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    16 September 2022, Volume 55 Issue 18
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    QTL Mapping of Thousand-Grain-Weight and Its Related Traits in Zhou 8425B × Xiaoyan 81 Population and Haplotype Analysis
    LinHan ZOU,XinYing ZHOU,ZeYuan ZHANG,Rui YU,Meng YUAN,XiaoPeng SONG,JunTao JIAN,ChuanLiang ZHANG,DeJun HAN,QuanHao SONG
    Scientia Agricultura Sinica. 2022, 55(18):  3473-3483.  doi:10.3864/j.issn.0578-1752.2022.18.001
    Abstract ( 432 )   HTML ( 57 )   PDF (3981KB) ( 222 )   Save
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    【Objective】Zhou 8425B is one of the most important founder parents in China and Xiaoyan 81 is an elite cultivar with high yield and good quality. Thousand-grain weight (TGW) is an important factor that affects wheat yield. Identification of QTL associated with grain related traits from Zhou 8425B and Xiaoyan 81, and haplotype analysis of these QTL in wheat cultivars from different ecological regions would be beneficial for yield improvement by molecular marker-assisted selection.【Method】In this study, a RIL population (F8) derived from Zhou 8425B × Xiaoyan 81 was planted in Yangling during 2015 and 2016 cropping seasons to evaluate grain related traits. Using a high-density genetic map constructed by 90K SNP markers, QTL mapping of thousand-grain weight, grain length, grain width and thickness was performed under three environments. Simultaneously, the KASP markers linked to the identified QTL were developed and molecular detection was carried out among 479 wheat accessions worldwide. Moreover, haplotype analysis of target QTL was performed in 106 current wheat commercial cultivars from Yellow and Huai River Valley Winter Wheat Region selected from 479 wheat accessions.【Result】A total of 22 QTL on 8 chromosomes were detected, and the phenotypic variation explanation (PVE) ranged from 4.77% to 19.95%. Among them, 12 QTL are major QTL (PVE>10%) and Qkgw.nwafu-6B is a new QTL. QTL on chromosomes 4A, 6A, 6B, and 7D were detected in multiple environments, of which, the QTL on chromosomes 4A and 7D are same as previously reported ones. Compared to TaGW2-6A using molecular detection, both Zhou 8425B and Xiaoyan 81 carried the same allele of TaGW2. Based on haplotype result of Qkgw.nwafu-6A, Zhou 8425B and Xiaoyan 81 were placed in different groups. Therefore, Qkgw.nwafu-6A tends to be a new one. Haplotype analysis showed that there were five haplotypes for Qkgw.nwafu-6A and there were eight haplotypes for Qkgw.nwafu-6B. 6A_h1 and 6B_h6 accounted for over 20% in different ecological regions. In addition, a co-segregated KASP marker was developed for Qkgw.nwafu-6B and was significantly associated with the grain weight in the 479 wheat accessions.【Conclusion】Qkgw.nwafu-6A and Qkgw.nwafu-6B are possible new QTL associated with thousand-grain-weight, and 6A_h1 and 6B_h6 are likely favorable haplotypes. A molecular marker KASP_IWA349 co-segregated with Qkgw.nwafu-6B was developed and will be useful for marker assisted selection.

    Deciphering of the Genetic Diversity After Field Late Blight Resistance Evaluation of Potato Breeds
    XiaoChuan LI,ChaoHai WANG,Ping ZHOU,Wei MA,Rui WU,ZhiHao SONG,Yan MEI
    Scientia Agricultura Sinica. 2022, 55(18):  3484-3500.  doi:10.3864/j.issn.0578-1752.2022.18.002
    Abstract ( 323 )   HTML ( 50 )   PDF (3315KB) ( 192 )   Save
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    【Objective】To evaluate the late blight resistance in field. To use SNP markers to analyze the genetic diversity of late blight resistance of potato germplasm and discern the genetic segments that may affect the phenotype of potato late blight resistance, and to provide a theoretical basis for the innovation and utilization of potato late blight resistant germplasm.【Method】Field resistance to late blight was evaluated in potato germplasm at multiple locations and over the course of several years. SNP markers were detected using a dd-RAD simplified genome sequencing strategy. The population genetic structure was analyzed using Admixture, principal component analysis was conducted using GCTA, a phylogenetic tree was constructed using fastTree, population genetic diversity parameters were calculated using the populations command in the Stacks package, selective sweep parameters were calculated using vcftools, protein sequences were aligned using Clustal Omega, and a proteins phylogenetic tree was drawn using MEGA6. Genome-wide association analysis was constructed using GEMMA 0.98.1 and QQ and manhattan plots were drawn using CMplot.【Result】Through years of evaluation of late blight field resistance in multiple locations, potato germplasm of 101 late blight resistant varieties (lines) and 21 susceptible varieties were obtained. A total of 8 697 602 relatively evenly distributed SNPs were obtained using dd-RAD simplified genome sequencing on these germplasms. This germplasm can be further divided into 6 populations through structural analysis, principal component analysis and phylogenetic analysis. The average nucleotide diversity (π) within the 6 populations ranged from 0.2055 to 0.2572 and the fixation index (Fst) among the six populations ranged from 0.156909 to 0.187336, revealing a relatively large genetic diversity for these germplasms. The expected heterozygosity (He) within the 6 populations ranged from 0.187 to 0.2297 and the observed heterozygosity (Ho) ranged from 0.0829 to 0.1186. The values of Ho were less than those of He in all six populations. Meanwhile, the inbreeding coefficient (Fis) for the six populations ranged from 0.2412 to 0.3554, indicating inbreeding events during the breeding process. To identify the genetic segments that may affect the phenotype of potato late blight resistance, π ratios and Fst among different late blight resistance germplasms in the whole potato genome were calculated using 20 kb as the window length and 5 kb as the step length. The 745 genetic segments which had a π ratio value in the lowest 5% and a Fst value in highest 10% were further analyzed by performing a selective sweep analysis. These selected segments contain a total of 507 genes, including 4 NBS-LRR genes. A genome-wide association analysis was also conducted, yielding 9 SNP highly associated with late blight resistance. Of the 69 genes located in the genome within 50 kb around the 9 SNPs, 15 genes were predicted to be involved in stress response, and 12 genes were predicted to be involved in removing peroxide radicals.【Conclusion】Large amount of SNPs, which are relatively evenly distributed in the potato genome, can be genotyped by dd-RAD simplified genome sequencing. Potato late blight field resistance germplasm has a large genetic diversity, but had inbreeding events in the process of breeding. Population structure analysis can reveal the genetic relationship between potato germplasms, which can further provide a theoretical basis for parental selection in breeding. Selective sweep and genome-wide association analyses help to isolate genetic segments that may affect late blight resistance traits.

    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    Effects of Multiple Cropping Green Manure After Wheat Harvest and Nitrogen Application Levels on Wheat Photosynthetic Performance and Yield in Arid Irrigated Areas
    BiJiao MA, ZhiWen GOU, Wen YIN, AiZhong YU, ZhiLong FAN, FaLong HU, Cai ZHAO, Qiang CHAI
    Scientia Agricultura Sinica. 2022, 55(18):  3501-3515.  doi:10.3864/j.issn.0578-1752.2022.18.003
    Abstract ( 290 )   HTML ( 8 )   PDF (1129KB) ( 206 )   Save
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    【Objective】The aim of this study was to explore the effects of returning green manure to soil on wheat yield under different nitrogen application levels in oasis irrigation areas, which had important guiding significance for establishing a green wheat production pattern based on the combined application of green manure and chemical nitrogen fertilizer. 【Method】 From 2018 to 2020, a split plot experiment was conducted in the Hexi oasis irrigation areas of Gansu province. The main plot was set up with two planting patterns of sole wheat (W) and multiple cropping hairy vetch after wheat harvest (W-G), and the sub-plot had 5 nitrogen levels, including no nitrogen application (N0), 55% N (N1), 70% N (N2), 85% N (N3), 100% N (N4), among which 100% N was the conventional nitrogen application rate of 180 kg·hm-2. The leaf area index (LAI), leaf area duration (LAD), net assimilation rate (NAR) and grain yield at maturity stage and its constituent factors were measured during the whole growth period of wheat, in order to provide the basis for optimizing planting pattern and nitrogen application level in this area.【Result】Compared with W, W-G significantly increased the mean LAI and LAD during the whole growth period of wheat by 9.5%-19.7% and 9.7%-21.0%, respectively; The moderate reduction of nitrogen fertilizer was beneficial to increase the mean LAI and LAD of wheat, with N3 being the most prominent. Compared with W-N3 and W-N4, W-G-N3 increased mean LAI by 4.1%-15.4% and 8.8%-17.5%, respectively, and the total LAD increased by 4.6%-9.2% and 16.8%-18.8%, respectively. W-G reduced the mean NAR of the whole growth period of wheat, which was 17.7% and 17.8% lower than that of sole wheat, respectively. Compared with W-N3 and W-N4, W-G-N3 reduced mean NAR by 16.4%-17.5% and 26.5%-40.1%, respectively. Planting over-pressing hairy vetch after wheat harvest and moderately reducing nitrogen fertilizer increased the grain yield of wheat. Compared with W-N3 and W-N4, W-G-N3increased the yield by 6.9%-16.7% and 7.9%-13.6%, respectively. Grey correlation analysis and correlation analysis showed that multiple planting hairy vetch after wheat harvest and appropriate nitrogen fertilizer yield high crop yields were mainly due to the synergistic increase of mean LAI, total LAD and yield components.【Conclusion】The treatment of multiple cropping hairy vetch after wheat harvest combined with 85% N fertilizer treatment promoted the growth of vegetative organs, which was conducive to the establishment, expansion and enrichment of the grain bank, thereby obtaining high yields. Therefore, W-G-N3 was an ideal planting pattern and nitrogen application level for optimizing wheat yield performance indicators in oasis irrigation areas to obtain high yields.

    Combined Effects of High Temperature and Drought on Yield and Photosynthetic Characteristics of Summer Maize
    XiaoFan LI, JingYi SHAO, WeiZhen YU, Peng LIU, Bin ZHAO, JiWang ZHANG, BaiZhao REN
    Scientia Agricultura Sinica. 2022, 55(18):  3516-3529.  doi:10.3864/j.issn.0578-1752.2022.18.004
    Abstract ( 406 )   HTML ( 17 )   PDF (587KB) ( 268 )   Save
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    【Objective】Climate change has led to global warming, with frequent occurrences of high temperatures and droughts, and high temperatures often accompany droughts during production. This study aimed to explore the physiological mechanism of the compound stress of high temperature and drought in different growth periods affecting the yield and photosynthetic characteristics of summer maize.【Method】DH605 was selected as the experimental hybrid. High temperature treatment (T), drought treatment (D) and the compound stress (T-D) treatment were set in different growth periods. In 2019, it was carried out at the third leaf stage (V3), the sixth leaf stage (V6) and flowering stage (VT); In 2020, it was carried out at the third leaf stage (V3), the twelfth leaf stage (V12) and flowering stage (VT). This experiment took natural temperature and normal moisture treatment as control (CK). The effects of high temperature and drought compound treatments on yield, photosynthetic characteristics, dry matter accumulation and distribution of summer maize were studied, and the differences of photosynthetic characteristics and yield between single treatment and compound treatment were compared.【Result】 After the combined stress treatment of high temperature and drought in different growth stages, the LAI and SPAD of summer maize decreased significantly, which affected the net photosynthetic rate (Pn) and it decreased significantly. Among that, the compound stress during the VT period had the most significant effect on Pn. The Pn under T-D treatment in the VT period decreased by 39.0% on average compared with CK, while the net photosynthetic rate of summer maize after the combined treatment of high temperature and drought decreased more than that under single stress such as high temperature and drought. The combined treatment of high temperature and drought caused the photosynthetic performance of summer maize to decrease, and it led to the decrease of dry matter accumulation capacity and distribution ratio of summer maize to grains, which in turn led to a significant decrease in yield. In 2019, the output of T-D at V3, V6, and VT decreased by 27.4%, 18.3%, and 66.5%, respectively, compared with CK; in 2020, the output of T-D treatment at V3, V12, and VT decreased by 14.5%, 14.6% and 68.7%, respectively, compared with CK.【Conclusion】After the combined stress of high temperature and drought, the leaf area index and chlorophyll content was decreased, gas exchange was inhibited, leading to the decrease of photosynthetic performance, and thus hindered the accumulation and distribution of photosynthetic compounds, resulting in a significant yield reduction of summer maize. The combined stress of high temperature and drought during the flowering stage had the greatest impact on the yield and photosynthetic characteristics of summer maize, and the combined stresses had greater impacts than that of single stress.

    Simulation of Canopy Silking Dynamic and Kernel Number of Spring Maize Under Drought Stress
    MengQi WANG, Na MI, Jing WANG, YuShu ZHANG, RuiPeng JI, NiNa CHEN, XiaXia LIU, Ying HAN, WangYiPu LI, JiaYing ZHANG
    Scientia Agricultura Sinica. 2022, 55(18):  3530-3542.  doi:10.3864/j.issn.0578-1752.2022.18.005
    Abstract ( 198 )   HTML ( 4 )   PDF (700KB) ( 139 )   Save
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    【Objective】In order to improve simulation accuracy of maize kernel number under drought stress, the study simulated canopy silking dynamic of maize under drought stress and developed the relationships among anthesis-silking interval (ASI), canopy silking percentage and maize kernel number per unit area. 【Method】Firstly, this study measured the average plant growth rate (PGR) of maize around anthesis, daily canopy silking percentage of maize, ASI, the biomass accumulation of ear after anthesis, and the kernel number per plant under different treatments of soil water content based on drought stress controlling experiment at Jinzhou Agrometeorological Experimental Station. Secondly, the parameters of maize canopy silking dynamic model were determined with experimental data. Sensitivity analysis was conducted to investigate the impact of changes in average plan growth rate (PGRAVE) and standard deviation (PGRSD) on simulated canopy silking percentage. Thirdly, based on the ear biomass accumulation dynamic, the canopy silking dynamic was simulated under different drought stresses before and after anthesis by considering the differences in PGR among individual plants in the canopy. The quantitative relationship between ASI and kernel setting rate (the percentage of kernel number per plant to the maximum potential kernel number per plant) was developed based on experimental data. Finally, based on simulated canopy silking percentage after anthesis, maximum potential kernel number per plant, and the kernel setting rate by canopy silking dynamic model, the maize kernel number model was developed and validated under drought stress. 【Result】Sensitivity analysis of change in canopy silking percentage in response to changes in PGRAVE and PGRSD showed that PGRAVE had a greater impact on canopy silking percentage than PGRSD. The larger the PGRAVE and the smaller the PGRSD, the shorter the time for the canopy reaching silking percentage of 50%. The maize canopy silking dynamic model could accurately simulate daily silking percentage after anthesis under drought stress, and the coefficient of determination (R2), the root mean square error (RMSE), and the normalized mean square error (NRMSE) between simulated and observed canopy silking percentage ranged from 0.88 to 0.98, from 4% to 12%, and from 8% to 27%, respectively. Maize kernel number model could accurately simulate the kernel number of maize per unit area under drought stress, and R2, RMSE, and NRMSE between simulated and observed kernel number was 0.85, 185 kernel/m2, and 10%, respectively. 【Conclusion】By introducing the canopy silking dynamic model, the study could simulate the key phenology (silking time, anthesis-silking interval, and silking percentage) and kernel number per unit area under drought stress. The result was an important foundation for the simulation of maize yield based on canopy silking dynamic under drought stress.

    PLANT PROTECTION
    Cloning, Expression and Anti-TMV Function Analysis of Nicotiana benthamiana NbMBF1c
    YuXia WEN,Jian ZHANG,Qin WANG,Jing WANG,YueHong PEI,ShaoRui TIAN,GuangJin FAN,XiaoZhou MA,XianChao SUN
    Scientia Agricultura Sinica. 2022, 55(18):  3543-3555.  doi:10.3864/j.issn.0578-1752.2022.18.006
    Abstract ( 451 )   HTML ( 29 )   PDF (5496KB) ( 128 )   Save
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    【Objective】Tobacco mosaic virus (TMV) is an important virus that harms crops such as Solanaceae, Cruciferae and Cucurbitaceae, causing great losses to agricultural production. The objective of this study is to obtain Nicotiana benthamiana multiprotein bridging factor 1c (MBF1c) by molecular cloning, clarify the antiviral function and mechanism of NbMBF1c by bioinformatics, cell biology and molecular biology methods, and to provide a theoretical basis for the antiviral breeding of crops.【Method】Based on the full-length NbMBF1c sequence reported in Sol Genomics Network, primers were designed to clone the full-length sequence of NbMBF1c. GeneDoc and MEGA X were employed to align the homologous protein sequences of NbMBF1c and other species. The gene characteristics and protein structure of NbMBF1c were analyzed by bioinformatics. The tissue expression and its expression after TMV infection were detected by real-time fluorescence quantitative PCR (qRT-PCR). Tobacco rattle virus (TRV)-induced gene silencing technology was used to silence NbMBF1c and determine its effect on TMV-GFP infection. The transient overexpression vectors of pART27-GFP-NbMBF1c and pART27-Myc-NbMBF1c were constructed and the fusion protein GFP-NbMBF1c was expressed in N. benthamiana leaf to observe its subcellular localization under confocal microscope. Myc-NbMBF1c was expressed and the changes of TMV-GFP infection after NbMBF1c expressed were observed. qRT-PCR was employed to detect the expression of hormone related genes after NbMBF1c silencing, and the mechanism of NbMBF1c affecting virus infection was analyzed.【Result】NbMBF1c, which is 441 bp in full length, encodes 146 amino acids and contains a conserved domain HTH (helix-turn-helix). Phylogenetic analysis showed that NbMBF1c had the most closely relationship with Nicotiana tomentosiformis MBF1c (XP_009614458.1). NbMBF1c was localized in the cytoplasm and nucleus and had specific tissue expression, with the highest expression in roots, followed by stems, leaves and flowers. Silencing of NbMBF1c in N. benthamiana significantly reduced the tobacco plant height and promoted TMV-GFP infection, on the 5th day after TMV-GFP inoculation, the virus content of new leaf in the silencing treatment was higher than that in the control group. Transient overexpression of Myc-NbMBF1c suppressed TMV-GFP infection, after inoculation with TMV-GFP, virus infection was reduced. However, NbMBF1c did not interact with TMV components, but silencing NbMBF1c up-regulated the expression of abscisic acid (ABA) related gene NCED3, PYL1, ABAI, and SnRK2E and jasmonic acid (JA) synthesis pathway AOS1, while down-regulated the expression of JA signaling pathway receptor COI1.【Conclusion】NbMBF1c is mainly located in cytoplasm and nucleus, and acts as a positive regulator to inhibit TMV infection in N. benthamiana. NbMBF1c does not inhibit viral infection by directly interacting with TMV components, but by regulating phytohormone production and signal transduction.

    Monitoring of Resistance Dynamics of Phytophthora infestans to Cymoxanil and Control Efficacy Validation of Cymoxanil-Containing Fungicides Against Potato Late Blight
    Fen LU,RunJie MENG,Jie WU,JianJiang ZHAO,Yang LI,QiuYan BI,XiuYing HAN,JingHua LI,WenQiao WANG
    Scientia Agricultura Sinica. 2022, 55(18):  3556-3564.  doi:10.3864/j.issn.0578-1752.2022.18.007
    Abstract ( 244 )   HTML ( 33 )   PDF (589KB) ( 112 )   Save
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    【Objective】The objective of this study is to clarify the temporal and spatial dynamics of resistance to cymoxanil of Phytophthora infestans from potato of Hebei Province, Inner Mongolia Autonomous Region and Jilin Province, assess the control efficacy of cymoxanil and the mixed fungicides containing cymoxanil against potato late blight, and to provide the clues for the management of resistance of P. infestans to cymoxanil and the rational use of cymoxanil for control of potato late blight.【Method】The sensitivities to cymoxanil of 824 single-sporangial strains of P. infestans collected from main potato production areas of Hebei Province, Inner Mongolia Autonomous Region and Jilin Province from 2011 to 2019 were measured by the mycelial growth inhibition test. Also the control efficacies of cymoxanil and the mixed fungicides containing cymoxanil to potato late blight were evaluated in the fields.【Result】The population of P. infestans of Hebei Province, Inner Mongolia Autonomous Region and Jilin Province slowly developed low resistance to cymoxanil. The average EC50, average resistance factor, resistance frequency and resistance index of P. infestans to cymoxanil fluctuated over years of monitoring and locations of sampling, and tended to keep low-level and even. The average EC50, average resistance factor and resistance index of P. infestans strains over years to cymoxanil were 0.06-0.44 μg·mL-1, 0.29-2.18, and 0.25-0.47, respectively. Only sensitive strains were detected in 2011 and 2017. The resistance frequency of P. infestans to cymoxanil from 2012 to 2016 was 1.90%-58.57%. The resistance frequency of P. infestans to cymoxanil in 2018 and 2019 was 20.83% and 88.14%, respectively. The average EC50, average resistance factor, resistance frequency and resistance index of P. infestans strains in different provinces (Autonomous Region) to cymoxanil were 0.17-0.18 μg·mL-1, 0.85-0.91, 16.61%-22.22%, and 0.29-0.31, respectively. The result of field trials on control efficacy of fungicides showed that cymoxanil 20% SC, cymoxanil·mancozeb 72% WP and famoxadone·cymoxanil 52.5% WG provided the control efficacies of 81.5%-87.1% against potato late blight, which were significantly higher than those (61.8%-69.3%) given by mefenoxam·mancozeb 68% WG.【Conclusion】P. infestans population of Hebei Province, Inner Mongolia Autonomous Region and Jilin Province remained sensitive to cymoxanil on the whole, currently. Cymoxanil 20% SC and the mixed fungicides cymoxanil·mancozeb 72% WP and famoxadone·cymoxanil 52.5% WG still had high efficacies in control of potato late blight. Thus, in the areas where P. infestans widely developed high resistance to metalaxyl and mefenoxam, cymoxanil 20% SC, cymoxanil·mancozeb 72% WP and famoxadone·cymoxanil 52.5% WG could be used instead of the phenylamide fungicides. But it was necessary to monitor the dynamics of resistance to cymoxanil of P. infestans, and limit the application times of cymoxanil singly or in mixtures within twice per growth season and apply the fungicides containing cymoxanil in rotation or in mixture with the fungicides of different action modes.

    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Overview of Soil Survey Works in Main Countries of World
    WeiLi ZHANG,H KOLBE,RenLian ZHANG,DingXiang ZHANG,ZhanGuo BAI,Jing ZHANG,HuaDing SHI
    Scientia Agricultura Sinica. 2022, 55(18):  3565-3583.  doi:10.3864/j.issn.0578-1752.2022.18.008
    Abstract ( 345 )   HTML ( 38 )   PDF (549KB) ( 238 )   Save
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    The reviewing soil surveys and soil investigations in different countries shows that in order to understand the soil resources and soil quality, the soil surveys and investigations carried out by different countries since the end of the 19th century can be divided into four categories, namely: survey for soil classification, investigation and evaluation of farmland fertility, testing of soil chemical properties for fertilizer recommendation and soil investigations related to environmental issues. Among the four categories, soil classification is the earliest and the most widely implemented survey works carried out in the world. Main purpose of the classification survey is to clarify the differences in soil types and their spatial distributions caused by various soil formation processes. Since soil formation processes under natural conditions can reach tens of thousands of years, the main outputs of the classification survey—the soil maps and the records of soil profiles, expressing the physicochemical properties of various soil types, have a long timeliness and are widely used in various research areas. After the completion of nation-wide soil classification surveys in the last century in developed countries, the updated survey was not conducted in these countries. In the second national soil survey in China, large-scale soil maps covering the whole country were completed by relatively higher profile sampling density. The physicochemical properties of 100 000 soil profiles, representing different soil types, were also recorded. Both the map scale and the richness of soil profile data from the survey surpassed the outputs from classification surveys in many developed country. Soil classification data obtained from this survey are valid for a long time to different disciplines and sectors. In order to meet needs for easy-to-understand farmland fertility grades by land management sectors and farmers, investigation and evaluation of farmland fertility have been carried out in Central European countries, where per capita arable land resources have been in shortage. Through high-density profile samplings, soil fertility evaluation for each farmland plot with an independent cadastral code was carried out. An officially certified fertility grades in percentile index and the records of soil profiles were established and archived for each plot. The investigation results have been widely used in farmland management, taxation, agricultural subsidies, farmland leasing, trading, lending, insurance and other sectors, and have become the indispensable and centennial basic information for farmland quality. China's per capita arable land is much less than that of Central European countries, but so far there has been still in shortage of soil fertility data for farmland management. For preservation of arable land resources with high fertility and ensure food safety, it is essential and also urgent to establish precise and reliable fertility archives for each farmland plot in China. Also more efficient investigation and evaluation approaches, which are compatible with China's economic and social condition should be developed. The review of testing soil chemical properties to make fertilizer recommendation shows that such testing has been incorporated into the technical supporting system for best farming management practices in the developed countries. To improve farmers' fertilization techniques, it is more important to establish an efficient technical supporting system and maintain its running, in comparison to nation-wide soil nutrient census of farmland. In the past, there was continuous weakening of applied researches and extension works for fertilization technological chain from basic research to farmers' application. Due to inadequate research works, the technical indexes for fertilization, that should be differentiated to and compatible with the various regional soil and climate conditions and hence easy to be followed by farmers of different regions, have not been issued yet in China. Also the intelligent tools that can reach and provide targeted guidance to farmers were not available. Unbalanced and excessive fertilization was common in vegetable, fruit and other cash crop growing area, which accounted for 23.6% of the total cropping area in China. This resulted in reduction of crop yield and quality as well as benefit of farmers and also leaded to agricultural non-point source pollution. Making up for the shortcomings in the technical supporting system has become the key to improve both crop yield and soil fertility. Since the end of the last century, soil investigations related with environment issues have been carried out in different countries. Main purposes of these investigations are to clarify the status of environmental pollution and changing, to develop control strategy and to check effects of countermeasures. With the quick progresses of GIS-, GPD-, RS- and big data techniques, the traditional mapping approaches have be replaced by digital soil mapping techniques. An exact defining of investigation objectives and a comprehensive reviewing of relevant research progress as well as available auxiliary data are essential for drafting sampling design and achieving investigation tasks finally.

    Research Progress of Direct Straw Return in Anhui Province over the Last Decade
    Chao MA,YuBao WANG,Gang WU,Hong WANG,JianFei WANG,Lin ZHU,JiaJia LI,XiaoJing MA,RuShan CHAI
    Scientia Agricultura Sinica. 2022, 55(18):  3584-3599.  doi:10.3864/j.issn.0578-1752.2022.18.009
    Abstract ( 371 )   HTML ( 42 )   PDF (3456KB) ( 199 )   Save
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    The returning straw directly to the field is the most widely adopted approach of straw utilization. Anhui is an important agricultural province with abundant main grain crop straw resources. In recent years, the direct straw return was actively promoted and a series of relative research were conducted in Anhui Province. In order to provide reference for the development of more in-depth research and promote the efficient utilization of straw resources, the recent important advances in straw direct returning technology and the effects of straw incorporation on soil fertility, crop production and agricultural eco-environment in Anhui Province were reviewed in this paper. The research progress of direct straw return in Anhui Province over the last decade were mainly embodied in the following aspects. (1) The technology of straw direct returning was optimized from manner of straw returning and combined application of chemical fertilizers. (2) The introduction of chemical structure of soil organic matter and soil microbial community structure broadened the research field concerning the effects of direct straw return on soil physical, chemical and biological properties. (3) The impact of straw incorporation on crop diseases, insect pests and weeds was beginning to be explored, and the increasingly attention was being paid to the environmental effects of straw return on nitrogen and phosphorus loss and greenhouse gas emission. Therefore, it could be concluded that the research on direct straw return in Anhui Province presented the transformation from a focus on crop production to more attention on eco-environmental effects. In the future, the study on straw directly returning to the field in Anhui Province should be strengthened from the following aspects: improving regional weak links, expanding research contents, clarifying environmental effects, and deepening mechanism exploration.

    Effect of Nitrification on Ammonium Toxicity to Citrus in Acidic Soil
    ZiHan FAN,YaYin LUO,HuaYe XIONG,YuWen ZHANG,FuRong KANG,YuHeng WANG,Jie WANG,XiaoJun SHI,YueQiang ZHANG
    Scientia Agricultura Sinica. 2022, 55(18):  3600-3612.  doi:10.3864/j.issn.0578-1752.2022.18.010
    Abstract ( 212 )   HTML ( 25 )   PDF (784KB) ( 67 )   Save
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    【Objective】The response process of citrus to NH4+-N was elaborated by observing the impact of NH4+-N level on different pH soil solutions, combined with a study on the impact of orange rootstock seedling growth and physiological indicators, which provided theoretical support for the best citrus nitrogen management in acid soil. 【Method】 The experiment was designed as a two-factor experiment, with 2 soils as the main treatment and 5 N levels as the secondary treatment. Using acidic yellow soil and alkaline purple soil as the test soil, the seedlings of Xiangcheng (junos Sieb. ex Tanaka) were selected as the test material, and five NH4+-N levels of 0 (A0), 50 (A50), 100 (A100), 200 (A200), and 400 mg·kg-1 (A400) were set. The effects of ammonium application level on the concentration of ammonium nitrogen and nitrate nitrogen in soil solution and on the biomass, root morphology, nitrogen absorption, antioxidant system, nitrogen metabolism, MDA content and root activity of citrus were studied. 【Result】 Compared with calcareous soil, the nitrification process in acid soil was slowed down. The NH4+-N concentration and NH4+/ NO3- ratio in the soil solution of acid soil remained at a higher level at 30 days of the test. The root length of the citrus under A400 treatment was reduced by 13% when compared with the A0 treatment, and the root vitality was significantly negatively correlated with the ammonium application level. The MDA content of leaves and roots was positively correlated with the ammonium level, which stimulated the oxidative stress response of the roots, especially increased the POD enzyme activity of the leaves. Compared with calcareous soil, the total nitrogen accumulation of citrus on acid soil decreased by 17.6%, while the ratio of ammonium to nitrate in leaves and roots increased by 27.2% and 61.1%, respectively. Cluster analysis showed that citrus in acidic soil was toxic when N application rate exceeded 100 mg kg-1, while citrus in alkaline soil was not significantly stressed.【Conclusion】 In acidic soil, the excessive application of ammonium nitrogen caused the accumulation of ammonium nitrogen in soil solution for a long time, resulting in the increase of MDA content, cell membrane damage and nitrogen metabolism disorders as well as other ammonium toxic phenomena, indicating that ammonium toxicity in citrus was closely related to soil nitrification.

    HORTICULTURE
    Resistant Evaluation of 84 Apple Cultivars to Alternaria alternata f. sp. mali and Genome-Wide Association Analysis
    BaoHua CHU,FuGuo CAO,NingNing BIAN,Qian QIAN,ZhongXing LI,XueWei LI,ZeYuan LIU,FengWang MA,QingMei GUAN
    Scientia Agricultura Sinica. 2022, 55(18):  3613-3628.  doi:10.3864/j.issn.0578-1752.2022.18.011
    Abstract ( 311 )   HTML ( 39 )   PDF (1448KB) ( 127 )   Save
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    【Objective】 Alternaria alternata f. sp. mali is a main disease occurring in the apple orchards in China, which seriously threatens the yield and quality of apples. The aim of this research was to identify apple cultivars and genes responsible for high resistance to Alternaria alternata f. sp. mali, thus providing the basis for disease resistance breeding of apples.【Method】 In this study, the isolated Alternaria alternata f. sp. mali strain was used to inoculate leaves of 84 apple cultivars. To evaluate the resistance of apple cultivars to Alternaria alternata f. sp. mali, the cluster analysis was carried out from two aspects including lesion area and lesion area growth rate. The genome-wide association study was performed based on the lesion area. The disease spot area of the inoculated leaves was used as a phenotypic trait, and the 1243071 high-quality SNP loci obtained by whole genome deep resequencing were used as genetic markers. Genome wide association analysis was performed by EMMAX method.【Result】The statistical analysis of lesion area showed significant diversity of 84 apple cultivars in resistance to Alternaria alternata f. sp. mali. Most apple cultivars were susceptible or resistant to Alternaria alternata f. sp. mali, while only a few cultivars were highly resistant and highly susceptible. The disease resistance of Alternaria alternata f. sp. mali has the characteristics of normal distribution, showing the genetic characteristics of quantitative characters. The genome-wide association trait analysis finally obtained 6 SNP loci with a significant level of P≤0.0000001 (-LgP≥7), and in-depth analysis linked them to 7 key candidate genes, including Integrin-linked protein kinase, FMN-linked oxidoreductase superfamily protein, B-box zinc finger family protein, GATA type transcription factor, etc. The biological function of Integrin-linked protein kinase in disease resistance was further verified. 【Conclusion】After two years of comprehensive analysis of data, 7 stable resistant varieties and 2 stable susceptible varieties were identified from 84 apple cultivars. Through genome-wide association trait analysis, six SNP loci and seven genes significantly related to the disease resistance of apple leaf spot were identified, and function of one gene was verified.

    Phenotypic Diversity Analysis of Chinese Ornamental Herbaceous Peonies and Its Germplasm Resource Evaluation
    YingLing WAN,MengTing ZHU,AiQing LIU,YiJia JIN,Yan LIU
    Scientia Agricultura Sinica. 2022, 55(18):  3629-3639.  doi:10.3864/j.issn.0578-1752.2022.18.012
    Abstract ( 305 )   HTML ( 39 )   PDF (1956KB) ( 104 )   Save
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    【Objective】The objective of this study was to estimate the resource diversity of Chinese ornamental herbaceous peonies and to analyze the characteristics of being cut peonies, so as to provide a theoretical basis for germplasm utilization and innovation.【Method】Phenotypic characteristics data of 346 herbaceous peonies from four cultivation areas were collected. Then, according to the whole sample and the samples from different cultivation areas, the frequency distribution, coefficient of variation and diversity index were calculated. Additionally, the varieties were analyzed by clustering.【Result】The cultivars of Chinese ornamental herbaceous peonies were rich in the diversity of many characteristics, with diversity indices of binary, ordered and disordered multistate characteristics of 0.29-1.90 as well as numerical characteristics of 1.90-2.08. In binary, ordered and disordered multistate characteristics, flower color and flower type showed the highest diversity index, i.e., 1.90 and 1.88, respectively, followed by flower bud color (1.46) and degree of stamen petalization (1.42). In numerical characteristics, plant height and stem length had the highest diversity index, i.e., 2.08 and 2.07, respectively, followed by bud diameter (2.04), flower diameter (2.04) and flower height (2.01), and only the diversity indices of stem diameter (1.99) and pedicel length (1.90) were less than 2. Furthermore, the plant height was distributed in the range of 38.50-100.50 cm, stem length was 43.19-120.00 cm, and both were normally distributed. However, bud diameter, flower height, flower diameter, stem diameter and pedicel length showed skewed distributions. Among them, the bud diameter was 1.22-3.99 cm, showing a negatively skewed distribution. The flower diameter was distributed in the range of 3.11-19.50 cm, and the variation coefficient was only 15.65%, ranking it last among the numerical characteristics. The flower height (1.73-14.30 cm) was negatively distributed, with a variation coefficient of 32.55%, ranking it first among the numerical characteristics. Stem diameter (0.18-0.59 cm) and pedicel length (5.57-40.00 cm) were positively skewed. The nine groups were divided by clustering, and the most of the clustering groups included varieties from two to four cultivation areas. Moreover, there were no significant differences in bud diameter, flower diameter, stem diameter or pedicel length among the clustering groups. The plant height, stem length, flower height and degree of stamen petalization were different. Among the samples, 71.39% of the varieties had round flower buds and were nonchapped, which were presently popular in the Chinese market. Many varieties were the double flower type, but the hydrangea type preferred by the market accounted for only 4.9%. Meanwhile, 79.8% of the varieties had no obvious flower fragrance, 71.7% of the varieties had side buds, and 62.7% of the varieties bloomed during the middle flowering period, lacking early- or late-flowering varieties. There were no significant differences in plant height or stem length among the various cultivation areas, while the stem diameters of cultivars from Lintao and Heze were significantly larger than those from Yangzhou and Luoyang. The bud diameter and flower diameter of cultivars from Heze were also significantly larger.【Conclusion】A relatively high diversity of Chinese cultivated ornamental herbaceous peonies was observed, and as a kind of multifunctional plant, the diversity was worth preserving. However, the cultivars were insufficient as cut peonies, lacking cultivars with straight stems, fragrance, single buds and diverse flowering periods. The varieties from Lintao and Heze had some advantages in stem, bud and flower diameters for cut flowers. Cut peony selection and breeding in China was inadequate. Innovative germplasms need to be developed. It is urgent to introduce germplasms with the characteristics of long stems, single flower on one stem, fragrant flowers and late flowering periods for breeding.

    FOOD SCIENCE AND ENGINEERING
    Eating Quality Evaluation of Lamb in Different Postmortem Phases Based on Consumers’ Sensory Preferences
    Yue GE,DeQuan ZHANG,ShaoBo LI,Li CHEN,XiaoChun ZHENG,Ce LIANG,TongJing YAN,JinHuo LI,ZhenYu WANG
    Scientia Agricultura Sinica. 2022, 55(18):  3640-3651.  doi:10.3864/j.issn.0578-1752.2022.18.013
    Abstract ( 228 )   HTML ( 22 )   PDF (840KB) ( 70 )   Save
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    【Objective】The eating quality of lamb in pre-rigor, rigor mortis and post-rigor phases was investigated by using consumers’ sensory evaluation to understand the differences of eating quality characteristics of lamb in different postmortem phases under instant boiling, roasting, boiling and stir-frying conditions, which was expected to provide the reference for lamb quality evaluation and high-quality meat production.【Method】The oyster cut, short loin, knuckle and silverside in per-rigor, rigor mortis and post-rigor phases were obtained from small-tail Han sheep (male, 8 months old). The sensory attributes (including tenderness, juiciness, flavor and overall liking) of lamb after instant boiling, roasting, boiling and stir-frying were evaluated by consumers. The MQ4 (Meat Quality, 4 variables) equation and the grades boundaries were determined by linear discriminant analysis, which were used to analyze the preference score and eating quality grade of lamb.【Result】After instant boiling, the short loin and silverside in pre-rigor had higher tenderness, flavor and MQ4 scores than those in rigor mortis and post-rigor phases (P<0.05), while there was no significant difference in consumers’ scores of oyster cut and knuckle in different postmortem phases (P>0.05). After roasting, the oyster cut in per-rigor and the silverside in post-rigor phase had higher tenderness, juiciness, overall liking and MQ4 scores than those in rigor mortis (P<0.05), but there was no significant difference in consumers’ scores of short loin and knuckle in different postmortem phases (P>0.05). After boiling, the juiciness and overall liking scores of knuckle in post-rigor were higher than those in per-rigor and rigor mortis phases (P<0.05), and there was no significant difference in consumers’ scores of oyster cut, short loin and silverside in different postmortem phases (P>0.05). After stir-frying, the tenderness, overall liking and MQ4 scores of oyster cut in pre-rigor and post-rigor were higher than those in rigor mortis, while the tenderness, flavor, overall liking and MQ4 scores of short loin in pre-rigor were higher than those in rigor mortis and post-rigor (P<0.05). There was no significant difference in consumers’ scores of knuckle and silverside in different postmortem phases (P>0.05). The premium quality (4star) and good quality (5star) samples were higher in pre-rigor (5star: 0%-8.33%; 4star: 47.83%-72.22%) and post-rigor (5star: 0%-8.70%: 4star: 52.78%-58.33%). There were no unsatisfactory quality (2star) samples in pre-rigor and post-rigor phases, while 4.17% and 4.35% of the samples in rigor mortis were 2star after roasting and boiling, respectively. 【Conclusion】In general, the pre-rigor and post-rigor lamb had a superior eating quality to rigor mortis lamb based on consumers’ sensory evaluation, and the pre-rigor and post-rigor lamb were preferred by Chinese consumers.

    ANIMAL SCIENCE·VETERINARY SCIENCE
    Effects of IL-15 Overexpression on Myoblast Differentiation of Porcine Skeletal Muscle Cells
    MingJie XING,XianHong GU,XiaoHong WANG,Yue HAO
    Scientia Agricultura Sinica. 2022, 55(18):  3652-3663.  doi:10.3864/j.issn.0578-1752.2022.18.014
    Abstract ( 292 )   HTML ( 60 )   PDF (2950KB) ( 109 )   Save
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    【Objective】The aim of this was to investigate the effects of interleukin 15 (IL-15) as a myokine on the proliferation and apoptosis of porcine skeletal muscle myoblast, so as to provide a basis for further studying on the regulation of IL-15 in animal muscle quality and the treatment of skeletal muscle diseases. 【Method】In this study, the IL-15 overexpressed lentiviral vector GV-492-IL-15 was constructed, and the porcine skeletal muscle satellite cells were aseptically isolated and cultured in vitro, then skeletal muscle cells morphology were subjected to myogenic differentiation, and the differentiated myoblast were verified by immunofluorescence staining. After myoblast differentiation, the IL-15 overexpressed recombinant lentiviral vector was transfected. A blank control group (Control), a negative control virus transfection group (IL-15-) and a GV-492-IL-15 lentivirus transfection group (IL-15+) were set for the experiment (n=3). The cells were cultured for 72 h; after growing to a certain number, the cells and the culture supernatant were harvested. Real-time quantitative PCR (qRT-PCR) and Western Blot were used to analyze the expression of target genes and proteins. ELISA kit was used to analyze the content of IL-15 in the culture medium, CCK-8 kit was used to analyze cell viability, and the flow cytometry was used to analyze the results of cell cycle and apoptosis. Moreover, Western Blot was used again to detect changes of the level of caspase-3 protein in the cells, which was closely related to apoptosis.【Result】(1) The identified plasmid was transfected with 293T cells, and as a result, a distinct green fluorescence could be observed in the cells, and a characteristic band near 20 KD could be observed by Western Blot. (2) Fusiform or fusiform porcine skeletal muscle satellite cells were obtained by microscopic observation and differentiated into tubular myoblasts after induction. The differentiated myoblasts were subjected to immunofluorescence staining with α-SMA monoclonal antibody. About 90% of the cells in the visual field were positive, and the cytoplasm stained red, indicating that the cultured cells were skeletal muscle myoblast cells. (3) After transfected with GV-492-IL-15 lentivirus, the relative mRNA and protein expression of IL-15 in myoblasts were significantly increased compared with the control group (P<0.001), however, the protein level of IL-15 in culture medium was not significantly changed (P>0.05). Compared with the control group, the early apoptosis rate of cells transfected with GV-492-IL-15 lentivirus was not significantly different (P>0.05), but the late apoptosis rate of cells was significantly decreased (P<0.05); there was a tendency for caspase-3 protein to decrease compared to the control group, but the overall difference was not significant (P>0.05). The CCK-8 assays showed that overexpression of IL-15 increased the ability of cell proliferation (P<0.05). In addition, the proportion of cells in G1 phase was significantly decreased by transfected with IL-15 overexpression lentivirus, while the proportion of cells in S phase and G2/M phase was significantly increased (P<0.05). 【Conclusion】Under the normal physiological conditions, IL-15 was localized in cells and played a role. The overexpression of IL-15 had no significant effects on the early apoptosis of porcine skeletal muscle myoblasts, but it could inhibit the late apoptosis and promote cell proliferation. This study provided a technical and theoretical basis for the positive regulation of IL-15 on pig skeletal muscle quality and the treatment of related muscle diseases.

    Effects of Circadian Rhythm on Rumen Fermentation and Nutrient Digestion of Mutton Sheep
    ChunTao ZHANG,Tao MA,Yan TU,QiYu DIAO
    Scientia Agricultura Sinica. 2022, 55(18):  3664-3674.  doi:10.3864/j.issn.0578-1752.2022.18.015
    Abstract ( 191 )   HTML ( 25 )   PDF (472KB) ( 80 )   Save
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    【Objective】The biological clock system generally exists at all levels of life, which is closely related to the digestive physiology and growth performance of animals. This experiment explored the relationship between circadian rhythm and rumen fermentation parameters, nutrient digestion and metabolism of Hu sheep, in order to explore the relationship between circadian rhythm and physiological digestion and metabolism of Hu sheep, so as to provide an important theoretical basis for improving the growth performance and nutrient utilization of fattening sheep.【Method】Forty-five healthy Hu sheep with body weight of 21.57 ± 0.77kg weight were randomly divided into 3 groups, with 15 sheep in each group. Each treatment group was used the same concentrate supplement and coarse feed. The three treatments were respectively set as follows: day treatment group(DH), that was, 70% of the total daily concentrate + 30% of the total daily coarse feed in the morning; in the evening treatment group(DL), 30% of the daily total concentrate + 70% of the daily total coarse feed were fed in the morning; the control group(CON), i.e. 50% of the daily total concentrate + 50% of the daily total crude feed in the morning and evening. After feeding for two months, the digestion and metabolism test was carried out. Total fecal collection and urine were used to determine nutrient apparent digestibility and metabolic rate. After the feeding experiment, the rumen fluid samples were collected from oral cavity 2 h before morning feeding and 2 h before evening feeding, respectively, for the determination of related indexes. 【Result】 ① The daily gain of DH was 215.00 g, the food intake was lower than that of DL and CON, and the feed conversion ratio was the best, with the value of 5.35, which was 11.19% and 16.04% better than the other two groups, respectively. ② Under different feeding modes, the digestibility of dry matter (DM), organic matter (OM), crude protein (CP), neutral detergent fiber (NDF) and acid detergent fiber (ADF) of sheep in DH group were higher than those in CON and DL groups. Compared with DL group, The DM digestibility, CP, NDF, and ADF was increased by 21.42%, 22.29%, 9.85%, and 28.69%, respectively. ③ The biological value of nitrogen in DH group was 20.31% higher than that of CON and 67.41% higher than that of DL (P>0.05). There was no significant difference in the intake of digestible nitrogen among all groups, and the total nitrogen excretion in DL group was significantly higher than that in DH group (P<0.05). The nitrogen deposition, nitrogen utilization and biological value in DH group were significantly higher than those in DL group (P<0.05) and CON group (P>0.05). ④ There were no significant differences in rumen microbial protein (MCP), acetic acid and total volatile fatty acid (TVFA) among groups with different treatments (P>0.05), however, the rumen pH, the ratio of acetic to propionic acid and butyric acid in DH group were significantly lower than those in DL group (P<0.05). With the alternation of day and night, rumen pH, NH3-N, acetic acid, propionic acid, total volatile fatty acid and the ratio of acetic to propionic acid were significantly changed (P<0.05), and the related parameters were higher in day than in night.【Conclusion】Growth performance and nutrient digestibility of sheep could be improved by changing the ration of concentrate to forage in the early and late days under the same nutrient content in the diet, and the feeding high concentrate ratio in the morning and low concentrate ratio in the afternoon. Rumen fermentation was affected by day and night, and feeding high concentrate diet in day promoted the contents of TVFA, NH3-N and MCP in rumen. In a word, it is better to increase the proportion of concentrate in the morning, which is more in line with the rhythm of digestion and absorption of animal body, and has better effect on the utilization of feed.

    Molecular Mechanism of Regulation by H-NS on IncFⅡ Plasmid Transmission of Multi-drug Resistant Chicken Escherichia coli
    YaTing JIA,HuiHui HU,YaJun ZHAI,Bing ZHAO,Kun HE,YuShan PAN,GongZheng HU,Li YUAN
    Scientia Agricultura Sinica. 2022, 55(18):  3675-3684.  doi:10.3864/j.issn.0578-1752.2022.18.016
    Abstract ( 197 )   HTML ( 25 )   PDF (854KB) ( 65 )   Save
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    【Objective】The objective of this study was to investigate the molecular mechanism of H-NS regulating conjugation of the IncFⅡ plasmid from a clinically isolated multi-drug resistant Escherichia coli from chicken, so as to provide a theoretical basis for controlling the rapid spread of IncFⅡ plasmid mediated multidrug resistance genes.【Method】The growth curves of Escherichia coli ATCC25922 and four recombinant strains (F25922, pBAD25922, FΔhns and FΔhns/phns) were determined to compare the influence of hns on different strains. The conjugation experiments were conducted with F25922, FΔhns and FΔhns/phns as donors and Escherichia coli J53 as recipient, then the conjugation frequency was calculated. The mRNA expression levels of IncFⅡ plasmid conjugation transfer related genes (traM, traJ and traY) in each recombinant strains (F25922, FΔhns and FΔhns/phns) were detected by RT-qPCR. The LacZ reporter strains F25922/PM(PJ/PY), FΔhns/PM(PJ/PY) and FΔhns/phns/PM(PJ/PY) were constructed to determine the β-galactosidase activity of three promoters of tra genes (traM, traJ and traY). The H-NS protein was purified by Ni-NTA resin affinity chromatography. The DNA sequences of three promoters of tra genes were amplified by PCR. The mechanism of H-NS regulating IncFⅡ plasmid transmission was identified by EMSA, and the binding sites of H-NS to different promoters were predicted and further verified by ESMA.【Result】The growth of recombinant strains F25922 and pBAD25922 were not significantly different from that of Escherichia coli ATCC25922, while the growth rate of deleted recombinant strain FΔhns and complemented strain FΔhns/phns were significantly lower than that of the control strain F25922. The results showed that the absence of hns could make the adaptability of strains worse, but did not affect the survival of the strains. The results of the conjugation test showed that the conjugation frequency of IncFⅡ plasmid in FΔhns was 1 279.33 times higher than that of the control strain F25922 (P<0.001), and the FΔhns/phns conjugation frequency of the supplementary strain was significantly lower than that of FΔhns, although it did not completely recover to the level of the control strain F25922. Similarly, the mRNA expression levels of these tra genes (traM, traJ and traY) were significantly higher in the deletion mutant FΔhns. The mRNA expression level of traJ was the highest in FΔhns, which was 1 510.14 times that of F25922, followed by traY and traM, which were 448.14 times and 81.54 times that of F25922, respectively. Compared to the deletion strain FΔhns, expression levels of the tra genes (traM, traJ and traY) in the complemented strain FΔhns/phns were significantly decreased. The β-galactosidase activities of promoters PM, PJ and PY in the reporter strains FΔhns/PM (PJ/PY) were 5.66, 10.45 and 21.91, respectively, which were significantly higher than that of the corresponding promoters of F25922/PM (PJ/PY) (P<0.001). The activities of promoters PM, PJ and PY in the complement reporter strains FΔhns/phns/PM (PJ/PY) were significantly lower than that of FΔhns/PM (PJ/PY), and there was no significant difference with the control strain F25922/PM (PJ/PY). EMSA results showed that H-NS protein could block the DNA migration of three promoters of tra genes, indicating that H-NS could directly bind to the three promoters. By predicting the binding sites and further verified by EMSA, it was confirmed that H-NS protein could bind directly to the AT enrichment region of the promoters of the three genes (traM, traJ and traY).【Conclusion】H-NS protein could bind directly to the AT enrichment region of the promoter region of IncFⅡ plasmid conjugation transfer related genes (traM, traJ and traY), and negatively regulate the conjugation transfer of IncFⅡ plasmid by inhibiting the activity of promoter.