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    15 April 2012, Volume 45 Issue 8
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Study on the Renewed Tendency and Key Backbone-Parents of Inbred Rice Varieties (O. sativa L.) in China
    TANG Sheng-Xiang, WANG Xiu-Dong, LIU Xu
    Scientia Agricultura Sinica. 2012, 45(8):  1455-1464.  doi:10.3864/j.issn.0578-1752.2012.08.001
    Abstract ( 721 )   PDF (375KB) ( 1511 )   Save
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    【Objective】The objective of the study is to explicit and understand the renewed tendency and key backbone-parents of inbred rice varieties (O.sativa L.) since 1949. 【Method】 The present study tracked and analyzed the blood relationship of 3 656 inbred rice varieties bred and extended in China. 【Result】The study indicated that during the latest period of 60 years, by dwarf breeding, inbred rice varieties have been renewed in mass scale for 8 times, among which 3 times appeared in the years of 1953-1977 and another 5 times in 1978-2010. The blood relationship analysis of variety genealogy indicated that there are 35 most important key backbone-parents (19 indica and 16 japonica ones) have been widely and long utilized in rice breeding for structuring new varieties with high yielding potential and ideal traits. On the other hand, the study also showed that though there was a rapid increase in terms of per unit yield and total rice production, but the rate of rice germplasm innovation and utilization slowed down, and the extension area and occupied proportion of modern inbred rice varieties with high yield potential reduced in the past 30 years. 【Conclusion】The release and mass extension of more than 4 000 modern inbred rice varieties as well as varieties renewed for 8 times in mass scale during 1949-2010 are the major factors in increase of inbred rice production in China. However, challenges will be faced in rice breeding programs: how to strengthen exploration, introduction and utilization of excellent rice germplasm, and how to overcome germplasm bottleneck for continuously releasing new rice varieties.  
    Creative Process of a New Restorer Material with Ogu CMS in Brassica nupus
    CHEN Wei-Jiang, LI Mei, WANG Tong-Hua, HUI Rong-Kui, TU Jin-Xing, FU Ting-Dong
    Scientia Agricultura Sinica. 2012, 45(8):  1465-1474.  doi:10.3864/j.issn.0578-1752.2012.08.002
    Abstract ( 743 )   PDF (731KB) ( 689 )   Save
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    【Objective】 The objective of this study was to explore the radish restore gene transferring and the stable restorer line screening for Ogu CMS in rapeseed.【Method】 The Raphanobrassica (2n=58) as the restoring gene donor and grafting as the restoring gene importing tool was used to obtain the hybrid of Ogu CMS in B. napus. And the stable restoring  materials were bred through selection of the seed setting rate in single plant and the restoring ratio in populations from every selfing and backcross generations. Also the selected materials were detected by the cytological observation and the molecular marker screening. 【Result】With the high strength selection, a stable homozygous restoring line named as CLR650 of Ogu CMS in B. napus was selected. This line can restore the male fertility in both progenies of self-pollination and testcross at a rate of 100%. Although it’s somatic chromosome numbers were in 2n= 38 to 40 and some abnormal phenomena such as chromosome bridge and lagged chromosomes in meiosis were observed, the pollens still could be normally formed without the influence on the normal seed setting. It was proved that the Ogu CMS restoring gene existed in CLR650 and the length of radish fragment was shorter than the Ogu CMS restoring line R113 by detecting the molecular markers which closely linked with radish restoring gene (Rf0). 【Conclusion】 The restoring line CLR650 could be a new restoring type for the Ogu CMS in B. napus.
    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    超级稻|产量|时期|硅|硅素吸收、利用效率Effect of Application of Silicon at Different Periods on Grain Yield and Silicon Absorption, Use Efficiency in Super Rice
    GONG Jin-Long, HU Ya-Jie, LONG Hou-Yuan, CHANG Yong, GE Meng-Jie, GAO Hui, LIU Yan-Yang, ZHANG Hong-Cheng, DAI Qi-Gen, HUO Zhong-Yang, XU Ke, WEI Hai-Yan, LI De-Jian, SHA An-Qin, ZHOU You-Yan, LUO Xue-Chao
    Scientia Agricultura Sinica. 2012, 45(8):  1475-1488.  doi:10.3864/j.issn.0578-1752.2012.08.003
    Abstract ( 763 )   PDF (353KB) ( 767 )   Save
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    【Objective】The objective of this study was to identify the best period for application of Si and the mechanisms of efficient Si absorption and use efficiency in super rice. 【Method】A field experiment was conducted with two japonica super rice varieties Wuyunjing24 and Huaidao9 planted on a large scale in Jiangsu region as materials exposed to applied Si at different periods. Grain yield and its components, Si accumulation at the main growth and development stage, periodic Si accumulation, periodic Si uptake rate, Si use efficiency, etc, were investigated. Furthermore, correlation analysis of grain yield and its components and traits associated with Si uptake and use efficiency was studied. 【Result】 Applied Si increased grain yield and its components, Si accumulation at the main growth and development stage, periodic Si accumulation, periodic Si uptake rate and partial factor productivity of applied Si, but it decreased Si use efficiency for grain production, significantly. It not only promoted Si supplies, but also improved the ability of providing Si of the soil. Morover, it raised available Si level in the soil and promoted Si absorption of rice. With the delaying periods of applied Si, grain yield, panicles, grains per panicle, total amount of Si absorption, Si uptake per 100kg of grain, Si recovery efficiency, Si agronomic efficiency and partial factor productivity of applied Si increased firstly and then decreased, while Si use efficiency for grain production showed an opposite trend, with applied Si at effective tillering reaching a peak for panicles and critical leaf-age for productive tillers for others. However, seed-setting rate and 1000-grain-weight followed an ascendent tendency accordingly, with 1000-grain-weight of applied Si at heading reaching a significant level, whereas Si physiological efficiency was not significantly correlated with the periods of applied Si. Si accumulation rate of rice were 20.69-29.02% from transplanting to panicle initiation, 47.46-59.65% from panicle initiation to heading, 17.99-25.52% from heading to maturity, panicle initiation-heading > transplanting-panicle initiation > heading-maturity, significantly, and the periodic Si uptake rate followed the same trend as it. Correlation analysis showed that, grain yield components of rice grew together with Si absorption in phase and there existed significant positive correlations between grain yield and total amount of Si absorption. The increase of periodic Si uptake rate and periodic Si accumulation from panicle initiation to heading improved by applied Si was the physiological basis of increasing grain yield improved by large panicle. 【Conclusion】Increase of periodic Si uptake rate and periodic Si accumulation, especially at panicle initiation-heading stage, could increase Si accumulation at heading and maturity satges and Si use efficiency. By achieving large panicle based on stabilizing and raising panicles, and promoting the enrichment of population sink, grain yield could be further improved by application of Si.
    Effects of Different Day and Night Temperature Enhancements on Wheat Grain Yield and Quality after Anthesis under Free Air Controlled Condition
    BIAN Xiao-Bo, CHEN Dan-Dan, WANG Qiang-Sheng, WANG Shao-Hua
    Scientia Agricultura Sinica. 2012, 45(8):  1489-1498.  doi:10.3864/j.issn.0578-1752.2012.08.004
    Abstract ( 736 )   PDF (390KB) ( 878 )   Save
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    【Objective】 The study is to explore the effects of free air temperature enhancement on wheat grain yield and quality after anthesis and provides a theoretical basis for global warming affecting on wheat of Jianghuai region.【Method】Field experiments of Yangmai 16 under the warming treatments with different times (all day time, daytime and nighttime warming) and different ranges (1.5 and 3℃) by using free-air temperature enhancement (FATE) technique were conducted in Danyang to simulate the effects of warming climate on the yield and quality including protein and starch in 2010 and 2011. 【Result】 Although the actual temperatures were below the anticipated ones, compared to the control, the results showed that a downturn of yield resulted from the decrease of grain number per spike and grain weigh, the former and latter were sensitive to night and day warming, respectively. The decrease amplitude aggravated accompanied with the increasing warming levels. The result also showed that yields of all-day warming treatment lost most at different warming levels among the three warming scenarios. If the mean daily temperature increased by about 1.8℃ on the basis of mean temperature of all-day time, daytime and nighttime at about 20.8, 23.7, 16.9℃, yield losses could be up to 22.3%. Temperature enhancement after anthesis increased the content of crude protein and wet gluten, compared to other treatments, the treatment 6 (night warming 3℃) increased most of two years results and the increasing ranges were respectively 7.9%(2010) and 13.2%(2011). The effect of warming on components of protein was complex. But that on glutenin was greater than that on albumin, globulin and gliadin, the synthesize and accumulation of glutenin could be easily controlled by the temperature. Meanwhile, temperature enhancement after anthesis had no significantly effect on the content of starch. 【Conclusion】 Temperature enhancement resulted in yield reduction of the wheat without changing the growing process before anthesis, meanwhile, the constitution of grain would also be significantly changed, which led to poor quality.
    PLANT PROTECTION
    Spatio-Temporal Localization Changes of Endogenous Hydrogen Peroxide During Cell Division Cycle of Xanthomonas oryzae pv. oryzae
    LI Xin, LI Xin-Ling, PANG Xin-Yue, ZHU Wen-Xue, FAN Jin-Ling, LUO Lei, DU Lin, WANG Na, WANG Li-Ping
    Scientia Agricultura Sinica. 2012, 45(8):  1499-1504.  doi:10.3864/j.issn.0578-1752.2012.08.005
    Abstract ( 739 )   PDF (699KB) ( 691 )   Save
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    【Objective】The objective of this study is to investigate the changes of spatio-temporal localization of endogenous hydrogen peroxide (H2O2) in strains of Xanthomonas oryzae pv. oryzae, and to illustrate the physiological function of H2O2 in bacterial proliferation. 【Method】 Histochemical methods through transmission electron microscope were used to observe localization of endogenous H2O2. 【Result】 Even though the levels of endogenous H2O2 production were different among various strains, the produced H2O2 was localized in the cell wall of all Xanthomonas oryzae pv. oryzae strains tested. Interestingly, the accumulated H2O2 was also localized in the mesosome-like structure and nucleoids during the cell division cycle of strains. Furthermore, results revealed the quantitative and dimensional changes of H2O2 accumulation in the two additional locations. 【Conclusion】Additional locations of the accumulated H2O2 are closely associated with the process of cell division of all Xanthomonas oryzae pv. oryzae strains investigated. Spatio-temporal localization changes of endogenous H2O2 should be an ubiquitous phenomenon during cell division cycle of bacterium. H2O2 may play key roles in bacterial proliferation.
    Cloning and Analysis of StLTPa7 from Solanum torvum
    XIE Chao, YANG Qing, SHI Ce, JUE Deng-Wei, ZHU Yan-Ping, LIU Shui-Ping
    Scientia Agricultura Sinica. 2012, 45(8):  1505-1512.  doi:10.3864/j.issn.0578-1752.2012.08.006
    Abstract ( 770 )   PDF (783KB) ( 748 )   Save
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    【Objective】The objective of this study is to clone non-specific lipid transfer protein gene StLTPa7 from Solanum torvum and to analyze the function of the gene. 【Method】 Primers were designed according to homologous cloning method. In order to clone StLTPa7, the gene cDNA was amplified by RT-PCR. Over-expression transgenic tobacco plants were generated via Agrobacterium-mediated transformation. Analysis on the inhibitory activity of proteins extracted from transgenic tobacco plants was done using mycelia growth rate method.【Result】StLTPa7 cDNA contained an ORF of 345 bp long and encoded a putative protein of 114 amino acids with a molecular weight of 11.42 kD and a theoretical pI of 9.01. StLTPa7 was induced to express by both salicylic acid (SA) and Verticillium dahliae and the level of transcript was the highest 24 to 48 h after treatment. To analyze the resistance of StLTPa7 to V. dahliae, the plants over-expressing StLTPa7 were generated by Agrobacterium-mediated transformation and 4 transgenic lines were identified by PCR. Quantitative RT-PCR indicated that the gene over-expressed in transgenic lines L5 and L7. Anti-fungal assay revealed that the inhibitory rate of the proteins extracted from the transgenic line L5 to V. dahliae was 2.5 times compared to the control.【Conclusion】StLTPa7 exhibits associated to inhibitory effect of V. dahliae growth, suggesting that it may be involved in plant defense against V. dahliae.
    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Effects of Straw Returning on Characteristics of Soil Aggregates in Semi-arid Areas in Southern Ningxia of China
    ZHANG Peng, JIA Zhi-Kuan, WANG Wei, LU Wen-Tao, GAO Fei, NIE Jun-Feng
    Scientia Agricultura Sinica. 2012, 45(8):  1513-1520.  doi:10.3864/j.issn.0578-1752.2012.08.007
    Abstract ( 999 )   PDF (651KB) ( 1010 )   Save
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    【Objective】 A field experiment was conducted to determine the effects of straw returning systems on soil aggregate of millet or corn under different straw returning supplies. in order to provide a theoretical basis for establishment of a fertilization system and crop management for semi-arid areas of southern Ningxia.【Method】The effects of straw returning, taking no straw returning as the control, on soil aggregate indexes by dry and wet sieving under different straw returning treatments (3 000 kg•hm-2 (L), 6 000 kg•hm-2 (M) and 9 000 kg•hm-2 (H) of millet straw and 4 500 kg•hm-2 (L), 9 000 kg•hm-2 (M) and 13 500 kg•hm-2 (H)    of corn straw) were studied for four years (2007-2010).【Result】After the millet harvest (2010), dry sieving results showed that DR0.25, DMWD and DGMD of straw returning treatments higher than CK, the fractal dimension lower than CK in 0-40 cm soil  layer, significantly. Wet sieving results showed that straw returning treatments had different effects on the WGMD, WMWD and fractal dimension of 0-40 cm soil layer, which followed the order from high to low: H,M>L>CK. There is a ‘Z’ tendency of PAD and ELT in 0-40 cm soil layer for each treatment, and CK is significantly higher than straw returning treatments(P<0.05). 【Conclusion】 The content of soil aggregates could be improved efficiently by using straw returning technology in semi-arid areas of Southern Ningxia.
    Effect of P Deficiency on the Emergence of Astragalus L. Root Exudates and Mobilization of Sparingly Soluble Phosphorus
    LAN Zhong-Ming, LIN Xin-Jian, ZHANG Wei-Guang, ZHANG Hui, WU Yi-Qun
    Scientia Agricultura Sinica. 2012, 45(8):  1521-1531.  doi:10.3864/j.issn.0578-1752.2012.08.008
    Abstract ( 872 )   PDF (478KB) ( 745 )   Save
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    【Objective】 The objective of this study was to investigate the mobilization and uptake capacity of sparingly soluble phosphorus by root exudate of Astragalus L., and genotypic variation of different Astragalus L. in activation of sparingly soluble phosphorus. 【Method】 Hoagland nutrient solution culture was carried out to collect root exudate and test the mobilization of sparingly soluble phosphorus via concentrated vacuum rotary evaporator, and the content of organic acids was analyzed by high performance liquid chromatography (HPLC). 【Result】 Under the condition of P-deficiency stress, root radius from different genotypes of Astragalus L. reduced, while the root-shoot ratio and root surface area were significantly higher than P-supply. Root exudates were oxalic acid, tartaric acid, citric acid, malic acid and so on, but it mainly exuded oxalic acid. Under the condition of P-deficiency, secreation of oxalic was significantly different among variou genotypes, and the amout of organic acid secretion was significantly higher than P-supply. Minzi 1, Zhezi 5 and Minzi 6 in organic acid secretion were significantly higher than Yijiangzi, Yujiangdaye. Root exudates on insoluble Al-P and Fe-P had certain activation capacity under P-deficiency and P-supply. The activation P values for Al-P and Fe-P were 36.40-157.39 μg•g-1, 32.20-139.42 μg•g-1, and P-deficient root exudates on sparingly soluble phosphate was higher than P-supply, and activation capacity of Al-P was slightly higher than Fe-P. Through the activation of simulation experiment, it was confirmed that organic acids could mobilize sparingly soluble Al-P and Fe-P, and the difference was significant.【Conclusion】 Under P-deficiency stress, Astragalus L. exudated organic acids, which could significantly mobilize sparingly soluble phosphate. There were obvious differences among various genotypes, showing differences between Astragalus L. varieties. Cultivating Astragalus L. could be used to improve soil P-deficiency nutrient cycling in red soils region of southern China, but further research should be carried out to scientifically evaluate organic acids on the activation capacity of Al-P and Fe-P.
    Growth Promotion Potential and Distribution Features of Nitrogen-Fixing Bacteria in Field Environments
    SUN Jian-Guang, HU Hai-Yan, LIU Jun, CHEN Qian, GAO Miao, XU Jing, ZHOU Yi-Qing
    Scientia Agricultura Sinica. 2012, 45(8):  1532-1544.  doi:10.3864/j.issn.0578-1752.2012.08.009
    Abstract ( 897 )   PDF (798KB) ( 873 )   Save
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    【Objective】 The objective of this experiment is to determine the dominant species and phylogenetic position of nitrogen fixing bacteria in crop rhizosphere and field environments, and to screen for nitrogen-fixing bacteria with growth promotion potential. 【Method】Nitrogen-free medium was used to culture nitrogen-fixing bacteria and nitrogenase activity was determined with acetylene reduction assay. Confrontation method was used to test antagonistic bacteria against plant pathogenic fungus. ACC (1-aminocyclopropane-1-carboxylate) nitrogen source was used to test ACC deaminase. 16S rDNA was amplified by PCR and 16S rDNA sequence was analysed with MEGA software. 【Result】 Nitrogenase activity of the 94 tested strains ranged from 0.99 to 180.59 nmol C2H4/h•mg protein. Fourty-two strains accounting for 44.7% of the tested strains showed nitrogenase activity above 10 nmol C2H4/h•mg protein. Paenibacillus and Bacillus were main groups, and their percentages were 33.0% and 26.6% of the tested strains, respectively. Six strains accounting for 6.4% of the tested strains showed antagonistic actions against plant pathogenic fungi Sclerotinia sclerotiorum, Gibberella zeae and Verticillium dahliae with the inhibitory rate from 23.9% to 65.9%. Twenty strains accounting for 21.3% of the tested strains produced ACC deaminase with activity ranging 0.33 to 21.98 µmol α-ketobutyric acid/h•mg protein, which were identified as Bacillus, Paenibacillus, Rhizobium, and extensively distributed in crop rhizosphere and field environments.【Conclusion】The dominant species of nitrogen-fixing bacteria in crop rhizosphere and field environments belong to the genus Paenibacillus and Bacillus. Most of the nitrogen-fixing bacterial strains possess strong potential for nitrogen fixation, part of them possess potential for growth promotion, and a few of them possess potential for disease resistance. The potential strains belong to Paenibacillus, Bacillus and Rhizobium, and extensively colonized in crops with no specificity.
    HORTICULTURE
    Progress in Research on the Metabolic Regulation and Molecular Mechanism of Green Leave Volatiles (GLVs)  
    CHEN Shu-Xia, CHEN Qiao, WANG Cong-Ying, HAO Li-Ning, FANG Yu-Lin
    Scientia Agricultura Sinica. 2012, 45(8):  1545-1557.  doi:10.3864/j.issn.0578-1752.2012.08.010
    Abstract ( 778 )   PDF (821KB) ( 2401 )   Save
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    Plants are always attacked by herbivores, pathogens and sometime suffer mechanical wounding in complicated environments, and then the lesions are caused and the complex defensive responses are induced. One of the defensive responses is releasing the green leaf volatiles (GLVs). The green leaf volatiles (GLVs) of plants are the C6 aldehydes, alcohols and their esters which are usually formed from polyunsaturated fatty acid catalyzed by lipoxygenase (LOX) and hydroperoxide lyase (HPL) in the oxylinpins pathway. The green leaf volatiles are important signaling moleculars, which introduce defensive responses between plants and pathogens, plants and herbivores, within plants and between neighbor plants. It is very important to study the metabolic pathway and the regulatory mechanisms of GLVs for probing into the direct or indirect defenses to pathogens and insects, improving the flavor of crop and increasing the new resistant germplasm. It is also very important to guide the strategy of integrated controlling of crop diseases and pest insects in agro-forestry ecosystems.
    Construction of Cucumber-Sour Cucumber Chromosome Introgression Lines and Location of Fruit Related QTLs
    MENG Jia-Li, LOU Qun-Feng, ZHOU Xiao-Hui, SHI Jian-Lei, CHEN Jin-Feng
    Scientia Agricultura Sinica. 2012, 45(8):  1558-1567.  doi:10.3864/j.issn.0578-1752.2012.08.011
    Abstract ( 609 )   PDF (868KB) ( 435 )   Save
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    【Objective】 A set of cucumber-sour cucumber chromosome introgression lines were constructed by using cultivated cucumber (Cucumis sativus L., 2n=14) cultivar ‘Beijingjietou’ as recurrent parent and the wild ‘sour cucumber’ (C. hystrix Chakr., 2n = 24) as donor parent by the way of marker-assisted selection, and QTL controlling cucumber fruit shape was detected initially. 【Method】 Fristly, through interspecific-backcrossing-selfing approach, a lot of cucumber-sour cucumber chromosome introgression lines were obtained. Then polymorphism of the parents was detected by 298 pairs of SSR markers which uniformly distributed in the cucumber genome. And then the cucumber-sour cucumber chromosome introgression lines which contained the wild sour cucumber chromosome segments from the strains were screened by using the detected differences between parents SSR markers. QTL analysis of three main traits affecting cucumber fruit shape(fruit length, fruit diameter, fruit length/diameter ratio) was conducted using t-test compared with the recurrent parent. 【Result】 A set of 50 cucumber-sour cucumber chromosome introgression lines was constructed. There are 149 chromosome introgression segments containing 61 different introgression segments were identified in the lines. The total length of the different segments was 259.95 cM, and the coverage was 45.37% in the cucumber genome. The length of the introgression segments ranged from 1.65 cM to 15.4 cM with an average of 5.41 cM. They distributed in 7 chromosomes of cucumber. A total of 13 QTL affecting cucumber fruit shape were detected in CSIL population initially. 【Conclusion】 In this research, a set of cucumber-sour cucumber introgression lines were created and QTL affecting cucumber fruit shape was detected initially. It provided new resources for utilization of genes from C. hystrix and also the material basis for fine mapping of quantitative traits in cucumber.
    Isolation and Expression of Pathogenesis-Related Protein Gene MhPR8 from Malus hupehensis 
    ZHANG Ji-Yu, QU Shen-Chun, XUE Hua-Bai, GAO Zhi-Hong, GUO Zhong-Ren, ZHANG Zhen
    Scientia Agricultura Sinica. 2012, 45(8):  1568-1575.  doi:10.3864/j.issn.0578-1752.2012.08.012
    Abstract ( 598 )   PDF (765KB) ( 795 )   Save
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    【Objective】This study aimed to clone a full length cDNA and genomic DNA sequences of MhPR8 from Malus hupehensis and analyze the expression pattern of MhPR8 after treatment with the chemical reagents salicylic acid (SA), methyl jasmonate (MeJA), 1-aminocyclopropane-1-carboxylic acid (ACC), abscisic acid (ABA), abiotic stress NaCl and low temperature (4℃), and biotic factors apple ring spot pathogen (Botryosphaeria berengeriana) and apple aphids (Aphis citricota).【Method】The full length cDNA and genomic DNA sequences of MhPR8 were isolated from the M. hupehensis complete cDNA library treated by SA and genomic DNA using the method of PCR amplification, respectively. Real-time quantitative RT-PCR (qRT-PCR) was performed to determine the expression pattern of MhPR8 in M. hupehensis.【Result】The open reading frame of this gene was 897 bp, encoding 298 amino acids with a molecular weight of 31.67 kD and isoelectric point of 4.77. There was no intron in ORF of MhPR8. MhPR8 contains conserved construction domain GH18_hevamine_XipI_class_III, belongs to Class III chitinase. And this protein has six conserved cysteines, and aspartic acid and glutamic acid play a role of catalysis in plants class III chitinase, which are located in 151th and 153th amino acids, respectively. qRT-PCR results show that during the first 48 hours, MhPR8 was obviously induced by SA, MeJA, and ACC, but weakly induced by ABA. And this gene was clearly induced by low temperature, but weakly induced by NaCl. Infected with B. berengeriana resulted in obvious accumulation of MhPR8 transcripts during the first 96 h. The expression of the MhPR8 was clearly induced by the apple aphid (A. citricota) in leaves and stems of M. hupehensis. 【Conclusion】MhPR8 could be involved with the SA- signal pathway and the jasmonic acid (JA)/ ethylene (ET)-signal pathway in M. hupehensis. MhPR8 might be play a key role in resistance to biotic and abiotic stresses.
    STORAGE·FRESH-KEEPING·PROCESSING
    Effects of Alginate Oligosaccharides on the Accumulation of Glyceollins in Soybean
    HU Jia, SHI Bo, OjokohEromoselesolomon , LIANG Ping, LI Jing-Mei
    Scientia Agricultura Sinica. 2012, 45(8):  1576-1586.  doi:10.3864/j.issn.0578-1752.2012.08.013
    Abstract ( 746 )   PDF (511KB) ( 656 )   Save
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    【Objective】The objective of this study was to investigate the relationship among alginate oligosaccharides(AOS) elicitation and glyceollins accumulation in soybean.【Method】The preparative high performance liquid chromatography was selected to separate and purify the ethanol extracts of elicited soybeans seeds with AOS, the separation process was monitored by HPLC and UPLC-MS was used to detect their molecular weights. The optimal parameters were determined by investigating the effects of the concentration of AOS, the soaking time, the temperature, moisture and time course.【Result】AOS induced glyceollins synthesis in soybean seeds. The optimal conditions of elicitation were as follows: concentration of AOS 4%, presoaked in the sterile water for 5 h, stored at a controlled temperature of 25℃ and humidity of 60% in the dark for 4 d. Results from the optimal conditions in the study indicated that the maximum concentration of the phytoalexin glyceollins was 0.525 mg•g-1 fresh weight.【Conclusion】 These findings will give some science suggestions for soybean high value processing and the development of AOS.
    ANIMAL SCIENCE·RESOURCE INSECT
    Genetic Diversity of Mitochondrial DNA D-loop Sequences in Ningqiang Pony
    ZHANG Tao, LU Hong-Chao
    Scientia Agricultura Sinica. 2012, 45(8):  1587-1594.  doi:10.3864/j.issn.0578-1752.2012.08.014
    Abstract ( 880 )   PDF (682KB) ( 588 )   Save
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    【Objective】In order to provide a scientific and theoretical basis for genetic resource protection of Ningqiang pony, the genetic diversity of mitochondrial DNA (mtDNA) D-loop and maternal origin of Ningqiang pony was investigated.【Method】Twelve 600 bp mtDNA D-loop sequences were analyzed by PCR and sequencing techniques, and the phylogenetic trees were constructed based on the mtDNA D-loop sequences of Ningqiang pony samples and other equine D-loop sequence available from NCBI.【Result】There were 9 unique haplotypes with 34 polymorphic sites in Ningqiang pony, which account for 5.67% of 600 bp sequence analyzed. The haplotype diversity (h) and nucleotide diversity (π) were 0.978±0.054 and 0.01988±0.00818, respectively.Moreover, Ningqiang pony distributed in four branches, and are closely related to Hucul,Shetlan, Debao horse, Cheju horse and Mongolian horse , whereas they are less related to Thoroughbred, Yunnan horse and Jeju native horse. 【Conclusion】 Ningqiang pony have rather abundant genetic diversity and were greatly affected by other horse breeds in history.
    Effect of Thiamin on the Rumen Fermentation and Microbial Community of the Substrates in Rusitec
    WANG Hong-Rong, ZHANG Hong-Wei
    Scientia Agricultura Sinica. 2012, 45(8):  1595-1605.  doi:10.3864/j.issn.0578-1752.2012.08.015
    Abstract ( 882 )   PDF (595KB) ( 514 )   Save
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    【Objective】An experiment was conducted to determine the effect of thiamin on rumen fermentation and microbial community under different nitrogen levels to sulfur ratio in Rusitec. 【Method】 A 2×2 two factorial design of an experiment was adopted: one factor was different nitrogen levels to sulphur ratios (5﹕1, 15﹕1), the other was different substrates with and without adding thiamin (0 and 60 mg•kg-1). 【Result】 The results showed that there was no significant effect on pH and ammonia concentration in the substrates. The high sulfur level restrained the synthesis of bacterial protein and significantly decreased the ratio of protozoal protein and bacterial protein (P<0.05). There was no effect on the concentrations of acetate, propionate and the acetate/propionate ratio (P<0.05). The low sulfur level increased the concentration of butyrate, total VFAs and the proportion of butyrate, and the high S level reduced the total concentration of thiamin while the total thiamin was increased by adding thiamin. There was an obvious consequence that the high sulfur substrate declined the concentration of the lactate in Rusitec. Compared to the without adding thiamin group, the relative number of Strepococcus bovis in substrates with adding thiamin decreased by 5.90% (P<0.05) and 2.05% (P<0.01) under the same nitrogen/sulfur ratios, and there was no significant effect on the number of Butyrivibrio fibrisovens. 【Conclusion】 It was concluded that rumen fermentation was response to sulfur level in Rusitec, higher levels of sulfur in sulfur substrate could decline the concentration of the lactate and bacterial protein while protozal protein dropped in Rusitec. It was found that sulfur and thiamin play an important role in suppressing of subacute rumen acidosis (SARA). Adding thiamin in Rusitec resulted in the decrease of the relative number of S. bovis in substrates, but had no significant effect on the number of B. fibrisovens.
    Effects of Different Levels of Diet on Plasma Physiochemical Indexes and Folliculogenesis in Hu Sheep During the Luteal Phase
    YING Shi-Jia, NIE Hai-Tao, ZHANG Guo-Min, WU Yong-Cong, WANG Zi-Yu, PANG Xun-Sheng, WANG Chang-Long, HE Dong-Yang, JIA Ruo-Xin, WANG Feng
    Scientia Agricultura Sinica. 2012, 45(8):  1606-1612.  doi:10.3864/j.issn.0578-1752.2012.08.016
    Abstract ( 642 )   PDF (540KB) ( 593 )   Save
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    【Objective】 The present study was designed to investigate the effects of different plasma physiochemical indexes on folliculogenesis in Hu sheep fed with different levels of diet during the luteal phase.【Method】Twenty-eight multiparous Hu sheep were assigned into 3 groups and received a maintenance diet (C group), 0.5×M (R group) and 1.5×M (S group), respectively, for 6 days of their estrous cycle. Six ewes in each group were laughtered on day 12 of the estrous cycle. The remaining ewes were received a maintenance diet until the next estrus. Blood was sampled on days 7, 8, 10 and 12 of the estrous cycle.【Result】As dietary intake increased, the number of follicles of ≥3.5 mm was significantly increased (P<0.05), the number of follicles of 2.5-3.5 mm was significantly decreased (P<0.05) and the estrous cycle was shortened (P<0.05). Compared with R group, S group significantly decreased the plasma urea, total cholesterol, free fatty acid concentrations (P<0.05) and significantly increased the plasma triglyceride concentration (P<0.05). The activity of lactate dehydrogenase in S group was higher than C group (P<0.05). There was no significant effect of treatment on plasma uric acid, plasma ammonia, high density lipoprotein and low density lipoprotein concentrations and spartate transaminase, alanine transaminase activities (P>0.05), however, there was a significant effect of day on plasma uric acid, ammonia, low density lipoprotein concentrations and aspartate transaminase activity (P<0.05).【Conclusion】 Different nutritional requirements and metabolic features were present in different physiological periods during ovine luteal phase and the mechanism by which dietary restriction inhibited folliculogenesis may involve responses to the increased capacity of lipolysis and protein degradation and the decreased capacity of lipid and protein synthesis.
    Genetic Variation Analysis of 3′-UTR Region of Chicken Lpin1 Gene and the Potential Effect on miRNA Binding Sites
    LI Su-Ya, ZHANG Jian-Hong, CHEN Wen, HUANG Yan-Qun, SUN Gui-Rong, HAN Rui-Li, KANG Xiang-Tao
    Scientia Agricultura Sinica. 2012, 45(8):  1613-1620.  doi:10.3864/j.issn.0578-1752.2012.08.017
    Abstract ( 661 )   PDF (704KB) ( 678 )   Save
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    【Objective】 This experiment was conducted to study the genetic variation/haplotype of the 3′ -UTR of chicken Lpin1 gene and their distributions among breeds, and their potential effects on miRNA binding sites were predicted. 【Method】According to the chicken Lpin1 genome sequence ( GenBank accession: NC_006090), a pair of specific primers was designed to conduct the genetic variation/haplotype analysis in the 3′ -UTR of chicken Lpin1 gene in different breeds of chickens through direct PCR sequence combined with clone sequencing. 【Result】Eight variable sites were uncovered, these variation sites were all located at the 3′-UTR of chicken Lpin1 gene. Analysis showed the no variation sites were detected from Henan Game chicken in this region, however, six variation sites were detected from Gushi and Lushi chickens. Six variable sites (g.11A>T, g.77C>G, g.108_109delinsG, g.110_111delinsG, g.270A>G and g. 348G>T) with minor allele frequency ≥5% were selected for haplotype reconstruction, and six haplotypes were detected from six chicken breeds. P1 and P4 were the predominant haplotypes with frequencies >30%, respectively, only one haplotype was detected from Henan game chicken. Software prediction showed that g.77C>G variant could cause the increase/loss of several miRNA binding sites. 【Conclusion】 The results showed that the 3′-UTR of chicken Lpin1 gene was highly polymorphic, the distribution of variable sites and haplotypes in different breeds of chickens had clear difference.
    VETERINARY SCIENCE
    Modulation of Lactobacillus rhamnosus on Innate Immune Responses in Macrophages
    HUANG Qin, HUANG Yi, CUI Zhi-Wen, LI Ya-Li, LI Wei-Fen, YU Dong-You
    Scientia Agricultura Sinica. 2012, 45(8):  1621-1626.  doi:10.3864/j.issn.0578-1752.2012.08.018
    Abstract ( 976 )   PDF (324KB) ( 610 )   Save
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    【Objective】Lactobacillus rhamnosus (L. rhamnosas) was examined for its capacity to facilitate the innate immune responses of macrophage cell in vitro in terms of innate immune factor production. 【Method】 The murine macrophage cell line, RAW 264. 7, was treated with PBS (designated as group CT, control group), Escherichia coli K88(designated as group EC)and L. rhamnosus(designated as group LR)for 12 h,respectively, as well as treated with L. rhamnosus prior for 1 h, and then exposed to E. coli K88 for 12 h(designated as group LR-EC). After incubation, culture supernatants were collected for further analysis of TNF-α, IFN-γ, IL-1β, IL-6, IL-12, IL-8, IL-10, PGE2 and   using ELISA. 【Result】Results showed that TNF-α, IFN-γ, IL-8 and IL-10 production was observed in group CT (the control group),whereas the concentrations of IL-1β, IL-6 and IL-12 were below the detection limits. Production of TNF-α, IFN-γ, IL-8, IL-10, PGE2 and   in group EC increased significantly(P<0.01)compared to group CT, and higher concentrations of IL-1β, IL-6 and IL-12 were also detectd in group EC. Higher levels of TNF-α, IFN-γ, IL-10, PGE2,     (P<0.01)and IL-8(P<0.05)were determined in group LR while no detectable effect on IL-1β and IL-6 was found. Moreover,  the release of all tested proinflammatory cytokines(TNF-α, IFN-γ, IL-1β, IL-6, IL-12), chemotactic factor (IL-8) and PGE2 in  group LR-EC was up-regulated remarkably(P<0.01)while secretion of IL-10 and   was noticeably suppressed in comparison to group EC(P<0.01). 【Conclusion】 It was concluded that probiotics strain L. rhamnosus is capable of triggering a moderate innate immune response on direct contact with murine macrophage cell line RAW264.7. Furthermore, L. rhamnosus could enhance proinflammatory response to infected macrophages; meanwhile, it might prevent excessive inflammatory reaction in vitro.
    RESEARCH NOTES
    Effect of Phosphorus Additions on Grain Yield, Biomass and P and As Concentration in Different Organs of Three Rice Cultivars with Different P-efficiencies Under As-contaminated Conditions
    YANG Ling, LIAN Juan, GUO Zai-Hua, ZHANG Xiu, DU Shuang-Shuang, HE Xing-Long, ZHAO Zhu-Qing
    Scientia Agricultura Sinica. 2012, 45(8):  1627-1635.  doi:10.3864/j.issn.0578-1752.2012.08.019
    Abstract ( 804 )   PDF (309KB) ( 571 )   Save
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    【Objective】The objective of this study was to clarify the effects of P additions and P-efficiency on grain yield, biomass and quality of rice in As-contaminated soil, and discuss agronomic measures for mitigation of As accumulation in rice. 【Method】Low-P tolerant rice cultivars 99011 with high P-absorption-efficiency and 580 with high P-use-efficiency, and low-P sensitive rice cultivar 99056 were used in a pot experiment to study the effects of P additions on grain yield, biomass and P and As concentration in the straw, husk and brown rice in moderate and high As-contaminated soils.【Result】Compared to no As-contamination treatment, 50 mg?kg-1 of As increased the biomass of three supplied rice slightly, but decreased grain yield significantly. Both the biomass and the grain yield increased significantly with the increase of P supply. As at 100 mg?kg-1 decreased both the biomass and the grain yield significantly. The biomass increased obviously with increasing P supply, however, the grain yield was the highest at 30 mg?kg-1 of P level but the lowest (0) at 150 mg?kg-1 of P level. Under As-contaminated conditions, the As concentration in different organs were straw >> husk >> brown rice, and they increased with increasing P and/or As additions. In As stress or P and As double stress soils, the grain yield, biomass and the P concentration in straw, husk and brown rice were all 99011>580>99056, with remarkable difference among three rice cultivars, moreover, the difference among the three rice cultivars increased with the reduce of P supply and the increase of As supply. The As concentrations in straw, husk and brown rice were all 99056>580>99011, with prominent difference among the three rice cultivars, moreover, the difference among the three rice cultivars increased with the increase of P and/or As supply. Under the conditions of As addition was ≤50 mg?kg-1and P supply was ≤30 mg?kg-1, low-P tolerant rice cultivar 99011 could get considerable grain yield and the As concentration in brown rice was lower than the national food safety standard. 【Conclusion】Under As-contaminated conditions, the grain yield and the quality of rice are closely related to the P additions and the uptake ability of rice to low concentration of P. Using rice cultivars with high P-absorption-efficiency and reducing P fertilizer input properly are promising rotes for getting considerable production with reduced grain As in moderatly and slightly As-contaminated soil.
    Improvement Efficiency of Recurrent Selection for Starch Content in Maize Population Yu Syn5
    KU Li-Xia, WANG Fu-Juan, GUO Shu-Lei, XIE Li-Li, LIU Hai-Jing, ZHANG Jun, TIAN Lei, CHEN Yan-Hui
    Scientia Agricultura Sinica. 2012, 45(8):  1636-1643.  doi:10.3864/j.issn.0578-1752.2012.08.020
    Abstract ( 818 )   PDF (513KB) ( 513 )   Save
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    【Objective】 The objective of the study is to evaluate the improvement efficiency in maize population Yu Syn5 so as to provide reference for creating high-starch content germplasm resources, breeding high-starch content hybrids and keeping on improving starch content of Yu Syn5. 【Method】 The improvement for starch content in maize population Yu Syn5 was made for 4 cycles using near infrared analytical technology. Twenty testcrossing combinations made by Huangzao 4, Dan 340, Qi 319, Ye 478 and Mo17 and crossed for four cycles to improve the populations according to NC II design were evaluated for the starch content and the combining ability in two environments. 【Result】 The starch content of Yu Syn5 gradually increased from 69.39% of C0 to 72.31% of C4 with increase of improvement cycles, which the actual gain of 2.92% was obtained after four cycles improvement. But the protein content and lysine content decreased with increase of improvement cycles. The starch content of testcrossing combinations also increased with increase of improvement cycles. The results showed that the recurrent selection of nondestructive starch content of single grain could effectively improve the starch content of Yu Syn5. Regarding the general combining ability (GCA), the value of starch content GCA was improved in each cycle for the populations. The values of special combining ability (SCA) between Huangzao4 and the improved populations were increased per cycle, while the values of SCA between other 4 testers and the populations appeared fluctuation. 【Conclusion】 It was efficient to improve the starch content of Yu Syn5 by the recurrent selection of nondestructive starch content of single grain. The starch content of populations gradually increased with increase of improvement cycles. 
    Cloning and Characterization of Drought-stress Responsive Gene GhGR in Gossypium hirsutum L.
    SONG Gui-Fang-., FAN Wei-Li, WANG Jun-Juan, WANG De-Long, WANG Shuai, ZHOU Kai, YE Wu-Wei
    Scientia Agricultura Sinica. 2012, 45(8):  1644-1652.  doi:10.3864/j.issn.0578-1752.2012.08.021
    Abstract ( 782 )   PDF (1036KB) ( 593 )   Save
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    【Objective】The objective of this study is to clone and make bioinformatics analysis of  the sequence of the glutathione reductase gene from G. hirsutum L.【Method】The full length of GhGR was cloned by RACE and RT-PCR techniques,the gene sequence obtained and the putative amino acid sequence were analyzed by bioinformatics software. Its expression patterns were analyzed by real-time PCR and instantaneous expression. 【Result】The GhGR full length cDNA sequence contained 1 035 bp, and ORF was 792 bp, which encoded 263 amino acid residues. Bioinformatics analysis showed that GhGR had high similarity of 90%, 91% and 91% to Populus trichocarpa (XP_002299276.1), Ricinus communis (XP_002518118.1) and Vitis vinifera (CAN74593.1), respectively. Phylogenetic analysis showed that GhGR was much closer to Vitis vinifera, the expression protein of GhGR was located in the cell membrane and cell nucleus membrane, and its expression could be induced by drought.【Conclusion】The glutathione reductase gene GhGR was cloned for the first time from G. hirsutum L. The further studies of GhGR will have an important significance in drought- tolerance for cotton breeding.
    Over-Expression of the Arabidopsis CBF1 Gene in Dongguandajiao (Musa spp.ABB group) and Detection of Its Cold Resistance
    LIU Kai, HU Chun-Hua, DU Fa-Xiu, ZHANG Yu-E, WEI Yue-Rong, YI Gan-Jun
    Scientia Agricultura Sinica. 2012, 45(8):  1653-1660.  doi:10.3864/j.issn.0578-1752.2012.08.022
    Abstract ( 788 )   PDF (423KB) ( 665 )   Save
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    【Objective】To elucidate the effects of over-expression of AtCBF1 gene on tolerance to low-temperature in Musa ABB cv. Dongguandajiao and establish a foundation for cloning cold resistance relative genes from Dongguandajiao.【Method】Transgenic plants transformed by AtCBF1 gene were generated based on Agrobacterium tumerficiens-mediated transformation of embryogenic cell suspensions (ECS) of Dongguandajiao. The transgenic plants were confirmed via GUS staining, PCR, RT-PCR and real-time fluorescence quantitative RT-PCR(RT-qPCR). The cold tolerance of transgenic plants were evaluated by the comparisons of plant morphology and physiological parameters such as chilling injury symptom, superoxide dismutase (SOD) activities, malonaldehyde (MDA) content and so on among the transgenic lines and non-transformed plants (CK) after low-temperature treatment.【Result】Six resistant lines were initiated in this experiment. Results from GUS staining showed that all resistant plants were positive except T1 line. PCR showed that AtCBF1 gene was detected in the six transformed lines, but gus gene not in T1 line. Expression of AtCBF1 gene in the six transgenic lines was confirmed using RT-PCR. Difference existed among T1, T2 and T3 for AtCBF1 expression in RT-qPCR analysis. After low-temperature stress treatment, the relative electric conductivity and MDA content of leaves of transgenic lines were lower than that of non-transformed plants. However, the superoxide dismutase (SOD) activities was higher in transgenic lines. The chilling injury symptom of leaves of transgenic lines were much slighter than that of the CK under low-temperature treatment.【Conclusion】 Over-expression of AtCBF1 gene increased SOD activities and decreased MDA content and electrolyte leakage rate caused by low -temperature. Thus the degree of over-oxidation of the cellular membrane is lessenned and the low-temperature stress tolerant capacities in transgenic plantain plants are improved.
    Effects of Maturity Stage on Fermentation Quality of Whole Crop Wheat Silage
    QIN Meng-Zhen, SHEN Yi-Xin
    Scientia Agricultura Sinica. 2012, 45(8):  1661-1666.  doi:10.3864/j.issn.0578-1752.2012.08.023
    Abstract ( 753 )   PDF (248KB) ( 663 )   Save
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    【Objective】 The changes in fermentation quality and feeding value of whole crop silage of wheat at different maturity stages were studied.【Method】Three wheat cultivars (Xinmai 18, Zhoumai 22, and Huaimai 22) were ensiled by polyethylene bag at milk and dough ripening stages to test the fermentation quality and feeding value.【Result】Whole crop of wheat ensiled at two maturation stages both provided good silage, but the fermentation quality and feeding value of the silages were significantly different between the maturity stages (P<0.05). The pH, volatile fatty acid (VFA) including acetic acid (AA), prop ionic acid (PA), and butyric acid (BA) concentrations in the silage ensiled at milk ripening stage were significantly lower than that at dough ripening stage, while the lactic acid (LA) content was significantly higher. It indicates that fermentation quality of silage ensiled at the milk ripening stage was better than that at the dough ripening stage. The concentration of neutral detergent fiber (NDF), acid detergent fiber (ADF), water soluble carbohydrates (WSC) and crude protein (CP) of the silage ensiled at dough ripening stage were significantly lower than that at milk ripening stage, but the starch content was significantly higher. Then, feeding value of whole crop silage of wheat at dough ripening stage was better.【Conclusion】At the milk ripening stage, the fermentation quality of whole crop silage was better owing to the higher WSC content, while at the dough ripening stage, the feeding value of whole crop silage was better owing to seeds increasing and starch accumulation.