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Table of Content

    10 June 2009, Volume 42 Issue 6
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Analysis of Relationship Between RVA Profile and Amylose Content of the Transgenic Rice with Antisense Waxy Gene
    WANG Xin-qi,YIN Li-qing,SHEN Ge-zhi,XU Li,LIU Qiao-quan
    Scientia Agricultura Sinica. 2009, 42(6):  1881-1888 .  doi:10.3864/j.issn.0578-1752.2009.06.001
    Abstract ( 1098 )   PDF (315KB) ( 782 )   Save
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    【Objective】 Previous studies focused on the analysis of RVA profile in rice at the level of the subspecies or varieties (Oryza sativa L.) in which there were differences in genetic background. In this study, antisense waxy homozygous transgenic lines were used to analyze the differences in RVA characteristic values among the lines with different amylase contents in an effort to eliminate the influence of different genetic backgrounds and to more clearly understand the relationship between the amylose content and RVA characteristic value. 【Method】 Anther culture pure lines containing antisense waxy gene were generated by Agrobacterium tumefaciens mediated co-transformation and followed by anther culture in transgenic rice variety. The amylose content and RVA profile were determined in the anther culture lines, and the effects of the amylose content on RVA characteristic value were analyzed under the same genetic background conditions through the RVA profile graphic comparison and the significant determination of RVA eigen value. 【Result】 Seventy-seven pure lines of anther culture that contained only antisense waxy gene but no hygromycin resistance marker were obtained from the transgenic plant offspring of Wu Yunjing No.7 rice variety (O. sativa L. subsp. japonica cv.). Through the dertermination of the amylose content to the T1H1 and T1H2, it was showed that 19 lines had the levels of amylose content similar to the control (16.0%), and that 50 lines were 1%–5% lower than that of control in the amylose content, and some of 30 lines had the amylose content within 11.0%﹣13.9%, and that 8 lines had the amount of amylase content reduced to 3.1%–4.0%. The results of the analysis of the rice starch RVA profile in the pure lines with different amylose contents indicated that there were two kinds of RVA profile: the lines with 3.1%﹣4.0% of amylose content were distinctly different from other lines and conventional glutinous rice. The results of comparison of the lines with same RVA profile to the lines with different amylose content, also showed that the lines with lower amylose content had a distinguished RVA profile with the curve of increasing gradually in finally, but no exceeding the first apex. In significance tests for the characteristic values of RVA profile in the lines with different amylose content, there were five characteristic values of RVA remarkably different among the lines with 3.1%–4.0% of amylose content compared with two other groups, and that there were three characteristic values remarkably different among the lines wiht 11.0%﹣13.5% of amylose content compared with control group. An optimized mathematical model, which characterized the relationships between the amylose content and RVA eigenvalue, was established in this study. From this model, it clearly indicated that the parameters PeT and SBV were more related to the amylose content. 【Conclusion】 Introduction of antisense waxy gene could lead to a reduced amylose content in rice. Anther culture could result in producing pure lines more quickly. The amylose content differences not only affected the characteristic value of RVA but caused different RVA profile as well.

    The Large Kernel Number in the Novel Wheat-Agropyron Germplasm 3228 and Its Inheritance Analysis
    WANG Jian-sheng,WANG Hui,LIU Wei-hua,WU Jun,LI Li-hui
    Scientia Agricultura Sinica. 2009, 42(6):  1889-1895 .  doi:10.3864/j.issn.0578-1752.2009.06.002
    Abstract ( 983 )   PDF (316KB) ( 1017 )   Save
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    【Objective】 This research was conducted to study the utility value and the inheritance of the kernels per spike in the novel wheat-Agropyron germplasm 3228. 【Method】 A field experiment in five wheat regions was designed to explore the stability of kernels per spike of wheat-Agropyron germplasm 3228. Inheritance analysis of the F2 population derived from the cross between 3228 and Jing4839 was carried out using mixed major gene plus polygenes inheritance model of quantitative traits. 【Result】 Results of field experiment showed that the kernels per spike of 3228 were more 90 and has the desirable stability in different planting regions. The analysis result of the F2 population in which the 78.11% individual plants have more kernels per spike than Jing4839 indicated that the kernels per spike of the 3228 was mainly controlled by one major gene, the heritability value and the additive effect of the major gene were estimated as 27.81% and 9.33, respectively. 【Conclusion】 The large kernels number per spike of 3228, which mainly controlled by one major gene, has the better environmental stability, which suggested that the kernel gene would play an important role in improving wheat yield through increasing the kernels per spike.

    Inheritance of Fertility Restoration for Cytoplasmic Male Sterility in a New Gossypium barbadense Restorer
    ZHANG Xiao-quan,WANG Xue-de,JIANG Pei-dong,ZHU Wei
    Scientia Agricultura Sinica. 2009, 42(6):  1896-1900 .  doi:10.3864/j.issn.0578-1752.2009.06.003
    Abstract ( 1148 )   PDF (262KB) ( 835 )   Save
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    【Objective】 In order to clarify inheritance mechanism of fertility restoration for cytoplasmic male sterility (CMS) in a new Gossypium barbadense restorer line ‘Hai R’which was found in the fertility test crossing of G. hirsutum CMS lines with G. barbadense germplasms. 【Method】 Twenty three fertility segregation populations of F2 and backcrossing were used to analyze the inheritance of fertility restoring gene(s) of ‘Hai R’. 【Result】 The result showed that ‘Hai R’ had one major dominant gene (RfB) to control the CMS fertility restoration and this fertility restoration gene functioned at the sporophytic level. The sterile cytoplasm background might not only influence the transmission rate of male gamete but also that of female gamete when the restorer gene was recessive. 【Conclusion】 It could be deduced that this fertility restoration gene might come from G. harknessii cotton, ‘Hai R’ is of value in the application of cotton interspecific hybrid breeding.

    Analysis of Genetic Structure Differentiation of Released Soybean Cultivar Population and Specificity of Subpopulations in China
    ZHANG Jun,ZHAO Tuan-jie,GAI Jun-yi
    Scientia Agricultura Sinica. 2009, 42(6):  1901-1910 .  doi:10.3864/j.issn.0578-1752.2009.06.004
    Abstract ( 1016 )   PDF (430KB) ( 1021 )   Save
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    【Objective】 The objective of the present study was to explore the differentiation of genetic structure of the general population of released soybean cultivars in China and the genetic diversity, genetic specificity and genetic relationship of the geo-ecological subpopulations and released period subpopulations for serving the genetic improvement of soybeans in China with relevant genetic background information in selecting key parental materials from the population. 【Method】 A total of 64 simple sequence repeat (SSR) markers scattered on the whole genome were used to analyze the differentiation of genetic structure of the population with a sample of 378 out of total 1 300 soybean cultivars released during 1923-2005 in China under the software Structure Version 2.2, to analyze the genetic differentiation of the population into geo-ecological subpopulations and released period subpopulations and to analyze the genetic diversity, specificity and relationship of the subpopulations. 【Result】 The Chinese soybean released cultivar population was composed of seven groups of germplasm and obviously differentiated into geo-ecological subpopulations and released period subpopulations with different composition of germplasm groups. Each geo-ecological subpopulation held its own specifically existent, deficient and complementary alleles, which reflected ecological specificity of its genetic source. Along with the advance of released period, new germplasm was introduced into the subpopulations, which made specifically existent, deficient and complementary alleles added to the subpopulations and, therefore, made the new subpopulations with their new specificity. In comparison with the Chinese wild soybean population and landrace population, the Chinese released cultivar population has relatively narrow genetic background due to the bottle-neck effect of a limited number of ancestors. Among the province subpopulations, the number of total alleles and that of complementary alleles in Heilongjiang and Jiangsu subpopulations were the first and second largest due to a relatively wide source of ancestors and broad genetic background. The number of total alleles in the 1923-1970 subpopulation was the least; it increased along with the advance of released period and that in the 2001-2004 subpopulation was the largest even only a short period involved, indicating its relatively high genetic diversity and broad genetic background. 【Conclusion】 The present results demonstrated that geo-ecological differentiation and released period differentiation existed in Chinese released soybean cultivar population, which caused the subpopulations having their own genetic specificities characterized by their germplasm composition and specifically existent, deficient and complementary alleles. All the subpopulation specificities have laid the genetic bases of germplasm or gene exchange among the subpopulations in future soybean breeding.

    Analysis of Genetic Variations of Pollen and Ovule in Several Subspecies and Varieties of Medicago sativa
    LI Xiao-xia,JIN Liang,WANG Xiao-juan
    Scientia Agricultura Sinica. 2009, 42(6):  1911-1917 .  doi:10.3864/j.issn.0578-1752.2009.06.005
    Abstract ( 1303 )   PDF (303KB) ( 1051 )   Save
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    【Objective】 The aim of the present study was to reveal the genetic variation of fertility and the relationship with seed set of three subspecies and one varieties of Medicago sativa, offering theoretical reference for selecting and breeding of superior species. 【Method】 In this paper, genetic variations of pollen vigor and ovule numbers in nine 9 varieties of Medicago sativa belong to M. sativa subsp. falcate, M. sativa subsp. sativa, M. sativa subsp. caerulea and M. sativa var. media were studied. From 2007 to 2008, the pollen vigor, number of ovules per flower and their relationships with seed set were investigated respectively. 【Result】 There existed significant difference (P<0.05) in pollen vigor and ovules per flower among these Medicago sativa plants, and the highest pollen vigor and ovules per flower were observed in Hungary (M. sativa subsp. sativa) with 85.48% and 13, respectively. According to the correlation analysis, the seeds set under self-pollination showed a significant linear correlation with pollen vigor (P<0.01), while seed set under artificial-pollination showed a significant linear correlation with ovules per flower (P<0.05). The variations of seed set among-varieties were larger than those of within-variety under self-pollination and artificial pollination, but variations of seed set within-varieties were larger than those among-varieties under open pollination. The results indicated that the ways of pollination had significant effects on seed set in Medicago sativa plants, and the number of seeds per inflorescence was self pollination<artificial pollination<open pollination. 【Conclusion】 The high genetic polymorphisms of pollen vigor and ovules per flower in Medicago sativa were observed. There was a positive correlation between seed set and pollen vigor under self pollination as well as ovules per flower under artificial pollination.

    Transient Expression of Transferred Gene Induced by Polyethylenimine in Plant

    LI Ying,CUI Hai-xin,SONG Yu,LI Yao,HUANG Jin-li
    Scientia Agricultura Sinica. 2009, 42(6):  1918-1923 .  doi:10.3864/j.issn.0578-1752.2009.06.006
    Abstract ( 1172 )   PDF (466KB) ( 2216 )   Save
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    【Objective】 The experimental research was carried out to investigate the possibility and method of gene delivery into plant cells by nano-vector polyethylenimine (PEI) and obtain transient expression of gene, based on some research results that PEI can condense DNA into granules of nanometer size for delivering gene into cells in the field of medical science. 【Method】 PEI/DNA complexes were prepared. The ability of PEI to combine and protect DNA was investigated by agarose gel electrophoresis retardation assay. The surface characteristics of PEI/DNA complexes were observed with transmission electron microscope. For Arabidopsis thaliana protoplasts, transformation efficiency of PEI/DNA at different charge ratios (N/P) was analyzed and compared with PEG. 【Result】 When the quality rate of PEI and DNA is in range of 5﹕1-1﹕4, PEI could bind and condense DNA, and form stable PEI/DNA complexes with nanoparticles diameter in 100-200 nm. Transformation efficiency of PEI/DNA can be improved with the increase of N/P ratios and reached highest at N/P=5. There was higher transformation efficiency of PEI/DNA complexes than PEG at N/P=5, but began to decrease at N/P>5 with higher toxicity to cells. 【Conclusion】 The experimental results confirmed that PEI is a new kind of efficient gene vector for transferring genes into Arabidopsis thaliana protoplasts.

    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY

    Effects of Sulfur on Rice Quality of Japonica Rice Under Solution Culture Conditions

    LIU Lu-lu,LIU Zheng-hui,DING Yan-feng,WANG Shao-hua,WANG Qiang-sheng,LI Gang-hua
    Scientia Agricultura Sinica. 2009, 42(6):  1924-1932 .  doi:10.3864/j.issn.0578-1752.2009.06.007
    Abstract ( 945 )   PDF (274KB) ( 1184 )   Save
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    【Objective】 The study was conducted to investigate the improvement of sulfur nutrition on rice quality of japonica rice cultivar and find out the change tendency of japonica rice cultivar's quality with sulfur concentration increasing in solution culture. And it was also hoped it could make people give enough attention to sulfur fertilizer and give guidance for sulfur fertilizer' accurate and rational application. 【Method】 Solution culture method was used to study the effect of sulfur on physiochemical properties of rice grain of two japonica rice cultivars, Wujing15 and Wuyujing3. 【Result】 The results showed that Wujing15 and Wuyujing3 rice quality were improved significantly under rational sulfur concentration. Milling quality and appearance quality were improved remarkably, under 200 μmol S?L-1 treatment for Wujing15 and 260 μmol S?L-1 for Wuyujing3, in comparison with 80 μmol S?L-1 treatment. In comparison with 80μmol S ?L-1 treatment, the gel consistency increased by 12.1mm and 8.4mm for Wujing15 and Wuyujing3, respectively. For Wujing15, the peak viscosity increased by 143 cP, setback decreased by 290 cP. For Wuyujing3, the peak viscosity increased by 158 cP, setback value reduced by 34 cP, in comparison with 80 μmol S?L-1 treatment. Additionally, the protein contents and total amino acid contents increased with S concentration increasing for both cultivars, in comparison with 80 μmol S?L-1 treatment. The two rice cultivars' four protein fractions content of the two rice cultivars had positive relationship with sulfur concentrations on the whole. 【Conclusion】Under 200 μmol S?L-1 treatment for Wujing15 and 260 μmol S?L-1 for Wuyujing3, the physiochemical properties of grain of the two cultivars had substantial improvement.

    Effects of Enhanced UV-B Irradiance and Drought Stress on the Growth, Production, and Biomass of Spring Wheat
    GAO Tian-peng,AN Li-zhe,FENG Hu-yuan
    Scientia Agricultura Sinica. 2009, 42(6):  1933-1940 .  doi:10.3864/j.issn.0578-1752.2009.06.008
    Abstract ( 1002 )   PDF (257KB) ( 891 )   Save
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    【Objective】 The effects of enhanced UV-B radiation and drought stress on growth, production, and biomass of three spring wheat varieties were studied to provide a basis for breeding spring wheat varieties of high yield and tolerance to drought and anti UV-B radiation. 【Method】 Drought, enhanced UV-B radiation, and interaction between drought and enhanced UV-B radiation were set to analyze the morphologic index such as flag leaf area, leaf thickness, leaf area index, biomass, root to shoot ratio, and production harvest index, and other biological indicators of three spring wheat cultivars in northwestern China as well. 【Result】 The results showed that enhanced UV-B radiation restrained the node-tillering, resulting in decreased number of tillering nodes, and that moderate drought increased the number of tillering nodes. Under the enhanced UV-B radiation and drought, the biomass of Dingxi-24, Longchun-8139 and Heshangtou reduced by 22%, 21% and 15%, respectively, the average reduction of biomass was 20%. Drought had a less adverse effect on harvest index and yield than enhanced Ultraviolet-B radiation and the interaction between them did. Drought stress increased the harvest index of Heshangtou and Dingxi-24 by 10%, Longchun-8139 by 23%. In contrast, enhanced ultraviolet radiation-B decreased the harvest index of Heshangtou by 10%, and it increased the harvest index of Dingxi-24 and Longchun-8139 by 10%. This indicated that different species allocated inequable proportion of biomass to procreation growth under enhanced ultraviolet-B radiation, drought and the interaction between them. 【Conclusion】 There is a difference among varieties of spring wheat under enhanced ultraviolet-B radiation and drought. Response trend of the three species to enhanced ultraviolet-B radiation and drought is by and large accordant. Drought and enhanced ultraviolet-B radiation had a restraining effect on the three species, but sensible difference amongst species. Longchun-8139 showed a lower number of tillering nodes, highest harvest index and yield in contrast to Heshangtou and Dingxi-24 under enhanced ultraviolet-B radiation and drought. Therefore, Longchun-8139 had more advantages over Heshangtou and Dingxi-24 under enhanced ultraviolet-B radiation and drought.
    Evolution and Development of Maize Production Techniques in China
    LI Shao-kun,WANG Chong-tao
    Scientia Agricultura Sinica. 2009, 42(6):  1941-1951 .  doi:10.3864/j.issn.0578-1752.2009.06.009
    Abstract ( 1037 )   PDF (380KB) ( 1351 )   Save
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    To study the approaches for maize yield increase, this paper has analyzed the change and development of maize production techniques in the past 5 decades and more in China from the perspectives of main measures, technical characteristics and mechanisms for per unit area yield increase. The results showed that maize yield per unit area in China increased by 4 205.1 kg or 437.3% from 1949 to 2007, with annual growth rate of 85.84 kg?hm-2, and the total yield increased from 12.418 million tons to 152.30 million tons, an increase of 12.3 times, of which 68.4% was contributed by yield increase per unit area and 31.6% by planting area expansion. Total yield increase was mainly relied on yield increase per unit area. Maize production techniques have been developed in breeding and planting. Maize breeding and popularization experienced four stages: selection and popularization of open-pollinated varieties, intervarietal hybrid, double-cross hybrid and triple-cross hybrid, single-cross hybrid. After heterosis was extensively used, disease-resistant gene selection, plant type improvement, green-keeping and late-maturing have become main development trends of breeding techniques to increase maize yield potential. In future, the development trend of maize varieties is close planting, adversity resistance, high adaptability, high yield and adaptability for mechanized operation. Planting techniques has shifted from popularization of individual techniques and improvement of farmland conditions to technical integration and modeled cultivation. In future, varieties suitable for high-density and with high adversity resistance and simplified high-yield and high-effective planting techniques with mechanization as carrier will be the development trend. With constant increase of planting density, the mechanisms for maize yield increase have shifted from yield increase per plant to improvement of plant types, colony enlargement, green keeping, late maturing and increase of colony yield via extension of photosynthesis and filling time, improvement of colony uniformity, and high effective carbohydrate production and translocation after florescence. In addition, higher requirements have been set for lodging resistance, disease resistance and adaptability of colony. Based on the above analysis, it is proposed the main technical requirements and measures for maize yield increase in the near future in China.

    Photosynthesis Characteristics of Super-High-Yield Hybrid Cotton in Xinjiang

    DU Ming-wei,LUO Hong-hai,ZHANG Ya-li,YAO Yan-di,ZHANG Wang-feng,XIA Dong-li,MA Li,ZHU Bo
    Scientia Agricultura Sinica. 2009, 42(6):  1952-1962 .  doi:10.3864/j.issn.0578-1752.2009.06.010
    Abstract ( 960 )   PDF (511KB) ( 1160 )   Save
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    【Objective】 To explore photosynthesis characteristics, summarize the principle of yield formation, and propose yield components and photosynthetic physiological indexes, experiments were conducted with super-high-yield cotton in field. It was of great significance to excavate yield potential and construct super-high-yield cultivation technique system. 【Method】 One hybrid cotton cultivar Biaoza A1 and four traditional cultivars including Xinluzao 13, Xinluzao 26, Wanshi 217 and Wanshi 315 were selected as the experimental materials. Leaf area index, chlorophyll SPAD value, photosynthetic rate, photosynthate accumulation and distribution were measured at different growth stages. 【Result】 Compared to high yield traditional cultivar, super-high-yield hybrid cultivar (lint 3 500 kg?hm-2) had higher leaf area index, chlorophyll SPAD value and photosynthetic rate, all of which had longer duration during growth stage. Furthermore, the ratio of canopy respiration rate to total canopy apparent photosynthesis rate which maintained at a low level at the middle and late growth stage, was significantly lower than that of high yield traditional cultivar especially at early boll opening stage. Biaoza A1 had longer and earlier linear increase stage of vegetative organs photosynthate accumulation, which led to greater photosynthate accumulation. However, the linear increase stage of reproductive organs and total photosynthate accumulation and the peak of accumulation appeared comparatively later than those of high yield traditional cultivar. 【Conclusion】 The indexes of yield components and photosynthetic physiology of super-high-yield Biaoza A1 (lint 3 500 kg?hm-2) were proposed, i.e., total number of bolls>150×104?hm-2, boll weight >5.5 g, lint percentage >44%; leaf area index at early full bolling stage 4.9-5.2, at early boll opening stage above 3.3; chlorophyll SPAD value at full bolling stage 65.4-66.5, at early boll opening stage above 64.8; net photosynthetic rate at flowering and boll forming stage 32.2-36.5 μmol?m-2?s-1, at early boll opening stage above 22.2 μmol?m-2?s-1; canopy apparent photosynthesis rate at early full bolling stage approximately 43.4 μmol?m-2?s-1, at early boll opening stage above 16.3 μmol?m-2?s-1; total photosynthate accumulation approximately 26 345.4 kg?hm-2, economic coefficient no less than 31%.

    Differences of Cell Wall Polysaccharides in Border Cells and Root Apices of Pea (Pisum sativum) Under Aluminium Stress
    LIU Jia-you,YU Min,LIU Li-ping,XIAO Hong-dong
    Scientia Agricultura Sinica. 2009, 42(6):  1963-1971 .  doi:10.3864/j.issn.0578-1752.2009.06.011
    Abstract ( 990 )   PDF (485KB) ( 1347 )   Save
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    【Objective】 Root border cells are released from root apices of most plant species, and the root tip is the target of A1 toxicity, thus it was presumed that border cells might be more sensitive to A1 toxicity than root cells. The objective of the present study is therefore to investigate the differences in cell wall polysaccharides between border cells and root apices of pea (Pisum sativum) under A1 exposure. 【Method】 The root border cells of pea (Pisum sativum) were cultivated by mist containing different levels of Al in 24°C growth chamber for 24 h after seed soaking. 【Result】 Results showed that the viability of border cells was significantly decreased by the 4.0 mmol?L-1AlCl3 treatment, and the inhibitory rate of the viability of border cells was 19.2%and the inhibitory rate of relative root elongation was 10.1% after 24 h treatment of 4.0 mmol?L-1AlCl3. The uronic acid and sugar contents of pectin and cellulose in border cell walls were significantly (P<0.05) higher than in the root segments of 0-10 mm and 10-20 mm at the absence of Al. The uronic acid content of pectin in border cell walls increased obviously related to the root segments of 0-10 mm and 10-20 mm at 4.0 mmol?L-1 A1C13. The sugar content of pectin in border cell walls was increased significantly (P<0.05), but decreased seriously in hemi-cellulose and cellulose at 4.0 mmol?L-1A1C13. The Al content of pectin, hemi-cellulose and cellulose in the border cell walls were significantly (P<0.05) higher than in the root segments of 0-10 mm and 10-20 mm. Al contents of pectin 1 in the border cell walls were 14.4 and 17.4 fold to that in the root segments of 0-10 mm and 10-20 mm at 4.0 mmol?L-1 A1C13, respectively, and the Al content in the root segments of 0-10 mm was higher than in the 10-20 mm; at the same time,. hemi-cellulose and cellulose, the major compositions of cell wall, contained more Al than pectin. 【Conclusion】 These results indicated that the contents of Al binding to cell walls is related to the contents of pectin in cell walls and the border cells were more sensitive to Al than the root segments of 0-10 mm and 10-20 mm. Cell wall polysaccharides including pectin, hemi-cellulose and cellulose might play important roles in binding Al in border cell walls.

    PLANT PROTECTION
    Molecular Mapping and Monosomic Location of Stripe Rust Resistant Gene in N.Strampelli
    HU Mao-lin,CHEN Jie,CHENG Jing-jing,HOU Lu,LI Qiang,WANG Bao-tong,JING Jin-xue
    Scientia Agricultura Sinica. 2009, 42(6):  1972-1979 .  doi:10.3864/j.issn.0578-1752.2009.06.012
    Abstract ( 1208 )   PDF (433KB) ( 757 )   Save
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    【Objective】 N. Strampelli is a very important source of durable resistant material. Inheritance, location and molecular mapping of stripe rust resistance genes in N. Strampelli is important for revealing the resistance to stripe rust and developing durable cultivars. 【Method】 To confirm the resistant genes, N. Strampelli was crossed with susceptible cultivar MingXian169, Chinese spring and then backcrossed with them. The gene’s chromosome location was determined by the monosomic analysis of Chinese spring, which were then backcrossed among them. The microscope observation of F1 was performed to assess whether they were monosomic materials. To test molecular markers for N. Strampelli, 208 SSR pairs of primers, the PCR amplification, non-denatured PAGE and silver staining are used for detecting polymorphic bands. 【Result】 The resistance to SU-4 was dependently or overlappedly controlled by two recessive genes. The resistance to CYR31 was overlappedly controlled by two recessive genes. There was one recessive resistant gene controlled when tested with CYR29-mut3. It was temporarily named YrN.S. Xgwm499, Xwmc415 and Xwmc537 were located on chromorome 5BL. The map distance between Xgwm499, Xwmc415 and Xwmc537 were 7.6 cM, 5.4 cM and 10.7 cM, respectively. 【Conclusion】 The result indicated that YrN.S is a novel resistant gene which is different from other stripe rust resistance genes.

    Development of SCAR Markers for Sugarcane Mosaic Virus Resistance in Maize
    Lü Xiang-ling,WANG Bang-tai,SHI Li-yu,SHI Hong-liang,XIE Chuan-xiao,LI Xin-hai,ZHANG Shi-huang
    Scientia Agricultura Sinica. 2009, 42(6):  1980-1987 .  doi:10.3864/j.issn.0578-1752.2009.06.013
    Abstract ( 1105 )   PDF (504KB) ( 854 )   Save
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    【Objective】 Maize dwarf mosaic disease caused by sugarcane mosaic virus (SCMV) is one of the important virus diseases in China and European countries. Marker development and markers-assisted selection (MAS) is an efficient way for disease-resistant breeding. 【Method】Resistant and susceptible DNA bulks were composed by using DNAs of 10 resistant and 12 susceptible inbred lines, respectively. Polymorphic AFLP markers were screened between two bulks and then transformed into SCAR (sequence characterized amplified region) markers. These SCAR markers associated with SCMV resistance was analyzed with disease incidence of 100 inbred lines. 【Result】 Two polymorphic AFLP markers, P66M38-220 and P55M51-240, were identified, and P66M38-220 was transformed into SCAR112, which was validated to be highly associated with SCMV resistance. 【Conclusion】 Improved BSA method is an efficient way for developing markers tightly linked with target traits. The SCAR112 developed in this study could be used for MAS of sugarcane mosaic virus resistance in maize.

    Eukaryotic Expression and Functional Analysis of Pcpme3 From Phytophthora capsici
    JIA Yong-jian,FENG Bao-zhen,LI Pei-qian,ZHANG Xiu-guo,FU Hong-bo
    Scientia Agricultura Sinica. 2009, 42(6):  1988-1993 .  doi:10.3864/j.issn.0578-1752.2009.06.014
    Abstract ( 1242 )   PDF (459KB) ( 816 )   Save
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    【Objective】 The Pcpme3 gene of Phytophthora capsici was expressed in Eukaryotic expression system to obtain purified protein and specific antiserum, and to detect the function of Pcpme3 in phytophthora-infected pepper through Western blot. 【Method】 Mature peptide of Pcpme3 obtained by RT-PCR was cloned into pPIC9K, and the eucaryotic expression plasmid was constructed. The recombinant plasmid was transformed into Pichia pastoris GS115 by electroporation. After induced by methanol, the culture supernatant was analyzed by SDS-PAGE, and the antiserum was prepared. 【Result】 The high copy transformants with Mut+/His+ phenotype were selected by screening on MD, MM and G418 medium and PCR. The culture supernatant was analyzed by SDS-PAGE after induced by 1% methanol, and the results showed that a specific protein of about 43 kD was expressed. The specific antiserum was produced after the rabbit was immunized with recombinant protein, and Western blot indicated that Pcpme3 could be highly expressed in phytophthora-infected peppers, and the expressed level became stronger with the symptom became heavier. 【Conclusion】 Antiserum prepared from expressed protein can be used to detect the function of Pcpme3 in the pathogenesis of Phytophthora capsici.

    Isolation and Structural Indentification of Herbicidal Toxin Fractions Produced by Pythium aphanidermatum
    XU Jiao,XU Wen-chao,KANG Zhan-hai,LIU Bao-feng,ZHANG Jin-lin
    Scientia Agricultura Sinica. 2009, 42(6):  1994-2001 .  doi:10.3864/j.issn.0578-1752.2009.06.015
    Abstract ( 966 )   PDF (375KB) ( 658 )   Save
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    【Objective】 In order to understand the compsition and structure of herbicidal component from Pythium aphanidermatum, the isolation and structural indentification was researched. 【Method】 The culture filtrate was extracted by ethyl acetate, petroleum, chloroform with the same volume respectively and the activity of the crude toxin was bioassayed, the toxin was separated by using the method of TLC, then the main fraction was separated by HPLC, and the structure was analyzed by the sepctrum of IR,13C-NMR and 1HNMR. 【Result】 The results showed that the ethyl acetate extracts had the strongest herbicidal activity. Using the method of TLC, the bioassay results showed that the extracts with Rf 0.19 had strongest effect on weeds and the inhibition to Digitaria sanguinalis L. and Amaranthus retroflexus L. reached five degree, the component was got using analysis, also was proved to be dimethyl o-phthalate from the sepctrum of IR, 13C-NMR and 1HNMR. 【Conclusion】 Dimethyl o-phthalate was one of the components from the toxin, and it had herbicidal activity.

    Molecular Identification of Badnavirus in Dracaena sanderiana from Hubei of China
    CHEN Xiu,RUAN Xiao-lei,ZHAO Qin,LI Hua-ping
    Scientia Agricultura Sinica. 2009, 42(6):  2002-2009 .  doi:10.3864/j.issn.0578-1752.2009.06.016
    Abstract ( 1177 )   PDF (360KB) ( 960 )   Save
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    【Objective】 In order to identify badnavirus in Dracaena sanderiana with typical mottle symptom from Hubei of China, and to compare the genome molecular variation between isolates of Dracaena sanderiana badnaviruses, the complete genome sequence of Dracaena sanderiana badnavirus Hubei isolate was obtained. 【Method】 The complete genome sequence was got through subsection cloning and sequencing, and the sequences were analyzed with BLAST and other softwares. 【Result】 The complete circular genome nucleotide sequence comprised 7 522bp, which contained 7 open reading frames (ORFs) on the plus-strand that potentially encoded proteins of 17.58 kD, 14.93 kD, 214.77 kD, 11.86 kD, 11.31 kD, 16.12 kD and 11.00 kD. The genome organization was highly similar with the newly reported badnavirus Dracaena mottle virus (DrMV). The size of Hubei isolate was 9 nucleotides shorter than DrMV. They shared 99.7% identities over the entire nucleotide sequences, and 99.3%, 100%, 99.2% over the presumed amino acid sequences of ORFs 1-3, respectively. 【Conclusion】 The genome organization and the comparison analysis of sequences between the genomes of badnaviruses revealed that Dracaena sanderiana with typical mottle symptom from Hubei of China contained badnavirus, and Dracaena sanderiana badnavirus Hubei isolate and DrMV belonged to the same species, and the genome molecular variation between isolates of Dracaena sanderiana badnavirus was very small, which was much different from previous results that significant molecular variation existed in badnavirus isolates from different plants.
    Preparation of Lambda-Cyhalothrin Microemulsion with Biodiesel as Substitute Solvent and Study of Its Performance
    LI Gang,ZHANG Zheng-qun,MA Chao,MU Wei,LIU Feng
    Scientia Agricultura Sinica. 2009, 42(6):  2010-2018 .  doi:10.3864/j.issn.0578-1752.2009.06.017
    Abstract ( 1440 )   PDF (416KB) ( 1042 )   Save
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    【Objective】 The objective was to study the application of environment-friendly solvent, biodiesel as substitute solvent, in preparing the lambda-cyhalothrin microemulsion, and to provide a reference for biodiesel widely used in green pesticide preparations. 【Method】 Using phase transfer method, three 2.5% lambda-cyhalothrin microemulsions with different solvents of biodiesel, cyclohexanone and 0# diesel were prepared. The phase behavior, toxicity to Aphis gossypii Glover and larva of Pieris rapae, and the safety to cotton of the formulation with different solvents were compared. 【Result】 Biodiesel had good dissolving ability to Lambda-cyhalothrin, could reach 326 g?L-1 under 25℃. The microemulsion with biodiesel as solvent had transparent appearance, good low-temperature performance, and the decomposed ratios of active ingredients only 2.0% after 14 days storage at 54℃. The LC50 values to A. gossypii Glover and 3rd larva of P. rapae had no difference among the three formulations prepared with different solvents, and had no effect on the shoot height and total chlorophyll content of cotton seedlings. 【Conclusion】Good lambda-cyhalothrin microemulsion formulation could be prepared with biodiesel as the solvent, the biodiesel has no negative effects on the activity of active ingredients, and is comparatively safe to cotton seedlings.

    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Fuzzy Synthetic Evaluation of Soil Quality in Coastal Reclamation Region of North Jiangsu Province
    YAO Rong-jiang,YANG Jing-song,CHEN Xiao-bing,YU Shi-peng,LI Xiao-ming
    Scientia Agricultura Sinica. 2009, 42(6):  2019-2027 .  doi:10.3864/j.issn.0578-1752.2009.06.018
    Abstract ( 1000 )   PDF (714KB) ( 663 )   Save
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    【Objective】 This study was to systematically comprehend soil quality status and its spatial pattern in coastal saline reclamation region of north Jiangsu Province, and acquire management measures to improve soil fertility and quality and restrain soil salinity. 【Method】 With the application of fuzzy synthetic evaluation method and GIS technology, fuzzy synthetic evaluation and classification of regional soil quality was conducted, and spatial patterns of synthetic evaluation results of regional soil quality was obtained to provide a theoretical reference to scientific amelioration and field management of moderate-and-low-yield farmlands. 【Result】 The results indicated that soil quality status across the study area was generally low, soil organic matter (SOM) and N nutrient was in severe deficiency, and it is particularly the case for available N. Soil bulk density showed week variation and other evaluation indices all exhibited moderate variation. SOM was the most important evaluation factor of soil quality, while soil salinity and groundwater mineralization were limiting factors of soil quality. Soil quality status differed significantly at different parts of the study area, and the spatial pattern of soil quality was similar to that of SOM, while it was opposite to that of salinity. Soil quality in the cotton-growing field was superior to that in the rice-growing field generally. The area of moderate soil quality was the most in the study area, followed by that of low soil quality, and the area of high soil quality was the least. 【Conclusion】 The soil quality map can be used for the improvement of moderate-and-low-yield farmlands, comprehensive treatment and rational fertilization of saline-alkali soil in our study area. The field irrigation and drainage installations should be improved in the area of low soil quality, while agro-biological treatments should be employed in the area of moderate soil quality.

    Study on Abundance and Deficiency Indices of Soil Available P, K and B for Winter Rapeseed in Yangtze River Valley Based on ASI Method
    ZOU Juan,LU Jian-wei,CHEN Fang,LI Yin-shui
    Scientia Agricultura Sinica. 2009, 42(6):  2028-2033 .  doi:10.3864/j.issn.0578-1752.2009.06.019
    Abstract ( 1172 )   PDF (227KB) ( 691 )   Save
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    【Objective】 To provide a theoretical basis for soil testing and fertilizing recommendation for rapeseed production in the region of Yangtze River valley. 【Method】 Field experiments were conducted in 10 main rapeseed production provinces in the valley during 2004-2006. A series of soil samples were taken before rapeseed transplanting and analyzed by ASI (Agro Services International) method. The abundance and deficiency indices were determined based on the relationship between crop relative yield and available soil nutrient values. Compared to the complete treatment, the relative yields of 60%, 75%, 90% and 95% obtained from the -P, -K and -B treatments were selected to establish the abundance and deficiency indices of soil available P, K and B for winter rapeseed. 【Result】 The extreme deficiency, deficiency, slight deficiency, optimum and abundance indices for soil available P were <5.5, 5.5-12.5, 12.5-28.5, 28.5-38.0 and >38.0 mg P?L-1, respectively. The extreme deficiency critical values for soil available K and B were not appeared in this research. The deficiency, slight deficiency, optimum and abundance critical values for soil available K and B were <30, 30-75, 75-100 and >100 mg K?L-1; and <0.25, 0.25-0.95, 0.95-1.50 and >1.50 mg B?L-1, respectively. 【Conclusion】 The relative yields of the -P, -K and -B treatments were significantly correlated with soil available P, K and B contents. The soil available P, K and B critical indices established using the ASI systematic approach and field experiments can be used as guidance for soil testing and fertilizing recommendation for winter rapeseed production in the valleys.

    Study on Nitrogen Management and Irrigation Methods of Greenhouse Tomato
    GAO Bing,LI Jun-liang,CHEN Qing,LIU Qing-hua,WANG Jing
    Scientia Agricultura Sinica. 2009, 42(6):  2034-2042 .  doi:10.3864/j.issn.0578-1752.2009.06.020
    Abstract ( 1007 )   PDF (283KB) ( 940 )   Save
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    【Objective】Nitrogen management and irrigation model of greenhouse tomato were studied. 【Method】 Four treatments, control (CK), conventional nitrogen (CN),optimized nitrogen (ON) and recommended nitrogen (RN) management treatments were designed to investigate tomato yield, N application rate, loss of N, the agronomic efficiency of chemical N fertilizer and irrigation water. 【Result】 Tomato yield of CK reached a higher level although any chemical N fertilizer was not used, the phenomenon of yield reduction appeared with the increasing of the rate of N sidedressed in winter-spring season. The rate of N sidedressed of CN is 600 kgN?hm-2, and the irrigation water is about 7 500 m3?hm-2, there are 1 416 kgN?hm-2 has been lossed as the result of the unreasonably water and N management; Compared with CN, the loss sidedressed N and the loss of total N decreased by 50% and 32.2% in RN; The rate of N sidedressed of ON is 314 and 124 kgN?hm-2 in two seasons respectively, and the irrigation water is 3 900 and 4 550 m3?hm-2, with the total loss of N reduced by 38.6%. The agronomic efficiency of chemical N fertilizer in CN, ON and RN treatments in the whole year is 0, 24.9 and 0.3 kgFW?kg-1N, respectively. Tha agronomic efficiencies of irrigation water of the conventional and optimized irrigation treatments are 12.2 and 23.2 kgFW?m-3, respectively. The ON can reduce 4 000 yuan/hm2 by decreasing N fertilizer and irrigation water input. 【Conclusion】 The rate of N sidedressed is not the leading factor restricting the increase of tomato yield. Water and N management mothods of optimizing N sidedressing and irrigation by employing bubbler irrigation system during the growing season, and applying calcium cyanamide mixed with wheat straw in summer-fallow are considered as the better water and N management.

    Isolation and Biological Characteristic Investigation on Efficient Nitrogen-Fixing Bacilli
    SUN Jian-guang,ZHANG Yan-chun,XU Jing,HU Hai-yan
    Scientia Agricultura Sinica. 2009, 42(6):  2043-2051 .  doi:10.3864/j.issn.0578-1752.2009.06.021
    Abstract ( 1330 )   PDF (590KB) ( 1760 )   Save
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    【Objective】 Efficient nitrogen-fixing bacilli were isolated, and the relationships between microbial nitrogenase activity, competition ability and inoculation effect were investigated. The influence of inoculating nitrogen-fixing bacillus on the bacterial community in crop rhizosphere was explored. 【Method】 Nitrogen-free medium was used to isolate nitrogen-fixers and heat treatment was used for the nitrogen-fixing bacilli. Nitrogenase activity was determined with acetylene reduction assay. Microbial competition ability and inoculation effect were tested with pot culture in greenhouse. Strain identification was carried out based on the morphology, physiology, biochemical test and 16S rDNA sequences analysis. 【Result】 Three strains termed as GD062, GD082 and GD282 with relative high nitrogenase activity and strong nitrogen-fixing ability were isolated, which were identified as Bacillus spp. and Paenibacillus sp. respectively. Further result showed that strain GD082 have strong competition ability, produced an inoculation effect as well as the chemical fertilizer treatment did in the natural soil. Result also indicated that no correspondence between nitrogenase activity and inoculation effect was found. Strain competition ability influences the inoculation effect. Rhizosphere bacterial community increased significantly with the inoculation of nitrogen-fixing bacilli. 【Conclusion】 It is meaningful in agricultural practice to offer crop nitrogen nutrient through inoculation of nitrogen-fixers. Selection of efficient nitrogen fixer is a complexed process. There is no correspondence between nitrogenase activity and inoculation effect. Strain competition ability influences the inoculation effect even stronger. A nitrogen-fixing bacillus GD082 with high nitrogenase activity, strong nitrogen-fixing ability and competition ability was obtained, which is possible to be further developed as an excellent strain used in the microbial fertilizer production.

    Methane and Nitrous Oxide Emissions from Rice-Fish and Rice-Duck Complex Ecosystems and the Evaluation of Their Economic Significance
    YUAN Wei-ling,CAO Cou-gui,LI Cheng-fang,ZHAN Ming,CAI Ming-li,WANG Jin-ping
    Scientia Agricultura Sinica. 2009, 42(6):  2052-2060 .  doi:10.3864/j.issn.0578-1752.2009.06.022
    Abstract ( 1363 )   PDF (418KB) ( 1626 )   Save
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    【Objective】 The rice-duck and rice-fish ecological systems are the major complex planting and breeding models of rice paddy fields in Southern China. Studying on methane and nitrous oxide emissions and their economic value from these ecosystems can provide a theoretical and practical basis for further development and utilization of these classical agricultural techniques. 【Method】 Methane and nitrous oxide emissions from rice-duck (RD) and rice-fish (RF) ecological systems were measured in situ by using static chambers technique. Using global warming potentials (GWPs), the greenhouse effects of methane and nitrous oxide and the economic value were assessed. 【Result】 Results shown that the peak of methane emission fluxes appeared at full tillering stage and complete heading stage, and the average emission fluxes from RD and RF were significant lower than that from CK. Nitrous oxide fluxes remained low in flooding irrigation and high in draining of water. Compared with CK, the total amount of nitrous oxide emissions from RD significantly higher and RF lower. In 2006 and 2007, the total greenhouse effects of methane and nitrous oxide were 4 728.3 kg CO2?hm-2 and 4 611.0 kg CO2?hm-2, 4 545.0 kg CO2?hm-2 and 4754.3 kg CO2?hm-2, respectively. So the cost of greenhouse effect was 970.89 yuan/hm2 and 946.81 yuan/hm2, 933.25 yuan/hm2 and 976.23 yuan/hm2, respectively, which were significant lower than that from CK (5 997.6 yuan/hm2 and 5 391.5 yuan/hm2). Except for the environment cost of methane and nitrous oxide, the achieved economic benefits from RD and RF were higher than that from CK (2 210.64 yuan/hm2 and 4 881.92 yuan/hm2; 3 798.37 yuan/hm2 and 5 310.64 yuan/hm2, respectively). 【Conclusion】 Rice-duck and rice-fish complex ecological planting and breeding models can effectively decrease and control methane and nitrous oxide emissions, and they are effective strategies to reduce greenhouse gas from rice paddy fields. Thus, they are worthy to be populated by farmer in view of their economic benefit.

    HORTICULTURE
    Chromosomal Location and AFLP Marker Screening of Genic Male Sterile Gene in Chinese Cabbage
    YUAN He,ZHANG Cheng-he,LIU Hai-he,XUAN Shu-xin,LI Xiao-feng,SHEN Shu-xing
    Scientia Agricultura Sinica. 2009, 42(6):  2061-2067 .  doi:10.3864/j.issn.0578-1752.2009.06.023
    Abstract ( 1009 )   PDF (447KB) ( 1127 )   Save
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    【Objective】 Chromosome localization of genes is an important work for genetic study and breeding practice. In this paper Dayang AB line and trisomics of Chinese cabbage were used for assigning the male sterile gene to a particular chromosome and screening the molecular markers linked to the locus. 【Method】 Locating the male sterile gene to a particular chromosome was conducted by primary trisomic analysis and χ2 test, and screening the molecular markers linked to the gene locus by AFLP techniques.【Result】 All F1 progenies in the nine combinations derived from sterile line × Tri-1 to sterile line × Tri-9 were male ferterile. This indicated that the male fertility was dominant to the sterility in Chinese cabbage Dayang AB line. Selecting the trisomic(2n+1) plants from each F1 progenies and being selfed or test-crossed to the sterile line, and the results showed that only Tri-4 combination displayed 15.5﹕1 segregation proportion of male fertile plants to male sterile plants in F2 and 3.24﹕1 in Tc (test-cross), which disobeyed Mendelian segregation ratios for a monogenetic factor, while other combinations varied from 2.60:1 to 3.76﹕1 in F2 and 0.81﹕1 to 1.48﹕1 in Tc, which fitted Mendelian segregation proportions (3﹕1 and 1﹕1) for a monogenetic factor. These results indicated that the gene controlling the male sterility of Chinese cabbage Dayang AB line was associated with trisomic 4, i.e. the gene was located on the chromosome 4. Further trisomic analysis based on the chromosomal segregation (assuming the gene closely linked to the centromere ) and on the chromatid segregation(assuming the gene apart from the centromere ) showed that the gene was closer to the centromere . Furthermore, an AFLP marker (E34M49) linked to the sterile gene was preliminarily screened out from 124 random primers. 【Conclusion】 Using primary trisomic analysis and molecular screening techniques, the sterile gene of Chinese cabbage was assigned to chromosome 4 and an AFLP marker (398 bp) linked to the locus was obtained preliminarily.

    Isolation and Functional Characterization of BrARGOS Gene from Chinese Cabbage
    WANG Bao
    Scientia Agricultura Sinica. 2009, 42(6):  2068-2075 .  doi:10.3864/j.issn.0578-1752.2009.06.024
    Abstract ( 918 )   PDF (583KB) ( 828 )   Save
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    【Objective】 A new homolog of Arabidopsis ARGOS was isolated from leaves of Chinese cabbage inbred line Rikaze. Alignment of amino acid sequences and expression pattern were analyzed. The function of the gene was investigated by overexpressing it in Arabidopsis. The relation between the expression quantity of BrARGOS and the leaf heterosis of Chinese cabbage was analyzed. These results have provided data for investigating the functions of BrARGOS in Chinese cabbage. 【Method】 The cDNA sequence homologous to ARGOS was isolated from leaves of Chinese cabbage by RT-PCR. The secondary structure of the protein was predicted by DNAMAN PROGRAM. Expression pattern of BrARGOS was analyzed in different organs of Chinese cabbage by semiquantitive RT-PCR. The function of BrARGOS was investigated by transforming it into Arabidopsis. The relation between the expression quantity of BrARGOS and the leaf heterosis of Chinese cabbage was analyzed by semiquantitive RT-PCR. 【Result】 Using the Arabidopsis ARGOS sequence, a Blast search was performed against DNA sequences. The nucleotide sequence in BAC bank of Chinese cabbage was obtained and specific primers were designed. A new homolog with ARGOS was isolated from leaves of Chinese cabbage by RT-PCR and named BrARGOS. Alignment of predicted amino acid sequences of BrARGOS, Arabidopsis and tomato showed the proteins all contained Leu-rich C terminuses. The secondary structure of BrARGOS contained random coils, extended strands and alpha helixes. BrARGOS was expressed in all organs of Chinese cabbage, at high levels in young siliques and cotyledons, whereas at lowest level in cauline leaves. The result that NPTⅡ was transformed into Arabidopsis genome by examining genome DNA of positive Arabidopsis transformants confirmed foreign BrARGOS was transformed into Arabidopsis genome. Compared with vector control plants, the transgenic Arabidopsis had larger leaves and flowers, higher leaf fresh weight and plant height. 【Conclusion】 A new homolog with ARGOS was isolated from leaves of Chinese cabbage inbred line Rikaze. Expression pattern and function of BrARGOS were analyzed. Overexpression of BrARGOS in Arabidopsis increased aerial organ size. The Chinese cabbage BrARGOS gene is related to regulating plant organ size.

    Adavances in Genetic Engineering of Floral Scent
    XIANG Lin,CHEN Long-qing
    Scientia Agricultura Sinica. 2009, 42(6):  2076-2084 .  doi:10.3864/j.issn.0578-1752.2009.06.025
    Abstract ( 1067 )   PDF (370KB) ( 980 )   Save
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    Floral scent plays an important role in the reproductive processes of many plants and also enhances the aesthetic properties of ornamental plants. In the past decade, investigators have witnessed significant progress in both identifying genes and enzymes involved in the biosynthesis of floral scent volatiles. Survey of the latest advances in the clone of floral scent- related genes, regulation of floral scent production and metabolic engineering of floral scent is made. Finally, the perspectives and prospects of research on floral scent are presented.

    Cellular Mechanism of Reproductive Barrier During Cross Breeding Between Dendranthema grandiflorum cv. Aoyuntianshi and D. japonense
    SUN Chun-qing,CHEN Fa-di,FANG Wei-min,LIU Zhao-lei,MA Jing,TENG Nian-jun,SHE Lin-fang
    Scientia Agricultura Sinica. 2009, 42(6):  2085-2091 .  doi:10.3864/j.issn.0578-1752.2009.06.026
    Abstract ( 1044 )   PDF (728KB) ( 1111 )   Save
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    【Objective】 Our aim is to reveal the factors resulting in low seed set during chrysanthemum breeding and seriously influencing utilization of its wild germplasm.【Method】 The interspecific distant hybridization was carried out between Dendranthema grandiflorum cv. Aoyuntianshi (female parent) and D. japonense (male parent). In addition, pollen viability of male parent was tested in culture medium just before pollination, and germination behavior of pollen grains on the stigma after pollination was examined using aniline blue staining method. In addition, embryo development after fertilization was observed with technique of paraffin section. 【Result】 Pollen viability was 10.0% just before pollination. The average number of pollen grains germinating on stigma increased slowly between 0.5 h and 8 h after pollination, and 16.1 pollen grains germinated on each stigma at 8 h after pollination. But after that, a large number of pollen grains germinated, and the number was 40.2 at 12 h after pollination. At 24 h after pollination, the number reached the highest value, 51.5. Subsequently, the number began to decrease and dropped to 42.3 at 48 h after pollination. During the process of embryo development, the percentage of normal embryos decreased gradually due to degeneration of many embryos and the values were 26.7% at stage of globular embryo, 18.3% at stage of heart embryo, 14.4% at stage of torpedo embryo, and 10.6% at stage of cotyledon embryo, respectively. Seed set in the artificial crossing was 7.6%, but the number under open pollination conditions was 51.4%. 【Conclusion】 Pollen viability of male parent and germination behavior of pollen grains on stigmas have no significant effect on seed set in the artificial crossing. And hybrid inviability, i.e. the degeneration or abortion of a large number of embryos during their developmental processes, may be the main factor leading to low seed set during cross breeding between D. grandiflorum cv. Aoyuntianshi and D. japonense.

    STORAGE·FRESH-KEEPING·PROCESSING
    Effects of Propylene and 1-Methylcyclopropene on Expressions of ACC Synthase and ACC Oxidase Genes in Persimmon Fruits
    LIU Le,RAO Jing-ping,CHANG Xiao-xiao,YI Shun-chao
    Scientia Agricultura Sinica. 2009, 42(6):  2092-2097 .  doi:10.3864/j.issn.0578-1752.2009.06.027
    Abstract ( 1233 )   PDF (347KB) ( 1031 )   Save
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    【Objective】 The effects of postharvest treatment with propylene and 1-MCP on ethylene production and expressions of ACS and ACO genes in Fupingjianshi persimmon (Diospyros kaki L.) fruit were investigated. 【Method】 Fruits were treated with propylene and 1-MCP, than stored at 20℃, ethylene released rate of the treated fruits were measured at regular intervals and RNA was extracted for Northern blotting analysis. 【Result】 Treatment with propylene accelerated the expression of ACS and ACO genes and enhanced the ethylene biosynthesis, while treatment with 1-MCP inhibited the expression of two genes and their ethylene biosynthesis. Furthermore, different effects on expressions caused by treatments with propylene and 1-MCP existed in various fruit tissues. The inhibitory effect on ACS and ACO genes by 1-MCP was the strongest in pulp and core tissues, followed by pericarp and the area near fruit stalk while the enhanced effect on ACS and ACO genes by propylene increased from pulp, core, and pericarp to the area near fruit stalk. Expression of each member of ACS and ACO families in various tissues were also completely different, ACS3 gene just expressed in the area near fruit stalk and didn’t express in other tissues, but ACS2 gene expressed in four tissues by treatment with propylene.【Conclusion】 Treatment with propylene accelerated the expression of ACS and ACO genes, while treatment with 1-MCP inhibited the expression of ACS and ACO genes. Different effects on expressions caused by treatments with propylene and 1-MCP existed in various fruit tissues. Expression of each member of ACS and ACO families in various tissues were also completely different.

    Heat-Induced Gelation of Myofibrillar Proteins as Affected by pH——A Low Field NMR Study
    HAN Min-yi,FEI Ying,XU Xing-lian,ZHOU Guang-hong
    Scientia Agricultura Sinica. 2009, 42(6):  2098-2104 .  doi:10.3864/j.issn.0578-1752.2009.06.028
    Abstract ( 1121 )   PDF (368KB) ( 863 )   Save
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    【Objective】 The objective of this study is to discuss the effect of pH on the water holding capacity (WHC) and water mobility of pork myofibrillar protein heat-induced gelation. 【Method】 Myo?brillar proteins were extracted from porcine muscle, and their heat-induced gelation water properties were characterized using low-?eld nuclear magnetic resonance (NMR) T2 relaxometry. The WHC of the system was determined by centrifugation method. 【Result】 The NMR data was fitted to four components, which were merged into three states of water, i.e. immobile, mobile and free water. T2 relaxation time, peak area, and WHC which attribute to mobile water increased as pH away from pI, the principal components analysis result showed that the sample near pI clustered differently from others. 【Conclusion】 Improvement of WHC could be ascribed to the increasing of the mobile water, and the possible increase of the myo?brilar protein gel network pore diameter may leads to more water could be entrapped.

    Isolation and Structure Analysis of the Components of Anti-Gram- Negative Bacteria by the Strain CP7 of Paenibacillus polymyxa
    CHEN Hai-ying,LIAO Fu-ping,LIN Jian-rong
    Scientia Agricultura Sinica. 2009, 42(6):  2105-2110 .  doi:10.3864/j.issn.0578-1752.2009.06.029
    Abstract ( 1182 )   PDF (428KB) ( 1041 )   Save
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    【Objective】 The components of anti-gram-negative bacteria by the strain CP7 of Paenibacillus polymyxa were purified, and the structure was identified. 【Method】 The antibacterial components from CP7 strain were purified by the solid phase extraction, cation-exchange chromatography, gel-filtration chromatography and HPLC. The components were analyzed with the technique of the ultraviolet-visible spectrum, circular dichroism spectroscopy, IR and MALDI-TOF-TOF-MS. 【Result】Three components of anti-gram-negative bacteria, B, C1 and C2, were purified from Paenibacillus polymyxa CP7 broth. Their molecular weight was 1 155 D, 1 169 D and 1 191 D, respectively. The molecular formula of C1 was C53H100O13N16, which was composed by six DAB, two threonine and two leucine, with the same structure as polymyxin E1. 【Conclusion】Three kinds of small molecule peptide of anti-gram-negative bacteria were isolated from the CP7 fermentation broth. It is speculated that B was also a member of the polymyxin family, and C2 may be a new family member.

    ANIMAL SCIENCE·RESOURCE INSECT
    Genetic Variation of Porcine Sperm Adhesion Molecule 1 (SPAM1) Gene and Its Association with Litter Size in a White Duroc×Erhualian Resource Population
    WU Yan-bo,CHEN Cong-ying,ZHANG Zhi-yan,GUO Yuan-mei,GAO Jun
    Scientia Agricultura Sinica. 2009, 42(6):  2111-2117 .  doi:10.3864/j.issn.0578-1752.2009.06.030
    Abstract ( 932 )   PDF (301KB) ( 700 )   Save
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    【Objective】 Porcine sperm adhesion molecule 1 (SPAM1) gene plays an important role in fertilization. It is an important candidate gene for porcine litter size. In this study, identification of genetic variation of SPAM1 gene and association study with litter size were carried out. 【Method】 Detection of polymorphisms of intron1 g298 C>T, intron1 g357 C>T and intron3 g380 C>T and construction of haplotypes were carried out in 305 F2 sows from a white Duroc × Erhualian resource population, whose litter size was recorded in detail in 3 subsequent parities. Association study of genotypes of each SNP and haplotypes with total number born (TNB), number born alive (NBA) and number stillborn (NSB) was also performed. 【Result】 The results showed that, compared to individuals of CT and TT at SNP intron1 g298 C>T, individuals of CC had significantly less TNB and NBA (P<0.05). Sows of CT had significantly higher number of NSB than sows of TT (P<0.05). For SNP intron1 g357 C>T, the CT sows produced significantly more NSB compared to CC and TT individuals. And for SNP intron3 g380 C>T, CC individuals had significantly less NBA and more NSB than CT individuals. Six haplotypes were constructed in this study. Haplotype TTT showed significantly higher TNB and NBA than the other 5 haplotypes. Individuals of TCT and TTC had significantly less NSB than individuals of CTC, TTT and TCC (P<0.05). 【Conclusion】 SPAM1 gene has significant association with porcine litter size.

    Polymorphisms of TLR1 Gene and Their Relationship with Somatic Cell Score in Holstein Cows
    LI Chun-miao,SHI Wan-hai,CHU Ming-xing,AN Yong-fu,CHEN Hong-quan,DI Ran,FANG Li
    Scientia Agricultura Sinica. 2009, 42(6):  2118-2125 .  doi:10.3864/j.issn.0578-1752.2009.06.031
    Abstract ( 986 )   PDF (574KB) ( 783 )   Save
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    【Objective】 The objectives of the present study were to elucidate the relationship between Toll-like receptor 1 (TLR1) gene and mastitis resistance of Holstein cows and to provide a scientific basis for marker-assisted selection for mastitis resistance of dairy cattle. 【Method】 Polymorphisms of all exons of TLR1 gene were detected in 208 Holstein cows by PCR-SSCP and PCR-RFLP. The relationship between polymorphisms of TLR1 gene and somatic cell score (SCS) in Holstein cows was analyzed by least squares linear model. 【Result】 Four polymorphic sites, G1409A, A1475C, G1550A and G1596A, were found, and G1596A mutation resulted in an amino acid change of isoleucine→valine. For G1409A polymorphic site, two genotypes (AA and AB) were found in 208 Holstein cows by digesting the PCR products with restriction endonuclease BsiYⅠ, and the frequency of AA and AB genotypes was 0.375 and 0.625, respectively. Chi-square test showed that Holstein cows were not in Hardy-Weinberg equilibrium. Least squares mean of SCS for AA was not significantly lower than that for AB (P>0.05). For A1475C polymorphic site, two genotypes (CC and CD) were found in 208 Holstein cows by digesting the PCR products with restriction endonuclease BstXⅠ, and the frequency of CC and CD genotypes was 0.178 and 0.822, respectively. Holstein cows were not in Hardy-Weinberg equilibrium. Least squares mean of SCS for CC was not significantly lower than that for CD (P>0.05). For G1550A polymorphic site, three genotypes (EE, EF and FF) were found in 208 Holstein cows by digesting the PCR products with restriction endonuclease BsiYⅠ, and the frequency of EE, EF and FF genotypes was 0.236, 0.370 and 0.394, respectively. Holstein cows were not in Hardy-Weinberg equilibrium. The difference of the least squares means for SCS between EE, EF and FF was insignificant (P>0.05). For G1596A polymorphic site, three genotypes (GG, GH and HH) were found in 208 Holstein cows by digesting the PCR products with restriction endonuclease BclⅠ, and the frequency of GG, GH and HH genotypes was 0.260, 0.567 and 0.173, respectively. Holstein cows were in Hardy-Weinberg equilibrium. Least squares mean of SCS for HH was significantly lower than that for GH (P<0.01) or GG (P<0.0001), least squares mean of SCS for GH was significantly lower than that for GG (P<0.01). 【Conclusion】 HH was the most favorable genotype, GG was the most unfavorable genotype for mastitis resistance. The results preliminarily indicated that allele H of G1596A polymorphic site of TLR1 gene is a potential DNA marker for improving mastitis resistance in dairy cattle.

    Quantity of Microorganisms Associated with Solid and Liquid Phases of Rumen Contents Using RT- PCR
    YANG Jin-li,HOU Xian-zhi,GAO Ai-wu
    Scientia Agricultura Sinica. 2009, 42(6):  2126-2132 .  doi:10.3864/j.issn.0578-1752.2009.06.032
    Abstract ( 1232 )   PDF (318KB) ( 1014 )   Save
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    【Objective】 Using Real-time PCR (RT-PCR)assays to quantify the microbes associated with particular phases of rumen contents. 【Method】 Whole rumen contents were sampled from three Mongolian sheep and partitioned into solid- and liquid-associated microorganisms. 【Result】 On the basis of 16S or 18S rRNA gene copy number analysis, the number of general bacteria, general anaerobic fungi and three representative ruminal cellulolytic bacteria (Ruminococcus albus, Ruminococcus flavefaciens, Fibrobacter succinogenes) associated with rumen solid content were 7.22-fold (P<0.05), 56.85-fold (P<0.01), 2.69-fold (P<0.05), 4.19-fold (P<0.01) and 7.59-fold (P<0.01) as those associated with rumen liquid content. The abundance of F. succinogenes in the solid phase was 0.55%, which was higher than that of R. flavefaciens (0.53%) and R. albus (0.09%). However in the liquid phase R. flavefaciens accounted for 0.91% of the bacterial rRNA gene copies, which was more abundant than F. succinogenes (0.52%) and R. albus (0.24%). 【Conclusion】 RT-PCR method could accurately detect the distribution and abundance of microorganisms associated with particular phase of rumen content.

    Polymorphic and Linkage Analysis of Microsatellite OarJL36 and FecB Gene in Sheep
    CHU Ming-xing,ZHANG Bao-yun,WANG Ping-qing,FANG Li,DI Ran,MA Yue-hui,LI Kui
    Scientia Agricultura Sinica. 2009, 42(6):  2133-2141 .  doi:10.3864/j.issn.0578-1752.2009.06.033
    Abstract ( 1028 )   PDF (472KB) ( 624 )   Save
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    【Objective】 The objectives of the present study were to elucidate the linkage relationship between microsatellite locus OarJL36 and FecB gene of Small Tail Han sheep and to provide a scientific basis for marker-assisted selection for high prolificacy in sheep. 【Method】 Genetic polymorphisms of microsatellite locus OarJL36 which was the closest linked marker to the ovine prolificacy major gene FecB were detected in high fecundity sheep breeds (Small Tail Han and Hu sheep) and low fecundity sheep breeds (Texel and Dorset). The linkage disequilibrium between microsatellite locus OarJL36 and FecB gene of Small Tail Han sheep was also analyzed. 【Result】 There was a same FecB mutation (A746G) of BMPR-IB gene in both Small Tail Han and Hu sheep as that in Booroola Merino ewes. The same mutation did not exist in both Texel and Dorset sheep. The frequency of BB, B+ and ++ genotype was 0.500, 0.386 and 0.114 in Small Tail Han sheep, and was 0.800, 0.200 and 0.000 in Hu sheep, respectively. Ten alleles and 31 genotypes were detected at OarJL36 in 463 individuals from four sheep breeds, in which 160 bp was the shortest allele and 200 bp was the longest allele. The 182 bp was the predominant allele in Small Tail Han (n=308) and Hu sheep (n=60), of which frequency was 0.696 and 0.925, but 0.085 and 0.125 in Texel (n=47) and Dorset (n=48). Especially, the 182 bp allele frequency was 0.915 and 0.938 in BB genotype group from Small Tail Han and Hu sheep, but 0.129 in ++ genotype group from Small Tail Han sheep. The polymorphism information content (PIC) was 0.475, 0.139, 0.661, 0.768, 0.153, 0.588, 0.843, 0.116 and 0.212 in Small Tail Han, Hu, Texel, Dorset, BB group from Small Tail Han, B+ group from Small Tail Han, ++ group from Small Tail Han, BB group from Hu sheep, B+ group from Hu sheep, respectively. In Small Tail Han sheep, linkage analysis indicated that there were fervent linkage disequilibrium as well as certain recombination between 182 bp allele of microsatellite locus OarJL36 and B allele of FecB gene (D′=0.614) , and complete linkage between 160 bp,192 bp, 196 bp, 200 bp alleles of microsatellite OarJL36 and + allele of FecB gene (D′=1.000). 【Conclusion】 These results preliminarily showed that 182 bp allele of microsatellite locus OarJL36 is a genetic marker closely linked to B allele of the prolificacy major gene FecB in Small Tail Han sheep.

    Research on the Efficiency of Goat Cumulus Cell Nuclear Transfer Technique
    LI Jun-jie,LI Jun,SUN Jie,SANG Run-zi
    Scientia Agricultura Sinica. 2009, 42(6):  2142-2148 .  doi:10.3864/j.issn.0578-1752.2009.06.034
    Abstract ( 1057 )   PDF (309KB) ( 672 )   Save
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    【Objective】 This study was performed to increase the efficiency of goat cumulus cell nuclear transfer. 【Method】Based on the problem of low efficiency of goat somatic nuclear transplantation (SNT), the application of demecolcine was used to increase goat oocytes enucleation rate, the effects of 5-aza-dC and Trichostatin A (TSA) were also investigated when cumulus cells were treated with them on the development rate of goat cloned embryos. 【Result】 The optimum concentration of demecolcine for chemically assisted enucleation was 0.5 μg?mL-1 and the extrusion cones rate of goat oocytes was 91.67% (P<0.05). Compared with blind enucleation method, the enucleation rate of demecolcine assisted-enucleation method was 100% and increased significantly (P<0.05). Furthermore, the morula rate and blastula rate of cloned embryos reconstrusted by demecolcine assisted-enucleation method were 25.23% and 13.08%, which were significantly higher than that of embryos reconstrusted by blind enucleation method (P<0.05). At the same time, it was found that the blastula rate and morula rate of goat cloned embryos reconstructed by cumulus cells treated with 0.01μmol?L-15-aza-dC was 16.96% and 30.41%, which were extremely higher than that of other groups (P<0.05). When the goat cloned embryos reconstructed by cumulus cells treated with 400 nmol?L-1 TSA, the rate of morula and blastula was 31.52% and 15.22%. 【Conclusion】The enucleation rate of chemically assisted enucleation method was significantly higher than that of blind enucleation method. The effect of goat cloned embryos reconstructed by cumulus cells treated with 0.01 μmol?L-1 5-aza-dC or 400 nmol?L-1 TSA was the best.

    Expression and Activity Analysis of Non-Structural Protein 2 (NS2) of Bombyx mori Densovirus Zhenjiang Strain
    ZHAO Pan,TANG Shun-ming,LIU Ting,QIN Guang-xing,GUO Xi-jie
    Scientia Agricultura Sinica. 2009, 42(6):  2149-2155 .  doi:10.3864/j.issn.0578-1752.2009.06.035
    Abstract ( 922 )   PDF (518KB) ( 689 )   Save
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    【Objective】 The non-structural protein 2 (NS2) gene of Bombyx mori densovirus Zhenjiang strain (BmDNV-Z) was cloned and expressed so as to test the biological activities of the expressed recombinant protein. 【Method】 The gene of the non-structure protein 2 (NS2) was cloned by PCR from the genome of Bombyx mori densovirus Zhenjiang strain, inserted into prokaryotic expression vector pET28a to construct recombinant plasmid pET28a-NS2 and then expressed in bacteria E. coli BL21 (DE3). The expressed recombinant protein was identified by SDS-PAGE and Western blot analysis. Then the recombinant protein was purified by Ni-NTA column, renatured and tested for enzyme activities. 【Result】 The gene of BmDNV-Z NS2 was cloned and successfully expressed in bacteria E. coli, and the expressed target protein NS2 was purified by Ni–NTA affinity chromatography. The purified NS2 protein exhibited a helicase activity unwinding double-stranded DNA substrates into single-strand primers, and higher unwinding activity to polarity substrate. At the meantime, the purified NS2 protein possessed a ATPase activity and its enzyme activity was 0.276 μmol?μg-1?h-1 in this study. 【Conclusion】 The non-structure protein which encoded by the gene of BmDNV-Z NS2 possesses the biological activities of helicase and ATPase, and meanwhile the helicase prefers to polarity substrates. Based on these results, it is speculated that the gene of BmDNV-Z NS2 plays an important role in the viral DNA replication.

    VETERINARY SCIENCE
    Evolution Analysis of the PB1-F2 Genes of Influenza A Viruses Isolated from China
    HUANG Yan-yan,HU Bei-xia,WEN Xin-tian,CAO San-jie,XU Dong,ZHANG Xiu-mei
    Scientia Agricultura Sinica. 2009, 42(6):  2156-2163 .  doi:10.3864/j.issn.0578-1752.2009.06.036
    Abstract ( 1077 )   PDF (305KB) ( 755 )   Save
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    【Objective】 The molecular characterization of PB1-F2 gene of chicken H9N2 subtype avian influenza viruses(AIVs) isolated from China was analyzed and the prevalence of the PB1-F2 genes of influenza A viruses (IAVs) prevailed in China was further investigated. 【Method】 Fourteen H9N2 viruses were isolated from sick chickens in China from 1998 to 2008. The sequences of the PB1-F2 genes of these isolates were determined by RT-PCR and sequencing. Then, 337 PB1 genes of IAVs derived from avian (H5N1, H9N2 subtype), pig (H5N1, H9N2, H1N1 and H3N2 subtype) and human beings (H5N1, H9N2, H1N1 and H3N2 subtype) in China were downloaded from GenBank database and analyzed using molecular biological software. 【Result】 Phylogenic inference drawn from PB1-F2 sequences of the above IAVs showed six distinct clusters. The deduced sequences of PB1-F2 proteins have multifold lengths and the ratio of PB1-F2 genes encoding full-length PB1-F2 protein differed in the HA subtype and host origin of IAV isolates. 【Conclusion】 Through this study, the molecular epidemiologic characteristics of PB1-F2 gene of IAVs in China was illuminated, thus providing a basic foundation of PB1-F2 gene function research in the infection process of IAVs.

    Immune Enhancement Effect of Mixed Herbal Extracts in Chicks Immnunosuppressed by Reticuloendotheliosis Virus Infection
    LIU Feng-xiang,CUI Zhi-zhong,GUO Hui-jun
    Scientia Agricultura Sinica. 2009, 42(6):  2164-2171 .  doi:10.3864/j.issn.0578-1752.2009.06.037
    Abstract ( 895 )   PDF (239KB) ( 729 )   Save
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    【Objective】 To research immune enhancement effect of mixed herbal extracrs in chicks immunosuppressed by reticuloendotheliosis virus infection and establish an experiment model for determination of immune enhancing effects of some herbs.【Method】 Immune enhancement of herbal extracts was determined in clinical health chicks and chicks with immunosuppression induced by reticuloendotheliosis virus (REV) infection. 【Result】 Hemagglutination inhibition (HI) antibody titers against Newcastle disease virus (NDV) and H5 avian influenza virus (H5-AIV) after vaccination were not influenced by supplement with the extract of mixed herbs, Radix Astragali, Radix Codonopis, Herba Epimedii and Radix Glycyrrizae in drinking water in clinical health chicks. In chicks immunosuppressed by reticuloendotheliosis virus (REV) infection, however, supplement of the same herbal extract in drinking water significantly increased HI antibody titers to NDV and H5-AIV when compared to the immunosuppressed control group (P<0.01). 【Conclusion】 It is concluded that chicks with REV-induced immunosuppression could be used as an experiment model for determination of immune enhancing effects of some herbs.

    Establishment of Hamster Model for Taenia solium
    ZHANG Yan,ZHAI Jun-jun,CAI Xue-peng,JING Zhi-zhong,LUO Xue-nong
    Scientia Agricultura Sinica. 2009, 42(6):  2172-2177 .  doi:10.3864/j.issn.0578-1752.2009.06.038
    Abstract ( 919 )   PDF (382KB) ( 657 )   Save
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    【Objective】 To carry out the observation of pathogen, biologic characteristics of Taenia solium and solve the problem of material source for Taenia solium, experimental model of Taenia solium in hamsters was established. 【Method】 The immune-suppressed hamster was infected orally with Cysticercus cellulosa in order to observe the biological characteristics of Taenia solium in hamsters such as infectivity, growth and development and parasitic site, and the morphological structure of detected cestode in hamsters was obersved using the squash technique and tissue section technique. 【Result】 At the 5mg dose of the immunosuppressant, the hamster was infected only one time, and Cysticercus cellulosa could grow to adult. At the same time, the detectable rate of Taenia solium was very high, the cestode containing somite could be detected in the hamste, after 80d infection, gravid proglottid could be detected. The most polypide adsorbed in ahead of 1/3 small intestine, some were also found in gall bladder and bile duct, even some escaped form gall bladder and parasited in liver. Four suckers and two cycles of granular structure were found in scolex. Compared with human-taenia solium, mature proglottids were fewer. 【Conclusion】 The experimental model of Taenia solium in hamsters has been successfully developed in this experiment. This made it becomes possible to successfully solve the problem of material source for Taenia solium, and has also laid a foundation for the further research of Taenia solium.

    Effects of Baishao Preparation on the Functions and Phosphodiesterase Activity of Neutrophils
    JIANG Dai-xun,CHEN Wu,HOU Xian-tao,YU Tong-quan,LU Ping,MU Xiang
    Scientia Agricultura Sinica. 2009, 42(6):  2178-2183 .  doi:10.3864/j.issn.0578-1752.2009.06.039
    Abstract ( 891 )   PDF (325KB) ( 777 )   Save
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    【Objective】 The objective of this experiment is to study the influence of Baishao (Paeonia lactiflora) preparation on respiratory burst and elastase release of neutrophils with porcine neutrophils as the target cells. 【Method】 after the separation and purification of neutrophils, respiratory burst of neutrophils was detected with cytochrome C reduction method, elastase release was indicated with the substrate reduction of elastase, and phosphodiesterase (PDE) activity was calculated by cAMP change examined with HPLC before and after the reaction. 【Result】 Baishao preparation could significantly inhibit neutrophils respiratory burst dose dependently (P<0.001), the inhibition ratios of three different concentrations were 33.15%, 45.96% and 62.22%, respectively; the adjustment of Baishao preparation on elastase release was duplicate, having inhibition in low concentration, inhibition ratio was 23.22%, while having promotion in high concentration. Baishao preparation could obviously inhibit PDE activity in neutrophils, and the inhibition ratios of low and high concentration were 13.24% and 25.87% (P<0.01), respectively. Analysis of HPLC finger print showed that there were five ingredients in Baishao preparation. 【Conclusion】 Baishao preparation has significant inhibition on respiratory burst of neutrophils, and has obvious inhibition on elastase release at low concentration, the mechanism of which possibly has a close relation with the inhibition on PDE activity.

    Ultrastructural Characteristics of Neurons and Varicosis in Intestinal Nerve of Remak of Chicken
    LIU Jin-xiong,YU Zu-gong,XU Chun-sheng,FENG Ya-mei,YANG Ping,QIN Jun-hui,CHEN Qiu-sheng
    Scientia Agricultura Sinica. 2009, 42(6):  2184-2191 .  doi:10.3864/j.issn.0578-1752.2009.06.040
    Abstract ( 1179 )   PDF (1291KB) ( 1175 )   Save
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    【Objective】 The purposes of this study are to make up a deficiency of ultrastructures of neurons and varicosis in intestinal nerve of Remak (INR) of chicken, and to provide a rational foundation for future physiologic function researches of INR.【Method】 Neurons and varicosis in INR of chicken were observed with transmission electron microscope. 【Result】 Considerable number of large neurons and a little small intensely fluorescent cells were found in INR. Neurons were enclosed by scattered satellite cells, which didn’t form integrity envelopes. Sinapsic contacts between neurons and satellite cells were also found. Because of lack normal basal lamina, the cell membrane of neurons directly contacted with intercellular substance. The shape of nucleus in neurons was round, and the nucleus had smooth surface. Loosen and clear chromatins, rhoptries and nucleoli with integrity structure were found in nucleus of neurons. While, in cytoplasm, many special organelles distributed in it. These organelles included the copious microtubules, active Golgi apparatuses, active rough endoplasmic reticulum which often contained inflated endoplasmic reticulum cisterna, fenestrated dense granules which could be neuropeptide, plenty of free ribosomes, mitochondria with diversified shapes and some of which were pyknosis to some extent. A little clumped small intensely fluorescent cells with high electron density cytoplasm and nucleus distributed under capsule of INR, which can form asymmetry synapses each other. According to the different kinds of synaptic vesicle they contained, 4 kinds of varicosis were found in the INR, which formed different function synaptes with surrounding structures. 【Conclusion】 The ultrastructural characteristics of INR neurons suggested that the INR posses actively synthesis and secretion function, 4 kinds of varicosis and their different synaptic connections indicated the diversity and comlexity of INR functions.
    RESEARCH NOTES
    Identification of Maize Drought-Tolerance at Seedling Stage Based on Leaf Temperature Using Infrared Thermography
    LIU Ya,DING Jun-qiang,Subhash Chander,LIAO Deng-qun,ZHAO Jiu-ran,LI Jian-sheng
    Scientia Agricultura Sinica. 2009, 42(6):  2192-2201 .  doi:10.3864/j.issn.0578-1752.2009.06.041
    Abstract ( 1064 )   PDF (307KB) ( 1161 )   Save
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    【Objective】 One efficient approach to enhance plant drought tolerance is to decrease water loss during leaf transpiration through stoma regulation, which results in the variation of leaf temperature. The objectives of this study were to detect the relation between leaf temperature variation and biomass accumulation to serve as an evidence for screening the germplasms tolerant to drought at maize seedling stage under moderate drought stress. 【Method】 Eighty- three elite maize inbred lines from China were used to identify the variation of seedling leaf temperature under moderate drought stress. An infrared thermography was employed to measure leaf temperature. Leaf stomatal conductance and transpiration rate were determined by porometer. Plant biomass was measured by electronic balance. 【Result】 Among the measured maize inbred lines at seedling stage under drought stress, relative biomass weight ranged from 0.271 to 0.997 (relative fresh weight), and from 0.338 to 0.969 (relative dry weight), leaf temperature difference varied between -0.1-+0.5 ℃, which reached to significant levels. Under the drought condition, leaf temperature of drought tolerance lines rose significantly, while those of drought sensitive had not obvious variation. Leaf temperature difference between drought stress and well-watered conditions showed a significant positive correlation with relative fresh biomass (0.283*, 0.288**, n=83) and relative dry biomass (0.239*, 0.273**, n=83). Significant correlations of leaf temperature difference with stomatal conductance and transpiration rate were also detected. 【Conclusion】 Transpiration rate was one of the main causes affecting leaf temperature variation and stomatal behaviour served as an important regulation in leaf temperature response to water stress. Under drought stress, leaf temperature difference can reflect the capability of drought tolerance of maize significantly. It was feasible that infrared thermography could be adopted in screening drought-tolerance germplams at maize seedling stage and accelerates the breeding effect for drought-tolerance as an assistant means.

    Study on the Relationship Between the Toxin of Phytophthora Infestans and Resistance of Potato
    YANG Yan-li,XIAO Lang-tao,HU Xian-qi
    Scientia Agricultura Sinica. 2009, 42(6):  2202-2210 .  doi:10.3864/j.issn.0578-1752.2009.06.042
    Abstract ( 1046 )   PDF (571KB) ( 950 )   Save
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    【Objective】 This study is to make clear the poisoning effect of phytophthora infestans toxins for potato, and to discover the toxicity different from physiological race to potato verieties.【Method】 Putative P. infestans toxins were prepared by culturing in liquid medium ( 60 g rye and 10% tomato juice per litre) for 1 month at 17℃, filtering through 4 layers of cheesecloth, and being precipitated by ammonium sulfate. The resulted putative toxin solutions were used to examine the reactions of 3 potato varieties (both leaf and tuber tissues) to the putative toxins. 【Result】 The results show that potato leaves and tubers exhibit symptoms similar to the late blight resulted from P. infestans infection. Potato varieties reacted differentially to both the toxin dilutions and toxins produced by different P. infestans isolates, suggesting the presence of toxin-mediated specificities between potato and P. infestans. Potato leaf and tuber tissues have different and contrary reactions to the toxin. 【Conclusion】 Toxin-mediated specificities are likely present between potato and P. infestans.
    The Isolation of Yeast-Like-Symbiots in the Brown Planthopper and the Sequences Analysis of Its 26S rDNA
    ZHANG Jue-feng,CHEN Jian-ming,CHEN Fa-jun,ZHENG Xu-song,CHEN Lie-zhong,YU Xiao-ping
    Scientia Agricultura Sinica. 2009, 42(6):  2211-2216 .  doi:10.3864/j.issn.0578-1752.2009.06.043
    Abstract ( 928 )   PDF (296KB) ( 1349 )   Save
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    褐飞虱;类酵母;离体培养;26SrDNA;解脂假丝酵母;嗜盐梗孢酵母
    Acaricidal Bioactivity of Artemisia annua Extracts Against Panonychus citri (Acari: Tetranychidae)
    ZHANG Yong-qiang,DING Wei,TIAN Li,ZHAO Zhi-mo
    Scientia Agricultura Sinica. 2009, 42(6):  2217-2222 .  doi:10.3864/j.issn.0578-1752.2009.06.044
    Abstract ( 1181 )   PDF (265KB) ( 783 )   Save
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    【Objective】 To determine the acaricidal bioactivities of Artemisia annua against Panonychus citri,so to provide a scientific basis for the comprehensive development and utilization of A. annua. 【Method】 Collect The whole A. annua plants were collected in June and July respectively, and the whole plants were cut into root, stems and leaves, petroleum ether (30-60℃), petroleum ether (60-90℃), ethanol, acetone and water were used and extracted by the parallel and sequenced solvents extraction methods, finally 54 kinds of extracts were obtained and their indoor acaricidal bioactivity against P. citri were determined. 【Results】 The results showed that the biological activity of the A. annua plant in July was better than in June and the acetone parallel extract of leaf in July was stronger than other extracts. The LC50 of the acetone parallel extract of leaf in July was 0.4222 mg?ml-1, which was only 44% of 0.9489 mg?ml-1 in June. With the bio-guided isolation method, the acaricidal activity of different components isolated from A. annua July leaf parallel acetone extracts by column chromatography were determined in the laboratory. The 20 kinds of components were obtained, the acaricidal activity of component 17 was the strongest. There was a significant difference between component 17 and other components (P<0.05). 【Conclusion】 The acetone parallel extract of leaf in July showed a strongest biological activity against P. citri. And separated by column chromatography, 20 kinds of components were obtained, component 17 exhibited a highest acaricidal bioactivity and certain acaricidal bioactivity compound would be identified in further studies.

    Effect of Long-Term Application of Organic Fertilizer on Structural Characteristics of Humin in Black Soil ——A Solid-State 13C NMR Study
    ZHANG Jin-jing,DOU Sen,ZHU Ping,GAO Hong-jun,SONG Xiang-yun,WANG Li-chun
    Scientia Agricultura Sinica. 2009, 42(6):  2223-2228 .  doi:10.3864/j.issn.0578-1752.2009.06.045
    Abstract ( 1225 )   PDF (354KB) ( 1145 )   Save
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    【Objective】 In order to provide a theoretical basis for illustrating the role of humin (HM) in soil fertility, the structural characteristics of HM were studied after long-term application of organic fertilizer (pig manure) under continuous cultivation of corn in agricultural black soil, and the structural difference of HM was compared between different dosage and duration of organic fertilizer applied. 【Method】 The experiment was carried out based on a long-term fertilizer experiment of “Black Soil Fertility and Fertilizer Efficiency Long Term Monitoring Base (founded in 1980)” at Gongzhuling, Jilin Province, China. Soil (0-20 cm) was sampled at 1992 and 2005, respectively. The treatments included: CK (no fertilizer), O1 (low level of organic fertilizer at a dosage of 30 t?hm-2?a-1) and O2 (high level of organic fertilizer at a dosage of 60 t?hm-2?a-1). Soil HM were separated with NaOH-Na4P2O7 mixture, and then dashed with HF-HCl mixture. Solid state 13C cross polarization magic angle spinning nuclear magnetic resonance (13C CPMAS NMR) spectroscopy was used to characterize the chemical structure of the HM isolates. 【Result】 Compared with CK, the organic carbon contents of HM increased after application of organic fertilizer; alkyl C also increased while O-alkyl C and aromatic C decreased; the ratios of alkyl C/ O-alkyl C, aliphatic C/ aromatic C and hydrophobic C/hydrophilic C all increased. The above change tendency was more obvious with the increase in the dosage and the duration of organic fertilizer applied. With the prolonged duration for the CK treatment, the organic carbon content of HM decreased. And the change tendency of alkyl C, O-alkyl C, aromatic C, alkyl C/ O-alkyl C ratio, aliphatic C/ aromatic C ratio and hydrophobic C/hydrophilic C ratio of HM from CK were all identical with that of HM from O1 and O2 with duration, but the change extents of theirs were lower in the CK treatment. 【Conclusion】The long-term application of organic fertilizer makes the HM structure tend to become more alkyl and hydrophobic, and thus is beneficial for maintaining soil fertility.

    Effect of Harvesting Period on Yield and Fuel Quality of Corn Stover
    LIU Ji-li,CHENG Xu,XIE Guang-hui,XIONG Shao-jun,ZHU Wan-bin
    Scientia Agricultura Sinica. 2009, 42(6):  2229-2236 .  doi:10.3864/j.issn.0578-1752.2009.06.046
    Abstract ( 1199 )   PDF (302KB) ( 1139 )   Save
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    【Objective】 The objective of this experiment is to evaluate the variation in yield and fuel quality of corn stover under different harvesting periods and to determine appropriate harvesting periods. 【Method】 Using Zhengdan 958 as research material, the effect of harvesting periods on yield and fuel quality of corn stover was studied under field conditions in Baicheng Academy of Agricultural Sciences during 2007-2008. 【Result】 Results showed that with the delayed harvesting period, yield of corn stover decreased, and the proportion of leaves reduced gradually. Moisture, ash, and mineral elements contents of corn stover significantly decreased, cellulose and lignin contents increased, but hemicellulose content decreased. Calorific value of corn stover was maximum at filling stage and minimum at maturity, it was slightly higher after maturity. Calorific value was significantly correlated with cellulose and lignin contents. Ash content was significantly positively correlated with mineral elements contents, and was significantly negatively correlated with Si/K ratio. 【Conclusion】 Delayed harvesting reduced the yield of corn stover, but significantly improved its fuel quality. Corn stover fuel quality was better when harvested in winter and spring.

    The Difference Between Starch Chain Length Distribution and Main Quality Characteristics of High Resistant Starch Lines of Japonica Rice
    ZHANG Shu-mei,ZHANG Jian-ming,LEE Jung-ro,LI Mao-bai,WANG Hui,PIAO Zhong-ze,ZOU De-tang
    Scientia Agricultura Sinica. 2009, 42(6):  2237-2243 .  doi:10.3864/j.issn.0578-1752.2009.06.047
    Abstract ( 1079 )   PDF (329KB) ( 1508 )   Save
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    【Objective】 Providing a theoretical and practical basis for breeding and germplasm enhancement by means of genetic analysis on resistant starch content in japonica rice. 【Method】 Using japonica rice lines with significant differences in resistant starch content , this paper analyses differences and correlation of amylopectin chain length distribution, RVA spectral properties value, dietary fiber and amylose content and other quality characteristics between them. 【Result】 The chain length distribution of amylopectin shows significant differences between high and low resistant starch content strains . For the high resistant starch content strains, the relative percentage of the degree of short chain polymerization 5 to 12 about amylopectin content, which is obviously lower than low resistant starch content strains. However, the degree of polymerization in long chain (particularly above 13 proportions) is obviously higher than the low ones. Dietary fiber and amylose content of high resistant starch content is higher than that of low ones. And the peak viscosity, cool paste value, hot paste viscosity and breakdown value of high resistant starch content is significantly or very significantly lower than that of the low ones. The polymerization degree (DP≤12) of short chain amylopectin shows very significantly negative correlation to resistant starch content. On the contrary, the very significantly positive correlation between resistant starch content and the proportion of polymerization degree (12<DP≤36) of long chain amylopectin, and the amylose content and the dietary fiber content was observed. 【Conclusion】 The resistant starch content is closely related to amylopectin chain length distribution, amylose content, RVA spectral properties value. Therefore, it could be used as a breeding index.

    Oxidative Damage of Heat-Stressed Mouse Oocytes and Preimplantation Embryos and Their Environments
    LI Hui-hui,TIAN Wen-ru,ZHANG Zhi-hong,ZHANG Qi-yao,GAO Shan-song
    Scientia Agricultura Sinica. 2009, 42(6):  2244-2249 .  doi:10.3864/j.issn.0578-1752.2009.06.048
    Abstract ( 1186 )   PDF (405KB) ( 813 )   Save
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    【Objective】 This study was conducted to determine the effects of heat stress on the oxidate damage of organism(liver), genital system, oocytes and preimplantation embryos, and to observe oocytes’ and embryos’ subsequently developmental competence. 【Method】 TGSH and MDA in the oocytes, preimplatation embryos and several organs were determined by using total glutathione and malonaldehyde assay kit, respectively. The subsequently developmental competence of heat-stressed oocytes and embryos was determined. 【Result】 The subsequently developmental competence of the oocytes (6 h) and embryos (12 h) on day 1 to day 3 (before morula) in the mice heat-stressed at 35℃ for 6 h or 12 h decreased significantly. The TGSH content decreased and MDA output increased significantly in heat-stressed embryos on days 1, 2 and 3, but not in heat-stressed oocytes. However, nothing changed in embryos on days 4 and 5 of age cultured in vivo and the embryos on days 1 to 5 of age heat-stressed at 39℃ for 12 h cultured in vitro. The TGSH content increased and MDA output decreased significantly in liver and oviduct with the increased grading of heat stress and prolonged time, and in the ovary and uterus with 41℃ heat stress only.【Conclusion】The results demonstrate that the loss of developmental competence of early embryos is associated with oxidate damage of maternally heat-stressed enbryos before morula, but neither of maternally heat-stressed oocytes nor in vitro cultured 1 to 5d embryos.

    Identification of Brood Pheromone in Chinese Honeybee [Apis cerana cerana (Hymenoptera: Apidae)]
    YAN Wei-yu,Yves LE CONTE,Dominique BESLAY,ZENG Zhi-jiang
    Scientia Agricultura Sinica. 2009, 42(6):  2250-2254 .  doi:10.3864/j.issn.0578-1752.2009.06.049
    Abstract ( 1018 )   PDF (240KB) ( 1071 )   Save
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    【Objective】 To analyze the brood kairmone and pheromone of different stage brood in Apis cerena cerena and compare it with Apis mellifera. 【Method】 Worker larvae (2-day old, 6-day old and 7-day old) and drone larvae (2-day old, 7-day old and 8-day old) of Apis cerena cerena were extracted with hexane by crushing, the supernatant were then fractioned by chromatography on a column of silica gel and tested by gas chromatogram. 【Result】 It is the first time to identify the components of brood kairmone and pheromone from Apis cerena cerena are methyl palmitate, methyl oleate, methyl stearate, methyl linoleate, methyl linolenate, ethyl palmitate, ethyl oleate, ethyl stearate, ethyl linoleate and ethyl linolenate. 【Conclusion】 The kairmone and 10 aliphatic esters in brood have been found in both Apis mellifera and Apis cerana cerana, while the content and distribution of the two pheromone are different at different brood stages.