Effects of forage type and gibberellic acid on nitrate leaching losses

Nitrogen (N) leaching from soil into water is a significant concern for intensively grazed forage‐based systems because it can cause a decline in water quality and is a risk to human health. Urine patches from grazing animals are the main source of this N. The objective of this study was to quantify the effect that forage type and gibberellic acid (GA) application had on N leaching and herbage N uptake from urine patches on perennial ryegrass–white clover (RGWC), Italian ryegrass and lucerne. A lysimeter study was conducted over 17 months to measure herbage growth, N uptake and N loss to water beneath each of the three forage types with the following treatments: control, urine (700 kg N/ha) and urine with GA (8 g GA active ingredient/ha). Compared with RGWC (205 kg N/ha), N leaching losses were 35.3% lower from Italian ryegrass (133 kg N/ha) and 98.5% higher from lucerne (407 kg N/ha). These differences in leaching loss are likely to be due to winter plant growth and N uptake. During the winter months, Italian ryegrass had higher N uptake, whereas lucerne had lower N uptake, compared with RGWC. The application of GA had no effect on N leaching losses, DM yield or N uptake of forage treated with 700 kg N/ha urine.     

水稻高产、优质和氮高效协同的氮素调控研究

以超级粳稻‘新稻18号’为试验材料,大田条件下,研究不同施氮量(0、232.5、255.0、277.5、300.0、322.5和345.0kg/hm2纯氮水平)对水稻产量、品质及氮肥吸收利用效率的影响,以明确高产、优质和氮高效协同的适宜氮素水平。结果表明,随氮素水平提高,水稻产量、稻米品质和氮肥吸收利用效率均呈先增后降趋势。施氮量为255.0kg/hm2纯氮处理下,产量最高为9 878.11kg/hm2,稻米的糙米率、精米率和整精米率较高,分别为84.81%、72.85%和66.94%,垩白粒率和垩白度较低,分别10.00%和2.86%,氮肥吸收利用效率较高,为41.34%。低氮肥和高氮肥处理,产量、品质和氮肥利用效率相对较低。说明,在适宜的氮肥水平下,水稻高产、优质和氮高效可达到协同一致。     

Nitrogen use efficiency of cotton (Gossypium hirsutum L.) as influenced by wheat–cotton cropping systems

Wheat–cotton rotations largely increase crop yield and improve resources use efficiency, such as the radiation use efficiency. However, little information is available on the nitrogen (N) utilization and requirement of cotton under wheat–cotton rotations. This study was to determine the N uptake and use efficiency by evaluating the cotton (Gossypium hirsutum L.) N use and the soil N balances, which will help to improve N resource management in wheat–cotton rotations. Field experiments were conducted during 2011/2012 and 2012/2013 growing seasons in the Yangtze River region in China. Two cotton cultivars (Siza 3, mid-late maturity with 130 days growth duration; CCRI 50, early maturity with 110 days growth duration) were planted under four cropping systems including monoculture cotton (MC), wheat/intercropped cotton (W/IC), wheat/transplanted cotton (W/TC) and wheat/direct-seeded cotton (W/DC). The N uptake and use efficiency of cotton were quantified under different cropping systems. The results showed that wheat–cotton rotations decreased the cotton N uptake through reducing the N accumulation rate and shortening the duration of fast N accumulation phase as compared to the monoculture cotton. Compared with MC, the N uptake of IC, TC and DC were decreased by 12.0%, 20.5% and 23.4% for Siza 3, respectively, and 7.3%, 10.7% and 17.6% for CCRI 50, respectively. Wheat–cotton rotations had a lower N harvest index as a consequence of the weaker sink capacity in the cotton plant caused by the delayed fruiting and boll formation. Wheat–cotton rotations used N inefficiently relative to the monoculture cotton, showing consistently lower level of the N agronomic use efficiency (NAE), N apparent recovery efficiency (NRE), N physiological efficiency (NPE) and N partial factor productivity (NPFP), particularly for DC. Relative to the mid–late maturity cultivar of Siza 3, the early maturity cultivar of CCRI 50 had higher N use efficiency in wheat–cotton rotations. An analysis of the crop N balance suggested that the high N excess in preceding wheat (Triticum aestivum L.) in wheat–cotton rotations led to significantly higher N surpluses than the monoculture cotton. The N management for the cotton in wheat–cotton rotations should be improved by means of reducing the base fertilizer input and increasing the bloom application.     

Achieving legislation requirements with different nitrogen fertilization strategies: Results from a long term experiment

The Nitrates Directive (91/676/EEC, Anonymous, 1991) was developed in Europe to limit environmental threats from intensive livestock farming and N fertilizer applications to crops. It imposed several rules on farmers and public bodies, one of which was nutrient fertilization plan adoption. Here we use results from the Tetto Frati (Northern Italy) Long-Term Experiment to verify the terms and coefficients in the official Italian guidelines and evaluate the limitations imposed to organic fertilization amounts. For this purpose, we mined long-term experimental data of crop yield, N uptake, N use efficiency, and soil organic matter content from miscellanea cropping systems fertilized with farmyard manure (FYM) and bovine slurry (SLU), typical of a dairy farm in Northern Italy. N fertilization efficiency indicators (Removal to Fertilizer ratio, Apparent Recovery and Nitrogen Fertilizer Replacement Value) indicated that in the long run, FYM behaved similarly to urea, and better than SLU. Even N supply rates as high as 250 kg N ha⁻¹ were justified by high rates of crop removal. In fact, among the terms of the mass-balance equation, SOM mineralization was found to be most relevant, followed by meadow rotation residual effects. We conclude that a revised Nitrates Directives application scheme could be more relaxed in its application limit of manure-N, but should be more ambitious in setting efficiency coefficients for manure fertilization.     

Level and Timing of Nitrogen Fertilizer Application to Early and Semi-early Potatoes (Solanum tuberosum L.) Grown with Irrigation on Light Soils in Norway

Trials were performed with early and semi-early potatoes to test the effects of nitrogen (N) fertilizer level (0-160 kg N ha^-1) and timing (all at planting versus half then and half either soon after emergence or 3 weeks later). All seven trials with earlies were irrigated as required, whilst different irrigation regimes (moderate versus intensive) were compared in two trials with semi-earlies. No benefit was derived from splitting the N application. Haulm growth and N uptake increased in all cases almost linearly up to the highest N level, but tuber yield did not respond in the same way. The optimum N level was 80 kg N ha^-1 for a yield of 15 Mg ha^-1, rising to 120 kg N ha^-1 for a yield of 40 Mg ha^-1. Tuber quality was lowered by the use of excess N fertilizer, particularly in the case of earlies. The quantity of mineralised N present in the soil after harvest rose sharply with above optimum fertilizer use, and the amount of N present in crop residues also increased. The likely leaching after early potatoes was estimated to be up to 80 kg N ha^-1. The proportion of fertilizer N which was not accounted for in either tuber yield, crop residues or mineral N in soil was 26% in earlies and 38% in semi-earlies.     

An experimental study of charcoal degradation in a boreal forest

Degradation rates of pyrogenic carbon (PyC) under natural environmental conditions are largely unknown. Here we present results from a field experiment monitoring the change in mass, C- and N concentrations of a variety of charcoal types in a Norwegian boreal forest over a period of 20 months. The charcoal types represent different feedstock tree species, production temperature regimes, and placements in the forest, i.e. above ground, in the humus layer or in contact with the mineral subsoil. The types of charcoal had different initial C concentrations mainly depending on their production temperature. Nevertheless, all types of charcoal at all placements in the forest showed an initial drop in their C concentrations, which subsequently rose back to reach near initial values in part of the charcoal types. In part of the charcoal types, N concentrations decreased throughout the experiment, exhibiting considerable variation among feedstock species, production temperature regime, and placements in the forest. C/N ratios rose especially in charcoal made from wood of Scots pine (Pinus sylvestris L.), and charcoal that had been stored in contact with the mineral subsoil showed the most rapid mass gain. Our results confirm the important influence of production temperature and feedstock type on the degradation of charcoal, but they also show that microbial activity and environmental conditions play significant roles in charcoal degradation and thus for the fate of pyrogenic carbon under natural conditions.     

猪急性腹泻综合征冠状病毒SYBR Green荧光定量PCR检测方法的建立及应用

为建立快速、灵敏且特异的检测猪急性腹泻综合征冠状病毒（swine acute diarrhea syndrome coronavirus,SADS-CoV）检测方法,本试验扩增SADS-CoV N基因保守区域将其克隆至pMD18-T载体。所构建的重组质粒pMD18-T-SADS-qN作为阳性质粒标准品,以其为模板建立一种SYBR Green荧光定量PCR检测方法。结果显示,所建立方法在3.31×10¹~3.31×10⁷拷贝·μL^-1模板量时,呈良好的线性关系,相关系数（R²）为0.997,斜率为-3.318。该方法特异性检测SADS-CoV;而猪流行性腹泻病毒（PEDV）、猪传染性胃肠炎病毒（TGEV）、猪德尔塔冠状病毒（PDCoV）和猪繁殖与呼吸综合征病毒（PRRSV）检测结果均为阴性。所构建的标准品检测灵敏度下限可以达到3.31×10¹拷贝·μL^-1,组内和组间变异系数均小于1%,表明其具有良好的灵敏性和重复性。用该方法检测SADS-CoV感染IPI-2I和IPEC-J2细胞后不同时间点和不同接毒剂量的复制情况,结果显示,SADS-CoV感染细胞后2 h病毒含量较低,在12~36 h病毒含量迅速增长,36 h后增长速度减缓且病毒含量维持在较高水平。分别用0、0.1、1 MOI SADS-CoV感染细胞结果显示病毒的mRAN转录水平呈现剂量依赖性增加,当MOI为1时,IPI-2I和IPEC-J2细胞病毒含量分别为10^6.7、10^5.3拷贝·mL^-1。进一步利用所建立的方法对经口服攻毒SADS-CoV仔猪的临床样本进行检测,结果发现病毒在空肠回肠含量较高,表明病毒主要定殖于空肠和回肠。综上表明,本研究建立SYBR Green荧光定量PCR检测方法能灵敏特异地检测SADS-CoV,为SADS-CoV的诊断和病毒相关基础研究提供可靠的检测手段。     

绵羊BMP15基因B2突变可视化检测技术的建立

骨形态发生蛋白15（bone morphogenetic protein 15,BMP15）基因突变对绵羊排卵率、产仔数以及繁殖力有很大影响,本研究旨在建立快速可视化检测BMP15基因B2突变的技术。将改良的扩增阻滞突变系统（amplification refractory mutation system,ARMS）与核酸染料SYBR Green Ⅰ结合,针对BMP15基因B2突变设计ARMS特异性引物,使引物下游3'端的最后一个碱基为A,并在下游引物3'端的第3位碱基处设计额外的错配,以提高引物的特异性;经ARMS PCR扩增后,向产物中加入SYBR Green Ⅰ,通过肉眼观察PCR管内的颜色变化来检测BMP15基因的B2突变。经测序发现所采集的样本均为野生型,因此,利用重叠延伸PCR构建BMP15基因B2突变型模板,测序结果显示BMP15基因B2突变型模板构建成功。ARMS PCR结果显示,ARMS特异性引物能够只扩增突变型模板,而不会扩增野生型模板,且扩增效果良好。ARMS PCR扩增后加入SYBR Green Ⅰ发现已知野生型模板与阴性对照一致,呈SYBR Green Ⅰ原始颜色橙黄色,而已知突变型模板呈亮绿色,且色差明显,肉眼可辨。用建立的可视化检测BMP15基因B2突变的技术检测50个小尾寒羊基因组DNA （随机向其中20个样本中加入构建的BMP15基因B2突变型模板）,将可视化检测的突变型样本编号与混合样本时记录的编号对比,结果表明该技术能够准确检测绵羊BMP15基因的B2突变,且准确率高达100%。将构建的BMP15基因B2突变型的模板进行梯度稀释,对本试验可视化检测体系的灵敏度进行检测,发现ARMS可视化检测技术的灵敏度达到0.006 ng/μL。因此,本研究建立的技术能够可视化地检测绵羊BMP15基因B2突变,且准确率高,有望为高繁殖力绵羊的选育提供技术支持。     

Cloning and Sequence Analysis of HA and NA Genes of One Strain of H6N6 Subtype Avian Influenza Virus Isolated from Guizhou Province

To investigate the epidemic situation of H6N6 subtype avian influenza virus (AIV) in Guizhou province,A/duck/Guizhou/013/2014 was isolated from Sansui duck in live poultry market of Guizhou in 2014,the hemagglutinin (HA) and neuraminidase (NA) genes of DK/GZ/14 were subjected to clone and sequence analysis.The results showed that HA gene had the highest nucleotide homologies (97.5%) with the duck-origin H6N6 subtype AIV isolated from Eastern China in 2009,and the strains of HA gene proteolytic cleavage sites was P-Q-I-E-T-R-G,which accordeol with the molecular characteristic of low pathogenic AIV (LPAIV).However,NA gene of A/duck/Guizhou/013/2014 had the highest nucleotide homologies (98.2%) with the duck-origin H6N6 subtype AIV isolated from Fujian in 2007.The phylogenetic tree showed that A/duck/Guizhou/013/2014 and Hunan strains located in the same branch,while three duck-origin H6N6 subtype AIV isolated from Guizhou in 2007 and A/duck/Guizhou/013/2014 located in the different branch for HA and NA genes in genetic evolution,which suggested that A/duck/Guizhou/013/2014 was far with the local H6N6 subtype.The results also clearly indicated that duck-origin H6N6 subtype AIV had genetic diversity in duck population in Guizhou.