泛素激活酶抑制剂对猪精子获能状态、膜重构及体外受精的影响

【目的】评估泛素激活酶（E1）对猪精子获能、膜重构和体外受精的影响。【方法】选取5头长白猪采集精液，以硫醇酯法评估泛素激活酶（ubiquitin activating enzyme，E1）抑制剂（TAK-243）对精子中泛素（ubiquitin，Ub）与E1结合的影响；将精子分为鲜精组（Fresh）、DMSO组（DMSO）、获能组（Capacitated）及2.4、4.8和7.2 nmol/L TAK-243组，鲜精组用2 mL PBS重悬精子沉淀；获能组用2 mL获能液重悬精子沉淀；DMSO组及2.4、4.8和7.2 nmol/L TAK-243组分别用2 mL含0.07% DMSO及2.4、4.8和7.2 nmol/L TAK-243的获能液重悬精子沉淀，使用微生物动（静）态图像检测系统检测精子的动力学参数；以Western blotting法检测精子的酪氨酸磷酸化水平；Zn²⁺荧光探针检测精子的Zn²⁺含量；花生凝集素染色检测精子的顶体膜重构；免疫荧光法检测顶体表面精子黏附蛋白（AQN-1、AWN）的表达；体外受精法检测TAK-243对精卵结合和卵裂率的影响。【结果】硫醇酯分析结果表明，TAK-243处理组精子未形成Ub-E1复合物，说明TAK-243抑制了E1对Ub的激活作用。与鲜精组相比，获能组精子的曲线速度（VCL）和平均鞭打频率显著升高（P<0.05）。3个浓度TAK-243组与获能组精子的各项动力学参数差异均不显著（P>0.05）。与获能组相比，3个浓度TAK-243组酪氨酸磷酸化水平均显著降低（P<0.05），且随着TAK-243浓度的增加，精子蛋白的酪氨酸磷酸化水平逐渐降低。后续试验使用7.2 nmol/L TAK-243进行精子中Zn²⁺水平和顶体膜重构的研究，其结果表明，与获能组相比，7.2 nmol/L TAK-243组Zn²⁺水平、顶体膜完整性及AQN-1和AWN蛋白的表达水平均显著增加（P<0.05），精子与卵母细胞的黏附率和卵裂率均显著降低（P<0.05）。【结论】7.2 nmol/L TAK-243显著降低了精子获能期间蛋白酪氨酸磷酸化水平、精卵结合率和卵裂率，显著增加了Zn²⁺含量、顶体膜完整率及顶体表面蛋白AWN和AQN-1的表达，说明泛素-蛋白酶体信号通路参与了精子的体外获能。     

施肥方式对不同密度下玉米光合生理的影响

为进一步研究玉米品种登海605不同密度和施肥方式对光合生理的影响，挖掘其增产潜力，建立高产栽培模式。本试验设计5个密度处理（D1、D2、D3、D4、D5）、3种施肥方式处理（S1：农大缓释肥、S2：分次施肥、S3：一次性施肥），并对不同处理的光合指标、叶面积指数、光合有效辐射、透光率等指标进行测定，结果表明：不同的施肥方式上层、中层、下层光合指标变化规律表现一致，各层次表现为上层>中层>下层，S2>S1>S3，玉米植株上层无论哪种处理都能获得足够的光能和CO2，光合指标相差不大，适宜的密度和施肥方式将获得更多的光能、有效辐射，合成有机物，形成更多的干物质积累，从而提高产量。     

不同施肥模式下的稻-克氏原螯虾田块水体菌群初探

为研究不同施肥模式对稻田养殖克氏原螯虾(Procambarus clarkii)田块水体细菌的影响,通过采用Illumina高通量测序技术针对施用化肥(CF)、有机肥(OF)和有机肥加腐熟鸡粪(OM)等3种施肥模式的水体细菌进行检测。高通量测序结果显示在3种不同施肥模式下的稻田中主要细菌门类均为变形菌门(Proteobacteria)、放线菌门(Actinobacteria)、拟杆菌门(Bacteroidetes)、蓝藻门(Cyanobacteria)和厚壁菌门(Firmicutes)。主要优势菌属为Sporichthyaceae hgcl_clade、Limnohabitans、Polynucleobacter、Alpinimonas、Comamonadacea和Hydrogenophaga。在3种模式中,施用有机肥的田块细菌丰度显著高于施用化肥和有机肥加腐熟鸡粪的田块,细菌物种多样性显著高于施用有机肥加腐熟鸡粪田块,与施用化肥田块差异不显著。结果表明在施用有机肥模式下,水体微生物具有更好的多样性,生态系统更加稳定。有机肥与腐熟鸡粪配施虽然会降低水体细菌的物种多样性,但是腐熟鸡粪与有机肥配施会大大降低蓝藻细菌的丰度。聚类分析结果显示有机肥组与有机肥加腐熟鸡粪组水体细菌群落相似度更高。对稻田水体细菌与水体理化因子关联分析发现,对水体细菌影响最主要的环境因子是溶氧、总氮、pH和总磷。研究结果为不同施肥处理对稻田水体生态环境的影响以及为稻田综合种养施肥模式优化提供理论依据。     

夏休闲期复种饲料油菜提升后作冬小麦产量品质、土壤肥力及周年经济效益研究

为明确晋中麦区夏休闲期复种饲料油菜的可行性,试验于2015-2016年采用大田试验法,研究了夏休闲期复种饲料油菜对后作冬小麦生长、麦田土壤养分及周年经济效益的影响。前茬小麦收获后复种油菜,供试油菜和小麦品种分别为华油杂62和CA0547,设置3个处理:以不种油菜的休闲田作为对照(CK₀),油菜地上部被收获为CK₁,油菜被全量还田为T。结果表明,与CK₀和CK₁相比,T有利于增加后作冬小麦抽穗前单株绿叶数,提高生育中后期植株干物质及N、P积累,保持灌浆期旗叶面积,进而增加穗粒数,提高籽粒产量,提高籽粒蛋白质及醇溶蛋白和谷蛋白含量;同时T有利于提高后作冬小麦020 cm土层土壤有机质含量、040 cm土层土壤脲酶和蔗糖酶活性。此外,从油菜-小麦周年经济效益来看,CK₁较CK₀和T分别高出128.9%和260.0%。综合分析可知,夏休闲期复种饲料油菜并且全量还田可以提高小麦产量,提升籽粒品质,改良土壤有机质含量和土壤酶活性;夏休期复种饲料油菜并收获可以获得更高的周年经济效益。本研究结果为合理利用夏休闲期以提升小麦产量与品质、改良土壤、提高年经济效益提供了理论依据。     

北方夏玉米滴灌施肥一体化技术应用效果

为有效提高夏玉米水肥利用效率,通过田间试验对滴灌施肥一体化高效调控与利用技术进行系统的探究。结果表明,与常规漫灌(T1)相比,采用滴灌施肥一体化技术(T2和T3)可显著提高夏玉米大喇叭口期至灌浆期的叶面积指数;显著促进吐丝期至成熟期的干物质积累;在产量构成因素方面显著增加穗粗和穗粒数,并在一定程度上降低玉米穗的秃顶长度,进而显著提高夏玉米的产量。与T1相比,T2和T3增产5.32%~6.13%;水分利用效率提高36.10%~39.42%;氮、磷、钾肥利用率分别提高37.56%~35.29%、23.09%~30.30%、106.21%~121.51%。此外,采用滴灌施肥一体化技术可显著提高夏玉米生产经济效益。与T1相比,T2和T3可有效降低投产比,收益率平均提高4.71个百分点。在化肥用量较常规灌溉模式减少20%条件下,采用滴灌施肥一体化技术既能显著提高夏玉米产量,又能提高经济效益,是适宜在潮土区推广的一种高效节本增收的灌溉模式。     

缓释肥侧位深施及用量对油菜产量和肥料利用率的影响

为明确红壤稻田直播油菜缓释肥（N-P2O5-K2O:25-7-8）侧位深施效果及适宜用量，该研究连续2 a在两熟制和三熟制2种种植模式下开展缓释肥侧位深施效果对比试验（设置不施肥、土表撒施和侧位深施3个施肥方式）和施用量试验（设置0、300、450、600、750和900 kg/hm2 共6个施肥水平），研究缓释肥侧位深施及不同用量对油菜产量形成和肥料利用率的影响。结果表明，施肥方式对红壤稻田油菜产量形成和肥料利用率均有显著影响，且对两熟制油菜影响更为显著。相比传统土表撒施，侧位深施显著促进了油菜产量和肥料利用率的提高。缓释肥侧位深施明显提高了各时期油菜干物质量，尤其是显著增加了初花期至成熟期的干物质积累量，促进了花后根部与地上部干物质同步增长；促进了根系对N、P、K的吸收，提高了油菜产量和肥料利用率。菜籽产量与缓释肥用量呈线性加平台关系，适宜施肥量可保证较大的收获密度，并协同产生较多的每株角果数和每角粒数，从而提高籽粒产量、肥料利用率和经济效益。两熟制和三熟制油菜缓释肥侧位深施的适宜用量分别为715和585 kg/hm2，产量潜力可分别达2 450和1 700 kg/hm2。研究表明，侧位深施适量缓释肥可显著提高红壤稻田直播油菜生产力，建议结合机械化种植因地制宜推广应用。     

长期施肥下黄土旱塬黑垆土磷平衡及农学阈值

为研究长期施肥条件下土壤磷平衡和土壤有效磷对磷盈亏的响应,明确土壤有效磷(Olsen-P)的农学阈值及合理磷肥施用量,依托甘肃平凉肥料长期定位试验(始于1979),分析了黄土旱塬黑垆土36 a土壤磷盈亏动态、累积磷盈亏与有效磷的响应关系以及土壤磷残余,通过Mitscherlich方程模拟作物相对产量对土壤有效磷的响应关系,计算黄土旱塬黑垆土小麦和玉米的土壤有效磷农学阈值。结果表明:不施肥(CK)和单施氮肥(N)处理土壤磷始终亏缺,N处理每亏缺磷100 kg·hm~(-2),有效磷含量下降1.05 mg·kg~(-1);施磷处理土壤磷当季盈余4.3～207.9 kg·hm~(-2),累积盈余154.9～7 483.6 kg·hm~(-2),其中有机、无机配施(MNP)处理磷盈余最大;土壤累积磷盈余与土壤有效磷增量呈线性正相关,土壤中每盈余100 kg·hm~(-2)磷,秸秆还田配施化肥(SNP,磷肥隔年施)、氮磷配施(NP)、有机肥(M)和有机、无机配施(MNP)处理土壤有效磷分别增加7.55、2.47、0.28 mg·kg~(-1)和0.46 mg·kg~(-1);黄土旱塬黑垆土农田有效磷的小麦和玉米农学阈值分别为22.05 mg·kg~(-1)和13.96 mg·kg~(-1),MNP处理土壤有效磷含量已高于作物农学阈值,NP和SNP处理土壤有效磷含量达到小麦农学阈值分别需要21 a和24 a,达到玉米农学阈值分别需要2 a和8 a,M处理土壤有效磷含量已高于玉米农学阈值,还需要3 a可达到小麦农学阈值。当磷投入量每年平均达22.9 kg·hm~(-2)时,土壤磷呈持平状态;当磷用量达33 kg·hm~(-2)时,不仅作物产量较高,而且磷肥当季利用率也较高;当磷用量增加到233 kg·hm~(-2)时,作物产量对增加磷投入无响应,土壤磷残余超过90%,大量磷素累积在土壤中,增加了土壤磷素的流失风险。黄土旱塬小麦玉米一年一熟轮作黑垆土农田土壤有效磷农学阈值为13.96 mg·kg~(-1)(玉米)和22.05 mg·kg~(-1)(小麦),秸秆还田可促进旱地农田耕层土壤有效磷含量的增加。     

Dense planting with less basal nitrogen fertilization might benefit rice cropping for high yield with less environmental impacts

Dense planting and less basal nitrogen (N) fertilization have been recommended to further increase rice (Oryza sativa L.) grain yield and N use efficiency (NUE), respectively. The objective of this study was to evaluate the integrative impacts of dense planting with reduced basal N application (DR) on rice yield, NUE and greenhouse gas (GHG) emissions. Field experiments with one conventional sparse planting (CK) and four treatments of dense planting (increased seedlings per hill) with less basal N application were conducted in northeast China from 2012 to 2013. In addition, a two-factor experiment was conducted to isolate the effect of planting density and basal N rate on CH₄ emission in 2013. Our results show that an increase in planting density by about 50% with a correspondingly reduction in basal N rate by about 30% (DR1 and DR2) enhanced NUE by 14.3–50.6% and rice grain yield by 0.5–7.4% over CK. Meanwhile, DR1 and DR2 reduced GWP by 6.4–12.6% and yield-scaled GWP by 7.0–17.0% over CK. According to the two-factor experiment, soil CH₄ production and oxidation and CH₄ emission were not affected by planting density. However, reduced basal N rate decreased CH₄ emission due to it significantly reduced soil CH₄ production with a smaller reduction in soil CH₄ oxidation. The above results indicate that moderate dense planting with less basal N application might be an environment friendly mode for rice cropping for high yield and NUE with less GHG emissions.     

Design and Experiment of Slave Computer Control System for Applying Variable-rate Liquid Fertilizer

In order to increase the applying rate of liquid fertilizer and reduce environmental pollution, a slave computer control system for applying variable-rate liquid fertilizer was designed. The system used SMC as core processor and electrically controlled pressure regulator as execution component. The characteristic equation of the system was obtained by using classical control theory. Results indicated that the characteristic equation met the requirements of routh-criterion, which indicated the working process of the system was stable. Performance of the slave computer was verified via bench tests. Results demonstrated that there was no significant influence on the response from interclass error. The fertilization error was less than 0.9, and the fertilization accuracy was larger than 97%. The liquid fertilizer emitted by the fertilizing devices had no significant difference in uniformity, which met the demands of the slave computer control system for applying variable-rate liquid fertilizer.     

Long-term fertilization effects on active ammonia oxidizers in an acidic upland soil in China

The effects of long-term fertilization of acidic soils on ammonia-oxidizing archaea (AOA) and bacteria (AOB) communities and its ecological implications remain poorly understood. We chose an acidic upland soil site under long-term (27-year) fertilization to investigate ammonia oxidizer communities under four different regimes: mineral N fertilizer (N), mineral NPK fertilizer (NPK), organic manure (OM) and an unfertilized control (CK). Soil net nitrification rates were significantly higher in OM soils than in CK, N or NPK soils. Quantitative analysis of the distribution of amoA genes by DNA-based stable isotope probing revealed that AOA dominate in CK, N and NPK soils, while AOB dominate in OM soils. Denaturing gradient gel electrophoresis and clone library analyses of amoA genes revealed that Group 1.1a-associated AOA (also referred to as Nitrosotalea) were the most dominant active AOA population (>92%), while Nitrosospira Cluster 3 and Cluster 9 were predominant among active AOB communities. The functional diversity of active ammonia oxidizers in acidic soils is affected by long-term fertilization practices, and the responses of active ammonia oxidizers to mineral fertilizer and organic manure are clearly different. Our results provide strong evidence that AOA are more highly adapted to growth at low pH and low substrate availability than AOB, and they suggest that the niche differentiation and metabolic diversity of ammonia oxidizers in acidic soils are more complex than previously thought.