不同温度下TMV侵染枯斑三生烟的LncRNA差异表达

【目的】 筛选不同温度下烟草花叶病毒（Tobacco mosaic virus,TMV）侵染后枯斑三生烟（Nicotiana tabacum var. Samsun NN）差异表达的长链非编码RNA（long non-coding RNA,lncRNA）,研究lncRNA在枯斑三生烟抗性反应中的作用。【方法】 N基因的温度敏感性使枯斑三生烟在25℃时具备对TMV的抗性、在31℃抗性丧失,在这两个温度条件下对枯斑三生烟接种TMV和磷酸盐缓冲盐水（phosphate buffered saline,PBS）,48 h后提取系统叶总RNA,构建链特异性文库后进行深度测序。对测序结果进行过滤后利用HTSeq将有效数据与近缘品种TN90（N. tabacum var. TN90）基因组比对,筛选得到lncRNA后利用FPKM法估计lncRNA的表达水平。通过edgeR筛选差异表达lncRNA（differentially expressed lncRNA,DElncRNA）,并利用qRT-PCR技术对这一结果进行验证。通过共定位及共表达分析预测DElncRNA的靶基因,通过参考基因组注释、GO和KEGG富集分析研究靶基因的功能。【结果】 4个处理共12个样本经lncRNA-seq各测得约8 000万条clean reads,共获得4 737条已知lncRNA、40 169条新lncRNA。其中64个lncRNA在不同温度条件下TMV侵染后存在差异表达,qRT-PCR测定结果显示这些lncRNA的测序正确率在80%左右,表明本研究所得测序数据具备较高的可信度。对DElncRNA进行靶基因预测,发现一些基因同时被25℃下调和31℃上调的DElncRNA靶向。靶基因注释功能丰富,主要参与植物抗病、激素和代谢等生理过程。部分可能与激素通路相关的lncRNA,在25℃下TMV侵染时呈现下调趋势,而在31℃下TMV侵染则呈现上调趋势。GO富集分析显示靶基因主要参与构成膜、囊泡等组分,具备钙、钾离子通道抑制剂活性等分子功能,使相应离子得以转运引发随后的反应,同时也参与发病、抗原加工和呈现、细胞分裂素代谢等生理过程。KEGG分析发现靶基因显著富集在植物激素信号转导通路,25℃下调和31℃上调的DElncRNA靶基因同时富集在激素信号传导、ABC运输蛋白、苯丙烷类生物合成等通路。【结论】 不同温度（25℃和31℃）条件下TMV侵染枯斑三生烟后,长链非编码RNA差异表达,DElncRNA通过作用于激素信号传导、物质转运等过程参与寄主系统获得性抗性反应。研究结果可为揭示植物系统获得性抗性中lncRNA的调控功能以及新型抗病毒技术开发提供依据。     

Mass balanced based LCA of a common carp-lettuce aquaponics system

An aquaponics system (AS) is an integrated system that combines a recirculating aquaculture system and a hydroponics system (HS). It is designed to recover nutrients released from fish and transfer them to plants to provide a system more environmentally-friendly than the two systems working separately. As a result, several AS are under development, but little information is available about their overall performances. The aim of this study was to assess nutrient-use efficiency and environmental impacts of an AS, specifically a common carp-lettuce AS located in a greenhouse. Nutrient budgets of nitrogen (N) and phosphorus (P) were calculated and Life Cycle Assessment (LCA) was performed for the AS and for a lettuce Individual Hydroponics System (IHS), similar to the HS of the AS, operating in the greenhouse at the same time. The experiment was performed over a 52-day cycle, which corresponds to the growing time required to harvest marketable lettuce. The nutrient budgets were well balanced, with 24.6% of the N unaccounted-for, most likely due to N2 gas emission, and 6.6% of the P unaccounted-for, most likely due to having underestimated the quantity of sediment. At the beginning of the experiment, N represented 55.9%, 37.1% and 0.1% of the total N input in the formulated feed, stocked fish and lettuce seedlings, respectively. At the end of the experiment, N represented 47.6% and 0.4% of the total N input in the harvested fish and lettuce, respectively. At the beginning of the experiment, P represented 56.94%, 40.20% and 0.03% of the total P input in the formulated feed, stocked fish and lettuce seedlings, respectively. At the end of the experiment, P represented 51.52% and 0.42% of the total P input in the harvested fish and lettuce, respectively. LCA clearly indicated two environmental impact hotspots: the origin of nutrients and energy use. One kg of lettuce growth in the AS clearly had lower environmental impacts than that in the IHS for climate change, acidification, eutrophication, land competition and cumulative energy demand; however, a decrease in water dependence was not observed. The indicator for net primary production use highlighted the dependence of the AS on natural resources, especially fish meal and fish oil. Compared to the use of chemical nutrients in the IHS, the use of nutrients from formulated feed in the AS decreased climate change impact but increased the use of natural resources.     

Can differences in 15N natural abundance be used to quantify the transfer of nitrogen from legumes to neighbouring non-legume plant species?

Natural variations in the stable isotope ¹⁵N are often exploited in studies of N cycling in ecosystems. Lower ¹⁵N natural abundance in non-legume plants growing in association with legumes, compared with the non-legume grown alone in pure stands have been observed in cropping, forage, and agroforestry systems. Such observations have frequently been attributed to the transfer of biologically-fixed nitrogen (N) from the legume to the companion non-legume, and various methodologies have been employed to calculate the extent of the N transfer. While some of these ¹⁵N natural abundance-based estimates of N transfer were within the range previously reported using equivalent ¹⁵N-enriched techniques (<20% of non-legume plant N and <10 kg N ha⁻¹ derived from fixed N contributed by neighbouring legumes), many of the values obtained using natural abundance were much higher (30%–83% of the non-legume N derived from fixed N representing up to 30–40 kg N ha⁻¹) than generally measured by ¹⁵N-enriched methods; with even greater estimates being determined where data were available to allow N transfer to be re-calculated on the basis of total legume N rather than fixed N (42% to >100%, and up to 110 kg N ha⁻¹ per year). This review raises concerns about the assumptions behind the natural abundance approach, and provides some alternative interpretations for the observed differences in natural ¹⁵N abundance between plants grown in the presence and absence of legumes. It was concluded that simple comparative measures of non-legume δ¹⁵N alone cannot provide a quantitative estimate of N transfer between plant species if the dominant source and the isotopic identity of the transferred N cannot be validated, and if the extent of any isotopic fractionation associated with relevant N transformations occurring during transfer cannot be defined. To date this information is not forthcoming. There is a need to greatly improve our understanding of the transfer processes before the real value of the δ¹⁵N technology can be realized. In the first instance this will primarily be achieved by carefully executed experiments under controlled conditions, and in the field, employing both ¹⁵N natural abundance and enrichment approaches so estimates of transfer can be compared, and the data interrogated using modelling approaches to explore isotopic fractionation.     

国家重点实验室专项经费绩效预算管理方式应用于其他财政专项管理的思考

绩效预算是我国部门预算管理改革的重要内容。国家重点实验室专项经费绩效预算安排机制，是财政科研经费绩效预算的典型示例。文章通过剖析国家重点实验室专项经费绩效考评管理机制，对国家重点实验室绩效预算机制推广应用于其他财政专项的范围、考评指标、考评工作方法、考评结果应用等方面提出了意见建议。     

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.     

工程造价控制的几个阶段

随着社会的发展,基础设施投资力度不断加大,工程项目的建设在不断扩展,作为贯穿项目的始终的工程造价控制也势在必行,本文着重介绍了工程造价控制的几个阶段。     

吉林莫莫格白鹤春季行为研究

白鹤属大型涉禽,仅分布于亚洲地区,其东亚种群数量为3 750只左右;由于环境变迁,种群数量越来越少,已被列为国家Ⅰ级重点保护动物。为阐明在沼泽湿地生态系统环境中白鹤春季的活动规律以及各种行为的时间分配,于2010年4月15日～5月13日,在吉林莫莫格国家级自然保护区对白鹤春季迁徙期行为进行了初步研究。白鹤的各种行为的时间分配依次为:觅食占64.34%、静栖占4.42%、警戒占13.92%、游走占4.57%、理羽占11.51%、其他(包括打斗、飞行、呜叫等情况)占2.56%。成体与亚成体白鹤、雄鹤与雌鹤在行为和时间分配上有明显差异,在觅食时间上雌鹤高出雄鹤10%左右,雄鹤的警戒时间高于雌鹤。