Yu-Ling ZHANG Yu-Long ZHANG Xiu-Li DANG Na YU Masahiro Okamoto
    Scientia Agricultura Sinica. 2008, 41(5):  1383-1389 .  doi:10.3864/j.issn.0578-1752.2008.05.016
    Abstract ( 1286 )   PDF (438KB) ( 797 )   Save
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    The nitrogen liquation of coated urea(LP30)were resolved based on reaction rate theory with culture experiment method under different temperatures. The results showed that the nitrogen liquation of coated urea(LP30)could be obeyed to the first-order reaction model. Liquation rate constant(k)was dependent on temperature. When the liquation rate constants changed from 0.0081~0.0425, activation energy was 75733 J.mol-1 in range of 10℃~25℃.The days of temperature switch method could apply and estimate the liquation amount of coated urea based on reaction rate theory in natural conditions.
    The Synergistic effect between vegetation recovery and soil quality on Abandoned Arable Land in Eroded Hilly Loess Plateau
    Dai Quan Hou Liu Guo Bin Xue Sha Lan Xue Yu Na
    Scientia Agricultura Sinica. 2008, 41(5):  1390-1399 .  doi:10.3864/j.issn.0578-1752.2008.05.017
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    This research regarded vegetation recovery on abandoned arable as subject investigated and adopted correlation analysis and regressional analysis method to study the synergistic effect between vegetation and soil environmental factor. The reseach indicated that vegetation biomass had a significant positive relationship with vegetation coverage. They had a significant negative correlation with bulk density of soil, and positive correlation with soil total porosity, macroaggregate, water stability macroaggregate, organic matter content, total nitrogen content, hydrolyzable nitrogen content, available kalium content, microbial biomass, and microbial respiration. However, they had not significant relationship with total phosphor and availabal phosphor. At the same time, the vegetation biomass and vegetation coverage correlated soil antierodibility closely. They had significant positive correlation with soil aggregation degree and structure coefficient,and negative correlation with soil dispersive coefficient and structural damage rate. Bulk density of soil had negative correlation with soil total porosity, macroaggregate content and water stability macroaggregate content, and the soil total porosity had positive correlation with macroaggregate content and water stability macroaggregate content. The soil quality was hightened, which promoted vegetation succession in the process of vegetation recovery on abandoned arable land in eroded hilly loess plateau. Simultaneity, the vegetation succession and recovery improved soil quality. Therefore, there is synergistic effect between them.
    The Influencing Factors of Soil Anti-scourability of Tree-crop Intercropping Land in Purple Soil Hilly Region
    Scientia Agricultura Sinica. 2008, 41(5):  1400-1409 .  doi:10.3864/j.issn.0578-1752.2008.05.018
    Abstract ( 1517 )   PDF (266KB) ( 918 )   Save
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    This paper studied on the influencing factors of soil anti-scourability of tree-crop intercropping land in purple soil hilly region by used the methods of undisturbed soil sample washing and soil physicall properties analysis indoor. The result showed that: (1)Soil bulk density had remarkable positive correlation with the index of soil anti-scourability and soil porosity had remarkable negative correlation with the index, and this result showed that the soil bulk density more big ,the soil porosity more small, and the soil will be more solid ,the soil anti-scourability will be improved under the short time and high intensity washing condition;(2) The fine sand(0.25~0.05mm)content had remarkable positive correlation with the index of soil anti-scourability ,and which showed that the coarse granule could enhance soil anti-scourability ;(3)The >5mm water stable aggregates content had great remarkable positive correlation with the index of soil anti-scourability and the >0.25mm water stable aggregates content also had remarkable positive correlation with the index, which showed that big water stable aggregates content was more, the soi structure will be finer, the soil anti-scourability will be enhanced ; (4)The steady water infiltration rate had great remarkable positive linear correlation with the index of soil anti-scourability and the initial water infiltration rate had remarkable positive linear correlation with the index ,too; (5)The root biomass in different diameters all had great remarkable positive correlation with the index of soil anti-scourability,and the root density had remarkable positive correlation with the index of soil anti-scourability, the root length of <3mm root also had remarkable positive correlation with the index of soil anti-scourability. The root system can enhance soil anti-scourability effectually. Soil bulk density, fine sand(0.25~0.05mm)content, >5mm water stable aggregates content, steady water infiltration rate and the root biomass of <1mm root can forecast and evaluate the purple soil anti-scourability in this area.
    HORTICULTURE
    Expression of the Antisense BcMF12 Gene Driven by the BcA9 promoter in transgenic Chinese cabbage
    Jiang-Hua SONG Jia-Shu CAO Xiao-Lin YU Xun Xiang
    Scientia Agricultura Sinica. 2008, 41(5):  1410-1415 .  doi:10.3864/j.issn.0578-1752.2008.05.019
    Abstract ( 1059 )   PDF (466KB) ( 849 )   Save
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    【Objective】The study analyzed the expression of BcMF12 gene regulated by BcA9 promoter in the transgenic Chinese cabbage, and confirmed the effect of antisense BcMF12 gene on the pollen development.【Method】An conserved BcMF12 gene fragment was amplified from the cDNA of flower buds in Chinese cabbage (Brassica campestris L. ssp. chinensis Makino), and was fused to the anther specific BcA9 promoter. The plant antisense expression vector was constructed and then introduced into Chinese cabbage via Agrobacterium-mediated transformation. The transgenic plants were screened by antibiotics and molecular analysis.【Result】The analysis of GUS activity revealed that the expression level of antisense BcMF12-GUS fusion gene regulated by the BcA9 promoter was strongest in the anther, while very weak or no activity could be detected in other organs of transgenic plants. It showed that antisense BcMF12-GUS fusion genes could be tissue-specifically expressed in transgenic Chinese cabbage when driven by the BcA9 promoter. Northern blot suggested that the expression of BcMF12 gene in pollen was down-regulated significantly. The pollen germinated rate of transgenic plants contained the antisense BcMF12 gene decreased as compared with that of the control plants.【Conclusion】The expression of the gene BcMF12 related to the pollen development was inhibited by the antisense BcMF12 driven by the anther-specific BcA9 promoter, which affected the pollen development in Chinese cabbage.
    Effects of Phosphorus, Potassium and Calcium Distribution in Malus hupehensis Rehd under Zinc and Iron Stress
    Yan-An WANG Kun LI Di LIU Dequan LI Dianpeng DONG
    Scientia Agricultura Sinica. 2008, 41(5):  1416-1422 .  doi:10.3864/j.issn.0578-1752.2008.05.020
    Abstract ( 1201 )   PDF (275KB) ( 696 )   Save
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    【Objective】 In order to explain the stress mechanism under zinc and iron stress and the homeostasis mechanism among mineral elements in apple trees, the distribution characteristic and interaction relation of phosphorus, potassium and calcium of Malus hupehensis Rehd were studied.【Method】 Distribution characteristic of phosphorus, potassium and calcium, and their correlation in organs of apple stock—Malus hupehensis Rehd under zinc and iron interaction was analyzed, with solution culture and orthogonal design.【Results】 Under zinc deficiency, the concentrations of phosphorus, potassium and calcium significantly reduced in roots, while the concentration of potassium reduced in shoots, but the concentrations of phosphorus and calcium significantly increased. At excess zinc, the concentration differences of phosphorus, potassium and calcium in organs were not significant. Under iron stress, the concentrations of phosphorus and calcium reduced at excess and low concentration of iron in roots, the concentration of calcium increased at low concentration of iron in shoots. Zinc concentration was positively correlated with phosphorus, potassium and calcium concentrations in roots, while zinc concentration was negatively correlated with phosphorus and calcium concentrations in shoots.【Conclusion】Under zinc deficiency, potassium are excessively reduced in all organs; the phosphorus and calcium also significantly reduced in roots, however, the accumulation of phosphorus and calcium increased in shoots. At proper and excess zinc, the concentrations of calcium in organs decreased significantly when increased the iron concentration of solution.
    The Relation Study of Ramie (Boehmeria nivea (L.) Gaud) Sex and Ethylene
    Hu-Cheng XING zhanjun qin wei she Zhong-qin LUO yucheng jie
    Scientia Agricultura Sinica. 2008, 41(5):  1423-1428 .  doi:10.3864/j.issn.0578-1752.2008.05.021
    Abstract ( 1224 )   PDF (306KB) ( 826 )   Save
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    The ethylene release rate was mensurated in shoot tip of different sex ramie in different time, same node in female and hermaphrodite ramie in second crops and different sex single inflorescence in hermaphrodite ramie. Effects of two ethylene inhibitor on sex expression in ramie were done. The result indicated there was closed relation between sexual different in ramie and ethylene release rate. The female ramie could be induced by high ethylene release rate. The female flower could be inhibited by AVG and AgNO3,but AVG at a concentration of 300mg/L and were the best..
    The Grey Analysis, Kriging and Selection Index of Flower Yield in Rugosa Rose
    Zhen Feng Feng Lanyong Zhao Zhao
    Scientia Agricultura Sinica. 2008, 41(5):  1429-1435 .  doi:10.3864/j.issn.0578-1752.2008.05.022
    Abstract ( 1335 )   PDF (301KB) ( 991 )   Save
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    Abstract: [Objective]Seeking for the theory and techniques of selection breeding of high-yield rugosa rose cultivars, [method] the analysis of grey system, Kriging interpolation and integration selection index were employed to investigate the relationship between the flower yield per plant and 15 quantitative traits of 20 rugosa rose cultivars. [Results] The result showed that: The grey relational grade of the flower number/plant, the number of branch, the width of floral bud and the weight of single flower to the flower yield/plant were larger (> 0.5). Kriging interpolation simulation was applied to analyze the flower yield/plant. It was found that the value of target trait went up with increase of the number of flower/plant and the number of branch. Moreover, the indirect selection of either trait couldn’t get better improvement of flower yield/plant. [Conclusion] It’s necessary to improve flower yield/plant by multi-trait selection. The integration selection index I = + 0.3187x1 - 318.6 x2 + 670.1x4 + 6.3x8. Index heritability = 0.8014, selective response of the integration breeding value = 245.8811. This will provide theoretic base for genetic breeding of rugosa rose.
    Production and cytomorphological research of the male sterile lines of Zinnia elegans
    Scientia Agricultura Sinica. 2008, 41(5):  1436-1443 .  doi:10.3864/j.issn.0578-1752.2008.05.023
    Abstract ( 1035 )   PDF (562KB) ( 855 )   Save
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    【OBJECTIVE】In order to find out a new pathway for utilizing heterosis of Zinnia elegans and accelerate breeding process, the mechanism of anther development of a male sterile line was explored. 【METHOD】 Backcross, Sibmating, Selfing of fertile plants and testcross with inbred lines were analysed and identified in the field, and cytomorphology was observed.【RESULTS】Recessive nucleus male sterile line AH209AB that capable of working as a maintainer was obtained by successive backcrosses with male sterile plants and fertile F1 plants as male parents. Cytological and anatomical studies indicated that: (1) The wall of normal anther was constituted of four layers of cells such as epidermis, powder chamber wall, middle level and tapetum cells. The process in meiosis of pollen mother cell in Zinnia elegans was normal, cytoplasm divided simultaneity. Mature pollen grains were tricellular. (2) The petal of male sterile plant degenerated as a threadlike structure, the stamen were villiform in appearance and no pollens were formed. The result showed that the growth of anther of male sterile plant is inhibited at the stage of chear sporial cel1 or the stage of sporial mother cell. During bud development, the size of the aborted anther was increasing but did not differentiate the sporogenous cell and the pollen sac. There was no evident boundary between tapetum cell,middle lamella and inner wall of PMC. Tapetal cells did not develop from the very beginning. So the abortion type was completely structural male sterility. 【CONCLUSION】 The male sterile line belongs to non-sporange male sterile type and is of great use in F1 seeds production.
    STORAGE·FRESH-KEEPING·PROCESSING
    Advances in the study of Tomato Aroma Components
    Scientia Agricultura Sinica. 2008, 41(5):  1444-1451 .  doi:10.3864/j.issn.0578-1752.2008.05.024
    Abstract ( 1276 )   PDF (333KB) ( 1735 )   Save
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    This paper summarized the achievements of the study on aroma components of tomato. The kinds of tomato Aroma Components, the importance of components to flavor and the effects of varieties,growing stages,and different growth condition on aroma compounds were discussed. The major synthesizing pathway of flavor compounds, the key enzymes in biosynthesizing process and the factors affected the synthesizing were analyzed. The development of molecular marking in aroma components was also introduced. Finally, the separating and identification methods of tomato aroma components were also reviewed.
    Study on Aroma Components in Fruit from Three Different Satsuma Mandarins Varieties
    YU QIAO Bi-jun XIE Yan ZHANG Hai-yan ZHOU Si-yi PAN
    Scientia Agricultura Sinica. 2008, 41(5):  1452-1458 .  doi:10.3864/j.issn.0578-1752.2008.05.025
    Abstract ( 1243 )   PDF (404KB) ( 961 )   Save
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    【OBJECTIVE】Fruit aroma of Guoqing No.1, Miyagawa Wase and Owari from three different Satsuma mandarins varieties were investigated.【METHOD】Fruit aroma of different varieties were analyzed by headspace solid phase microextraction(HS-SPME) combined with GC-MS.【RESULTS】The results showed that there were 73, 71 and 66 aroma components in three varieties, and the total contents were 584.67μg•g-1,505.29μg•g-1,494.63μg•g-1, respectively. The main aroma components were D-Limonene, linalool, γ-terpinene, β-myrcene and octanal in three fruit. All 31 constituents were found common in three varieties. It was also found that Guoqing No.1, Miyagawa Wase and Owari had 12, 5 and 2 unique components, respectively. 【CONCLUSION】Differences in the composition of volatile molecules from various varieties were observed, and these data could be used for the characterization of each varieties.
    Screening and Identification of Aflatoxin B1 Degradation Strains
    Scientia Agricultura Sinica. 2008, 41(5):  1459-1463 .  doi:10.3864/j.issn.0578-1752.2008.05.026
    Abstract ( 1238 )   PDF (302KB) ( 1755 )   Save
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    Abstract:【Objective】The text was mainly to find the microorganism to degrade aflatoxin B1 and realized the application of biological degradation on AFB1.【Method】Using cumarin as the carbon source and energy,ten strains were first screened out of 1500,and the strain NMO-3 had high growth speed,its Morphology colony was big,the ability of utilizing cumarin was strong.We use aflatoxin B1 to have the second screening,through the ELISA method detecting AFB1 degradation ratio.【Results】The strain NMO-3 was screened out of ten strains, the degradation ratio of AFB1 reached 85.7%,it was prominent to the others(p﹤0.01).With the analysis of colony morphology,physiological and biochemistry experiments,and 16SrDNA gene sequence,the strain NMO-3 was identified as Stenotrophomnas sp.【Conclusion】The results of this text showed that using cumarin as the carbon source and energy to screen out the AFB1 degradation strains was feasible.The text firstly reported the strain’s detoxification function.
    ANIMAL SCIENCE
    Establishment of Real-time TaqMan-Fluorescence Quantitative RT-PCR Assay for Detection and Quantification of Porcine Lipoprotein Lipase mRNA
    Scientia Agricultura Sinica. 2008, 41(5):  1464-1469 .  doi:10.3864/j.issn.0578-1752.2008.05.027
    Abstract ( 1117 )   PDF (499KB) ( 1250 )   Save
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    bstract: Objective : Cloning porcine LPL cDNA , using it as the standard for real-time quantifying LPL mRNA and establishing TaqMan FQ-RT-PCR assay for detection of it. Methods : Total RNA extracted from longissimus dorsi of porcine was reverse transcribed to cDNA. LPL cDNA was ligated with pGM-T vector and transformed to bacterium TOP10. Plasmid DNA extracted from positive clones was verified by PCR amplification and sequenced. LPL was amplified by real-time fluorescence quantitative PCR from the plasmid DNA. The concentration of DNA template purified was detected by analysing absorbance in 260 nm and then was diluted to series standard concentrations of LPL αre combined plasmid for FQ-PCR. Results : The method of LPL mRNA real-time PCR was well established, which detected as low as 103 copies with the linear range from 103 to 1010 copies. The standard curves showed high correlations(R2=0.9871).Conclusion : A series of standards for real-time PCR analysis have been constructed successfully,and real-time TaqMan-Fluorescence Quantitative RT-PCR is reliable to quantitatively evaluate FQ-PCR mRNA in longissimus dorsi of porcine.
    Yak Lactate Dehydrogenase-A: Purification, Properties and cDNA Cloning and Sequencing
    Scientia Agricultura Sinica. 2008, 41(5):  1470-1475 .  doi:10.3864/j.issn.0578-1752.2008.05.028
    Abstract ( 1193 )   PDF (507KB) ( 947 )   Save
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    [Objective] In order to study the adaptation of yak (Bos. grunniens) to high altitude and low oxygen plateau at molecular level, lactate dehydrogenase A (LDH-A) from yak skeletal muscle was purified, and LDH-A cDNA was cloned. The kinetics and cDNA of yak LDH-A were compared with those of bovine LDH-A. [Methods] Dye affinity chromatography and DEAE-Sephadex ion-exchange chromatography were used to purify LDH-A from skeletal muscle of yak and bovine, and their enzyme properties were compared. The cDNA of yak LDH-A was cloned by RT-PCR methods and compared with that of bovine LDH-A in the GenBank. [Result] The relative activity of purified LDH-A was 103.9 U/mg protein, with purification factor of 18.2. Only one band was observed when the purified LDH-A was separated with SDS-PAGE or native PAGE. Kinetic analysis showed that Michaelis constants (Km) value for NADH was 0.097, and Km value for pyruvate was 1.897, both was higher than that of bovine. The Km value for pyruvate of yak LDH-A was about two folds that of bovine. Hg2+ inhibited LDH-A activity, and the inhibition effect was partly removed by adding β- mercaptoethanol. [Conclusion] The high Km for pyruvate of yak LDH-A can prevent from producing too much lactate in skeletal muscle, and might be a result of adaptive evolution.The two amino acid replacements are responsible for the increased Km value.
    The Preliminary Report on Rumen Protozoa Grazing Rate on Bacteria with a Fluorescence-labeled Technique
    WangMeng-zhi hongrong wang guoxiang li hengchun cao zhanjun lu
    Scientia Agricultura Sinica. 2008, 41(5):  1476-1481 .  doi:10.3864/j.issn.0578-1752.2008.05.029
    Abstract ( 1400 )   PDF (308KB) ( 1049 )   Save
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    Studies on the grazing rate of rumen protozoa were carried out under laboratory conditions using a technique of fluorescence-labeled bacteria, in Nov. 2006. Four Xuhuai goats were used in this experiment to provide rumen protozoa and bacteria. Two groups were designed as follows: one group was the whole bacteria which were marked using fluorescence through removing free bacteria from rumen fluid (WFLB); the other group was bacteria which were marked using fluorescence without removing free bacteria from rumen fluid (FLB). The result showed that, The grazing rates of rumen protozoa was 398.4cells/ (cell H) for the group with (WFLB), 230.4cells/ (cell H) for the group with (FLB); Conversed into bacteria-N, they were: 2.15pg N/ (cell H) for the group with (WFLB) and 1.24pgN/ (cell H) for the group with (FLB) respectively. Extrapolating the assimilation quantity of Nitrogen by ciliates on bacteria of Xuhuai goat, were 103.2 mg N/ (d capita) for the group with (WFLB), 59.5mg N/ (d capita) for the group with (FLB) respectively; It also could be estimated as protein loss, the protein micro-cycling were 0.645g Pr/ (d capita) for the group with (WFLB) and 0.372g Pr/ (d capita) for the group with (FLB) respectively, and finally fluorescence-labeled technique has a potential for research on the grazing rate of rumen protozoa.
    Study on Dynamic Expression of the Apoptotic Gene-Fas/FasL in Immunity Organs of Gushi Chickens and its Physiological Significane
    LI Kui Ying LIU Gui-rong SUN
    Scientia Agricultura Sinica. 2008, 41(5):  1489-1496 .  doi:10.3864/j.issn.0578-1752.2008.05.031
    Abstract ( 1097 )   PDF (233KB) ( 877 )   Save
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    Abstract: The dynamic expression of the apoptotic gene-Fas/FasL in immunity organs of Gushi chickens was studied by using immunohistochemistry technology and Leica Microsystems, and its physiological significane was disscussed. The results showed that the expression of Fas in different immunity organs were observed, however, the number of the positive Fas-cells were different between different developmental stages. The positive Fas-protein was found in cell membrane, cytoplasm, but not in nucleus of lymphocytes. The positive Fas-cells were dispersed (or in clusters), and distributed in different places for different immunity organs. The majority of positive Fas-cells, in bursa, were observed in lamina propria of mucous membrane near epithelium and areas between lymphoid nodules, edges of lymphoid nodules, and a few in inner of lymphoid nodules. In thymus, the positive Fas-cells localized mainly in thymic medulla, and few in thymic cortex. The positive Fas-cells, in spleen, were found predominantly in red pulp, marginal zones, and the surronding areas of splenic nodules and periarterial lymphatic sheath, few in splenic nodules and periarterial lymphatic sheath. The expression of FasL in immunity organs of Gushi chickens is similar to Fas, but, the positive FasL-cells is less than Fas. All results mentioned above indicate that the apoptotic gene-Fas/FasL participates in the regulation of apoptosis of thymocytes and lymphocytes in immunity organs of Gushi chickens, and it is important for the stability of immunity organs.
    The expresstional differences and developmental changes of bcl-2、P53 Gene in early embryos of generic hybrids of chicken-quail
    Scientia Agricultura Sinica. 2008, 41(5):  1497-1502 .  doi:10.3864/j.issn.0578-1752.2008.05.032
    Abstract ( 1207 )   PDF (514KB) ( 711 )   Save
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    Abstract:【OBJECTIVE】To investigate the influences of apoptotic factors bcl-2、P53 on early embryos of generic hybrids;【METHOD】we acquired the cross bred eggs of chicken(♂)-quail(♀)with artificial insemination,hatched the eggs in one batch according to the standard condition of chicken,collected the early living-embryos of 2.75,3.0,3.25,3.50,3.75,4.0,4.25,4.5,4.75 and 5.0 d at random.Adopted the method of RT- PCR,used multi-ply PCR to identify the embryo sex with primers of Wpkci and β-actin,then selected four embryos of female and male respectively of each period,taken β-actin for internal standard to determine the relative quantity of bcl-2、P53 mRNA of embryos;【RESULTS】1)The bcl-2 mRNA expression of 2.75~4.75 d of male embryos maintained lower level,declined on 4.0 d,then reached the initial level,rose distinctly on 5.0 d(P<0.01)and reached the peak;the bcl-2 mRNA expression of 2.75~4.5 d of male embryos maintained lower level,rose distinctly on 4.75 d(P<0.01),then maintained higher level and reached the peak on 5.0 d;the female bcl-2 mRNA expression of 4.75 d were higher distinctly than male of the same days(P<0.05); 2)The patterns of P53 mRNA expression of male and female embryos were basically consistent.They were higher on 2.75 d,declined on 3.0 d(P<0.05)and reached the lowest level,declined distinctly in male,but the whole differences weren’t significant in female,then reached and maintained the initial level.Comparing the expression between different sex, the differences weren't significant.【CONCLUSION】We found two abnormal points in gene expresstional periods in female(3.0 and 4.75 d),but one point in male(3.0 d).The disorder regulated by key factors of apoptosis may induce embryo death.This conclusion should be confirmed further after the research of bcl-2、P53 expression of chicken and quail's embryos of the same period.
    VETERINARY SCIENCERE·SOURCE INSECT
    Studies of immunogenicity responses of mice immuned with eukaryon expression plasmid on PRRSV SD2 E containing CpG motifs
    Scientia Agricultura Sinica. 2008, 41(5):  1503-1510 .  doi:10.3864/j.issn.0578-1752.2008.05.033
    Abstract ( 1342 )   PDF (385KB) ( 791 )   Save
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    The recombinant expression vector plasmid CpG-pVAX1-E was transfected into COS-7 cells by using lipofectamine methods.The ORF5 mRNA of transfected cells were detected by RT-PCR. The transient expression of PRRSV SD2 ORF5 nucleocapsid protein was detected by indirect immunofluorescence assay(IFA). In some transfected COS-7 cells, the green fluorescence was showed.The recombinant expression plasmid was inoculated into SPF mice.The antibody ies specific for E protein in serum were confirmed by ELISA. T lymphocyte proliferation test(MTT) were used to detect the number of CD4+ 、CD8 + and the T lymphocyte proliferation in peripheral blood of mice vaccinated. The results showed that the recombinant plasmid induced CpG-pVAX1-E could induce higher response of cellular immune response and specific humoral immune responses in mice . CpG-ODN was effective adjuvant which could significantly enhance the immunogenicity of PRRSV SD2 ORF5 DNA vaccine.
    The Study of Compound Chinese Medicine against the Anti-avian Influenza Virus
    Scientia Agricultura Sinica. 2008, 41(5):  1511-1518 .  doi:10.3864/j.issn.0578-1752.2008.05.034
    Abstract ( 1291 )   PDF (351KB) ( 951 )   Save
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    【Objective】】To research the inhibition effects of compound Chinese medicine on the anti-avian influenza virus in vivo and in vitro, its pharmacokinetics and acute toxicity test , its effects on immune function of SPF chicken.【Method】the effects of direct inactivation,preventive and therapeutic of compound Chinese medicine on the anti-avian influenza virus were studied on chicken embryo and MDCK cell. Its pharmacokinetics was studied by drug residual accumulation. The acute toxicity test of compound Chinese medicine was studied by oral administration and intraperitoneal injection on Kunming mice. The effects of compound Chinese medicine on immune function of SPF chicken was studied by means of the indexes of immune organs in chickens,the contents of CD3、CD4、CD8 andγδTCR by FAC and antibody titer of avian influenza virus detected on HI method.【Result】The result showed that the compound medicine had a strong action of direct inactivation,preventive and therapeutic on the anti-avian influenza virus in vitro. Furthermore it inhibited avian influenza virus adsorbed in cell surface and entered into intracellular. It could prolong the mean death time of chicken and took effect on the chicken challenged the subtype H5N1 of highly avian influenza virus in vivo. The pharmacokinetics parameter showed that its elimination was slow. It had a long time in the mouse and it had long effect in the mouse. The study of acute toxic experiments on mouse was as following: After oral administration the compound medicine was safe. After intraperitoneal injection adminwastration, the LD50 was 425.6mg/kg.b.w.It enhanced cellular and humoral immune function and the indexes of immune organs in chickens.【Conclusion】The compound Chinese medicine is preventive and safe , prolong the mean death time of chicken and strengthen immunity of chickens.
    The prelimilary study on the potential of duck hepatitis virus(DHV-Ⅰ) stimulated the body weight gain and the effects of Silybin on it in younger duck
    Scientia Agricultura Sinica. 2008, 41(5):  1519-1523 .  doi:10.3864/j.issn.0578-1752.2008.05.035
    Abstract ( 1566 )   PDF (304KB) ( 853 )   Save
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    [Aim]To evaluate the effects of duck hepatitis virus-Ⅰ(DHV-Ⅰ) on the body weight gain in duck and the effects of Silybin on it in vivo.[Methods] 86 10d old ducks,both male and female, were collected to be subjected to the test. The experiments were conducted in 7 groups: In geoup 1~3,the animals were inoculated with 1:105 diluted duck hepatitis virus infected allantoic fluids and given 0,30 and 50mg/kg.bw.d Silybin orally respectively. In group 4~6 ,the animals were inoculated with 1:5×105 diluted duck hepatitis virus infected allantoic fluids and given 0,10 and 30mg/kg.bw.d Silybin orally respectively.The group 7 was the control one treated by injecting stilled saline into the leg muscles. By the 4d after inoculation,the wein blood were drawed from the dorsal foot veins and the plasma samples were collectde and stored at -20℃ .The body weight gain were measured from 0 to 10 days after inoculation.The plasma IGF-Ⅰ,T3 and T4 consentrations were measured by RIA [Results]At the virus dose of 1:5×105 diluted virus infected allantoic fluids,the inoculations of the virus enhanced the body weight gain significantly compared with that of control (P<0.01). While 10~50mg/kg.bw.d Silybinin could counteract the effects of the virus on the body weight gain dose-dependently.The plasma IGF-Ⅰlevels showed no correlative with the body weight gain,but the T3 levels showed a same tropism with the body weight gain. [Conclution] Sublethal dose of duck hepatitis virus enhanced the body weight gain of younger ducks significantly,the Silybin could counteract this effect in vivo.
    Development of antibodies against Diethylstilbestrol
    Scientia Agricultura Sinica. 2008, 41(5):  1524-1529 .  doi:10.3864/j.issn.0578-1752.2008.05.036
    Abstract ( 1216 )   PDF (364KB) ( 923 )   Save
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    Four rabbit antisera against DES with high titers (>1∶1 000 000) were obtained by immunizing rabbits with DES immunizing antigen. One of them (No.4) was used to develop an indirect competitive inhibition enzyme-linked immunoassay (Ci-ELISA) for the determination of diethylstilbestrol (DES). Four calibration curves were made by KCjunior software according to 4-parameter model. In the linear range of 0.01-50μg/L, the IC50 was 0.71~1.0μg/L,R2 was 0.9952~0.9978。The coefficients of variation intra-assay and inter-assay were 0.52%-22.48%, 2.64%-19.51% respectively.The antiserum No.4 had cross reactivity of 8.5%, 38.5% with other two synthetic anabolic HEX, DIEN respectively. But had hardly cross reactivity with native estrogen E2, EE2 and Tyr.
    Nutritional Crossbreed of Honeybee and Breeding of Resistibility on Mite
    Xian-Bing XIE Wen-Jun PENG Zhi-Jiang ZENG
    Scientia Agricultura Sinica. 2008, 41(5):  1530-1535 .  doi:10.3864/j.issn.0578-1752.2008.05.037
    Abstract ( 1237 )   PDF (250KB) ( 1006 )   Save
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    【Objective】Mite(varroa jacobsoni)is one of the most serious parasite threat to the honey bee (Apis mellifera) reared in china. We mainly use the drug to control and kill the mite for these years, but the honey products may be contaminated and the mite is becoming drug-resistant. Our main idea is to research the possibility to rear the mite-resistant honeybee by the nutritional crossbreed. 【Method】Breed the larvae (Apis mellifera ligustica) with the royal jelly of Apis carana carana, and then measure the morphological index of the worker generation, genotypic frequency and gene frequency of the MHCⅡ, genetic resemblance and mite resistance..【Result】Compared with the parent workers the results showed: proboscis length, anterior wing area, the total length of 3rd and 4th dorsal plate of abdominal, the length of 4th dorsal plate tuberculum, the area of 6th abdominal segment, the area of wax mirrors are significant different, but the differences of the brachium index, dactylus index and wing claw are not significant. And there are some mutation in the genotypic frequency and gene frequency of the MHCⅡ. The mite resistance of the nutritional crossbreed worker are significantly higher.【Conclusion】We can change the morphological, physical and biochemical character, genetic resemblance and the mite resistant ability of the worker generation by the nutritional crossbreed. The nutritional crossbreed can be a new way to breed the honeybee.
    RESEARCH NOTES
    Quality Difference of Near-isogenic Lines of HMW Glutenin Subunits
    Scientia Agricultura Sinica. 2008, 41(5):  1536-1541 .  doi:10.3864/j.issn.0578-1752.2008.05.038
    Abstract ( 1333 )   PDF (230KB) ( 818 )   Save
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    High molecular weigh glutenin subunits (HMW-GS) 7+8* was introduced into Longmai 20(1,17+18,5+10) by 5 consecutive backcrosses with biochemical marker assisted selection from a Canadian extra strong wheat cultivar Glenlea which possessing an overexpression of the Bx7 subunit. The near isogenic lines (NILs) of HMW-GS were obtained and grown in the experimental field of Crop Breeding Institute in 2005. The experimental design was a randomized block with six replicates. As compared to NILs with 17+18 subunit, the average of quality data of NILs with 7+8* subunit were 5 %(P =0.012)、4%(P =0.018)、10% (P =0.013)、13%(P=0.258)、256%(P =0.029) and 86%(P =0.020) higher in flour protein content、dry gluten content、Zeleny sedimentation、development time、stability and breakdown time, respectively, and were 2%(P =0.033)and 35%(P=0.007)lower in wet gluten/dry gluten and degree of softening, respectively. The application of subunits 7+8* in wheat breeding for quality was discussed in this study.
    Differentiation of Host- and Geo-populations of Liriomyza sativae Blanchard Based on β-tubulin Gene Sequences
    Scientia Agricultura Sinica. 2008, 41(5):  1542-1547 .  doi:10.3864/j.issn.0578-1752.2008.05.039
    Abstract ( 1293 )   PDF (373KB) ( 817 )   Save
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    【Objective】To analysis the genetic differentiation of the host- and geo-populations of Liriomyza sativae Blanchard.【Method】Theβ-tubulin gene of 5 host-populations and 6 geo-populations of Liriomyza sativae Blanchard were sequenced. The identity and variation of β-tubulin gene sequenced were analyzed by biosoftware DNAStar and MEGA and the phylogenetic trees were constructed. 【Result】The results obtained by the biosoftware DNAStar and MEGA respectively were similar. The differences of β-tubulin gene among the host- and geo-populations of L.sativae were very low, with 8 variable sites and no insertions and deletions and the identities were all above 98%.【Conclusion】The trends of differentiation of host- and geo-populations of L. sativae were consistent with the hobby to hosts and the geographical distribution, respectively.
    Cloning and Expression of a Full-length cDNA of Expansin Gene from New Root of Malus hupehensis Rehd.
    Scientia Agricultura Sinica. 2008, 41(5):  1548-1553 .  doi:10.3864/j.issn.0578-1752.2008.05.040
    Abstract ( 1430 )   PDF (443KB) ( 832 )   Save
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    【Objective】Aim of this research is to clone and investigate the expression of the expansin gene from new root of Malus hupehensis (Pamp) Rehd., which will be helpful to study the action of expansin gene in roots development. 【Method】With the RT-PCR and RACE methods, a full length cDNA sequence of expansin gene was cloned in root of Malus hupehensis (Pamp) Rehd. The expression of expansin gene was analysed by Northern blot【Result】Sequencing and structural analysis showed that the full-length of MhEXP1 consisted of 1111bp with an open reading frame of 771bps that could code for a protein of 257 amino acids.The predicted MW and PI were 27.8kD and 8.9. The polypeptide had a N-terminal signal peptide of 23 amino acids like many other expansins. Essential features of the protein such as the eight cysteine residues and four tryptophan residues near the C-terminal end are conserved in the sequence. The HFD sequence, which has been shown to be a part of the endoglucanase active site is also present in MhExp1. Northern blot showed that the expression of MhEXP1 gene was more significant in roots than in leaves. The transcriptional level of the MhEXP1 gene in the roots of Malus hupehensis (Pamp) Rehd was most highly by 10μmol/L IBA treatment for 6 h.【Conclusion】All of the results indicated that the obtained gene is a new member of expansin gene family. The transcription of MhEXP1 was regulated by IBA and it played an important role in the development of Malus hupehensis (Pamp) Rehd. roots.
    Study on effects of alfalfa meal on production performance and carcass quality and serum items of fatting pigs
    Scientia Agricultura Sinica. 2008, 41(5):  1554-1559 .  doi:10.3864/j.issn.0578-1752.2008.05.041
    Abstract ( 1149 )   PDF (290KB) ( 950 )   Save
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    40 fatting pigs, body weight about 50kg for each one,were randomly divided into 5 groups,4 replicates for each group and 2 for each replicate.One of these groups was used as the contol group,other groups were used as test groups,groupI,groupII,groupIII and groupIV.The control group was fed basic diet,groupI、II、III were fed the diets containing alfalfa meal in different level (10%、15%、20%) and groupIV was fed the diet 20% alfalfa meal and 01% cellulase.The experimental period was 60d.The results showed:(1)Three test groups,groupII,groupIIIand groupIV,had higher daily intake and daily gain than the control group,and groupII and groupIV had higher feed conversion rate than the control group.(2)Compared with the control group,there were more apparent digestibility of nutrients for groupI、II,however,groupIII、IV had similar digestibility with the control group.The digestibility of groupIV was higher than groupIII one,showed that it was useful to add 0.1% cellulase enzyme for diet of higher amount of alfalfa meal.(3)There were no significant difference in eye muscle area,meat colour and marbling among groups,however,because of adding suitable amount of alfalfa meal,these ones,back fat thichness was dropped , killing out percentage and thin meat ratio were improved.(4)Adding alfalfa meal in fatting pig,s dietary,groupI、II、III、IV had significant improvement in GH、Ins、IGF-1 comparing with the control one,but there were significant derease in GLUand CHU.The study showed that it was best to add 15% alfalfa meal in diets of fatting pigs among all groups,so it should be recommend in diet.
    Single Nucleotide Polymorphisms (SNPs) of hypocretin receptor1 (HCRTR1) Gene in Nanyang Cattle
    Scientia Agricultura Sinica. 2008, 41(5):  1560-1566 .  doi:10.3864/j.issn.0578-1752.2008.05.042
    Abstract ( 1050 )   PDF (583KB) ( 668 )   Save
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    The Single nucleotide polymorphisms (SNPs) of 122 Nanyang cattle were detected for coding region of hypocretin receptor1 (HCRTR1) gene. Eleven SNP loci including 322 : G/A; 384:G/C; 420: T/C; 423: C/T; 481:T/A; 510:C/A; 627: T/C; 631:C/T; 690:G/A; 714:G/A and 736: C/T were found. The discovered SNPs were deposited in GenBank with accession No.: DQ981401, DQ986909, DQ986910, DQ986911, DQ986912, DQ986913, DQ986914, DQ901743, DQ986915, DQ986916 and DQ901742. The variation at 322,481,631 and 736 bp caused amino acid mutation,variation at the other 7 SNP loci were synonymity. Polymorphism information content(PIC), number of effective alleles(Ne) and heterozygosity(h) were also analyzed. three SNP loci (322 bp,510 bp and 736 bp) were moderate polymorphisms(0.3731, 0.3190 and 0.2851 respectively) while the left SNP loci showed low polymorphism(<0.25). The trends of heterozygosities were consistent with that of PIC. The SNP loci showed higher PIC as well as the higher heterozygosities.