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。     

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.     

Mapping QTL of Fiber Yield and Quality Traits in F2 Populations of Chromosome Segment Substitution Lines from Gossypium hirsutum × Gossypium barbadense

In this study, F₂ populations from two chromosome segment substitution lines MBI7455 and MBI7358, were quantified using SSR to evaluate the parents' genotype and detect QTL related to fiber quality plus yield traits of cotton. Results show that the recurrent parent (CCRI45) hosted 96.70% and 95.60% of chromosome segment substitution in MBI7455 with 12 chromosome segments and in MBI7358 with 16 chromosome substitution segments of Gossypium barbadense, respectively. In the F₂ population, the average rate of chromosome substitution of the recurrent parent (CCRI45) was 96.44%, and the average segments of chromosome substitution of Gossypium barbadense was 13.42, with an average segments of homozygous donor chromosome value of 3.90. Analysis showed 19 fiber quality-related QTL with a phenotypic variance of between 2.52%-13.11% and seven yield traits-related QTL with a range of 2.93%-11.40% phenotypic variance, resulting in a total of 26 QTL. The CSSLs could be used to detect QTL for fiber yield and quality traits, which offer an important foundation for the cotton molecular-assisted breeding.     

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.