Table of Content

    01 October 2012, Volume 45 Issue 19
    Screening and Identification of Susceptibility and Fertility Related Mutants from Transgenic Rice IRBB13 (PXa13:GFP)
    HAN Kuan-De, KONG Ling-Guang, JU Yan-Hu, CHEN Ming, CAO Zhi-Gang, DING Xin-Hua, CHU Zhao-Hui
    Scientia Agricultura Sinica. 2012, 45(19):  3899-3908.  doi:10.3864/j.issn.0578-1752.2012.19.001
    Abstract ( 722 )   PDF (520KB) ( 725 )   Save
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    【Objective】The objective of this study is to establish a simple and efficient mode of rice mutants selection, and lay the first stone for cloning rice genes involved in regulating xa13/ Xa13 gene expression or their mediated disease resistant /susceptible and pollen development signal pathway. 【Method】Using EMS treatment, a rice mutation population was constructed with IRBB13 (PXa13:GFP) which carrying green fluorescent protein(GFP) derived with dominant Xa13 gene promoter. Then the mutants were screened out with the following three periods: At seedling stage, selecting mutants in which GFP gene expression decline either in root or bud or non-expression; At adult stage, selecting mutants restored susceptible to PXO99; At booting stage, selecting mutants reduced fertility. 【Result】In total 3 000 M2 lines, 14 and 9 mutants were identified for both root and bud fluorescent declined and fluorescent normal only in root at seedling stage. At adult stage, relying on phenotype of PXO99 inoculation, a total of 25 lines were characterized to restore the susceptibility to PXO99. At booting stage, 7 mutants were identified, of which fertility declined and the green fluorescence of the anthers also disappeared and one mutant was found to restore susceptibility.【Conclusion】A simple and efficient mode of rice mutants selection system was established. Combining the abnormal expression pattern of GFP mutants, restored susceptible phenotype mutants and reduced fertility mutants, it was preliminarily indicated that the two signal transduction pathways mediated by xa13/ Xa13 genes are not only completely independent but also partially crossing between resistance/susceptibility and pollen development. Furthermore, after analysis and cloned the genes from these mutants, they will help us better understanding the functional mechanism of xa13/ Xa13 genes.
    Identification of QTLs for Major Isoflavone Components mong Multiple Environments in Soybean Seeds
    ZHANG Jing-Ying, GE Yi-Nan, SUN Jun-Ming, HAN Fen-Xia, YU Fu-Kuan, YAN Shu-Rong, YANG Hua
    Scientia Agricultura Sinica. 2012, 45(19):  3909-3920.  doi:10.3864/j.issn.0578-1752.2012.19.002
    Abstract ( 710 )   PDF (481KB) ( 1145 )   Save
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    【Objective】The QTLs for major isoflavone components were identified among multiple environments by different QTL mapping methods in soybean seeds, in order to provide a theoretical basis for soybean isoflavone marker-assisted selection. 【Method】 In this study, a recombinant inbreed line population (RIL, F5∶7-8) was developed by using the cross between the isoflavone contrasting cv. LHD2 with high-isoflavone concentration (3 697.24 μg•g-1) and cv. NHZ with low-isoflavone concentration (1 816.67 μg•g-1). The polymorphism of SSR markers among the RIL population was analyzed and the major isoflavone components were determined by HPLC. 【Result】The results showed that a soybean linkage map with a total distance of 3 546.54 cM was constructed using 161 polymorphism SSR molecular markers in the RIL population. A total of 14 QTLs associated with the major isoflavone components were found by ICIM, IM, and SMA methods in the ICIMapping 3.2 software under the four environments. 【Conclusion】The three QTLs flanked by the marker interval Sat_003-Satt306, Satt070-Satt122 and Satt571-Satt270 were detected among multiple environments in more than two methods.
    Validation of QTL for Oil Content in a Population of Worldwide Rapeseed Cultivars by Association Analysis
    SUN Zhong-Yong, CHENG Shuang, WANG Ji-Bian, HUANG Ji-Xiang, CHEN Fei, NI Xi-Yuan, ZHAO Jian-Yi
    Scientia Agricultura Sinica. 2012, 45(19):  3921-3931.  doi:10.3864/j.issn.0578-1752.2012.19.003
    Abstract ( 783 )   PDF (394KB) ( 1022 )   Save
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    【Objective】 The objective of this study is to validate the genetic stability of 7 major QTL for oil content,which were detected in authors’ previous researches, and to assess the feasibility of marker assisted selection for improving seed oil content using closely linked functional makers.【Method】Fourteen lipid related candidate gene markers located in the 7 QTL regions were chosen to amplify 81 core collections of Brassica napus and then a linkage disequilibrium was estimated by marker-trait association analysis. Meanwhile, the allelic distribution among 81 world-wide cultivars in each QTL was investigated.【Result】By association analysis, significant differences in oil content were detected between favorable and unfavorable allele genotypes in each of six linked markers flanking four QTL (OilA1, OilA5, OilA7 and OilC8-1), respectively. Over 5 percentage in oil content could be increased by assembling favorable alleles together through four QTL linked markers Ra2E04,ZAAS919,ZAAS828 and ZAAS441. According to the marker genotypes, in the loci of OilA1 and OilC8-1, higher than 80% of European cultivars have carried positive alleles from “Sollux”, while 80% and 60% Chinese materials contain unfavorable alleles. On OilA5 and OilA7, almost all the tested European varieties did not show favorable alleles from “Gaoyou” and for Chinese materials, also only around 20%-30% cultivars carrying these positive alleles.【Conclusion】Six orthologous gene markers (corresponding to At3g51830, At2g42450, At2g44620,At1g73600, At1g73480 and At1g13560), which closely link to OilA1, OilA5, OilA7 and OilC8-1 might be functional important in the accumulation of oil content in rapeseed. When favorable alleles from three QTL including OilA7 or all four selected QTL combined together, the oil content could be increased more than 5 percentages. The results showed that the positive alleles in screened out 4 QTL loci are potentially interesting for present breeding programs in China, while the Chinese alleles from OilA5 and OilA7 should be also attractive to rapeseed breeders in European countries.
    Effect of Planting Methods on Quality of Different Growth and Development Types of Japonica Rice Under High-Yielding ultivation Condition
    HUO Zhong-Yang, LI Jie, XU Ke, DAI Qi-Gen, WEI Hai-Yan, GONG Jin-Long, ZHANG Hong-Cheng
    Scientia Agricultura Sinica. 2012, 45(19):  3932-3945.  doi:10.3864/j.issn.0578-1752.2012.19.004
    Abstract ( 663 )   PDF (323KB) ( 782 )   Save
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    【Objective】The variations of rice quality among different planting methods of high-yielding cultivation conditions were investigated.【Method】A comparative experiment was carried out with early-maturing late japonica rice (EMLJ), late-maturing medium japonica rice (LMMJ) and medium-maturing medium japonica rice (MMMJ) (including Japonica conventional rice and japonica hybrid rice) as materials to study the difference in processing quality, appearance quality, cooking and eating quality, nutritional quality and RVA profile characteristics of artificial transplanting (AT), mechanical transplanting (MT) and direct seeding (DS) rice under high-yielding cultivation conditions. Furthermore, the correlation analysis between rice quality characters and temperature and light condition during grain-filling periods was studied. 【Result】 Brown rice rate was no significant difference among different planting methods. The order of milled rice and head milled rice rate among different planting methods was AT>MT>DS in EMLJ, while MT>AT>DS in LMMJ and MMMJ. Chalkiness rate was AT>MT>DS, and chalkiness size was MT>AT>DS. Chalkiness degree was AT>MT>DS in EMLJ and conventional LMMJ, while MT>AT>DS in MMMJ and hybrid LMMJ. Amylose content was DS>MT>AT and gel consistency was AT>MT>DS. Protein content between AT and MT was in generally quite, and DS was slightly lower than that of AT and MT, but there was no significant difference in addition to MMMJ. Peak viscosity, trough viscosity and breakdown was AT>MT>DS, setback was DS>MT>AT, peak time among different planting methods was no significant difference, final viscosity was AT>MT>DS in EMLJ and conventional LMMJ while DS>MT>AT in MMMJ and hybrid LMMJ, pasting temperature was DS>MT>AT in EMLJ and conventional LMMJ while AT>MT>DS in MMMJ and hybrid LMMJ.【Conclusion】Compared with AT and MT, the appearance quality of DS is slightly better, while processing quality, cooking and eating quality and nutritional quality become worse. The processing and appearance quality between AT and MT are different due to varieties types. The cooking and eating quality of AT is better than that of MT. The nutritional quality between AT and MT is in generally quite. Therefore, it is beneficial to obtain good quality characters to choose reasonable planting methods and supporting appropriate rice varieties types.
    Effect of Fenlong Cultivation on Yield and Quality of Rice
    WEI Ben-Hui, LIU Bin, GAN Xiu-Qin, SHEN Zhang-You, HU Bo, LI Yan-Ying, WU Yan-Yong, LU Liu-Ying
    Scientia Agricultura Sinica. 2012, 45(19):  3946-3954.  doi:10.3864/j.issn.0578-1752.2012.19.005
    Abstract ( 803 )   PDF (317KB) ( 1000 )   Save
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    【Objective】The feasibility of the new farming method of Fenlong cultivation for rice, and its effects on yield and quality of rice were studied.【Method】YLiangyou1 and Shanxiaozhan were used as materials in the test, the traditional ploughing with rake was used as CK, and smash-ridging cultivated early rice and no-tillage of late rice were carried out to investigate the related characters, yield and quality of rice. The yield of the third time planted rice after Fenlong and soil compactness were investigated.【Result】Compared with the traditional cultivation, Fenlong planted early rice increased the plough layer by 7 cm, most indicators of soil available nutrients increased by more than 10%, total number of root and white roots increased by 14.83%-25.36% and 20.18%-97.78%, respectively, the plant biological yield increased by 29.96%-31.89%, seed setting rate increased by 7.62%, yield increased by 23.87%, the whole head rice rate and protein increased by 15.95% and 14.61%, respectively; In no-tillage and seedling-throwing late rice, compared with CK, Fenlong total number of root and white roots increased by 6.02%-22.75% and 48.03%-65.91%, and yield increased by 9.25%, the whole head rice rate and protein increased by 7.43% and 5.61%. The early and late rice total yield increased by 16.48%, income increased by 6 706.5 Yuan/hm2. Compared with the CK, the third time planted rice after Fenlong farming, soil compactness reduced by 68.00%-333.33%, and yield increased by 7.97%. 【Conclusion】Fenlong cultivation of early rice and no-tillage late rice are feasible and it can increase rice production, improve rice quality, and increase  net income. Fenlong still can increase the yield of the third time planted rice, the rice paddy field possess the sustainable yield increasing ability after Fenlong farming.
    Effects of Soybean Continuous Cropping on Microbial and Soil Enzymes in Soybean Rhizosphere
    GU Yan, QIU Qiang, WANG Zhen-Min, CHEN Xi-Feng, WU Chun-Sheng
    Scientia Agricultura Sinica. 2012, 45(19):  3955-3964.  doi:10.3864/j.issn.0578-1752.2012.19.006
    Abstract ( 946 )   PDF (469KB) ( 1236 )   Save
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    【Objective】The objective of the experiment was to determine the effects of continuous and alternate cropping on microbe community structures and soil enzyme activities in soybean, and to provide a scientific basis for the obstacles of continuous cropping of soybean. 【Method】Rhizosphere soils of different growth stages was used to characterize the community structure of soil microbe based on phospholipid fatty acid (PLFA) method, and the contents of soil microbial biomass carbon and enzyme activities were determined. 【Result】In the soils of alternate and continuous cropping, the total PLFAs biomass, the microbial biomass carbon contents, the activities of urease and invertase all decreased, and the ratios of fungi and bacteria were enhanced, and the activities of catalase had no significant difference among the treatments. There were different influences on microbial and soil enzymes activities in different years of continuous cropping and growth stage, which closely related with the number and type of root exudates and straw decomposing matter. The correlation analysis showed that there was a positive relationship significantly between the total PLFAs biomass and microbial biomass carbon content and the activity of soil urease. There was no relationship between the total PLFAs biomass and the activity of soil catalase and invertase.【Conclusion】The microbial community structures and enzyme activities were changed by continuous and alternate soybean. The obstacles of continuous soybean were the result of the comprehensive effect of soil micro-ecological system and environmental factor.
    Effect of Hyperosmotic Stress on the Growth, Development and STK1 Expression of Setosphaeria turcica
    WANG Mei-Juan, LI Po, WU Min, FAN Yong-Shan, GU Shou-Qin, DONG Jin-Gao
    Scientia Agricultura Sinica. 2012, 45(19):  3965-3970.  doi:10.3864/j.issn.0578-1752.2012.19.007
    Abstract ( 575 )   PDF (490KB) ( 564 )   Save
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    【Objective】The objective of this study is to measure the effect of hyperosmotic stress on the growth and development of Setosphaeria turcica, to analyze whether or not the glycerol is one of the compatible solutes, and to detect the expression characteristic of STK1 when the pathogen is cultured under hyperosmotic stress. 【Method】 The growth and development of S. turcica, which cultured under 2 different hyperosmotic stresses, were observed. By analyzing the change of glycerol content in mycelium cells, it was confirmed that glycerol was one of the compatible solutes. The expression characteristic of STK1 was detected using semiquantitative RT-PCR method.【Result】Isotonic solution concentration of hyphal cells of S. turcica was 0.78 mol•L-1. The colony color changed distinctly under hyperosmotic stress. The colony cultured on PDA medium containing 1 mol•L-1 NaCl was reddish-brown in colour, the growth rate of the colony was repressed remarkably, cultured on PDA medium containing 1 mol•L-1 NaCl showed strong inhibition. The condensation was observed in the protoplasts when cultured in 1 mol•L-1 NaCl treatment. With the increase of hyperosmotic stress time and compatible solute concentration, glycerol content increased evidently. The expression level of STK1 enhanced distinctly within 48 hours under hyperosmotic stress.【Conclusion】Colony growth rate was inhibited, colony color turned obviously, protoplast became concentrated, some mycelium cells enlarged and changed into globosed under hyperosmotic stress in S. turcica. Glycerol was one of the compatible solutes in the mycelium cells of S. turcica. STK1 took part in regulating hyperosmotic stress responses in S. turcica.
    Rapid Detection and Identification of Fusarium oxysporum f. sp. cubense Race 1 and Race 4
    LI Min-Hui, YU Xiong-Tao, WANG Hong-Fei, ZHOU Jia-Nuan, XI Ping-Gen, JIANG Zi-De
    Scientia Agricultura Sinica. 2012, 45(19):  3971-3979.  doi:10.3864/j.issn.0578-1752.2012.19.008
    Abstract ( 1147 )   PDF (834KB) ( 1226 )   Save
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    【Objective】The objective of this study is to establish a method of rapid detection and identification of Fusarium oxysporum f. sp. cubense (FOC) race 1 and race 4 based on their respective specific sequence, avoid spreading and control banana Fusarium wilt, and further to provide a scientific basis for production and banana cultivar configuration. 【Method】DNAs of 6 FOC race 1 isolates, 7 FOC race 4 isolates, 7 F. oxysporum isolates and 2 outgrouped isolates were used as templates to screen the specific primers designed on the basis of candidate pathogenicity-relative gene sequences of FOC race 4 by PCR. 【Result】Two pairs of primers specific to FOC race 1 and race 4 respectively and one pair of universal primers of F. oxysporum were screened. Using the specific primers, pathogenic fungal DNA and diseased banana plant DNA and even contaminated soil DNA with the pathogen could be detected and identified. Meanwhile, the universal primers of F. oxysporum could also be used as control to test the DNA quality and avoid producing false negative and further to ensure the detected result. 【Conclusion】A rapid and accurate triple PCR detection method was established to identify FOC race 1 and race 4 in one PCR reaction, which is very useful to detect FOC races from banana tissue cultured plantlets and filed soil.
    Interspecific Competition Among Three Predacious Ladybirds (Coleoptera: Coccinelildae)
    WANG Su, TAN Xiao-Ling, XU Hong-Xing, ZHANG Fan
    Scientia Agricultura Sinica. 2012, 45(19):  3980-3987.  doi:10.3864/j.issn.0578-1752.2012.19.009
    Abstract ( 771 )   PDF (415KB) ( 635 )   Save
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    【Objective】The objective of this study is to determine the population dynamics of various predacious ladybird beetles after mixed releasing in the field, the behavioural bases in interspecific disturbance and the interspecific competition in niche level. 【Method】The population dynamics of the three predacious ladybird beetles, Harmonia axyridis, Propylea japonica and Hippodamia variegate were monitored after they were introduced into agricultural ecosystem. The cannibalism was observed with 1st instar larvae to eggs and 4th instar larvae each other among these three intraguild predators. Based on the field survey, the food prey of these three ladybirds as different resource levels were separated, and the niche breadth and niche overlap index were calculated. 【Result】The total scale of population showed linear increasing in all three ladybirds, and the rate in H. axyridis population was higher than others significantly. The eggs preyed in all ladybirds were tended to heterogenetic egg and larvae. The amount of egg consume in H. axyridis was significantly higher than other two and over 25% both. H. axyridis also showed the highest survival rate and the lowest damaged rate in survival samples, the damaged rates were only 22.3% to P. japonica and 29.8% to H. variegate. The field investigation showed that H. axyridis, P. japonica and H. variegate could prey 17 species (89.5% of the total), 12 species (63.2% of the total) and 9 species (47.4% of the total), respectively. The calculated niche breadths of H. Axyridis, P. japonica, H. variegate were 0.713, 0.393 and 0.304, rspectively. The niche overlap indexes were showed as H. axyridis to P. japonica was 0.992, P. japonica to H. variegate was 0.983 and H. axyridis to H. variegate was 0.964. 【Conclusion】H. axyridis showed extremely high competition with other predacious ladybird in field application. H. axyridis could obtain benefits via interspecific attacking under rare nutrition conditions. The wider niche breadth and higher niche overlap level of H. axyridis could influence the population development and niche separation of other predacious ladybird beetles.
    Effects of Bemisia tabaci (Homoptera: Aleyrodidae) Biotype B Infestation on Photosystem II in Nicotiana tabacum
    LI Qing-Liang, TAN Wei, XUE Ming
    Scientia Agricultura Sinica. 2012, 45(19):  3988-3995.  doi:10.3864/j.issn.0578-1752.2012.19.010
    Abstract ( 580 )   PDF (386KB) ( 744 )   Save
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    【Objective】The objective of this study is to define the effects of Bemisia tabaci biotype B infestation on the photosystem II (PSII) in tobacco, and to reveal the mechanism of B. tabaci infestation affecting photosynthesis of host plant. 【Method】Effects of B. tabaci infestation on the PSII performance in damaged leaves and systemic leaves of tobacco were investigated by measuring chlorophyll fluorescence transients and analyzing the related parameters using JIP-test.【Result】B. tabaci infestation significantly increased the maximal photochemical efficiency of primary photochemistry (φpo) and performance index on absorption basis (PIABS) both in local and systemic leaves (P<0.05), indicating that the original photochemical reaction was greatly damaged. Photosynthetic electron transport was inhibited and oxygen-evolving complexes (OEC) were severely damaged. B. tabaci infestation increased the energy absorption per active reaction centers (ABS/RC) and the energy dissipation per active reaction centers (DIo/RC) increase while decreased energy trapping per active reaction centers (TRo/RC) in systemic leaves. In damaged leaves, DIo/RC increased and TRo/RC decreased, however, the ABC/RC was not affected. B. tabaci infestation reduced the density of active reaction centers per excited cross-section (RC/CS) both in damaged and systemic leaves (P<0.05) while increased the closure degree of the PSⅡreaction centers by 69.83% and 142.58% (P<0.05), respectively, in damaged and systemic leaves.【Conclusion】B. tabaci infestation damaged the PSII of tobacco leaves. The reaction centers were damaged and the electron transporting was inhibited. The inhibited points of the electron transporting included the OEC and the electron transporting from QA to QB. Energy flux of PSII was also affected by the B. tabaci infestation. The damage on PSII caused by B. tabaci in tobacco leaves was systematically conductive.
    Cloning and Bioinformatics Analysis of Voltage-Gated Sodium Channel Gene cDNA in Bactrocera dorsalis (Hendel)
    JIANG Xuan-Zhao, WEI Dan-Dan, SHEN Guang-Mao, DOU Wei, WANG Jin-Jun
    Scientia Agricultura Sinica. 2012, 45(19):  3996-4003.  doi:10.3864/j.issn.0578-1752.2012.19.011
    Abstract ( 607 )   PDF (1248KB) ( 833 )   Save
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    【Objective】 The objective of this study is to clone voltage-gated sodium channel gene (VGSC) from Bactrocera dorsalis (Hendel), and to identify its typical hallmarks. It will provide basic molecular information for clarifying insecticide resistance mechanism of B. dorsalis.【Method】 The cDNA sequence was isolated using RT-PCR and PCR methods. Based on the sequencing results, the bioinformatics analysis of nucleic acid and putative amino acid was conducted.【Result】An almost full-length cDNA sequence (6 446 bp) of VGSC was obtained, including a complete open reading frame (ORF) of 6 405 bp, which encoded 2 134 amino acids and included all the typical hallmarks of VGSC. The amino acid shared 91.7%, 86.9% and 42.3% identity with sodium channel genes of Drosophila melanogaster (NP_001188635), Musca domestica (AAB47604), and Homo sapiens Nav1.2 (NP_066287), respectively. 【Conclusion】A complete ORF of VGSC was sequenced with clone strategy from oriental fruit fly for the first time. Abundant alternative splicing phenomena existed in the VGSC of B. dorsalis. Three potential mutation points, which may confer resistance to pyrethroids and DDT, were observed. It is suggested that VGSC would be used as a molecular marker in phylogenetic study of insect.
    The Effects of Drip Irrigation Uniformity on Spatial and Temporal Distributions of Water and Nitrogen in Soil for Spring Maize in North China Plain
    ZHANG Hang, LI Jiu-Sheng
    Scientia Agricultura Sinica. 2012, 45(19):  4004-4013.  doi:10.3864/j.issn.0578-1752.2012.19.012
    Abstract ( 628 )   PDF (366KB) ( 634 )   Save
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    【Objective】The objectives of this study were to investigate the effects of drip irrigation uniformity and soil properties on the spatial and temporal variability of water and nitrogen in the soil and to quantitatively evaluate the suitability of current design standard of drip irrigation uniformity.【Method】Field experiments were conducted on drip irrigated spring maize  (Zea mays L.) in north China plain. Three Christiansen uniformity coefficients (CU) of 0.66 (low), 0.81(medium), and 0.99 (high) and three levels of water application at 50%, 75%, and 100% of the irrigation requirement were used. Soil water and nitrogen content were periodically measured during the growing seasons of spring maize.【Result】The result indicated that the uniformity coefficient of soil water content was higher than 0.85 in 2009 and higher than 0.80 in 2010. The low uniformity treatment did not produce a uniformity coefficient of soil water content that was significantly different from the medium and the high uniformity treatments. For the low uniformity treatment, the uniformity of soil water content was considerably higher than its irrigation uniformity (0.66). The influence of the uniformity of initiate soil water content was more important than the uniformity of water application and irrigation amount. Drip irrigation uniformity imposed an insignificant influence on the uniformity coefficients of nitrate content.【Conclusion】The results suggested that drip irrigation uniformity was not the main factor influencing the spatial and temporal variability of soil water and nitrogen for spring maize in the semi-humid regions such as the north China plain. The current design standard of drip irrigation uniformity coefficient (CU≥0.80) could be fairly lowered to reduce the initial cost of drip irrigation systems.
    Characteristics of Annual Water Utilization in Winter Wheat- Summer Maize Rotation System in North China Plain
    QIN Xin, LIU Ke, ZHOU Li-Li, ZHOU Shun-Li, LU Lai-Qing, WANG Run-Zheng
    Scientia Agricultura Sinica. 2012, 45(19):  4014-4024.  doi:10.3864/j.issn.0578-1752.2012.19.013
    Abstract ( 661 )   PDF (363KB) ( 750 )   Save
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    【Objective】The objective of this experiment is to quantitatively study the characteristics of annual water utilization in winter wheat-summer maize rotation system in North China Plain.【Method】The experiment with four water treatments applied in wheat growing season including no irrigation (W0), irrigation at jointing (W1), irrigation at both jointing and anthesis (W2), and irrigation at regreening, booting, anthesis and grain filling (W4) was conducted for two continuous rotation cycles in 2008-2010 under field conditions.【Result】The highest yield of wheat and annual crops (wheat plus maize) were obtained in W1 and W2, respectively, and no significant differences were found in the yield of following maize. Water use efficiency (WUE) of wheat had a similar performance to its yield. The WUE of maize was significantly higher than that of wheat, and decreased significantly with the increase of irrigation amount. The highest annual WUE was obtained in W0 or W1, and then decreased significantly with the increase of irrigation amount. There was no significant difference in soil water storage of 2 m depth soil body in the treatment of W4 at different stages of wheat and maize. However, the soil water content reduced continuously following wheat growth in the other water treatments, and the less of irrigation amount, the bigger of decreasing range of soil water content, and it reached the lowest point at wheat harvest stage. At maize jointing stage, there was no difference in soil water content among all of the water treatments because of precipitation, and correspondingly, 178-188 mm (W0), 124-160 mm (W1), 38-93 mm (W2), and -30-21 mm (W4) rainfall were stored in 2 m depth soil body, respectively. Crop water consumption intensity and seasonal evapotranspiration (SET) increased as irrigation amount increased. There were no differences in the characteristics of water consumption and soil water content among all of water treatments after maize jointing stage. Drainage occurred because of precipitation, 163 mm (W0, W1), 181 mm (W2), and 217 mm (W4) in wet year and 13 mm (W0, W1, W2), and 45 mm (W4) in normal year were determined. There were 127 mm (W0) and 57 mm (W1) water entered groundwater in wet year.【Conclusion】Water-saving cultivation in wheat reduces the exploitation of groundwater obviously, and the use efficiency of rainfall is enhanced significantly, as a result, higher annual WUE and supplementary on groundwater by rainfall in wet year are implemented, so the practice of W1 in wet year and W2 in normal year is benefit to the combination of higher WUE and high-yielding, and it is very important to the sustainable development in agriculture in North China Plain.
    Effect of Topdressing Nitrogen Application on Leaf Senescence  and 15 N-Urea of Absorption, Distribution and Utilization in Potted ‘Fuji ’Apple
    DING Ning, JIANG Yuan-Mao, PENG Fu-Tian, CHEN Qian, WANG Fu-Lin, ZHOU 恩Da
    Scientia Agricultura Sinica. 2012, 45(19):  4025-4031.  doi:10.3864/j.issn.0578-1752.2012.19.014
    Abstract ( 644 )   PDF (338KB) ( 689 )   Save
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    【Objective】The aim of the experiment is to provide a theoretical basis for topdressing application in apple for delaying the leaf senescence.【Method】The effects on leaf senescence and the characteristics of absorption and utilization of potted two-year-old ‘Fuji’ apple/Malus hupenhensis in response to topdressing nitrogen were studied.【Result】Plant leaf area, SPAD and the total N content of leaves were obviously the highest in one-time topdressing application at full-bloom stage, and the lowest were found in three-time topdressing application. At flower bud differentiation stage, two-time topdressing application was the highest, and the lowest was in one-time topdressing application. At fruit maturity stage, three-time topdressing application were obviously the highest, while one-time topdressing application was the lowest. The leaf SOD, POD and CAT activities were obviously the highest in one-time topdressing application at full-bloom stage, the lowest were found in three-time topdressing application. At flower bud differentiation stage, two-time topdressing application was the highest, the lowest was in one-time topdressing application. At fruit maturity stage, three-time topdressing application was obviously the highest, while one-time topdressing application was the lowest. The Ndff value in different organs was significantly different and the value of three-time topdressing application was obviously higher than that of one or two-time topdressing at fruit maturity stage. Plant total nitrogen, 15N uptake and nitrogen ratio were the highest in three-time topdressing application fruit maturity stage.【Conclusion】Three-time topdressing application not only enhanced the absorption and transfer ability to N in apple tree, delayed leaf senescence, but also increased the nitrogen efficiency.
    Analysis of the Specificity of Rootstock and Scion Combinations of ‘Hamlin’ Sweet Orange Related to Fruit Oleocellosis by FTIR
    ZHENG Yong-Qiang, LIU Yan-Mei, HE Shao-Lan, YI Shi-Lai, DENG Lie, ZHOU Zhi-Qin, JIAN Shui-Xian, LI Song-Wei
    Scientia Agricultura Sinica. 2012, 45(19):  4032-4039.  doi:10.3864/j.issn.0578-1752.2012.19.015
    Abstract ( 738 )   PDF (596KB) ( 755 )   Save
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    【Objective】Fourier transform infrared (FTIR) spectrometry, which is accurate, simple and efficient, with high resolution, was used to investigate the molecular physiological mechanisms of the specificity of rootstock and scion combinations of ‘Hamlin’ sweet orange [Citrus sinensis (L.) Osbeck cv. Hamlin] related to on-tree oleocellosis.【Method】The influence of ‘Hamlin’ sweet orange on Goutouchen sour orange [C. aurantium (L.). GT], Rich trifoliate 16-6 [P. trifoliate (L.). LC] and Cleopatra mandarin [C. reticulate (L.). IO] on rates of oleocellosis (RO) and degree of oleocellosis (DO) were measured at harvest stage. And the leaf and fruit peel FTIR spectroscopy of ‘Hamlin’ sweet orange on GT, LC and IO were also determined at the sensitive period of on-tree oleocellosis.【Result】The results showed that rootstocks had a significant influence on RO and DO of ‘Hamlin’ sweet orange fruits, and the fruits of ‘Hamlin’ sweet orange on GT and LC had the highest DO and RO, respectively. In contrast, the fruits of ‘Hamlin’ sweet orange on IO had the lowest DO and second higher RO. And the leaf absorption peak intensity (API) at 3420, 2927, 1625 and 1069 cm-1 of ‘Hamlin’ sweet orange on GT, IO and LC decreased in order. The above results showed that the influence of high temperature on leaf carbohydrate synthesis and transport, cell walls composition, protein stability and leaf membrane lipid peroxidation balance of ‘Hamlin’ sweet orange on GT were minimal, next was Hamlin sweet orange on IO, and ‘Hamlin’ sweet orange on LC were maximum. However, the API of peel FTIR of ‘Hamlin’ sweet orange on IO and GT was significantly lower than that of ‘Hamlin’ sweet orange on LC. Whilst, the changes of RO and DO were consistent with leaf FTIR and fruit peel FTIR, respectively.【Conclusion】 RO may be related to the response of citrus leaves to high temperature stress, in contrast, DO is directly related to fruit peel metabolism. And it is feasible to predict RO and DO by leaves and peels FTIR analysis, respectively. At the same time, leaf and peel FTIR analysis is helpful to reveal the stock-scion specific molecular and physiological mechanisms of on-tree oleocellosis.
    Ultracytochemical Localization and Effects of Calcium on Isoenzyme in the Leaves of Cucumis melo L. Under the Stress of Powdery Mildew
    ZHONG Li, LI Guan
    Scientia Agricultura Sinica. 2012, 45(19):  4040-4049.  doi:10.3864/j.issn.0578-1752.2012.19.016
    Abstract ( 544 )   PDF (1773KB) ( 1041 )   Save
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    【Objective】The aim of this experiment is to study the uhracytochemical localization of calcium in mesophyll cells of Cucumis melo L. before and after inoculating Podosphaera xanthii and the effects of exogenous calcium on three isoenzymes of defense enzymes which will be useful for further elucidating the process of calcium signal transduction and resistance mechanism to powdery mildew of melon. 【Method】With the method of electron microscope and cytochemistry technology, a hydroponic experiment was carried out to explore the effects of different calcium treatments on isoenzymes of peroxidase (POD), catalase (CAT) and superoxide dismutase (SOD) in leaves of the melon seedlings under powdery mildew stress.【Result】The calcium in mesophyll cells of resistant variety ‘MR-1’ and in susceptible variety ‘JS’ gathered in cytoplasm and there were a remarkable increase of calcium level in cytoplasm and reduced greatly in vacuolate and intercellular space when inoculating P. xanthii for 2 d. The calcium localization in leaf cells of ‘MR-1’ trended to the status as before inoculating when the seedlings were treated for 6 d. However, larger calcium deposited and localized together in the cytoplasm of ‘JS’ and the structure of mesophyll cells were destroyed until to death and there were no recover of calcium from cytoplasm into vacuolate and intercellular space. Compared with the control, the application 6 mmol•L-1 CaCl2 in nutrient solution significantly alleviated the stress on the melon leaves system that the activities of SOD, POD and CAT isoenzymes were significantly improved, while application of 75 mmol•L-1 LaCl3 in nutrient solution significantly inhibited the three isoenzymes activities.【Conclusion】Calcium ditribution in leaf cells of C. melo L. changed greatly due to its resistance ability at the stress of powdery mildew. A lot of Ca2+was released from Ca2+ pool (vacuoles) into cytoplasm, then was pumped return vacuoles in mesophyll cells of resistance variety ‘MR-1’, but in mesophyll cells of susceptible variety ‘JS’, there only occurred the first stage. Exogenous calcium probably increased calcium transport to the plant, improved the signal transmission to the body, and enhanced the scavenging of reactive oxygen species in plant cell under stress.Thereby, exogenous calcium enhanced the ability of powdery mildew resistance of melon plants.
    Protoplasts Culture Isolated from Friable Embryogenic Callus of Cassava and Plant Regeneration
    WEN Feng, XIAO Shi-Xin, NIE Yang-Mei, MA Qiu-Xiang, ZHANG Peng, GUO Wen-Wu
    Scientia Agricultura Sinica. 2012, 45(19):  4050-4056.  doi:10.3864/j.issn.0578-1752.2012.19.017
    Abstract ( 890 )   PDF (540KB) ( 819 )   Save
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    【Objective】The objective of this study is to establish an efficient system of protoplast regeneration for further developing protoplast fusion and transformation in cassava. 【Method】 Protoplasts were isolated from suspension cultures derived from friable embryogenic callus (FEC) of cassava genotype TMS60444. The highest protoplast yield obtained was 3.5×106 protoplasts/g fresh weight. Viabilities of the protoplasts assessed by the fluorescein diacetate (FDA) were approximately 90%. Protoplasts were cultured in TM2G medium with liquid thin layer culture at densities of 5×105 p/mL or 2×105 p/mL. During the first 30 d, the medium was refreshed by 0.3 mol•L-1 TM2G fresh medium every 10 d. After that, the medium was refreshed by 0.25 mol•L-1 TM2G fresh medium every 10 d. After cultured for 45 d, calli of 1-2 mm were picked out and separately developed into embryos on MSN medium, into mature embryos on CMM medium, into shoots on CEM medium and into roots on MS medium. 【Result】It was showed that all protoplasts cultured at density of 5×105 p/mL developed into compact calli (could develop into embryos), protoplasts cultured at density of 2×105 p/mL developed into compact calli and vacuolar calli (could not develop into embryos). A total of 1 479 compact calli were picked out and developed into 757 cotyledon embryos and regenerated 186 plants in the experiment. 【Conclusion】The yield and viability of isolated protoplasts had been greatly increased, the bottleneck of predecessors mentioned was improved, and the efficiency of plant regeneration from protoplasts was promoted. 
    Distribution and Temporal Expression of Staphylococcus aureus Enterotoxin Genes in Foodborne Staphylococcus aureus Strains
    YANG Jing, YANG Jun, HUANG Ji-Chao, HE Wei-Ling, ZHANG Chi, HUANG Ming
    Scientia Agricultura Sinica. 2012, 45(19):  4057-4066.  doi:10.3864/j.issn.0578-1752.2012.19.018
    Abstract ( 530 )   PDF (599KB) ( 1313 )   Save
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    【Objective】 The objective of this study is to investigate the distribution of different serum enterotoxins in food-borne Staphylococcus aureus and to further analyze their temporal expression of mRNA.【Method】Fifty-one foodborne S. aureus isolates were separated, and the presence of 11 staphylococcal enterotoxins was identified by PCR technology. After the total RNA was extracted, the expression of enterotoxin mRNAs during the cell cycle were assayed in selected isolates by reverse transcriptional real-time PCR with internal control genes ftsZ and ropB. 【Result】 Eight SE genes were detected in the 51 food-borne S. aureus isolates. The most prevalent SE genes were seg and sei. During the growth cycle, the patterns of SE mRNA gene expression were similar in all of the selected isolates. The enterotoxin mRNAs were peaked in the post-exponential growth phase and then rapidly decreased. However, the relative expression level of different enterotoxin genes in the same isolate was different, and the same SE gene's expression level also varied among strains.【Conclusion】The distribution and temporal expression of foodborne enterotoxin genes were studied, which would be valuable to the study of staphylococcal food poisoning (SPF) mechanism and the control of food safety.
    Construction of Wine Yeast for Improving Cystathionine β-Lyase Activity
    MA Jie, LIU Yan-Lin
    Scientia Agricultura Sinica. 2012, 45(19):  4067-4074.  doi:10.3864/j.issn.0578-1752.2012.19.019
    Abstract ( 575 )   PDF (600KB) ( 710 )   Save
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    【Objective】 The wine yeast which exhibited higher cystathionine β-Lyase activity was contructed.【Method】Plasmid pAUR123 was used to express the tnaA gene of Escherichia coli in Saccharomyces cerevisiae strain LFP525 which is a indigenous wine yeast selected from China. Then the cystathionine β-Lyase activity of yeast transformants was detected. After that, model grape juice fermentation and white wine fermentation were carried out to evaluate the tenological properties and thiol-producing characteristics of them.【Result】Two engineering wine yeasts, TH1 and TH2, were obtained in this study. Compared with the host strain, their cystathionine β-Lyase activities were increased by 32.25% to 59.44%. The result of model grape juice fermentation showed that the fermentation duration of engineering yeast strains was one-day longer, the residual sugars and acetic acid were higher than their host strain, while the production of thiols increased obviously, which were 2.8-4.3 folds than that in the host strain. In Sauvignon Blanc wine making test, the technological characteristics of engineering yeast strains showed no significant difference with the host strain, whereas the production of thiols was 1.22 times higher than that of the host strains.【Conclusion】The cystathionine β-Lyase activity of wine yeast was enhanced successfully by using the gene modification method in this study. It will have great interests for improving flavor characteristics of wine in the future.
    Genome-Wide Association Study and Positional Candidate Gene Analysis on Age at Puberty of Gilts
    LI Ping-Hua, LI Jie, YANG Zhu-Qing, ZHANG Zhi-Yan, YANG Bin, CHEN Cong-Ying
    Scientia Agricultura Sinica. 2012, 45(19):  4075-4083.  doi:10.3864/j.issn.0578-1752.2012.19.020
    Abstract ( 633 )   PDF (565KB) ( 1217 )   Save
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    【Objective】Isolation and characterization of the causative gene or molecular marker for age at puberty will contribute to the selection and breeding of sows.【Method】The materials used in this study were F2 gilts from a white Duroc × Erhualian F2 resource population. F0, F1 and 316 well phenotyped F2 gilts were genotyped with Illumina porcine 60K SNP chip. Genome-wide association study (GWAS) and linkage and linkage-disequilibrium analysis (LDLA) were carried out to detect SNPs or haplotypes that were significantly associated with age at puberty of gilts. The associations of 3 SNPs within two candidate genes of Lin-28 Homolog B (LIN28B) and Trans-membrane Protein 38B (TMEM38B) with age at puberty of gilts were evaluated by standard association test, marker-assisted association test and F-drop test.【Result】The results from GWAS showed that the ASGA0032316 was the most significant SNP that was associated with age at puberty. It is located at 33.07 Mb of pig chromosome 7 (SSC7)  and exactly within the intron region of RAB23. At the same instant, several significant SNPs were also detected on chromosomes 1, 6, 12, 15 and 17. The results from LDLA analysis showed that all haplotypes that were associated with age at puberty at genome-wide significance level were located on SSC7. The most significant haplotype was located at 38.39-38.47 Mb and within the intergenic region of F1RVR7 and ZFAND3. In association analyses of positional candidate genes, although 3 SNPs from LIN28B and TMEM38B were significantly associated with age at puberty in standard association test (P<0.05), it was not achieved significance level both in the marker-assisted association test and F-drop test (P>0.05). 【Conclusion】The SNP and haplotype that were most significantly associated with age at puberty of gilts was detected on SSC7. Several other significantly associated SNPs were also detected on chromosomes 1, 6, 12, 15 and 17. The results from association tests suggested that LIN28B or TMEM38B gene on SSC1 is not the quantitative trait gene for age at puberty, or more SNPs from two genes need to be further analyzed.
    Positive Selection Driven the Differentiation of Melatonin Receptor MT1 and MT2 Genes
    HE Chun-Bo, JIANG Xun-Ping, LIU Gui-Qiong
    Scientia Agricultura Sinica. 2012, 45(19):  4084-4091.  doi:10.3864/j.issn.0578-1752.2012.19.021
    Abstract ( 765 )   PDF (311KB) ( 922 )   Save
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    【Objective】 This paper focused on molecular evolution of melatonin receptor (MTNR) genes for providing clues of gene biological function. 【Method】By using bioinformatics methods and softwares,  the phylogenetic tree, genetic similarity, selection pressures and selection direction of melatonin receptor 1a (MT1) and melatonin receptor 1b (MT2) genes from 11 species were analyzed.【Result】MT1 and MT2 genes might come from a duplication of a common ancestor gene, MT1 might be keeping most functions of the parental gene and MT2 might be a new functional gene. The genetic similarity of MT1 genes among species was significantly higher than MT2 (P<0.01). MT1 gene was undergone purifying selection while MT2 was under significant positive selection. Both of MT1 and MT2 were undergone strong selection pressure and there was no significant difference in selection pressure (P>0.05) between the two genes.【Conclusion】Purifying selection for MT1 gene could explain the discrepancy of the association of MT1 gene with seasonal reproduction in animals and positive selection for MT2 gene showed that positive selection might be the main force driving the differentiation of MT2 and MT1.
    Correlations Between SNP of MTNR Gene and Cashmere-Producing Traits of Inner Mongolia White Cashmere Goat
    LIU Bin, ZHAO Cun-Fa, LI Yu-Rong, GAO Feng-Qin, GAO Wa, CHENG Li-Xin, MENG Ke-Ba-Ya-尔, MA Yue-Jun, LIU Shao-Qing, LI Jin-Quan
    Scientia Agricultura Sinica. 2012, 45(19):  4092-4101.  doi:10.3864/j.issn.0578-1752.2012.19.022
    Abstract ( 662 )   PDF (665KB) ( 626 )   Save
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    【Objective】Correlations between SNP of cashmere goat melatonin receptor (MTNR) gene and cashmere-producing traits were studied to find molecular marker of MAS for molecular breeding.【Method】 Arbas white cashmere goats and partial sequences of MTNR1a and 1b genes were used as experimental samples and candidate genes, respectively. PCR-SSCP and DNA sequencing technologies were conducted to detect their single nucleotide polymorphism. Results were analyzed by least square mean (LSM) among production traits of genotypes.【Result】 There was no SSCP in the exons 1 and 2 of MTNR1a gene and the exon 1 of MTNR1b gene, while there was the SSCP in the exon 2 of MTNR1b gene and three of SNP were detected. P4 primer loci existed in C-G mutation of NM_001206907:g.648 bp which didn’t result in change of amino acid, and G-A mutation of NM_001206907:g.665 bp which led to CGC-CAC of the 223rd codon from Arg into His. The results showed that polymorphism loci had an significant effect on cashmere yield (P<0.05), and cashmere yield of AA genotype increased by 98.78 g and 148.23 g, as compared to AB and BB, respectively. Meanwhile, there was a significant correlation (P<0.05) in interaction effect between the polymorphism loci genotype and age cashmere thickness. P5 primer loci existed in G-A mutation of NM_001206 907:g.1015 bp which led to GGC-AGC of the 339th codon from Gly into Ser. The analyses indicated that interaction effect of the polymorphism loci genotype and age was the marginal significant correlation (P<0.1) with cashmere fineness. P4 and P5 primes locus were not significantly (P>0.05) with kidding number and birth weight.【Conclusion】P4-AA genotype can be used as molecular marker of MAS for cashmere-producing traits of Inner Mongolian Arbas white cashmere goats.
    Rescue and Identification of the Recombinant Bovine Rotavirus with Mutational NSP4 Gene
    YANG Shao-Hua, HE Hong-Bin, YANG Hong-Jun, CHEN Fang-Yuan, GAO Yun-Dong, ZHONG Ji-Feng
    Scientia Agricultura Sinica. 2012, 45(19):  4102-4108.  doi:10.3864/j.issn.0578-1752.2012.19.023
    Abstract ( 831 )   PDF (548KB) ( 752 )   Save
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    【Objective】The objective of the study is to rescue the attenuated rotavirus with mutational NSP4 gene by reverse genetic method and to purify further by RNAi technique and plaque clone method. 【Method】NSP4 gene of a wild type RV strain CHLY was cloned and five silent mutation nucleotides were introduced at 93 nt-109 nt and five missense mutations at 444 nt-453 nt, which resulted in M135L, I136T, A138P amino acid mutation. A recombinant plasmid △pT7-NSP4/89/D that contains T7 promoter and T7 terminator at 5′ and 3′ end of the manipulated NSP4 cDNA was constructed. △pT7-NSP4/89/D plasmid and expression vector pcDNA3.1/T7-RNAP carrying RNA polymerase in vivo was co-transfected MA104 cell layer that had been infected 24 h earlier with wild type RV as helper virus, and the transfected cell was cultured for further 24 h until harvest. The culture fluid colleted was subjected to passage on MA104 cell in the presence of Lentivirus-RNAi-H1-89, which could inhibit the amplification of helper virus specifically. The rescued virus was biologically cloned by five successive plaque purifications in MA104 cells.【Result】A RV strain with mutational NSP4 gene was successfully rescued. Compared to wild type RV, the rescued virus showed attenuated virulence to MA104 cell and mouse pups. 【Conclusion】A attenuated virulence RV was rescued by reverse genetic method and RNAi technique. The mutations of 135aa, 136aa and 138aa of NSP4 have effect on virus virulence.
    Genetic Mapping of the Dark Brown Fiber Lc1 Gene in Tetraploid Cotton
    LI Fu-Zhen, NING Xin-Min, QIU Xin-Mian, SU Cheng-Fu, YAO Jian-Qiang, TIAN Li-Wen
    Scientia Agricultura Sinica. 2012, 45(19):  4109-4114.  doi:10.3864/j.issn.0578-1752.2012.19.024
    Abstract ( 770 )   PDF (355KB) ( 719 )   Save
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    【Objective】The objective of the experiment is to study and understand the genetic law of brown cotton fiber, to find and locate the molecular markers linked with the locus of brown fiber gene for laying a foundation for fine-mapping, molecular cloning of the brown fiber gene and brown fiber quality improvement at the cotton genome level. 【Method】 Based on the upland cotton dominant multi-gene markers Line T586 (possesses the dark brown fiber gene Lc1) crossing with the sea island cotton Xinhai 16, the F2 population were constructed. Combined with fiber color classification and analysis by colorimeter, taking full advantage of the molecular marker genetic linkage map information of the cotton genome, the molecular markers tightly linked with the dark brown fiber Lc1 gene were mapped by screening of the polymorphism molecular markers and map-based cloning method. 【Result】 The F2 population of T586 crossing with Xinhai 16 were composed of 443 individuals, which of them were classified into the dark brown fiber, brown (middle color) and white by the fiber color. Finally, the Lc1 was located in the A subgroup of the cotton genome chromosome 7, which was flanked by the SSR marker NAU4030 and CGR5119 with a genetic distance of 8 cM, corresponding to the genetic distance of 5.1 cM and 2.8 cM, respectively. 【Conclusion】The trait of dark brown fiber is controlled by single gene, possesses the semi-dominant inheritance in tetraploid cotton. The information of molecular markers tightly linked with Lc1 locus could be used in marker-assisted selection in brown fiber cotton breeding.
    Effects of Different Treatments of Deep Tillage and Panicle Fertilization on Root Activity, Grain-Filling Dynamics and Yield of Rice (Oryza sativa L.)
    ZHANG Jing, ZHANG Zhi, DU Yan-Xiu, LI Jun-Zhou, SUN Hong-Zheng, ZHAO Quan-Zhi
    Scientia Agricultura Sinica. 2012, 45(19):  4115-4122.  doi:10.3864/j.issn.0578-1752.2012.19.025
    Abstract ( 576 )   PDF (309KB) ( 968 )   Save
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    【Objective】The effects of deep tillage and panicle fertilization on rice root activity, grain filling dynamics and yield were studied.【Method】A field experiment was carried out in a randomized complete block design with four treatments at top forth leaf stage. The treatments were broadcast-application without deep tillage, broadcast-application with deep tillage, broadcast- application with deep tillage, furrowing and ridging, and deep-application with deep tillage. 【Result】Rice yield and the bleeding intensity at the basal internode and neck-panicle node increased significantly in different treatments of deep tillage and panicle fertilization compared with the control. Deep-application with deep tillage showed the greatest positive effect, followed by broadcast-application with deep tillage, furrowing and ridging, and broadcast-application with deep tillage. Maximum grain weight, initial grain filling potential, relatively initial grain filling potential, maximum grain filling rate, and mean grain filling rate of inferior and superior grains were both increased under the different deep tillage and panicle fertilization patterns.【Conclusion】At the middle growth stage, deep tillage could increase the vigor of root and sink, improve grain-filling process, and then enhance grain yield of rice grown in sandy loam soil.