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Table of Content

    16 August 2019, Volume 52 Issue 16
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Development New Molecular Markers for Quantitative Trait Locus (QTL) Analysis of the Seed Protein Content Based on Whole Genome Re-Sequencing in Soybean
    WANG Jia, ZENG ZhaoQiong, LIANG JianQiu, YU XiaoBo, WU HaiYing, ZHANG MingRong
    Scientia Agricultura Sinica. 2019, 52(16):  2743-2757.  doi:10.3864/j.issn.0578-1752.2019.16.001
    Abstract ( 811 )   HTML ( 92 )   PDF (4355KB) ( 773 )   Save
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    【Objective】 Based on the results of genome-wide re-sequencing, molecular markers closely related to high protein, shade tolerance, lodging resistance and other traits were developed. At the same time, At the same time, genetic linkage maps were constructed using the developed molecular markers, and seed protein content was mapped by QTL, providing reference and molecular marker resources for subsequent research on high protein, shade tolerance and lodging resistance breeding. 【Method】 A F2 segregating population derived from the cross of Nandou 12 and Shiyuehuang consists of 672 individuals, and two parents were re-sequenced. With the published genome as a reference, the obtained data were assembled with BWA, and explored for the SNP and InDel by GATK and SV by Breakdancer. Carry out expression pattern analysis toward mutational storage proteins and genes related to environmental adaptation by combining with the transcriptome data obtained from different development stages and shade processing of seeds and qRT-PCR, At the same time, based on the resequencing data, excavate the SNP sites in the gene coding region between the parents, analyze the restriction enzyme cutting site and transform the SNP markers into CAPS or dCAPS markers. In addition, search the insertion/deletion mutation site and design primer development InDel marker in highly conserved regions on both sides of the insertion/deletion site. Perform polymorphism screening on the CAPS markers and InDel markers developed, select the CAPS molecular markers and InDel markers with polymorphism and carry out genotyping toward F2 materials. Utilize JoinMap 4.0 software to construct the genetic linkage map according to the genotyping result. Obtain the seed protein content data of F2 material according to the genetic map constructed by combining with the near-infrared analysis and use Windows QTL Cartographer V2.5 to carry out QTL analysis toward soybean seed protein content. 【Result】 The results showed that a large number of storage proteins and important genes or homologous genes related to environmental adaptation mutated in Nandou 12. The results of transcriptome data analysis showed that some variant genes showed different expression patterns and significant differences and the results were further validated by qRT-PCR analysis. In addition, 332 of the 540 CAPS molecular markers had polymorphic, and 201 of 300 pairs of InDel primers could amplify polymorphism. A genetic linkage map containing 20 linkage groups was constructed based on polymorphic molecular markers, covering 2973.87 cM with an average genetic distance of 5.58 cM. Using this map to map the seed protein content of soybean, six QTL loci were detected, which could explain 4.68%-18.25% phenotypic variation.【Conclusion】 Based on the variation loci among parents, 533 polymorphic molecular markers (including 8 gene-specific molecular markers) were developed. Six QTL loci were detected for seed protein content in soybean, including one major QTL locus (qSPC-6).

    Coefficient of Parentage Analysis Among Naked Barley Varieties in Qinghai-Tibet Plateau
    LI Jian, FENG XianHong, CAI YiLin
    Scientia Agricultura Sinica. 2019, 52(16):  2758-2767.  doi:10.3864/j.issn.0578-1752.2019.16.002
    Abstract ( 411 )   HTML ( 47 )   PDF (967KB) ( 377 )   Save
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    【Objective】 The present analysis of genetic diversity of bred naked barley varieties of the Qinghai-Tibet Plateau was conducted to evaluate their genetic diversity, to explore measures to broaden the genetic basis, and to promote the breeding of naked barley varieties. 【Method】 In total, 105 bred naked barley varieties grown commonly and broadly across Qinghai-Tibet plateau, which has been surveyed since 1950, and 3 introduced varieties were selected as study materials. The genetic diversity of the investigated varieties was estimated by coefficient of parentage (COP), and then based on these COP values, a clustering analysis was conducted using Lance and Williams distance and UPGMA. Ultimately, the core parents in Qinghai-Tibet plateau were determined by examining the cumulative direct coefficient of parentage (CD-COP). 【Result】Overall, 18.01% of variety combinations were genetically related. The COP of all combinations ranged from 0.000 to 0.750, with an average of 0.053, which was significantly higher than that of the main barley varieties of China and North American. Based on the released time, the investigated varieties were divided into seven decade-long periods since 1950. The average COP value of the combinations that was made by the investigated varieties bred in the 1970s and 1980s were higher, reaching 0.115 and 0.107, respectively, while this index was lower at the 1990s and the recent decade, reaching 0.040 and 0.032, respectively. The numbers of investigated varieties from the Tibet Autonomous Region, Qinghai Province, Sichuan Province, and Gansu Province were 48, 27, 17, and 12, respectively. The average COP of the combinations made by the varieties from Gansu Province was highest, reaching 0.220, while the average COP of the combinations made by the bred varieties from Tibet Autonomous Region, Qinghai Province, and Sichuan Province were similar, reaching 0.094, 0.122, and 0.138, respectively. Cluster analysis placed the investigated varieties into seven groups, of which 25 were in groups II and III, with a relatively low average COP of combinations within each group, while 83 were in the other five groups, which had higher average COP of combinations. The cumulative direct COP of 40 investigated varieties were more than 0; of these, 9 bred varieties were identified as important core parents in Qinghai-Tibet plateau, from which 67 varieties had been derived. 9 core parents and their 67 derivatives account for 72.38% of the number of investigated bred varieties. In the 1960s, the number of core parents and their derivatives accounted for 41.76% of the number of investigated varieties. For the next four decades, the proportions of core parents and their derivatives out of all investigated bred varieties in each period reached 80.00%, 80.77%, 76.47%, and 72.22%, respectively. In the last decade, the index has declined further, but still reaches 68.75%. In the Tibet Autonomous Region, Qinghai Province, Sichuan Province, and Gansu Province, the proportion of core parents and their derivatives out of the number of the investigated bred varieties reached 79.17%, 62.96%, 70.59%, and 75.00%, respectively. The total number of parents used in breeding was 118, including 97 local parents (82.20%), 15 introduced parents (12.71%), and 6 parents lacking source information. 【Conclusion】9 core parents and their 67 derivatives constitute the majority of naked barley varieties. The utilization of introduced barley germplasm has been inefficient. Consequently, the genetic diversity of naked barley varieties cultivated across the Qinghai-Tibet Plateau is relatively lower.

    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    Effects of Spectral Distribution on Photosynthetic and Chlorophyll Fluorescence Characteristics of Flag Leaves at Grain Filling Stage in Rice
    LI JiangPeng,LIU HaiJun,HUANG ZhiWu,LIU XiaoYing,YOU Jie,XU ZhiGang
    Scientia Agricultura Sinica. 2019, 52(16):  2768-2775.  doi:10.3864/j.issn.0578-1752.2019.16.003
    Abstract ( 406 )   HTML ( 39 )   PDF (456KB) ( 366 )   Save
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    【Objectives】 The study was aimed to explore the effects of different spectra on the photosynthetic and chlorophyll fluorescence characteristics of flag leaves in rice at grain filling stage. 【Method】 The rice (Oryza sativa L. cv. Huageng 5) was planted under four different light conditions, including red light of 660 nm (R660) and 630 nm (R630), blue light of 460 nm (B460) and 440 nm (B440) combined with a special plant lamp (W), and the photosynthetic and chlorophyll fluorescence characteristics of rice leaves were determined at grain filling stage. 【Result】 Compared with red light (R660+W and R630+W), blue light (B460+W and B440+W) could increase the maximum photochemistry efficiency (Fv/Fm), actual photochemical efficiency (ΦPSⅡ), electron transfer efficiency (ETR), non-photochemistry quenching (NPQ) and photochemical quenching (qP). The net photosynthetic rate in the late stage of grain filling under B460+W was higher than that under red light. Compared to those of other treatments, the panicles per unit area under R660+W were the greatest, but the 1000-grain weight and seed setting rate were both higher under B460+W. In addition, B460+W also increased the maximum net photosynthetic rate (Pnmax), initial slope of light response curve (α), quantum efficiency of dark respiration, compensation point quantum efficiency, eigen-absorption cross-section of photosynthetic pigment molecule (σik) and number of light-harvesting pigment molecule (No). 【Conclusion】 The stable in photosynthetic rate of flag leaves under the addition of 460 nm blue light conditions was mainly attributed to increase ФPSII, contributing to accumulate dry matter, finally improved the 1000-grain weight and seed setting rate of rice.

    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    Multi-View Geometric Reconstruction of Plant Based on Improved Region-Growing Algorithm
    XIAO ShunFu, LIU ShengPing, LI ShiJuan, DU MingZhu, Lü ChunYang, LIU DaZhong, YANG FeiFei, LIU Hang
    Scientia Agricultura Sinica. 2019, 52(16):  2776-2786.  doi:10.3864/j.issn.0578-1752.2019.16.004
    Abstract ( 323 )   HTML ( 12 )   PDF (4312KB) ( 246 )   Save
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    【Objective】 Three-dimensional reconstruction of three kinds of plants canopy with different levels of complexity was studied to provide a new method for more accurate acquisition of plants canopy phenotypic characteristics. 【Method】 In this paper, a sequence of pictures of three kinds of plant canopy with different levels of complexity were gotten by using a SLR camera, and dense point clouds of plants were gotten by using three-dimensional reconstruction method. Then, the original scale of dense point clouds of plants was restored, and noisy point of dense point clouds and segmenting plants canopy were filtered by using improved region-growing algorithm with the help of laser scanner, then the accuracy of leaves reconstructed from multi-view geometry method was evaluated from two-dimensional and three-dimensional aspects by manual measurement and laser scanning. Two-dimensional accuracy evaluation was carried out by statistical analysis of actual measured values of leaves length and width and comparing with leaves length and width reconstructed by multi-view geometry method and leaves length and width obtained by laser scanner, respectively. Three-dimensional accuracy was evaluated by using the traditional mesh comparison method Hausdorff distance and Geomagic Qualify software, which was a better accurate 3D accuracy comparison software in industrial meshes comparing. 【Result】 The results showed that the judgment coefficients (R 2) between the phenotypic information of plants leaves and manual measurements were higher than 0.96. The judgment coefficients (R 2) between the phenotypic information of plant leaves obtained by laser scanning and manual measurements were higher than 0.99. The proportion of leaves obtained by multi-view geometric reconstruction and laser scanning was more than 97% in the deviation range of 0-±1 mm. Taking the leaves mesh scanned by laser as a reference, more than 90% of the Hausdorff distance of the leaves mesh reconstructed by multi-view geometry was between 0-2 mm. It was proved that the combination of the multi-view geometric reconstruction method with the improved region-growing algorithm could achieve ideal reconstruction results for plants with different complexity. 【Conclusion】The reconstruction method combining multi-view geometry method with the improved region-growing algorithm proposed in this paper could make up for the shortcomings of region-growing algorithm. It had better segmentation effect on the surface of uneven plants canopy, and was suitable for three-dimensional reconstruction of plants with different complexity. It could provide some reference for breeding research to obtain plant phenotypes.

    PLANT PROTECTION
    Identification of the Pathogen Causing Cabbage Died in Gansu Province and Detection of Anastomosis Groups
    WANG Duo,XIE XueWen,CHAI ALi,SHI YanXia,LI BaoJu
    Scientia Agricultura Sinica. 2019, 52(16):  2787-2799.  doi:10.3864/j.issn.0578-1752.2019.16.005
    Abstract ( 325 )   HTML ( 8 )   PDF (1769KB) ( 224 )   Save
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    【Objective】 The objective of this study is to identify the pathogen causing cabbage died in Gansu Province, develop a molecular method for identification, and to provide a reference for prevention and control of the disease.【Method】Diseased samples collected from Gansu Province were purified and cultured, and isolates were characterized by colony morphology, it was initially identified the pathogen as Rhizoctonia solani. For accurate identification, the internal transcribed spacer region of the ribosomal DNA (rDNA-ITS) and translation elongation factor 1-alpha (TEF-1α) of isolates were amplified and sequenced. MEGA 7.0 was used to draw the phylogenetic tree of strains and other related sequences. Molecular identification of the pathogen was carried out by PCR amplification using a specific primer pair F-RS/R-RS. Pathogenicity on 20 cabbage cultivars was verified by spray inoculation and irrigating roots treatment. Based on the conserved region of rDNA-ITS, a pair of specific primer was designed and the SYBR Green I real-time PCR reaction system was established. The specificity, sensitivity and repeatability of conventional PCR and real-time PCR were also evaluated, and R. solani AG3-AG11, binulceate Rhizoctonia AG-A, Fusarium spp., Pythium spp. were used as control fungi. The established system was used to detect R. solani in the cabbage and rhizosphere soils. 【Result】 A total of 86 strains of R. solani were isolated. The results of conventional PCR specific test showed that most strains belonged to AG-2-1 (68/86) and others were AG-1-IB (18/86). The phylogenetic analyses showed that 50 strains were divided into AG-2-1 and AG-1-IB, and the homology of each strain in two groups was 100%. Pathogenicity test results showed that the two anastomosis groups could both cause lesions on stems, and AG-2-1 was not pathogenic to the leaves of Jinwawa and Xiaohuangbao. The pathogenicity of AG-1-IB was slightly stronger than that of AG-2-1. The primers were of great specificity, the specific PCR fragment was amplified from the DNA of R. solani strains, but not from the DNA of other fungal strains by conventional PCR. The real-time PCR assays also did not amplify DNA from control fungi. The sensitivity of conventional PCR was 8.41×10 5copies/μL plasmid, while the sensitivity of real-time PCR was 8.41×10 3copies/μL, which increased two orders of magnitude. The standard curve established by recombinant plasmid showed a fine linear relationship between threshold cycle and template concentration. The melt curve was specific with the correlation coefficient of 0.9983 and with high amplification efficiency (91%). The method could successfully detect the pathogens in cabbage and rhizosphere soils collected in fields. For the indoor potted experiments, the detection results of real-time PCR of infected cabbage samples inoculated after different days showed that the stems and rhizosphere soils had the largest amount of R. solani on the 5th day after inoculation. 【Conclusion】 Through molecular biological identification and pathogenicity test of pathogen of cabbage died, it was confirmed that the causal agent of cabbage died in Gansu were R. solani AG-2-1 and AG-1-IB. The developed real-time PCR system for R. solani has strong specificity and high sensitivity, it can realize the detection and monitoring of R. solani in plant and soil, and can provide technical support for early warning and management of diseases.

    Status of Major Diseases and Insect Pests of Potato and Pesticide Usage in China
    XU Jin, ZHU JieHua, YANG YanLi, TANG Hao, Lü HePing, FAN MingShou, SHI Ying, DONG DaoFeng, WANG GuiJiang, WANG WanXing, XIONG XingYao, GAO YuLin
    Scientia Agricultura Sinica. 2019, 52(16):  2800-2808.  doi:10.3864/j.issn.0578-1752.2019.16.006
    Abstract ( 731 )   HTML ( 63 )   PDF (545KB) ( 718 )   Save
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    As the fourth important staple crop just after rice, maize and wheat, potato plays a crucial role in food security, poverty alleviation, cropping structure adjustment and transformation and updating of agriculture in China. The total harvested area and production of potato in China rank first in the world. However, the yield is much lower than the average level of the world due to a variety of diseases and insect pests of potato. With the support of National Key Research and Development Program of China, the major diseases and insect pests as well as pesticide use in potato production were surveyed in the six main production areas. Twenty-seven major potato pests were found in China. Six diseases (i.e. late blight, early blight, black scurf, Fusarium wilt, potato black leg and potato common scab) and six insect pests (i.e. wireworms, chafer grubs, aphids, twenty-eight spot lady beetle, potato tuber moth and thrips) were proposed to be listed as the most important management target in all the production areas. Moreover, more attention should be paid to prevent and control of bacterial wilt, ring rot and powdery scab. On the basis of census data, the average number of pesticide application and usage per unit area of potato in China were 17 and 40.03 kg·hm -2, which were higher than the national average level of 4.16 and 3.49 kg·hm -2. The control target of current registered pesticides did not cover many increasingly severe potato pests, such as bacterial wilt, common scab, powdery scab, and so on. Adoption and spread of integrated pest management (IPM) were proposed to be the most important strategy for reducing pesticide use and costs in potato production. To make scientific pest-management and pesticide reduction strategy, a potato pest early warning system should be developed based on rapid, sensitive and reliable detection and monitoring method. As host plant resistance is the economically feasible options for pests managing, it is important to improve potato resistant variety breeding and application, optimize variety layout. Development and promotion of high-efficient pesticide application technology and environmentally friendly alternatives to chemicals are the core drivers to realize chemical pesticide reduction.

    One-Step Duplex Rapid Identification Technique of Frankliniella occidentalis Greenhouse and Lupin Races Based on Species-Specific COI Marker
    ZHANG Rong,WANG YuSheng,YANG LiMei,WAN FangHao,ZHANG GuiFen
    Scientia Agricultura Sinica. 2019, 52(16):  2809-2823.  doi:10.3864/j.issn.0578-1752.2019.16.007
    Abstract ( 300 )   HTML ( 5 )   PDF (993KB) ( 605 )   Save
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    【Background】Frankliniella occidentalis is one of the most important pests infesting agricultural and horticultural crops worldwide, also it is a national- and international-wide important quarantine pest. F. occidentalis has two races, i.e., greenhouse race (GR) and lupin race (LR). There are significant differences between the two races in host plant species, pesticide resistance, living environments, etc. However, it is difficult to identify these two races because of their high similarity in morphological characteristics.【Objective】In this study, a one-step duplex PCR procedure for rapid identification of F. occidentalis greenhouse race and lupin race was developed by using species-specific COI (SS-COI) marker based on mitochondrial DNA cytochrome c oxidase subunit I (mtDNA COI) gene, and other 14 common thrips species in the field were used as the control.【Method】The COI sequences of F. occidentalis greenhouse and lupin races, and other 14 thrips species were amplified using mtDNA COI gene universal primers. The race-specific primer combination system, TF6/GR32/LR12 (TF6, the universal forward primer; GR32, the greenhouse race-specific reverse primer; LR12, the lupin race-specific reverse primer), for one-step duplex amplification of the two races was established. The fragments amplified by the combined primers were 362 bp for the greenhouse race and 541 bp for the lupin race, respectively. The system and conditions (e.g., annealing temperature) were optimized. The species/race specificity and sensitivity/ detection threshold of the combined primers were evaluated.【Result】When the ratio of the combined primers (TF6/GR32/LR12) was 1.0/0.2/0.8 and the annealing temperature was 44℃, the amplification effect was the best. Species/race-specific tests performed with this set of combined primers indicated that all F. occidentalis greenhouse race and lupin race specimens were detected positively and no cross reaction with other 14 thrips species, including Frankliniella intonsa, Frankliniella tenuicornis, Thrips hawaiiensis, Thrips major, Thrips palmi, Thrips tabaci, Megalurothrips usitatus, Mycterothrips glycines, Sussericothrips melilotus, Fulmekiola serrate, Haplothrips aculeatus, Mesothrips jordani, Gynaikothrips ficorum, and Aeolothrips fasciatus, was identified. The system was tested on individual male and female adults, individual prepupae and pupae, individual 1st and 2nd instar larvae, and even single egg, and demonstrated to be applicable for all these stages of greenhouse and lupin races of F. occidentalis, either in single race or two races mixed together. The detection threshold for greenhouse race was 35.90 pg·μL -1, which was equivalent to 1/10 240 of a whole female adult. The detection threshold for lupin race was 146.95 pg·μL -1, which was equivalent to 1/2 560 of a whole female adult. Meanwhile, the system worked well in testing individuals from different geographic populations with different haplotypes within greenhouse race or lupin race of the thrips. 【Conclusion】The one-step duplex PCR system developed here for rapid identification of F. occidentalis greenhouse and lupin races could be useful in port quarantine and interception in national and international transportation of agricultural products, and should be significant in blocking further spreading and developing targeted prevention and control measures of F. occidentalis.

    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Nutrients Use Efficiency Change of Chemical Fertilizers for Spring Maize in a Typical Black Soil
    QIU ShaoJun, LI Ning, HE Ping, WEI Dan, JIN Liang, ZHAO ShiCheng, XU XinPeng, ZHOU Wei
    Scientia Agricultura Sinica. 2019, 52(16):  2824-2834.  doi:10.3864/j.issn.0578-1752.2019.16.008
    Abstract ( 382 )   HTML ( 15 )   PDF (489KB) ( 405 )   Save
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    【Objective】Reducing chemical fertilizer input and improving nutrients use efficiency are important agricultural strategies in China. In view of the declining soil fertility and acidification of black soil as the result of chemical fertilizers unbalanced application, the aim of this study was to explore reducing chemical fertilizers input and improving nutrients use efficiency in black soil, so as to promote the balanced utilization of chemical fertilizers in black soil in China.【Method】These treatments included no-fertilizer (CK), no N application (PK), no P application (NK), no K application (NP) and the combination of nitrogen, phosphorus and potassium treatment (NPK). Crop yield, nutrients uptake, soil inorganic nitrogen, Olsen-P and NH4OAc-K were determined in typical black soils from 2013 to 2016. 【Result】 The yield of spring maize in Heilongjiang was about 10 t·hm -2 per year. The yields and nutrients uptake by aboveground per year under NPK treatment were significantly higher than those under CK or PK treatments except for the year of 2013. The recovery efficiency, agronomic efficiency and partial productivity of N, P and K in NPK treatment increased year by year except for the P agronomic efficiency in 2016. During the period of 2013-2016 under NPK treatment, the averaged recovery efficiency of N, P and K was 45.8%, 6.1% and 3.5%, respectively; the averaged agronomic efficiency of N, P and K were 23.2 kg·kg -1, 7.2 kg·kg -1 and 5.0 kg·kg -1, respectively; and the averaged partial productivity of N, P and K are 58.3 kg·kg -1, 133.2 kg·kg -1 and 97.7 kg·kg -1, respectively. Soil mineral nitrogen test showed the alternative frozen and thawed promoted soil organic nitrogen mineralization from post-harvested to pre-sown in the following year. The averaged nutrients balances in the experimental four years showed that the rate of N or P in NPK treatment could meet the demand of P, K for spring maize, soil N or P was in the balanced status on the whole, and the luxury uptake of K by aboveground resulted in the loss status of soil potassium.【Conclusion】The continuous four years experiment in the typical black soil region showed that P and K rate greatly reduced and the use efficient of P and K increased with stable spring maize yield kept, while no chemical N fertilizer application only maintained maize yield in the first year, the decrease of maize yield and the increase of N use efficient occurred in the following years.

    Determination and Application of a Critical Nitrogen Dilution Curve for Direct-Sowing Winter Oilseed Rape in Central China
    LIU QiuXia, REN Tao, ZHANG YaWei, LIAO ShiPeng, LI XiaoKun, CONG RiHuan, LU JianWei
    Scientia Agricultura Sinica. 2019, 52(16):  2835-2844.  doi:10.3864/j.issn.0578-1752.2019.16.009
    Abstract ( 341 )   HTML ( 13 )   PDF (1053KB) ( 263 )   Save
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    【Objective】This study was conducted to determine the critical nitrogen (N) dilution curve for direct-sowing winter oilseed rape in Central China and its applicability, and the feasibility of N nutrition index (NNI) in evaluating N nutrition status of oilseed rape plant. 【Method】 Field experiments during 2015-2016 and 2016-2017 were conducted with different N application rates, including 0, 60, 120, 180 and 240 kg N·hm -2 during 2015-2016, and 0, 60, 120, 180, 240, 300 and 360 kg N·hm -2 during 2016-2017. The shoot dry matter and N concentration from seedling to flowering were studied. The model of the critical N concentration dilution curve of the shoot dry matter of direct-sowing winter oilseed rape was established. The NNI was calculated by using the model and plant shoot N concentration. The effects of N application rates on NNI at various stages were determined. The relationship between oilseed rape yield and NNI was investigated to determine the appropriate NNI in each period.【Result】Results showed that the N application significantly increased shoot dry matter and N concentration, and the difference between different N treatments was remarkable. The relationship between the shoot critical N concentration and shoot dry matter could be described by a power equation (Ncnc=3.49DM -0.26). This model could distinguish between N-limited and non-N limited group data from two independent experiments. The simulated N concentration was linearly correlated with the shoot actual N concentration, with the RMSE and n-RMSE of 0.37% and 13%, respectively, showing better stability of the model. The NNI increased as the N application rate increased within the range of the application rates in the study, and the response of NNI to N application rate was similar to the yield. The N application increased the seed yield significantly. Although there were differences in the response of N fertilizers to seed yield at varying experimental sites, the relative yield was closely related with the NNI of each experimental site. The NNI with the relative yield of 1 at the over-wintering, stem elongation and flowering stage were 1.35, 1.26 and 1.03, respectively. 【Conclusion】 The critical N concentration dilution curve Ncnc=3.49DM -0.26 was established, and the NNI could be used to evaluate plant N status and diagnose the N status of direct-sowing winter oilseed rape in Central China.

    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Effects of Natural Vegetation Restoration on Characteristics of Soil Aggregate and Soil Erodibility of Gully Heads in Gully Region of the Loess Plateau
    WANG WenXin,WANG WenLong,GUO MingMing,WANG TianChao,KANG HongLiang,YANG Bo,ZHAO Man,CHEN ZhuoXin
    Scientia Agricultura Sinica. 2019, 52(16):  2845-2857.  doi:10.3864/j.issn.0578-1752.2019.16.010
    Abstract ( 322 )   HTML ( 13 )   PDF (852KB) ( 342 )   Save
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    【Objective】 Gully head was the most active geomorphic part in gully region of the Loess Plateau, which was related to the soil erosion of the entire slope gully system. This research aimed at evaluating the effects of vegetation restoration on soil aggregate characteristics and soil erodibility of gully heads in gully region of the Loess Plateau. The research results could provide a scientific basis for regional ecological environment restoration and soil and water conservation benefit evaluation. 【Method】 With the farmland as control check, an investigation about soil aggregate characteristics and soil erodibility of different soil layers (0-10, 10-25, 25-40 cm) with different natural vegetation restoration ages (0-30 a) of gully heads was carried out. The aggregates amount, water-stable aggregate content (WR0.25), mean weight diameter (MWD), geometric mean diameter (GMD), destruction rate (PAD) and fractal dimension (D) were examined by dry and wet sieving methods. Soil mechanical composition and organic matter content were determined which were used for calculating soil erodibility factor K. 【Result】 (1) WR0.25, MWD and GMD of vegetation gully heads increased by 11.49%- 84.43%, 0.18-2.05 times, and 7.53%-108.62%, respectively, compared with the CK (farmland of gully heads). These three indicators linear increased with vegetation restoration years increasing (P<0.01), and decreased with the increase of soil depth; (2) PAD and D of soil aggregate of vegetation gully heads decreased with linear increase of vegetation restoration time (P<0.01), which was 3.81%-32.14% and 0.55%-6.63% lower than that of CK, respectively. Both of them increased with the increase of soil layer; (3) K decreased linearly with the increase of vegetation restoration period (P<0.01), which was 5.43%-14.44% lower than that of CK, and K increased with the increase of soil depth.【Conclusion】The increase of organic matter content under vegetation restoration conditions played an important role in the formation and stability of aggregate. The decrease of soil erodibility of gully heads was closely related to the increase of water-stable aggregate content and stability of aggregate. Under natural recovery conditions, soil aggregate stability and soil anti-erodibility of gully heads were significantly improved during the restoration period of 22-30 a.

    HORTICULTURE
    Identification of the Target Genes of VvmiR159s and Their Regulation in Response to GA in Different Tissues of Grape Berry
    ZHANG WenYing, HAN Xu, ZHU XuDong, XIE ZhenQiang, JIU SongTao, HUANG YuQing, JIA HaiFeng, FANG JingGui, WANG Chen
    Scientia Agricultura Sinica. 2019, 52(16):  2858-2870.  doi:10.3864/j.issn.0578-1752.2019.16.011
    Abstract ( 277 )   HTML ( 10 )   PDF (1824KB) ( 212 )   Save
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    【Objective】The main objective of the present research was to identify the miR159 family members and their target genes from grapevine genome, to validate the role of VvmiR159 family members and their target genes in response to exogenous GA in the development of different tissues of seedless grape berries. 【Method】The mature and precursor sequences of VvmiR159a/b/c were cloned and identified by miR-RACE and PCR techniques from grapevine cv. ‘Rosario Bianco’. The target genes of VvmiR159s were predicted by PsRNATarget software, and the phylogenetic and conserved domain analysis was performed by bioinformatics tools. The potential functions of VvmiR159s and their target genes were predicted by cis-elements analysis of their promoters. RLM-RACE and PPM-RACE verified the cleavage roles of four target genes by VvmiR159a/b/c, and the qRT-PCR method was used to detect their temporal and spatial expression patterns in different tissues of grape berry induced by exogenous GA application 【Result】The mature and precursor sequences of VvmiR159a/b/c were cloned from the ‘Rosario Bianco’ and further folded into the typical hairpin structures. Four target genes (VvGAMYB, VvMYB48, VvAIX15, and VvGRAS-1) of VvmiR159s were identified, of which VvmiR159s strongly matched with VvGAMYB the most and VvAIX15 was weak. Phylogenetic evolution and conserved domain analysis of target genes showed that the four target genes had high homology with other species. All VvmiR159s and four target genes had the main hormone related cis-elements responsive to GA and SA, indicating that they might be involved in the regulation of grape berry growth and development mainly by responding to corresponding hormones. The expression of VvmiR159s in different tissues was spatial-temporal-specific. The expression of VvmiR159c was deviated in berry pericarp and flesh, and the expression level of VvmiR159a/b/c was negatively correlated with VvAIX15 in the pericarp, however in flesh, VvmiR159a/b was contradictory to VvGAMYB and VvAIX15 expression pattern. In addition, GA treatment VvmiR159a/c was significantly down-regulated in grape pericarp and flesh, but VvmiR159b up-regulated in young fruit flesh. These results indicated that different members of the VvmiR159 family participated in the regulation of fruit development through different modes of response to GA signaling in different tissues of the fruit. 【Conclusion】The grape miR159 family contains three members of VvmiR159a/b/c; all of three members can cleave four target genes: VvGAMYB, VvMYB48, VvAIX15 and VvGRAS-l; VvmiR159a/b/c and their four target genes may participate in the regulation of grape pericarp and flesh development in different GA-responsive modes.

    Effects of ‘Tent’ Mulching on Soil Temperature and Grape Growth in The Yellow River Delta Saline-Alkali in Spring
    WANG Hui,ZHAO Shuo,YANG XingWang,JIN MengLing,DU YuanPeng,GUAN XueQiang,ZHAI Heng
    Scientia Agricultura Sinica. 2019, 52(16):  2871-2879.  doi:10.3864/j.issn.0578-1752.2019.16.012
    Abstract ( 335 )   HTML ( 12 )   PDF (600KB) ( 193 )   Save
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    【Objective】The objective of this paper was to explore the reasons of the delayed bud burst and to improve the growth of grapevine in saline land in spring.【Method】Three year old ‘Summer Black’ grape was used as test material, which was cultivated in Guangbei #2 field with saline land (1.5 m within rows and 3 m between rows, vertical shoot-positioning system, hedgerows, single stem and arm tree shape). Grapes were unearthed in March 2018, ‘tent’ was built after irrigating for the accelerating germination, namely, pulling a wire at a height of 50 cm, with it as the vertex, white plastic film with a width of 80 cm was superimposed on both sides with binding wire. The two sides were opened to the ground with an angle about 45 o, then soil was used to cover the edge, a small hermetically sealed triangle known as the ‘tent’ mulching. Two rows were laid for each treatment, plants without covering the ‘tent’ were used as control, and effects of ‘tent’ mulching on soil temperature and grape growth was studied.【Result】‘Tent’ mulching in the Yellow River delta saline land promoted the growth and development of grape and effectively increased the ground temperature in saline land. The average ground temperature of the 10 cm soil layer in the rhizosphere under ‘tent’ mulching was significantly increased by about 5℃, compared with the control. The ground temperature under ‘tent’ mulching increased earlier and retained longer time than that under the control treatment, which decreased the gap between ground temperature and air temperature. The phenological period of grapes bud burst under ‘tent’ mulching were 10-15 days earlier than that under the control, and the bud burst time was earlier and consistent. ‘Tent’ mulching significantly improved the growth quality of grape shoots, growth of new shoots (length), internode length and width of the third node, which were increased by 34.9%, 23.8% and 20%, respectively, compared with the control. Leaves area and weight under ‘Tent’ mulching were increased by 39.9% and 56.6%, respectively, while leaves thickness under ‘Tent’ mulching was increased but there was no significant difference, compared with the control. ‘Tent’ mulching significantly improved the leaves function of grape, leaves chlorophyll content, net photosynthetic rate (Pn) and maximum photochemical efficiency (Fv/Fm), which were increased by 27.6%, 30% and 6.8%, significantly, compared with the control. The photochemical quenching coefficient (qP) was also significantly increased by 21.9%, which indicated that the open degree of PSII reaction center in geothermal environment was large, the energy used in the photochemical pathway was increased, and photosynthesis was increased. In order to explore the causes of delayed temperature rise in saline land, the pot experiments were conducted under the same climatic conditions in Tai'an. The results showed as follows: the response of saline soil ground temperature to air temperature was significantly delayed about 3 h than that of brown soil. The largest differences about the two soil were salt content, soil conductivity, bulk density and porosity. Field measurements showed that saline soil was less aerated than brown soil. The bulk density of the 0-20 cm soil layer in saline soil was as high as 1.45 g?cm -3, which belong to a compaction condition. The bulk density was 11.7% higher than that of brown soil, soil porosity was 13.5% lower than that of brown soil, soil reoxidation reduction potential (Eh) and oxygen diffusion rate (ODR) were 49.9% and 13.8% lower than that of brown soil, respectively.【Conclusion】The ‘tent’ significantly increased the ground temperature of saline-alkali land, reduced the time-space difference of air and ground temperature, effectively improved the bud burst process of grape, advance the phenological period of bud burst, improved the growth quality of new shoots, and promoted the growth and development of grapevine in spring.

    ANIMAL SCIENCE·VETERINARY SCIENCE·RESOURCE INSECT
    The Variation/Heteroplasmy of Chicken Mitochondrial ND1 Gene and Its Association with Traits
    XU YuanYuan,HOU LingLing,JI JieFei,WANG HuanJie,LI SiLu,CHEN Wen,KANG XiangTao,HUANG YanQun
    Scientia Agricultura Sinica. 2019, 52(16):  2880-2890.  doi:10.3864/j.issn.0578-1752.2019.16.013
    Abstract ( 350 )   HTML ( 27 )   PDF (2113KB) ( 516 )   Save
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    【Objective】 The purpose of this study was to investigate the heteroplasmy of mitochondrial ND1 gene in chickens and to study the distribution of mitochondrial heteroplasmy among different breeds and Gushi chicken resource population, and furthermore, to analysis the effects of mitochondrial heteroplasmy on valuable economic traits. 【Method】 DNA extracted from blood samples of eight chicken breeds including Gushi chicken, Arbor Acres broiler chicken, Henan Cockfighting chicken, Lushi chicken, White Leghorn chicken, White Plymouth Rock chicken, Silkie chicken and Tibetan chicken, were used to study the distribution of mt.A4589G mutation/ heteroplasmy in natural populations by PCR-RFLP. In addition, DNA extracted from blood samples of Gushi chicken resource population constructed previously were used to study the distribution of mt.A4589G mutation/ heteroplasmy in Gushi chicken resource population by PCR-RFLP and to study the association analysis between mt.A4589G mutation/ heteroplasmy and F2 traits of Gushi chicken resource population, including carcass traits, growth traits, meat quality indicators and blood biochemical indicators, thereby grasping the heteroplasmy of chicken mitochondrial ND1 gene and its potential effects; 【Result】1) Among eight breeds, one synonymous mutation mt.A4589G was detected from the full length of mitochondrial ND1 gene, and its distribution showed obvious variety characteristics. The dominant alleles of exotic breeds such as broiler chicken, White Plymouth Rock chicken and White Leghorn chicken were A, while domestic local breeds such as Silky chicken, Henan Cockfighting chicken and Lushi chicken were G. (2) The heteroplasmy of mt.A4589G in different breeds and Gushi chicken resource population was detected by PCR-RFLP. Among eight breeds, only five AG heterogeneous individuals were detected in Gushi chicken, Henan Cockfighting chicken and broiler chicken. In the Gushi chicken resource population, mt.A4589G had a wider heteroplasmy and the frequencies of heteroplasmy in F0, F1 and F2 generations were 0.81, 0.41 and 0.70, respectively. (3) The correlation analysis between mt.A4589G heteroplasmy and F2 traits of Gushi chicken resource population showed that mt.A4589G heteroplasmy was significantly correlated with abdominal fat, sebum thickness eight-week-old shank length, diameter of leg muscle fiber and glucose levels in the blood (P<0.05). (4) The correlation analysis between the levels of heteroplasmy of mt.A4589G and F2 traits of Gushi chicken resource population indicated that the levels of heteroplasmy was significantly correlated with carcass traits, growth traits and serum biochemical indicators (P<0.05). 【Conclusion】 mt.A4589G is a heteroplasmic variation, and its heteroplasmic distribution exhibits obvious variety characteristics. The mt.A4589G heteroplasmy of the natural population occurs less frequently, while mt.A4589G has a wider heteroplasmy in the Gushi chicken resource population. Furthermore, through the correlation analysis between the heteroplasmy of mt.A4589G and the F2 traits of Gushi chicken resource population, it was found that the mt.A4589G mutation/heterogeneity was significantly correlated with the abdominal fat and the glucose level in the blood (P<0.05).

    Effects of Selenium on the Key Factors in Nod2/MAPK/mTORs Signaling Pathways in the bMECs Infected S. aureus
    BI ChongLiang,LIU JunJun,WANG Heng,WANG Juan,HAN ZhaoQing,GUAN LiZeng
    Scientia Agricultura Sinica. 2019, 52(16):  2891-2898.  doi:10.3864/j.issn.0578-1752.2019.16.014
    Abstract ( 345 )   HTML ( 14 )   PDF (930KB) ( 337 )   Save
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    【Objective】Whether selenium (Se) could regulate the inflammatory damage of bovine mammary epithelial cells (bMECs) induced by S. aureus through Nod2/MAPK/mTOR pathway remains to be further studied. So in the study, the effects of Se on the key proteins in the Nod2/MAPK/mTORs signaling pathway in the bovine mammary epithelial cells (bMECs) infected by S. aureus was studied in order to provide a theoretical basis for elucidating the immune regulation mechanism of Se.【Method】 Firstly, the bMECs were inoculated into the 6 well plates with 10 6cells/well. When more than 80% of the cells were confluent, the medium was replaced with the one containing different concentrations of Se (2, 4 and 8 μmol·L -1) and continued to culture for 12 h. Then after washing each well for 3 times with PBS, S.aureus was added into 6-well plates at a ratio of MOI=1:1 and continued to culture for 0.5 h. The bMECs were collected for further detection of related proteins expression. The experiment was divided into three groups: control (Con) group (bMECs), model (Mod) group (bMECs+S. aureus) and experimental group. The experimental group was divided into three sub-dose groups, namely Low group (bMECs+2 μmol·L -1 Se+S. aureus), Mid group (bMECs+4 μmol·L -1 Se+S.aureus) and Hig group (bMECs+8 μmol·L -1 Se+S. aureus), with three replicates each group. Total protein was extracted from the above bMECs using a bicinchoninic acid (BCA) protein assay kit. The expressions level of Nod2 and RIP2 and the phosphorylation level of JNK, AKT and mTOR proteins in bMECs were detected by Western blotting. The protein samples were loaded into 10% SDS polyacrylamide gel for electrophoresis, and the uniform volume of protein was 20 μg/hole. Then the protein was transferred to polyvinylidene fluoride (PVDF) membranes. The PVDF membranes were blocked with 5 mL 5% nonfat milk for 2 h, then skimmed the nonfat milk and washed the membranes with TBST, subsequently the membranes were incubated overnight with 5 mL primary antibodies including Nod2, RIP2, JNK, AKT, mTOR and β-actin. The primary antibodies were recovered, and 5 mL second antibodies were added to the membranes and incubated for 2 h at room temperature. Subsequently the second antibodies were recovered the membranes were washed with TBST for 5 times. Finally the membranes were developed with chemiluminescent substrate under darkroom conditions.【Result】S. aureus could significantly increase the expression of Nod2 and RIP2 proteins and the phosphorylation of JNK, AKT and mTOR proteins in bMECs (P<0.01). At 0.5 h after S. aureus infection, the level of Nod2 protein increased significantly (P<0.01). The expression of Nod2 protein was significantly inhibited by adding 2 μmol·L -1 Se to the medium (P<0.01), and the expression of Nod2 was significantly inhibited by adding 8 μmol·L -1 Se to the medium (P<0.05); at 0.5 h after S. aureus infection, RIP2 protein level was significantly increased (P<0.05), while the expression of RIP2 protein was significantly inhibited by adding 8 μmol·L -1 Se to the medium (P<0.05); at 0.5 h after S. aureus infection, the phosphorylation level of JNK protein in model group was significantly higher than that in control group (P<0.01). The phosphorylation of JNK protein was significantly inhibited by adding 4 μmol·L -1 Se to the medium (P<0.05), and the phosphorylation of JNK protein was significantly inhibited by adding 8 μmol·L -1 Se to the medium (P<0.01); at 0.5 h after S. aureus infection, the phosphorylation level of AKT protein in the model group was significantly higher than that in the control group (P<0.01). The phosphorylation level of JNK protein was significantly inhibited by adding 4 μmol·L -1 Se to the medium (P<0.01). The phosphorylation level of AKT protein was significantly inhibited by adding 8 μmol·L -1 Se to the medium (P<0.05). After 0.5 h of S. aureus infection, the phosphorylation level of mTOR protein was significantly increased in the model group (P<0.01). The phosphorylation of mTOR protein was significantly inhibited by adding 4 and 8 μmol·L -1 Se to the medium (P<0.05). 【Conclusion】Se could alleviate the inflammatory response of bMECs induced by S. aureus by inhibiting the protein expression of key factors in the bMECs Nod2/MAPK/mTORs signaling pathway.

    Antibacterial Activity in Vitro and Protection of Tildipirosin Injection Against Artificially Infected Haemophilus Paracoides in Piglets
    LI GuoJi, YAN ChaoQun, MA YuQiao, XIE Shun, GU Xin, CAO Ying, HUANG ShiXin, HUANG XianHui
    Scientia Agricultura Sinica. 2019, 52(16):  2899-2911.  doi:10.3864/j.issn.0578-1752.2019.16.015
    Abstract ( 454 )   HTML ( 12 )   PDF (2112KB) ( 492 )   Save
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    【Objective】The study was carried out to investigate the antibacterial activity in vitro and protection of tildipirosin injection against artificially infected Haemophilus Paracoides in piglets. 【Method】Minimal inhibitory concentration (MIC) of Tildipirosin injection against Haemophilus paracoides, Actinobacillus pleuropneumonia, Pasteurella multiflora, and Bronchial septicemic wave bacillus were determined by two-fold dilution method in vitro. In the protective effect of tildipirosin injection against Haemophilus paracoides in piglets, 0.5 mL·kg -1bw suspension of Haemophilus paracoides with a bacterial volume of 1 10 10 CFU·mL -1 was selected as the challenge dose for intraperitoneal injection to replicate the pathological model. All the therapeutic groups of pigs were medicated via intramuscular injection in single dose with tildipirosin injection (2, 4, 8 mg·kg -1bw), tulathromycin (2.5 mg·kg -1bw). In the process of isolating and identifying dead pig pathogens, isolated and purified Haemophilus paracoides colonies were selected for genomic DNA extraction. A pair of specific primers were designed for the 16s RNA sequence of Haemophilus paracoides, and the base sequence length of the target DNA fragment was tested after PCR amplification.【Result】The results of the antibacterial activity in vitro showed that MIC of Haemophilus paracoides, Septicopharynx, Actinobacillus pleuropneumoniae and Pasteurella multocida for tildipirosin injection was 0.06-8, 0.06-8, 0.25-1, and 2-32 μg·mL -1, respectively, which indicated the effect of Tildipirosin injection against Haemophilus paracoides and Septicopharynx was stronger than Actinobacillus pleuropneumoniae and Pasteurella multocida. PCR amplification results of the DNA target fragment showed that the length of the PCR amplification target fragment was consistent with the predicted band, and the specific fragment length was about 820bp, which indicated that all the dead pigs died of Haemophilus paracoides infection. The result of protective effect indicated that, in tildipirosin injection groups (2, 4, 8 mg·kg -1bw), the average gain of weight was (2.5±0.2), (2.9±0.2), (2.9±0.3) kg, respectively; the mortality rate were 40%, 0 and 0, respectively; the effective rate was 40%, 100% and 100%, respectively; the cure rate was 40%, 100% and 100%, respectively. In tulathromycin injection groups (2.5 mg·kg -1bw), the average gain of weight was (3.0±0.2) kg; the mortality rate were 10%; the effective rate was 90%; the cure rate was 80%. In infective control group, the average gain of weight was (2.1±0.1) kg; the mortality rate were 70%. The results of in vitro treatment showed that there was no significant difference between the high- and medium-dose group of tildipirosin and the tulathromycin group (P>0.05), which could alleviate the clinical symptoms rapidly and had significant therapeutic effects. The therapeutic effect of tildipirosin low-dose group was comparable to the non-administered group, and there was no significant difference (P>0.05). 【Conclusion】Through the above results, the product was recommended to be administered at a dose of 4 mg·kg -1bw, which could effectively treat respiratory diseases in pigs caused by Haemophilus Parasuis infection.

    Effects of Natural Bee Bread on Blood Lipid, Antioxidation and Immune Function in Rats with Hyperlipidemia
    LI Zhen,LIU ZhiYong,JIANG WuJun,HE XuJiang,YAN WeiYu,ZHANG LiZhen,ZENG ZhiJiang
    Scientia Agricultura Sinica. 2019, 52(16):  2912-2920.  doi:10.3864/j.issn.0578-1752.2019.16.016
    Abstract ( 455 )   HTML ( 24 )   PDF (2804KB) ( 392 )   Save
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    【Objective】Bee bread is that the worker bee stores the collected pollen mass in the nest room, then bites the pollen mass, spits honey moist, and ferments it for 2 to 3 weeks. Bee bread provides essential nutritional supply for the development of old larvae and young bees. Although the traditional view is that the nutritional value of natural bee bread is higher than that of bee pollen, but there is no scientific basis. In order to evaluate the nutritional effect of bee bread scientifically, the effects of natural bee bread on blood lipid, antioxidation and immune function in rats were systematically studied with bee pollen as control.【Method】The natural bee bread producer designed by the Honeybee Research Institute of Jiangxi Agricultural University was used to produce natural bee bread and pollen by honey bee workers (Apis mellifera) at the flowering stage of white lotus. The hyperlipidemia rats used in this study were established by standard method. A blank control group and a high-fat model control group were prepared. Three doses of bee bread and pollen (low-dose group of 80 mg?kg -1, middle-dose group of 400 mg?kg -1, and high-dose group of 800 mg?kg -1) were fed to hyperlipidemia rats, respectively. After intragastric administration natural bee bread and bee pollen for 60 days, the levels of blood lipids including triglyceride (TG), total cholesterol (TC), low density lipoprotein (LDL) and high density lipoprotein (HDL-C) were measured. Additionally, antioxidant indexes superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), malondialdehyde (MDA), total antioxidant capacity (T-AOC), immune factors interleukin 6 (IL-6) and tumor necrosis factor α (TNF-α) were detected in blood.【Result】Compared with the high-fat model group, low-dose bee bread treatment group significantly reduced the levels of TG, TC and LDL in the blood of hyperlipidemia rats. Middle-dose bee bread treatment group significantly increased the level of HDL-C. Low-dose bee pollen treatment group significantly reduced the levels of TG and TC, which indicated that both bee bread and bee pollen had a good effect on reducing hyperlipidemia. Additionally, low-, middle-, high-dose bee bread, as well as low- and middle-dose bee pollen significantly increased the content of TNF-α, which indicated that both bee bread and bee pollen enhanced immune responses compared with the high-fat model group. The SOD content in the blood of rats with high-dose bee pollen group increased significantly, but the differences among the three doses of bee bread groups, the low- and the middle-dose groups of bee pollen were not statistically significant compared with the high-fat model group. It suggested that the high-dose bee pollen enhanced antioxidant capacity. Compared with the high-fat model group, the contents of GSH-Px, T-AOC and MDA among the six experimental groups were not significantly different. Compared with the low- and middle-dose bee pollen groups, the middle-dose bee bread group significantly increased the content of HDL-C. Compared with the three groups of bee pollen, middle-dose bee bread group significantly increased the TNF-α content.【Conclusion】Combined with TG, TC, LDL, HDL-C and TNF-α, the effects of natural bee bread on reducing blood lipid and enhancing immunity were stronger than those of bee pollen. Natural bee bread is worthy of further development and utilization.