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Table of Content

    10 March 2008, Volume 41 Issue 3
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Genetic Diversity of Aromatic Rice Varieties Based on SSR Markers
    Scientia Agricultura Sinica. 2008, 41(3):  625-635 .  doi:10.3864/j.issn.0578-1752.2008.03.001
    Abstract ( 1499 )   PDF (756KB) ( 1727 )   Save
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    Abstract: Genetic diversity of 32 aromatic rice cultivars were analyzed by 60 SSR primers covered on 12 rice chromosomes. A total of 188 gene locus were detected, among of which 126 gene locus were effective number of alleles. The average number of gene locus per SSR locus was 3.13 with a ranged from 2 to 6. The value of allelic polymorphism information content (PIC) ranged from 0.116 to 0.744, on an average, 0.467±0.023 per SSR maker. The average similarities coefficient was 0.697±0.0035 with a ranged from 0.51 to 0.96. UPGMA cluster analysis showed that the 32 cultivars could be classified into indica and japonica distinct classes at similarities coefficient 0.58, which was generally coincident with pedigree analysis. The results indicated that SSR markers could be employed in diversity identification for aromatic varieties and in exploration for aromatic rice breeding.
    Isolation and chromosomal localization of cytokinin oxidase/dehydrogenase gene (TaCKX5) in wheat
    Scientia Agricultura Sinica. 2008, 41(3):  636-642 .  doi:10.3864/j.issn.0578-1752.2008.03.002
    Abstract ( 1419 )   PDF (588KB) ( 1177 )   Save
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    【Objective】Genetic manipulation of CKX activity holds the promise of improving agricultural traits in crops.【Method】and【Result】In this study, a strategy of homologous cloning combined with BAC library screening was used to isolate TaCKX5 gene from wheat. Sequence analysis of TaCKX5 revealed that TaCKX5 contains a FAD-binding domain which is well conserved in the N-terminal halves of all known CKX sequences. Using TargetP program, TaCKX5 was predicted to be a secretion pathway protein, and one N-glycosylation site (NFTE) was also found with NetNGly program, implied that TaCKX5 protein may be modified posttranslationally by glycosylation. Using nulli-tetrasomic stocks, TaCKX5 was located on wheat homoeologous group 3 chromosomes.【Conclusion】TaCKX5 was very closely related to its orthologous CKX gene (OsCKX5) in rice, so they might have similar characters and functions.
    Identification of Major Quality Genes in Cultivars from Yellow and Huai River Valley Wheat Region by Multiplex PCR
    Scientia Agricultura Sinica. 2008, 41(3):  643-653 .  doi:10.3864/j.issn.0578-1752.2008.03.003
    Abstract ( 1444 )   PDF (567KB) ( 944 )   Save
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    Abstract: 【Objective】Development of multiplex PCR for wheat quality traits is very important to improve the efficiency and reduce the cost in molecular marker assisted breeding. 【Method】 In the present study, according to genes controlling for pan bread and Chinese noodle quality, and the annealing temperatures of different markers, the molecular markers for important quality trait genes, i.e., high-molecular-weight glutenin subunit (HMW-GS) genes Ax2*, Bx14, Bx17 and Dx5, and low-molecular-weight glutenin subunit (LMW-GS) gene Glu-A3d, as well as the marker BDFL-BRD for Wx-B1, MAG269 for Wx-D1, ω-sec for 1BL/1RS translocation and PPO18 for polyphenol oxidase(PPO)activity, were selected and three combinations of molecular markers Ax2*/Bx17/Dx5, Bx14/Glu-A3d/ω-sec and BDFL-BRD/MAG269/PPO18 were tested. 【Result】Results indicated that the three multiplex PCRs, PCR-I (Ax2*/Bx17/Dx5), PCR-II (Bx14/Glu-A3d/ω-sec) and PCR-III (BDFL-BRD/MAG269/PPO18) can be used to identify the genes presented in wheat cultivars. A total of 141 wheat cultivars were evaluated with the multiplex PCRs, and the frequencies of HMW-GS Ax2*, Bx14, Bx17, Dx5 and LMW-GS Glu-A3d accounted for 4.3%, 7.1%, 1.4%, 17.7% and 27.7%, respectively. Frequencies of 1BL/1RS translocation and low polyphenol oxidase (PPO) activity genetypes were 44.0% and 51.8%, respectively. Wx-B1 null type accounted for 4.3%. Data obtained by SDS-PAGE for HMW-GS, LMW-GS and secalin for 80 genotypes correspond very well with those from the multiplex PCRs. 【Conclusion】This indicated that the multiplex PCRs developed can be steadily and efficiently used to detect the nine genes for the improvement of wheat quality in molecular marker assisted breeding.
    Mutagenic Effects of Mixed Particle Field Irradiation in Wheat
    Hui-Jun GUO Lu-Xiang LIU Wei-Bo HAN Shi-Rong Zhao Jia-Cai LI Lin-Shu Zhao Jing Wang
    Scientia Agricultura Sinica. 2008, 41(3):  654-660 .  doi:10.3864/j.issn.0578-1752.2008.03.004
    Abstract ( 1354 )   PDF (421KB) ( 860 )   Save
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    【Objective】The objective of the present study was to analyze the mutagenic effects and frequency of mixed particle field with high energy(CR) irradiation in winter wheat, and to compare with those of 60Co-γray irradiation. 【Method】Air-dried seeds of winter wheat cultivars ZY9 and ZH7 were used to irradiate by CR andγ-ray with the dosage of 0, 145Gy, 195Gy, 284Gy and 560Gy respectively. Irradiated seeds were germinated in the laboratory; mutants from M2 generation were screened and analyzed. 【Result】 There was obviously linear dose-effect relationship at inhibiting plant growth of these two genotypes, the damage effect of CR was higher than that of gamma rays, and the variety ZH7 was more sensitive than ZY9. The optimal irradiation dose of cosmic rays for wheat was 200 to 300Gy. Mutation phenotypes of CR treatment in M2 populations included plant height, spike type, heading date etc, the useful mutation frequency at the optimum or little higher dosage was high than that of gamma rays irradiation. The total mutation frequency and useful mutation frequency of CR treatment for ZY9 was 5.42~11.68% and 2.75~7.29%, for ZH7 was 10.35~22.12% and 6.62~10.49%, respectively, higher than that of gamma rays. 【Conclusion】These revealed the higher relative biological effect of CR treatment could induce than that of gamma rays; the more sensitive the variety was, the higher useful mutation frequency and the higher probability of screening improved genotypes would be.
    A Fast and High Efficient Technique for Agrobacterium-Mediated Transformation in Soybean (Glycine max L. Merrill)
    Scientia Agricultura Sinica. 2008, 41(3):  661-668 .  doi:10.3864/j.issn.0578-1752.2008.03.005
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    To develop a rapid and high efficient technique for soybean transformation, this report concentrated on resolving intractable problems, such as explants tissue culture, Agrobacterium tumeficiences infection and plant regeneration. Mature soybean seeds were germinated on MSB5-medium supplemented with 1 mg/L N6-benzylaminopurine (BAP) at 25~28℃ for 24 h and the explants were pre-cultured at the same condition as above. Explants were inoculated with Agrobacterium tumeficiences strain GV3101 harboring the vector pBI121 and cultured at 28 ℃ for 21 h in MSB5-medium suspension supplemented with 1mg/L N6-benzylaminopurine (BAP) and 100 μM acetosyringone. After co-cultivation for 3 days in the dark at 28 ℃, transformants were transferred to sterilized soil directly and incubated with suitable light and temperature conditions. The results of molecular test indicated that the regeneration ratio of this system was more than 90% (95% for Zhongpin661, 93% for Kennong18 and 90% for Lv75, respectively) compared to cotyledonary nodes no more than 50% and embryonic tips no more than 70%, and the transformation ratio was as high as 10% (Kennong18). Furthermore, as no complicated and time-consuming tissue culture operations were used in the latter cultivation for transformants, the whole regeneration period were shortened evidently. Therefore, this technique system for soybean transformation mediated by Agrobacterium tumeficiences was more fast and high efficient than others, and a powerful technical platform for Agrobacterium tumeficiences-mediated genetic transformation and mutant construction of genomic research in soybean.
    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY
    Progresses of Researches for Plant Morphological Structure Simulation and Visualization
    Hong-xin CAO Chun-Lin SHI
    Scientia Agricultura Sinica. 2008, 41(3):  669-677 .  doi:10.3864/j.issn.0578-1752.2008.03.006
    Abstract ( 1370 )   PDF (326KB) ( 2478 )   Save
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    At first, the thesis introduced social background, scientific values and economic significances of researches for plant morphological structure simulation and visualization, secondly, reviewed its progresses in found and development, collectivity trends and characteristics, institutions and scholars with world advance level and its sign, collectivity level and gap and causes analysis of formation in china, front fields and blank points and weakness taches, et al., lastly, its prospects and some policy suggestions were put forward.
    Tiller Pattern and Its Contribution to Yield of Rice Directly Sown after Winter Wheat Harvest in Huang-Huai Plain
    Guangcan Tao Guanghui Xie
    Scientia Agricultura Sinica. 2008, 41(3):  678-686 .  doi:10.3864/j.issn.0578-1752.2008.03.007
    Abstract ( 1515 )   PDF (434KB) ( 1269 )   Save
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    To research the tiller emergence frequency of rice cultivars sown directly under aerobic conditions after winter wheat harvest, field experiments were conducted in Mengcheng, Anhui province, in 2005 and 2006. The experiment included two adjacent fields: the submerged and non-submerged field. Five drought resistance rice cultivars were used. They were HanDao502, HanDao297, HanDao277, NingGeng16 and NingGeng35. The aboveground biomass and grain yields of rice cultivars at maturity were 6.3-11.3 t ha-1 and 1.9-5.4 t ha-1 in 2005, 6.6-9.5 t ha-1 and 3.6-4.6 t ha-1 in 2006, respectively. For each cultivar, the differences of aboveground biomass or yield between the non-submerged conditions and the submerged conditions were not significant. Under the both conditions, the tiller emerging potential of HanDao277 was the highest among the cultivars and the contribution of productive tillers to yield was about 50%. Though the tiller emergence frequency of the 3rd to 5th node for Handao502 and Handao297 was high before elongation, most of them were aborted progressively before heading, however. For NingGeng16 and NingGeng35, there were few tillers emerged from the plants before elongation. For all of the cultivars except HanDao277, the contribution of main stems to yield was above 80%.
    Starch Accumulation and Activities of Enzymes Involved in Starch Synthesis in Grains of Wheat Field-Grown in Two Water Regimes
    Scientia Agricultura Sinica. 2008, 41(3):  687-694 .  doi:10.3864/j.issn.0578-1752.2008.03.008
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    【Objective】The present study was carried out to characterize the changing activities of key enzymes during grain filling in two wheat cultivars differing in gluten under irrigated and rainfed conditions. This information will help to elucidate the enzymatic mechanisms of starch synthesis in wheat cultivars with different gluten. 【Method】The starch accumulation, sucrose metabolism and related enzymes activities were investigated in strong gluten type cultivar, JM20, and weak gluten type cultivar, BY535, in Taian and Dezhou experimental locations. 【Result】The soil water deficit led to an increase at early grain-filling period and decrease during late grain-filling, respectively, in starch accumulation rate and activities of key enzymes involved in starch synthesis, especially AGPase, SSS, and SBE. Water deficit enhanced grain starch accumulation in two wheat cultivars. It suggests that rainfed treatment increased the physiological activities during early grain filling, and promoted starch accumulation. Furthermore, the change of starch accumulation rate was consistent with SS,AGPase,SSS and GBSS. The results suggested that these enzymes play a key role in starch synthesis and the decrease of photosynthate produced in source organ is not the factor inhibiting starch accumulation.【Conclusion】The soil water deficit led to an increase at early grain-filling period and decrease during late grain-filling, respectively, in starch accumulation rate and activities of key enzymes involved in starch synthesis, especially AGPase, SSS, GBSS and SBE. It indicated that water deficit can increase the sink activities and promote starch synthesis and accumulation.
    Study on Integrated and Intelligent Crop Modeling System Based on Growth Meta-Model
    Hai-Yan JIANG Liang TIANG Yan ZHU Deng-Feng Hua
    Scientia Agricultura Sinica. 2008, 41(3):  695-703 .  doi:10.3864/j.issn.0578-1752.2008.03.009
    Abstract ( 1357 )   PDF (574KB) ( 878 )   Save
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    【OBJECTIVE】In order to explore the approach of integrated and intelligent crop modeling, the modeling tools were developed.【METHOD】By analyzing and extracting the crop (wheat, rice and cotton) growth models and simulation systems, the generic modeling flow, generic model framework, generic model interface and generic model template, and growth meta-model for agronomic crops were established. Based on crop model resource library, model components, model description script and model engine, the architecture and core parts of Integrated Crop Modeling Simulation System (ICMS) with layer and component characteristics were designed.【RESULTS】The prototype system was developed with C# based on .Net platform and SQL Server database. Further, a wheat growth model was reconstructed using the ICMS. The results shows that the model reconstructed by the ICMS could well simulate wheat growth processes and exhibit small errors as compared with wheat growth system based on code modeling approach. 【CONCLUSION】The ICMS realizes flowchart process of crop modeling,, industrialization of modeling method, generalization of interface and transparent of models. With ICMS, a crop model can be developed by scientists/modelers with no-code automatic approach through studying, customizing, designing, configuration, assembling, evaluation and extension.
    PLANT PROTECTION
    Controlling watermelon Fusarium wilt by intercropped with aerobically growing rice and its physiological mechanism
    Scientia Agricultura Sinica. 2008, 41(3):  704-712 .  doi:10.3864/j.issn.0578-1752.2008.03.010
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    Pot experiment was conducted to investigate the effect and physiological mechanism of alleviating watermelon wilt disease occurred in continuous cropping soil by intercropped with aerobically growing rice. The results showed that in mono-cropping system, 66.7% of watermelon was infected with wilt disease and 44.4% died after 40 days of transplanting, by contrast, watermelon grew normally in intercropping system. Intercropping significantly reduce occurrence of wilt disease and death rate of watermelon, with the results of lower malondialdehyd (MDA) in roots and leaves, higher soluble protein, lower activities of catalases (CAT), peroxidases (POD), polyphenoloxides (PPO) in roots and leaves and phenylalantine ammonia lyases (PAL) in leaves of watermelon, in comparison with monocropping. The number of Fusarium oxysporum in the rhizosphere soil of watermelon intercropped with aerobically growing rice was decreased by 91%, compared with those in monocropping. Intercropping changed microbiodiversity in rhizosphere soil by phospholipid fatty acids (PLFA), having more bacteria and actinomyces and less fungi compared to monocropping. This finding suggests that intercropping system decreased infection of watermelon wilt disease by restraining pathogen (Fusarium oxysporum) growth and regulating microbiodiversity in rhizosphere soil.
    Relationship between host plant preference of Liriomyza huidobrensis on different bean varieties and plant compound contents
    Scientia Agricultura Sinica. 2008, 41(3):  713-719 .  doi:10.3864/j.issn.0578-1752.2008.03.011
    Abstract ( 1115 )   PDF (271KB) ( 1167 )   Save
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    【Objective and Method】In order to investigate the host plant preference of Liriomyza huidobrensis on different bean varieties and its relationship with the compound contents in bean leaves, no-choice method was used to test the host plant preference, and acetone spectrophotometry, anthrone colorimetric method, coomassie brilliant blue (CBB) G250 staining, phosphor-molybdenum acid-phosphor-tungstenic acid colorimetry and Soxhlet extraction were applied to measure the contents of lamina’s chlorophyll, soluble sugar, protein, tannin acid and flavone, respectively.【Result】The preference of L. huidobrensis on different bean varieties was significantly different (F=4.678, P<0.0001). The clustering analysis divided all tested bean varieties into three groups according to the levels of the preference of L. huidobrensis. The most favourite varieties included Phaseolus vulgaris cv. Shuangfengerhao, Futianyihaojiadou, Jingpinjiadou, Zihuayoudou, Dachengdidou, Ribenwujinglv, Shuangfengjiadou, Shijijiangdou, Tai-guojiadouwang, Taikongqingmei and Wuchangdayoudou. The secondly favourite varieties were P. vulgaris cv. Jiucunlian, Tianma95-33, Jingxuan73-8, Tianmadidou, Taifengjiadouwang, Futian’erhaojiadou, Manjialian, Taiguojiadou 95-33, Lvlongjiadou, Qiuzidou and Qingdaojiadou. The lest were P. vulgaris cv. Junmadidou, Huajiangbahao, Tianmajialongwang, Chengpaijinglongwang and Xinxuanjiulibai. Chemical component analysis showed that the contents of chlorophyll, protein, soluble sugar, tannic acid and flavone among the tested varieties were significantly different. Correlative analysis showed that the host preference of L. huidobrersis was significantly negatively correlative with the contents of tannic acid and flavone but not with those of chlorophyll, soluble protein and soluble sugar in bean varieties.【Conclusion】 The tannic acid and flavone in the leaves of kidney bean significantly suppress oviposition and/or feeding of L. huidobrersis adults.
    Temporal and Spatial Dynamic Studies on Acarcidal Bioactivities of Artemisia annua L. against Tetranychus cinnabarinus Bois. (Acari: Tetranychidae)
    Yong-Qiang ZHANG Wei DING Zhi-mo ZHAO Jing WU Yu-Hu FAN
    Scientia Agricultura Sinica. 2008, 41(3):  720-726 .  doi:10.3864/j.issn.0578-1752.2008.03.012
    Abstract ( 1148 )   PDF (387KB) ( 982 )   Save
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    【Objective】This aim of this study was to determine the best extraction technique, the suitable solvent and the optimal plant parts and acaricidal activities temporal and spatial dynamic of Artemisia annua L.【Method】This paper dealt with the acarcidal bioactivities against Tetranychus cinnabarinus of total 135 extracts of petroleum ether 30-60℃, petroleum ether 60-90℃, ethanol, acetone and water parallel and sequenced extracts from the leaves, stems and roots of different period traditional Chinese medicine Artemisia annua L. in April, May, June, July and September 2006. All extracts were diluted to 5mg/ml, and examined their bioactivities against T. cinnabarinus in laboratory. 【Results】The results showed that at the concentration of 5mg/ml, the acaricidal bioactivities elevated as the development of A. annua plants, the general tendency exhibited July> June> May> April, but the September decreased comparing with the July. However, the most effective extracts in five months were all acetone parallel extract of A. annua leaf, and the corrected mortalities after 48h treatment were ranged from 74% to 100%. The median lethal concentrations (LC50) of acetone parallel extracts of A. annua leaves in September, July, June, May and April were 0.5986, 0.4341, 0.8376, 0.9443 and 1.3817mg/ml, respectively treated after 48h against T. cinnabarinus. The 13 groups were separated from acetone extracts of A. annua leaves in July by column chromatography, all of which the 11th and 12th groups exhibited strong activities. The median lethal concentrations of the 11th and 12th groups against T. cinnabarinus were 0.3683and 0.1586mg/ml, respectively. 【Conclusion】The acetone parallel extract of Artemisia annua leaf in July was most toxicity to T. cinnabarinus Bois. and the corrected mortality was 100% after exposure 48h. The No. 11 and No. 12 groups of acetone parallel extract was the most active components, could be acted as the emphases in further study.
    Use of Generic Hapten in the Production of Broad Specificity Anti-Organophosphorus Pesticides Antibody
    Scientia Agricultura Sinica. 2008, 41(3):  727-733 .  doi:10.3864/j.issn.0578-1752.2008.03.013
    Abstract ( 1605 )   PDF (387KB) ( 1142 )   Save
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    Diethylphosphono acetic acid (DPA) was used as a generic hapten in order to generate broad specificity polycolonal antibodies against a group of organophosphorus pesticides. Six New Zealand white rabbits were immunized with immunogens sysnthesised by the mixed-anhydried method and EDC method, the titers of antiserum had reached 25600 and 6400, respectively. The CI-ELISA was conducted with the antiserum, the detection limit for DPA was 3.536ng/mL and the I50 was 0.182μg/mL. The antiserum also showed high affinities to chlorpyrifos, diazinon, omethoate, parathion-ethyl and profenofos, the I50 to these pesticides were 0.12,0.15,0.21,0.88, 0.97 and 2.5μg/mL. It is concluded that this assay is suitable screening tool for quantities and semi-quantities detection of the five organophosphorus pesticides.
    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Estimate of Carbon Sequestration under Cropland Management in China
    Lin JIN Yu-e Li Qing-zhu Gao Yun-tong Liu Yun-fan Wan Xiao-bo Qin
    Scientia Agricultura Sinica. 2008, 41(3):  734-743 .  doi:10.3864/j.issn.0578-1752.2008.03.014
    Abstract ( 1308 )   PDF (481KB) ( 1307 )   Save
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    【Objective】For a long time, largely conventional tillage and burning straw cause much losses of Soil Organica Matter(SOM), which makes cropland be an emission source of Greenhouse Gas(GHG). In recent years, with no-tillage and straw returning being extended largely, as well as extensively using cropland management such as fertilizing and irrigating, organic carbon in cropland is rising again. It is supposed that cropland will be an important sink of GHG. By analyzing the change of soil organic carbon(SOC) under cropland management, carbon sequestration by cropland management in China was estimated and the character of carbon sink under cropland management in China was pointed.【Method】By reviewing lots of references and analyzing the change of SOC under cropland management in long-term experiment, we conceived cropland management scenarios. Meantime, according the areas partitioned by farming system in China, the quantity of carbon sink under cropland management was estimated and compared with the result of IPCC good practice guidance (GPG) Tier 2. Last, we used meta-analysis to estimate carbon sequestration by croplang management in China. 【Result】The total Chinese cropland carbon sink is 16.8~115.6Tg/a. It is different that SOC is influenced under different management in different area. Relationship between the annual ratio of SOC and original value in types of cropland management was analyzed. Both of them have distinct negative correlation. Saturation value of SOC is different under different managements.【Conclusion】The best management that increases SOC is using fertilizer and manure together. Next is straw returning, using manure and no-tillage. The last is using fertilizer, which even decreases the SOC. Last, We pointed the character of carbon sink under cropland management in China. Comparing with others, carbon sequestration of no-tillage, straw returning and using fertilizer and manure together is more.
    Effects of crop rotation and fertilization on soil organic C in semi-arid region
    Scientia Agricultura Sinica. 2008, 41(3):  744-751 .  doi:10.3864/j.issn.0578-1752.2008.03.015
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    【Objective】to better understanding on the mechanisms in crop rotation and fertilization on SOC in cropping soils in semi-arid regions. 【Method】Changes in soil organic C (SOC) (0-20 cm) and aboveground biomass from 1984 to 2002 was investigated in ten cropping systems in a long-term field experiment established in 1984 on the Loess Plateau. 【Result】For the continuous winter wheat cropping system (W/W), the above ground biomass without fertilization (W/W+CK) was 3.7 t hm-2, the above ground biomass in W/W+NP and W/W+NPM was 2.2 and 3.4 times that in W/W+CK. For the wheat- wheat+millet-pea rotation system (W/WM/P), the above ground biomass without fertilization (W/WM/P +CK) was 3.2 t hm-2, the above ground biomass in W/WM/P +NP and W/WM/P +NPM was 2.3 and 2.7 times that in W/W+CK. The above ground biomass in W/W/S +NP, G/G +CK, and G/G +NPM was 8.6, 25.5, and 30.5 t hm-2, respectively. After 18 years SOC in continuous winter wheat without fertilization was near to the value (6.5 g kg-1) at the experiment beginning. SOC in treatments applied with N and P fertilizers increased by 7-13% after 18 years, compared with control (the without fertilizer). SOC in the treatments applied with farmyard manure, N, and P fertilizer increased by above 47% after 18 years. Compared with continuous wheat, SOC with wheat-grass rotation or continuous alfalfa increased by 10-25% after 18 years. There was a highly significant correlation between SOC and soil total N ( ,R2=0.88**). 【Conclusion】The amount of returned root-C to soil plays an important role in SOC in the semi-arid region. Incorporation of crop straw into soil, crop rotation with grass, and conversion of cultivated land into grassland could promote SOC accumulation and sequestration of CO2 in farmlands in the semi-arid region.
    Study on response of rapeseed to boron application and critical level of soil available B in Hubei Province
    Scientia Agricultura Sinica. 2008, 41(3):  752-759 .  doi:10.3864/j.issn.0578-1752.2008.03.016
    Abstract ( 1494 )   PDF (254KB) ( 965 )   Save
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    【Objective】 In order to provide a scientific basis for balanced fertilization, the effect of boron fertilization on growth and yield of rapeseed was studied and soil critical available B was determined based on field trials in main rapeseed product region of Hubei Province. 【Method】30 field trials of B fertilizer application to rapeseed were conducted in 2005/2006 growth season. The soil critical value of B deficiency was determined according to effect of B fertilizer on seed yield and soil available B content.【Result】The results showed that B application promoted the nutritive growth and yield components of rapeseed. The average increment with B fertilization was 428 kg/hm2 and the increasing rate was 28.0% for all 30 trials. Compared to control (no B treatment), about 70% trails received significant increase by B application, and the percentage of yield increment more than 500 kg/hm2 was as high as 26.7% for all trials. The results indicated that the yield increment by B application had significantly negative relationship with soil available B content. Based on the relative yield of CK/+B at 90 percent level, the soil critical available B was 0.58 mg/kg which was higher than that in the 1980s determined by the second nationwide general soil survey achievement.【Conclusion】The yield-increasing effect of B fertilizer on rapeseed is notable and the soil critical available B is 0.58 mg/kg in Hubei province at present time. According to the soil B deficiency criterion, it is necessary apply B fertilizer to about 80% rapeseed cultivated field in the main product region of Hubei Province.
    Buffer capacity of the soil biota to soil nutrient and heavy metal
    Scientia Agricultura Sinica. 2008, 41(3):  760-765 .  doi:10.3864/j.issn.0578-1752.2008.03.017
    Abstract ( 1349 )   PDF (283KB) ( 1008 )   Save
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    The soil is a complex system with organic and abio-life-form. The soil-buffer is one of the important properties of the soil. The biodiversity and multifunctional nature of the soil improve the stability and harmony of the ecosystem. The soil biota plays an important role in the soil ecosystem. The author systematically described the function of the soil fauna (earthworms) and microbes in improving the soil nutrient buffer such as improving soil structure and controlling soil nutrients, the behavior in changing the soil physical and chemical properties through bio-sorption, enrichment, precipitation, dissolution and redox, as well as the influence to the bioavailability of heavy metals and the formation of heavy metal toxicity. The purpose is to provide a theoretical rationale for the modern agriculture, especially for organic agricultural.
    HORTICULTURE
    A Study on Formation and Transmission of n+1 Gametes of Primary Trisomics of Cabbage
    Scientia Agricultura Sinica. 2008, 41(3):  766-771 .  doi:10.3864/j.issn.0578-1752.2008.03.018
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    Abstract:【Objective】 Transmission rates of n+1 gametes is a important parameter for genetic analysis of trisomics. Determination of n+1 gamete transmission rate of the cabbage trisomics is significant for using this set of trisomics to carry out gene orientation and other genetic studies. 【Method】Each primary trisomics of the cabbage with their diploid parent-9601 was crossed reciprocally, and the hybrid seeds were cultured on MS medium to propagate into clones which were used for chromosome number identification by root tip squash method. The ratio of (2n+1)plants and total plants identified from progenies of 2n+1×2n was denoted as the female n+1 gamete transmission rate, and that from 2n×2n+1 was denoted as the male n+1 gamete transmission rate.【Result】The female n+1 gamete transmission rates were 15.28% for tri-1, 12.68% for tri-2, 12.31% for tri-3, 30.51% for tri-4, 22.81% for tri-5, 7.46% for tri-6, 5.36% for tri-7, 42.37% for tri-8, 9.23% for tri-9. The female n+1 gamete transmission rates were 12.12% for tri-1, 12.33% for tri-2, 7.81% for tri-3, 4.76% for tri-4, 8.93% for tri-5, 10.94% for tri-6, 1.54% for tri-7, 2.94% for tri-8, 13.04% fortri-9.The main affecting factors on male n+1 gamete formation and transmission were the rates of trivalent formation at prophaseⅠand 9/9/10/10 division at anaphaseⅡ, and pollen viability.【Conclusion】n+1 gametes of the cabbage trisomics can transmitted through female and male parents.
    Effects of Different Concentrations of Cu2+,Zn2+ and Mn2+ in nutrient solution on SOD Activity of Grafted and Own-Root Cucumber Seedling Leaves under Low temperature
    Scientia Agricultura Sinica. 2008, 41(3):  772-778 .  doi:10.3864/j.issn.0578-1752.2008.03.019
    Abstract ( 1370 )   PDF (382KB) ( 1050 )   Save
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    【Objective】The aim of this study is to investigate the functions of Cu2+, Zn2+and Mn2 on cold tolerance of cucumber seedlings. 【Method】The contents of Cu, Zn and Mn and the changes of Superoxide Dismutase(SOD) and its isoenzymes activities and electrolytic leakages of own-root and grafted cucumber seedlings leaves cultivated by different concentrations of Cu2+, Zn2+and Mn2+under low temperature were studied. 【Result】The contents of Cu, Zn and Mn ,the activities of SOD, Mn-SOD and Cu/Zn-SOD of own-root cucumber seedling leaves cultivated by high concentrations of Cu2+, Zn2+and Mn2+ in nutrient solution were higher than the control, while the Fe-SOD activity and electrolytic leakages were lower; When grafted cucumber seedlings were cultivated by low concentrations of Cu2+, Zn2+and Mn2+in nutrient solution, the results were contrary.【Conclusion】The functions of Cu2+, Zn2+and Mn2 on cold tolerance of cucumber seedlings were very important. And it was proved that the roots of grafted cucumber seedlings which could absorb more Cu, Zn and Mn after grafted by figleaf gourd was one of the reasons that the grafted cucumber seedlings could have more chilling tolerance than own-root cucumber seedlings. The contents of Cu, Zn and Mn and SOD activity of own-root cucumber seedlings both could be enhanced by increasing the contents of Cu2+,Zn2+and Mn2+in nutrient solution, so that the chilling tolerance also could been enhanced.
    Changes of Sucrose Content and Sucrose-metabolizing Enzymes Activities in the Florescence of Peach
    Scientia Agricultura Sinica. 2008, 41(3):  779-785 .  doi:10.3864/j.issn.0578-1752.2008.03.020
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    Abstract:【Objective】To study the law of changes about sucrose content ratio and sucrose-metabolizing enzymes specific activities in the florescence of peach.【Method】The content of sucrose, starch and the specific activities of sucrose-metabolizing enzymes, including acid invertase(AI), neutral invertase(NI), sucrose phosphate synthase(SPS) and sucrose synthase (SS) were studied in florescence(from the period of bud to wilt)of JingYu and Kubao peach.【Result】The rule of sucrose accumulation took a similar change in disparate parts of two varieties, and the content ratio was obviously higher in phloem than other parts. The content of soluble total sugar assumed descendent trend in the mass, and the content in phloem was higher than it in other parts except for the individual value in petal on the day of full flower (the 10th). At the same time the content of starch was elevatory continuously in ovary and leaf during florescence, and it reduced in phloem besides the content depressed then elevatory in petal. The specific activity of AI and NI kept infirm influence to the increase of sugar content, whereas sucrose accumulation accorded with SS-synthese and SPS specific activity. The accumulation and components of sugar were affected evidently by SS-disassemble activity.【Conclusion】Phloem was the fountainhead of carbohydrate-metabolizing in the florescence of peach. The space-time character expression of sucrose-metabolizing enzymes were regulated by correlative factors.
    Heredity of several flower characters in Dendranthema
    Scientia Agricultura Sinica. 2008, 41(3):  786-794 .  doi:10.3864/j.issn.0578-1752.2008.03.021
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    Abstract:【Objective】The study researched heredity of several flower characters in Dendranthema, that will provide theoretic foundation for breeding of interspecific cross.【Method】Several flower characters of wild diploid species D.lavandulifolium, D. nankingense , D. dichrum, tetraploid D.indicum, D.morifolium (hexaploid and its aueuploid) and their F1 generation were observed and analyzed.【Result】The hybrid’s mean inflorescence diameter, the number of ray florets and tubular florets were 112.4%,108.0%,118.6% of parent’s respectively, and the value of these three characters declined except for the number of tubular florets of F1 generation between wild diploid species when combination from between wild diploid species to between diploids and tetraploids, diploids or tetraploids and polyploid chrysanthemum varieties. 【Conclusion】The inflorescence diameter, number of ray florets and tubular florets appeared heterosis in F1 generation. Polyploid chrysanthemum varieties had stronger flower color genetic ability than diploid and tetraploid. the purple and white color of flowers were stronger than yellow. The heredity of flat plate shape was stronger than 1-3 cycle plate shaped flower. The heredity of double flowers was stronger than 1 cycle flowers.
    STORAGE·FRESH-KEEPING·PROCESSING
    Properties of Polyphenol Oxidase extracted from ‘zhonghuashoutao’ Peach Flesh
    Yu-Quan DUAN Wei DONG Ming-Jing ZHANG Shuang-Qing FENG Yu-Mei ZHAO
    Scientia Agricultura Sinica. 2008, 41(3):  795-799 .  doi:10.3864/j.issn.0578-1752.2008.03.022
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    Polyphenol oxidase purified from ‘ZHONGHUASHOUTAO’ peach flesh showed the protein as a single polypeptide with a molecular weight of about 66.0kD. The optimum pH for PPO activity was 6.5 with catechol. The enzyme was relatively temperature stable with maximum activity at 60℃. Chlorogenic acid, catechol, and p-hydroxybenzyl acid were good substrates for the enzyme. FeSO4, EDTA, and Vc markedly inhibited PPO activity, whereas CaCl2, CuSO4 and MnSO4 enhanced PPO activity.
    Influence of ionic environment on whey protein gels and gels microstructure
    Scientia Agricultura Sinica. 2008, 41(3):  800-807 .  doi:10.3864/j.issn.0578-1752.2008.03.023
    Abstract ( 1138 )   PDF (785KB) ( 1428 )   Save
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    The effects of ionic variety, ionic strength, heating temperature, pH value and protein concentration on texture properties,water-holding capacity and microstructure of whey protein gels were investigated. The influence of different concentration NaCl and CaCl2 on gels hardness,springiness and cohesiveness was studied by using TA-XT plus instrument;the microstructure was observed with scanning electron microscopy (SEM). The results showed that salts could greatly vary gels properties, it need higher concertation of NaCl than that of CaCl2 at the same gels conditions. The springiness and water-holding capacity (WHC) of NaCl gels was better than that of calcium chloride gels. The better texture would be obtained for protein concentration between 8~10%. The microstructure of NaCl gels was more compact than that CaCl2 gels, and it showed that NaCl gels have more hard texture and better water-holding capacity in macrostructure. Therefore, textural properties and functional properties of NaCl gels were better than that of CaCl2 gels.
    Response of Endogenous Salicylic Acid and Jasmonic Acid to Mechanical Wounding in Pea Leaves.
    Scientia Agricultura Sinica. 2008, 41(3):  808-815 .  doi:10.3864/j.issn.0578-1752.2008.03.024
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    Abstract: Effects of mechanical wounding on endogenous salicylic acid (SA) and jasmonic acid (JA) were investigated in pea (Pisum sativum L.) leaves. The results showed that endogenous JA rapidly accumulated within 30 min after wounding and then significantly decreased, which is called “JA burst”. The increase in the activities of both lipoxygenase (LOX) and allene oxide synthase (AOS) lagged behind JA burst. A second slight increase in JA level was observed at 24 h after wounding treatment, and at this moment, higher activities of LOX and AOS also were detected. Endogenous free SA content decreased accompanying with JA burst. Effects of exogenous JA application were similar to these of wounding treatment on endogenous SA level and phenylalanine ammonia lyase (PAL) activity, whereas exogenous SA application led to the significant inhibition of LOX and AOS activities and the decrease of endogenous JA level at the early stage of treatment. It is thus suggested that JA burst and SA decrease in early response to wounding may constitute an important mechanism by which plant starts the related defense reaction and adapts to wounding stress.
    Application of mixture design and optimization software to design of formulation of pure cultures in Tibetan kefir
    Scientia Agricultura Sinica. 2008, 41(3):  816-822 .  doi:10.3864/j.issn.0578-1752.2008.03.025
    Abstract ( 1417 )   PDF (491KB) ( 929 )   Save
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    【Objective】In order to obtain optimization formulation of pure cultures in Tibetan kefir. 【Method】The influence of the different mixture of five strains in pure cultures in Tibetan kefir on flavor components in fermented milk were studied using Mixture Design. Regression model on microorganism composition and main metabolites was established.【Results】The results suggested that the predictable production of lactic acid reached the maximum, 8.16g/L,the most predictable production of diacetyl was 77.23mg/L, the most predictable production of ethanol was 4259mg/L,and the production and CO2 was 2.12g/L. Based on these, response values satisfied all expectation were optimized, and the most excellent combination was Lactococcus lactis 27%, Leuconostoc mesenteroides 37%, Lactobacillus kefiri 11%, Lactobacillus casei 10% and Kluyveromyces marxianus 15%.【Conclusion】With the aid of analysis software (Design-Expert6.0.5), formulation of pure cultures in Tibetan kefir could be optimized for several responses and the best formulation could be gotten.
    ANIMAL SCIENCE·RESOURCE INSECT
    Mapping the GAS6 Gene of Porcine to SSC11q11-17 with SCHP
    Scientia Agricultura Sinica. 2008, 41(3):  823-827 .  doi:10.3864/j.issn.0578-1752.2008.03.026
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    Abstract: ObjectiveGAS6growth arrested-special 6is associated with adhesion, identification, survival, proliferation, differentiation, and growth of cell in human. However, little is known about gene of GAS6 of porcine. In order to map the position of the gene of GAS6 in chromosome in pigs and to found the basis of cloning this gene and studying the function of this gene, Methodwe used the Pig別rodent somatic cell hybrid panel(SCHP) and INRA-Minnesota porcine radiation hybrid panel(IMpRH). Results the isolated fragments GAS6 gene of swine was deposited into GenBank and accession number was AY880668. The results showed that the gene of GAS6 was located in the SSC11q11-17. The GAS6 gene were linkaged closely to the microsatellite SW1452 anchored on SSC11 with the rate of retension of 0.22, LOD score threshold were 16.88 and the distance in map of the radiation hybrid (RH) was 56 cR. ConclusionThat the position of GAS6 was on the SSC11q11-17 conformed to the conclusions which was drawn by means of IMpRH because the SSC11 is homologous with most of HSA11.
    Cloning and Sequence Analysis on Exon 9-15 of FAS Gene in Mammary Gland of Xinong Saanen Goat
    Scientia Agricultura Sinica. 2008, 41(3):  828-833 .  doi:10.3864/j.issn.0578-1752.2008.03.027
    Abstract ( 1536 )   PDF (619KB) ( 922 )   Save
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    The acetyl-CoA and malonyl-CoA transacylases region encoded exon 9-15 of goat FAS gene plays a central role in synthesis of short- and medium- chain fatty acids in goat milk. In the research, the mRNA from Xinong Saanen goat mammary gland on 28th day of lactation was isolated, and exon 9-15 of FAS gene in xinong saanen goat mammary gland and partial cDNA of 3’ end of exon 8 and 5’ end of exon 16 were amplified with RT-PCR using the mammary gland cDNA as the template by reverse transcription(GenBank accession no. DQ 915966), after sequencing we analyzed homology of sequence among species and speculated its function by bioinformatics. Sequencing of the fragments showed that the cDNA consisted of 1449 bp, and it contained 1338 bp exon 9-15 and 9 bp 3’ end of exon 8 and 52 bp 5’ end of exon 16, which encoded 483 amino acid including the 951 bp (454 nt-1404 nt) AT/MT domain;The homologies of nucleotide and amino acid sequence of exon 9-15 of Xinong Saanen goat mammary gland FAS gene with those of bovine (NM_001012669), human (NM_004104), rattus (NM_017332) and chicken (NM_205155) were 95%、85.7%、82.7%、73.2% and 92.9%、77.7%、82.3%、64.7% respectively; Compared with the other four species, an amino acid in exon 10 of goat FAS gene was absent.
    Bioavailability of intravenously injected manganese sources for broilers
    Scientia Agricultura Sinica. 2008, 41(3):  834-840 .  doi:10.3864/j.issn.0578-1752.2008.03.028
    Abstract ( 1570 )   PDF (318KB) ( 771 )   Save
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    [Objective] Differences in Mn metabolic utilization between organic and inorganic Mn sources or among organic Mn sources for broilers were detect. [Method] Broilers on 22 day-old were randomly allotted to one of five treatments and injected with 0.9% NaCl solution (Control), or the 0.9% NaCl solution dissolved with Mn sulfate, or organic Mn source with weak, moderate, or strong chelation stength for 20 days. [Result] On d 10 and 20 after injection, chicks injected with Mn containing solution had a significantly higher metatarsal bone ash Mn content, heart Mn content, heart MnSOD activity, and MnSOD mRNA level than those in control group (P < 0.03), but the indices except MnSOD mRNA level could not consistently detect the differences among Mn sources. Chicks injected with solution containing organic Mn sources with moderate chelation strength had higher heart MnSOD mRNA level than those injected with Mn sulfate (P < 0.01) and organic Mn sources with strong chelation strength (P<0.09), and chicks injected with solution containing organic Mn sources with weak chelation strength had higher heart MnSOD mRNA level than those injected with Mn sulfate (P < 0.06). [Conclusion] The results indicated that intravenously injected Mn could significantly affect heart MnSOD gene expression trancriptionally, and this functional index could more rapidly, sensitively and consistently detect the difference among Mn sources than other indices. Intravenously injected Mn from the organic Mn source with moderate and weak chelation strength was utilized and functioned in the sensitive target tissue of broilers more effectively than Mn from Mn sulfate, and the organic Mn source with the strong complex strength would be not good for tissue utilization.
    Establish and optimize the system of sexual identification of earlier embryos from generic hybridizationof chicken and quail
    Scientia Agricultura Sinica. 2008, 41(3):  841-845 .  doi:10.3864/j.issn.0578-1752.2008.03.029
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    [Abstract]We had selected the earlier embryos of generic hybridization of chicken and quail (2.75-5.0 d),designed primers from quail Wpkci and β- actin genes,and used RT-PCR method to establish a way to identify the sex of earlier embryos of hybridization of the chicken(♂) and quail(♀).Amplified cDNA of quails of known sex and hybridizations’embryos of unknown sex with Wpkci primers ,we had acquired two particular segments of 402 bp and 296 bp from the female embryos, but none from the male embryos. In order to verify credibility and stability of this system deeply, we had acquired three particular segments of 490 bp,402bp,296bp from the female embryos with multi-ply PCR, but only acquired the internal standard segment of 490 bp from male embryos.Based on the optimization of the key factors of the experiment, we had established a simple,fast,credible and stable method to identify the sex of earlier hybridizations 'embryos.
    VETERINARY SCIENCE
    The Determination of Specific Antibodies in Mice orally Immuned Lactobacillus casei expressing Porcine Parvovirus VP2 protein on cell surface
    Scientia Agricultura Sinica. 2008, 41(3):  846-851 .  doi:10.3864/j.issn.0578-1752.2008.03.030
    Abstract ( 1227 )   PDF (444KB) ( 903 )   Save
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    Lactobacillus casei strain 393 was selected as oral vaccine carrier for the expression of Porcine Parvovirus (PPV) main immune protective antigen VP2 protein. The constructed pPG-VP2 plasmid expressing PPV VP2 protein on cell surface was electroporated into Lactobacillus casei 393 generating recombinant strain pPG-VP2/L.casei393.The interest protein was detected by SDS-PAGE、Western-blot and indirect immunofluorescence after induced by 2% Lactose in MRS broth. Oral immunization of BALB/c mice were received with recombinant strain harboring pPG-VP2 and L.casei393 harboring pPG. Specific anti-PPV VP2 secret immunoglobulin A (sIgA) antibody was detected by indirect enzyme linked immunosorbent assay (ELISA) in the feces and intestines mucus after orogastric immunization, and anti-PPV VP2 immunoglobulin G (IgG) antibody was detected by indirect ELISA in the serum of immunized mice. The results indicated that the mice immunized with pPG-VP2/L.casei393 could produce clear anti-PPV antibody level. Our work establish important material basement for the development of PPV oral vaccine.
    The effect of high magnesium on urolithiasis in feedlot wethers
    Scientia Agricultura Sinica. 2008, 41(3):  852-860 .  doi:10.3864/j.issn.0578-1752.2008.03.031
    Abstract ( 1171 )   PDF (359KB) ( 937 )   Save
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    [Objective] The effect of high diatery magnesium on urolithiasis in feedlot wethers compared with high potassium and phosphate under the chinese traditional feeding style with cotton seed meal and rice straw diet. [Method] Eighteen wethers aged from 3-4 months were randomly allotted to three groups, as group A, B, C feeding for three months with cotton seed meal and rice straw diet comprised of P 0.40% ,K 1.24%, Mg 0.32% respectively. From the third month, KH2PO4 and K2HPO4 was provided to goats in group B by drinking water and so did MgO to goats in group C,. goats in group A still drink tap water, which means dietary P, K in group B was added up to 1.74% and 0.75%, and dietary Mg in group C to 0.56%. Blood and urine samples were collected to analyze the concentration of P, K, Mg, Ca ion, and the activity product of potassium magnesium phosphate was calculated to evaluate the supersaturation of urine before and after urolith formation. The composition of urinary sediments and stones was studied by chemical qualitative analysis,X-ray diffraction, X-ray energy dispersive spectrometry (EDS) and fourier transform infrared (FTIR) spectroscopy. [Result] The urolithiasis was found on day 5-8 in all 6 goats of group C after drinking water contains MgO, one urolithiasis case was found in group B and C on day 25,28 of the third month respectively. Synthetical analysis showed that the chemical composition of urinary stones and urinary sedimental crystals was potassium magnesiumphosphate and magnesium ammonium phosphate. P, K, Mg ion concentrations of urine decreased significantly, so did activity product of potassium magnesium phosphate. [Conclusion] High dietary levels of magnesium is a more important factor of urolithiasis in wethers compared with feeding with high P, K diets containing, and the effect of magnesium induced urolithiasis might be associated with the change of inhibitory activity of some inhibitor in urine.
    Research of Dissociation Culture of Chicken Trachea-bronchia Epithelium and Culture Characteristics of IBV in the Epithelium
    Scientia Agricultura Sinica. 2008, 41(3):  861-867 .  doi:10.3864/j.issn.0578-1752.2008.03.032
    Abstract ( 1637 )   PDF (435KB) ( 1346 )   Save
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    [Objective] The purpose of this study was to explore and find out appropriate technology of separation and culture for chicken trachea-bronchia epithelium and also to research culture characteristics of IBV in chicken trachea-bronchia epithelium. [Method] By the method of douche, chicken trachea-bronchia epithelium was digested overnight at 4℃ and separated. Then the test of immunohistochemistry of No.8/18 keratins of chicken trachea-bronchia epithelium was done to identify epithelium. The cultured epithelium was infected respectively by IBV-M41 strain(Style Breath ) and IBV-T strain(Style Kidney), and respectively at 12,24,32,48,60,72,84 and 96h after epithelium infected by IBV we observed morphologic changes of epithelium. At last the method of RT-PCR was used to detect IBV mRNA in each group of cells. At the same time the ability of haemagglutination and pathological changes of chicken embryo caused by IBV were tested. [Result] By the method of douche and digestion overnight chicken trachea-bronchia epithelium have been separated and have full morphologic structure and clearly visible cilia and high livingness. Through three-step purification the epithelium has good uniformity and grow faster. Respectively at 72h and 84h after each IBV strain and its corresponding epithelium co-culture, the infected epithelium appeared obvious cell pathologic effect(CPE), and the test of ability of haemagglutination and pathological changes of chicken embryo by IBV were both positive. [Conclusion] We have successfully separated and cultured chicken trachea-bronchia epithelium to fulfill the need of relative deeper research by providing chicken trachea-bronchia epithelium as research model. And both IBV-M41 strain and IBV-T strain may cause chicken trachea-bronchia epithelium to appear CPE, which provide nicer bases for future relative research about IBV laboratory diagnosis, medicine sensitivity test and pathology mechanism caused by IBV.
    Establishment of Indirect immunofluorescence assay technique to the detection for DSHDV in paraffin-embedded tissues and its application in the diagnosis and antigen localization
    Scientia Agricultura Sinica. 2008, 41(3):  868-874 .  doi:10.3864/j.issn.0578-1752.2008.03.033
    Abstract ( 1405 )   PDF (710KB) ( 913 )   Save
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    【Objective】To establish an Indirect Immunofluorescence assay (IFA) technique for the detection of Duck Swollen Head Haemorrhagic Disease Virus (DSHDV) in paraffin-embedded tissue,and provide an effective examination method for the diagnosis, subcellular antigen localization and dynamic distribution in vivo of DSHDV.【Method】Through differential centrifugation purified DSHDV,DSHDV-specofic antibodies were obtained from rabbits vaccinated with purified virus, and then purified through DEAE-SephadexA-50 columns. An IFA was established to detect the DSHDV antigen in paraffin-embedded tissue. then DSHDV-antigen was detected in each organ tissues of 28-day-old ducks which were experimentally infected with DSHDV.【Result】The optimum conditions of this IFA were as follows: Antigen was retrieved by microwave in 0.01 mmol/L citrate buffer solution(PH6.0) for twenty minutes, and blocked with 10% horse serum for 30min at 37℃.Slices were incubated overnight at 4℃ with diluted primary antibody(1:50) ,and then incubated for 30 min at 37℃ with diluted FITC-labelled-secondary antibody(1:100)which contain 0.01% Evans blue. The IFA was applied in detecting the DSHDV antigen in liver tissues of dead ducks, and the results were positive in DSHDV infected dead ducks and negative in dead ducks infected by Duck Plague Virus, Duck viral hepatitis Virus and Riemerella Anatipestifer. The IFA was also applied in detecting the virulent DSHDV antigen in different organs of artificially infected dead 28 old days ducks, and the viral antigen was detected in the heart、liver、spleen、pancreas、lung、kidney、bursa of fabricius、esophagus、trachea、muscle、thymus、duodenum、jejunum、ileum、cecum and rectum, while viral antigen was not detected in the harderian gland、brain、skin and proventriculus. The antigen distributes mainly in the cytoplasm of the infected cells. 【Conclusion】The establishment of IFA is a sensitive and specific method in detecting DSHDV antigen in paraffin-embedded tissue, and it’s a good method in diagnosis and antigen location of DSHDV. IFA can application in the experimentally diagnosis and study on the antigen distribution dynamics in duck tissue cells.
    Pharmacokinetics study of the berberine in ultra micro-powder Huanglian jiedusan on rabbits
    Scientia Agricultura Sinica. 2008, 41(3):  875-879 .  doi:10.3864/j.issn.0578-1752.2008.03.034
    Abstract ( 1337 )   PDF (310KB) ( 1329 )   Save
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    【Objective】This paper aimed at studying the effect of ultromicro-pulverization method on pharmacokinetics of the berberine in Huanglian jiedusan 【Method】HPLC method was applied to determine of the concentration of plamsa berberine in rabbits which were given the ultra micro-powder and the common powder of Huanglian jiedusan by po 4mg·kg-1 respectively. The plasma concentration-time curve of berberine was dealt with PKS pharmacokinetics software, then pharmacokinetics characteristics were compared between the ultra micro- powder and the common powder of Huanglian jiedusan. 【Result】The results showed that the best pharmacokinetics model of berberine in two groups were all two-compartment open model. The main pharmacokinetic parameters of berberine in ultra micro- powder were as below:A =0.672 (μg/mL), α=0.641(1/h) , B=0.015(μg/mL), β=0.0241( 1/h) , Ka=0.911(1/h), t1/2a=1.08(h),t1/2β=28.72(h),AUC0→∞=0.895(μg·h /mL),Vd =185.25(L/kg),Tpeak=1.48(h), Cmax=0.0913(μg/mL);The main pharmacokinetic parameters of berberine in common powder were as below:A=0.557(μg/mL) , α=0.520(1/h), B=0.0150(μg/mL), β=0.0367( 1/h) , Ka=0.617(1/h), t1/2a=1.33(h),t1/2β=23.56(h), AUC0→∞=0.613(μg·h /mL), Vd/F=178.02(L/kg),Tpeak=1.934(h),Cmax=0.0565(μg/mL). Compared to common powder, ultra micro- powder has shorter Tpeak and increased Cmax and the relative bioavailability of berberine in the ultramicro-powder was improved by 46.00%. 【Conclusion】the ultromicro-pulverization method could promote the absorption and distribution of HLJDS in body and then improve the bioavailability of berberine in Huanglian jiedusan significantly.
    Analysis of the Proteome of the Larvae of the Higher Royal Jelly Producing Worker Bees (Apis mellifera L.)
    Scientia Agricultura Sinica. 2008, 41(3):  880-889 .  doi:10.3864/j.issn.0578-1752.2008.03.035
    Abstract ( 1128 )   PDF (426KB) ( 713 )   Save
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    【Objective】The purpose was to investigate the protein composition of worker bees larvae of the higher producing royal jelly bees (Apis mellifera L.).【Method】Two-dimensional gel electrophoresis(2-DE) was employed to separate and quantify proteins of the larvae of the higher producing royal jelly worker bees (Apis mellifera L.) during their six days development.【Result】The results showed that 262, 418 and 194 proteins with a range of molecular weight 12.2-88.2 KDa and relatively narrow scope of pI 4.00-9.24 could be detected on day 2, day 4 and day 6 during the developmental process of the larvae, respectively. Meanwhile 84 protein spots were constantly detected during the larvae development, among them 33 percent were expressed in up trend along with the larvae development, 21 percent were in down trend and 46 percent were irregular. In addition, the specific proteins expressed on day 2, day 4 and day 6 were 88, 209, 63, respectively. Except the constant and specific expressed proteins, 84 proteins were expressed on day 2 and day 4 but silenced on day 6, 41 proteins wereexpressed on day 2 and day 4 but silenced on day 6, only 6 proteins were expressed on day 2 and day 6 but silenced on day 4.【Conclusion】The result indicates that the larvae development is a sequential and complex gene controlled process, while the proteins of larvae of day 4 express the most active. The eternal expressed proteins suggest that they are conservative and indispensable for this event. The specific proteins detected at different developmental stage suggesting it is indispensable to regulate the larvae’ growth.
    RESEARCH NOTES
    Genetic Diversity among Guangxi Local maize varieties and Canadian Maize Populations
    Scientia Agricultura Sinica. 2008, 41(3):  890-900 .  doi:10.3864/j.issn.0578-1752.2008.03.036
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    Abstract: In order to take the best of the Guangxi local maize varieties and broaden the genetic base of the Guangxi maize germplasm, the genetic diversity among 45 Guangxi local maize varieties (OPVs) and 15 Canadian maize populations, was analyzed used Bulk-SSR method. The total 240 DNA samples were extracted from 4 bulks of DNA for each population or OPV, and 10 random individual plants equally for each bulk. The results showed that 245 alleles were detected with 70 pairs of selected primers in the 240 bulks of DNA samples attracted from 60 OPVs or populations. The number of alleles per locus was on average 3.5 and ranged 2-6. The Clustering results with UPGMA method based on the genetic similarities of 60 populations showed that 45 Guangxi local OPVs and 15 Canadian populations were classified into 2 groups. Each group consisted of flint and dent subgroups. The waxy corn samples from Guangxi were not clustered as an independent group, but into flint or dent subgroups. Seven levels of allelic combinations among 245 were designed to analyze the least number of alleles required to reveal genetic relationship among the 60 populations. The results indicated that 160 alleles were enough to cluster the 60 germplasm in the current study. Diagnostic alleles were amplified with some certain primers in Guangxi and Canadian germplasm, respectively. Combined with the results of clustering analysis, these diagnostic alleles will be of great importance to find the useful maize germplasm in breeding efforts.
    Effects of Alkaline Stress on Biomass and Soluble Osmoticum Accumulations in Different Organs of Helianthus tuberosus L. Seedlings
    ChengLong Wu
    Scientia Agricultura Sinica. 2008, 41(3):  901-909 .  doi:10.3864/j.issn.0578-1752.2008.03.037
    Abstract ( 1126 )   PDF (444KB) ( 1006 )   Save
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    【Objectives】The purpose of this paper is to probe into the relationships between the resistance of Helianthus tuberosus L. Seedlings to alkaline stress and the accumulations of biomass and soluble osmoticum in different organs.【Method】with sandy culture in greenhouse,two Helianthus tuberosus L. genotypes Nanyu 8(Ht 1, alkaline-tolerant) and Nanyu 1(Ht 2, lower alkaline-tolerant) were exposed to nutrient solution with Na2CO3 added(0,12.5,25.0,37.5mmol mmol•L-1), and the biomass and osmoticum in different organs were determined.【Result】under alkaline stress, there were considerable differences in the biomass accumulations, the contents of Na+, K+, soluble sugar and proline in different organs between two genotypes. compared with each other it can be see that, Ht 1presented higher K+ in leaf and biomass invested in root under lower alkaline stress(12.5mmolL-1), higher soluble sugar in leaf and root and also higher K+ but lower Na+ in root under higher alkaline stress(25,37.5mmolL-1), whereas Ht 2 presented higher Na+ in root, proline in all organs and soluble sugar in stem in all designed concentrations.【Conclusion】The results indicated that considerable variations in the amount of biomass and soluble osmoticum among plant organs were related to Helianthus tuberosus L. genotypes in response to alkaline stress; Ht 1 had higher K+ in leaf and biomass invested in root under lower alkaline stress, higher soluble sugar in leaf and root and also higher K+ but lower Na+ in root under higher alkaline stress, which may ascribe to its higher alkaline tolerance. The accumulation of proline may be a damaged response of Helianthus tuberosus L .to alkaline stress.
    Inhibitory effects of persimmon tannins on enzyme activities from snake venoms and primary study on its mechanism
    Scientia Agricultura Sinica. 2008, 41(3):  910-917 .  doi:10.3864/j.issn.0578-1752.2008.03.038
    Abstract ( 1827 )   PDF (510KB) ( 1000 )   Save
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    【Objective】To study the inhibitory effects of persimmon tannins (PT) on the activities of proteolytic enzyme, phospholipase A2, L-amino acid oxidase, arginine esterase, acetylcholinesterase and thrombin-like enzyme from Agkistrodon halys Pallas, Agkistrodon acutus, Naja naja atra venom , and the action mechanism between venom and PT. 【Method】 Different amounts of PT were mixed with snake venoms, after incubation at 37℃ for 1h and centrifuge, the enzyme activities of proteolytic enzyme, phospholipase A2, L-amino acid oxidase, arginine esterase, acetylcholinesterase and thrombin-like enzyme from the supernatants were mensurated, and the precipitates , supernatants and venom were analyzed by SDS-PAGE.【Result】The enzyme activities of proteolytic enzyme, phospholipase A2, L-amino acid oxidase, arginine esterase, acetylcholinesterase, and thrombin like enzyme were inhibited completely when the rate of snake venom and PT reached 1:1 (w/w), and the inhibitory effect was in a dose-dependent manner; PT was found to have very high combinative ability with venom proteins in a non-specifically manner.,but with different selectivity for different protein.【Conclusion】It was suggested that the enzyme activities from snake venoms could be inhibited remarkably by persimmon tannin in vitro by non-specifically combination.
    Analysis of Off-odor Volatiles of Irradiated Vacuum-packaged Refrigerated Pork
    Scientia Agricultura Sinica. 2008, 41(3):  918-924 .  doi:10.3864/j.issn.0578-1752.2008.03.039
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    Abstract: [Objective] The purpose of this study was to lay a foundation for our study on the generation mechanism of off-odor in irradiated meat and the removal method. [Method] The main composition of off-odor volatiles of irradiated refrigerated vacuum-packaged pork is analyzed by gas chromatograph mass spectrometry (GC-MS). The off-odor volatiles were collected by a cooled via in liquid nitrogen, that via was connected to vacuum system. [Result] The main composition of off-odor volatiles are dimethyl disulfide, dimethyl sulfide, dimethyl trisulfide, S-methyl thioacetate and methanethiol. The off-odor volatiles come from Irradiated Cystine, Methionine and VB1. [Conclusion] The main composition of off-odor volatiles are S-containing composition that come from irradiated S-containing Amino acid and VB1.
    Study on Serum and Hypothalamic Hormone Levels Regulating Feed Intake in Different Genotypes of Newly-hatched Chickens
    Scientia Agricultura Sinica. 2008, 41(3):  925-932 .  doi:10.3864/j.issn.0578-1752.2008.03.040
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    Abstract: 【OBJECTIVE】This trial was conducted to study the importance of insulin, leptin, NPY and α-MSH in the regulation of feed intake in newly-hatched chicks, and mechanism of feed intake difference among different genotypes.【METHOD】Changes of feed intake, serum insulin and leptin, hypothalamic insulin, leptin, NPY and α-MSH during 11 days posthatching in Peking and AA chicks were observed.【RESULTS】Developmental changes of serum and hypothalamic hormone levels with age differed in genotypes and parameters. Serum insulin levels of Peking and AA chicks at 0 day were lower than that at 1~11 days. Serum leptin level of AA chicks at 0 day was also lower than that at 1 and 3 days; however in Peking chicks, serum leptin level at 0 day was similar to that at 1 day, and slightly higher than those at the other ages. Correlation analysis showed that ADFI showed high correlations with serum leptin, hypothalamic insulin, lepin and NPY levels, but there were no correlations between ADFI and serum insulin or hypothalamic α-MSH levels. Comparative results between lines showed that serum insulin level at 1 day, serum leptin levels at 1 and 3 days, and hypothalamic α-MSH level at 0 day of Peking chicks were significantly lower than that of AA chicks, but serum insulin levels at 0 and 11 days of Peking chicks were the opposite, and there were no differences in serum and hypothalamic hormone levels of the other days between Peking and AA chicks.【CONCLUSION】Insulin and leptin could play a role in regulation of feed intake in newly-hatched chick, but the parameters observed did not explain why there was significant difference in feed intake between Peking and AA chicks. Therefore, further studies are needed to elucidate mechanisms of regulating feed intake of newly-hatched chicks and feed intake difference between different genotypes.
    Cloning of cDNA encoding Odorant Binding Protein ASP2 in Working Bee’s Antenna of Apis cerana cerana and its Prokaryotic Expression
    Scientia Agricultura Sinica. 2008, 41(3):  933-938 .  doi:10.3864/j.issn.0578-1752.2008.03.041
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    【OBJECTIVE】Cloning and expression in prokaryotic system of a new cDNA, named Acer-ASP2, encoding odorant binding protein ASP2(antenna special protein 2) open reading frame(ORF).【METHOD】Based on the antenna materials of 13 ages in days from Apis cerana cerana, reverse transcription-polymerase chain reaction(RT-PCR) and pET-30a(+)/BL21(DE3) prokaryotic expression system, Acer-ASP2 was cloned and expressed. 【RESULTS】The full length of Acer-ASP2(GenBank loucs: DQ449667) was 429bp, encoding 142 amino acids and the predicted MW and pI were 15.7 kD and 4.36, respectively. Sequencing and analysis indicated that Acer-ASP2 was characterized by six conservative Cys, which shared typical feature of OBP from other insects. Further analysis showed Acer-ASP2 likely belongs to the family of GOBP. The molecular weight of recombinant protein of pET/Acer-ASP2 was about 21kD, induced by IPTG to highly express a protein of as an inclusion body and accumulated up to 59.7% of bacterial total protein.【CONCLUSION】We have cloned and expressed a new cDNA of Apis cerana cerana, for the further research to the molecular structure and function.