果梅幼树对春施~(15)N-硫铵的吸收与分配

以盆栽3年生细叶青梅/毛桃为试材,研究了早春施用~(15)N-(NH_4)_2SO_4条件下,果梅对~(15)N的吸收分配规律。结果表明:由于春季土温较低,限制了植株对肥料氮的利用率。在新梢旺长期,植株从肥料氮中吸收的氮素营养主要用于新生器官的建造,且新梢成为~(15)N的主要分配中心,其次即为果实,再其次为细根。至花芽分化期,植株的生长中心已发生转移和分散,但春施氮对促进当年生枝的花芽分化和维持叶片正常光合功能仍有重要作用,此期亦是根系生长的关键时期之一,且与贮氮相比,春施氮更有利于当年新根的萌发和根系的扩大。     

牛白细胞介素15基因的克隆及在大肠杆菌中的表达

根据GenBankTM中发表的牛IL-15基因序列设计并合成了特异性引物,以经LPS和ConA刺激的牛外周血单个核细胞提取的总RNA为模板,用RT-PCR方法扩增出长度约500 bp的目的基因片段,并将该基因克隆到pMD18-T载体上.酶切、PCR及序列测定结果表明,获得了牛IL-15全长基因的克隆.进一步通过PCR方法得到缺失其N端48个氨基酸的信号肽序列的牛IL-15成熟蛋白基因,将其亚克隆到原核表达载体pET-32a上,构建重组表达质粒pET-IL-15,转化大肠杆菌BL21(DE3),经IPTG诱导,该菌表达以包涵体形式存在的相对分子质量约为33 000的融合蛋白.用Ni螯合层析方法纯化表达的蛋白.Western-blot试验显示表达的重组融合蛋白能与抗6×His的单克隆抗体发生反应,同时也能与鼠抗猪IL-15的多克隆抗体发生明显交叉反应.     

梨幼树到结果初期春施15N–尿素的利用及其在土壤的残留与损失

2014—2016年,以‘黄冠’梨为材料,采用15N示踪技术研究了从幼树期到结果初期梨树对春季施用氮素的吸收利用及土壤残留与损失情况。研究结果表明,幼树期(2014—2015年)梨树生长以中心干和粗根等树体骨干结构建立为主,生长量相对较小;进入结果初期(2016年)后树体生长表现为树体骨干结构建立为主,枝梢等营养器官生长与产量形成并存,生长量大幅增加。整个试验期间,树体贮藏器官的标记氮素吸收量较大,其中幼树期中心干吸收量最大,结果初期粗根吸收量最大。0~100 cm土层标记氮素残留量随土层深度和施用年限增加逐渐降低,其中,施用标记氮素后第1年(2014年),土壤标记氮素残留量较高,残留率达63.61%,梨幼树对标记氮素利用率仅为3.25%。随后两年(2015—2016年)土壤残留量较低,树体对标记氮素利用率仅为0.51%和0.80%。试验结束时,幼树期到结果初期梨树对标记氮素的累计利用率为4.57%,土壤标记氮素残留量为20.34%,损失率达75.07%。     

食用蕈菌天然食品防腐剂发酵工艺的研究

将从食用蕈菌筛选出来的天然食品防腐剂生产菌Ｅ．Ｆ．１５和Ｅ．Ｆ．４２分别进行深层发酵,用既定筛选模型系统的跟踪模型逐一测试其发酵产物的抑菌效价。梯度试验结果表明,Ｅ．Ｆ．１５和Ｅ．Ｆ．４２的最适发酵周期,最适发酵温度,发酵液最适起始ｐＨ,摇床最适转速,发酵液最适装量和最适接种量（ｖ／ｖ）,分别为６ｄ,２５～３０℃,７．０～７．５,１５０ｒ／ｍｉｎ,２００ｍＬ（５００ｍＬ三角瓶）和１５％。     

Isotopic fractionation during N2 fixation by four tropical legumes

To quantify the contribution of biological nitrogen fixation (BNF) to legume crops using the ¹⁵N natural abundance technique, it is necessary to determine the ¹⁵N abundance of the N derived from BNF—the B value. In this study, we used a technique to determine B whereby both legume and non-N₂-fixing reference plants were grown under the same conditions in two similar soils, one artificially labelled with ¹⁵N, and the other not. The proportion of N derived from BNF (%Ndfa) was determined from the plants grown in the ¹⁵N-labelled soil and it was assumed that the %Ndfa values of the legumes grown in the two soils were the same, hence the B value of the legumes could be calculated. The legumes used were velvet bean (Mucuna pruriens), sunnhemp (Crotalaria juncea), groundnut (Arachis hypogaea) and soybean (Glycine max) inoculated, or not, with different strains of rhizobium. The values of %Ndfa were all over 89%, and all the legumes grown in unlabelled soil showed negative δ¹⁵N values even though the plant-available N in this soil was found to be approximately +6.0‰. The B values for the shoot tissue (B_s) were calculated and ranged from approximately −1.4‰ for inoculated sunnhemp and groundnut to −2.4 and −4.5‰ for soybean inoculated with Bradyrhizobium japonicum strain CPAC 7 and Bradyrhizobium elkanii strain 29W, respectively. The B (B_wp) values for the whole plants including roots, nodules and the original seed N were still significantly different between the soybean plants inoculated with CPAC 7 (−1.33‰) and 29W (−2.25‰). In a parallel experiment conducted in monoxenic culture using the same soybean variety and Bradyrhizobium strains, the plants accumulated less N from BNF and the values were less negative, but still significantly different for soybean inoculated with the two different Bradyrhizobium strains. The results suggest that the technique utilized in this study to determine B with legume plants grown in soil in the open air, yields B values that are more appropriate for use under field conditions.     

冬小麦对不同时期施用的尿素N的吸收利用

本文应用~(15)N尿素研究了不同施肥时期下尿素N在土壤中的转化和小麦对肥料氮的吸收利用。结果表明,尿素施入土壤后,硝态氮含量急剧增加。播前底施尿素有更多的硝态氮累积在20～40cm土层中。小麦对肥料氮的吸收高峰在4月13～18日,即拔节孕穗期,高峰期的吸氮量与小麦产量呈正相关,r=0.7715。     

根际注射施肥对黄土高原苹果氮素吸收利用及产量和品质的影响

【目的】在西北黄土高原地区,春季干旱少雨和肥料利用率低限制着生产优质苹果。国外通常应用水肥一体化技术来克服水肥利用率低的问题,但由于其硬件设施要求高,投资大,短期内在我国难以推广。近年来我国采用的根际注射施肥可利用施肥枪将肥料溶液直接注入根际土壤中,施肥成本低且技术简单。本研究利用同位素15N示踪技术,研究根际注射施肥对苹果氮素吸收利用及产量品质的影响,可为黄土高原果园水肥高效利用提供依据。【方法】以9年生富士/M26/新疆野苹果为试验材料,利用15N尿素标记肥料去向,最后通过MAT-251质谱计测15N丰度,得出果树各器官和土壤的肥料利用率。同时利用叶绿素仪(SPAD-502)测定标记叶片的SPAD值,用浸以磷酸甘油溶液的海绵进行田间原位测定,得到土壤氨挥发的量,用静态箱—气相色谱法测定土壤的N2O逸失量。综合对比分析黄土高原传统环状开沟撒施肥与根际注射施肥对苹果吸收利用氮素、肥料氮在土壤中残留及果实产量和品质的影响。【结果】黄土高原苹果园根际注射施肥的优越性体现在:1)施肥后一个月内,果园土壤的气态氮素损失发生变化,根际注射施肥比传统环状开沟施肥的氨挥发总量低54.9%,同时N2O的排放通量低5.0%。2)根际注射施肥后,促进了肥料在土壤中的扩散范围,扩大了根系肥水吸收容积,叶片和果实吸收的肥料氮比例(Ndff%)在整个生长季始终处于较高水平。生长季末期,根际注射施肥的整株氮素当季吸收率为53.04%,比环状开沟施肥提高12.25个百分点,表明根际注射施肥有利于氮素更快地被吸收利用,显著提高苹果树的氮素当季利用率。3)生长季末,在0—60 cm土层内,根际注射施肥的土壤氮素残留率为36.55%,而环状开沟施肥为43.13%,前者显著低于后者。4)在整个生长季内,根际注射施肥处理下的树体新梢叶片内叶绿素含量(SPAD)值一直高于环状开沟施肥。根际注射施肥能提高苹果单株产量和单果重,其单果重和单株产量分别比环状开沟施肥处理提高了3.8%和19.7%。【结论】黄土高原地区推广的果树根际注射施肥技术可以有效提高苹果树体氮素的利用率,降低了土壤中的氮素残留。此外注射施肥的深度、注射量、密度和时间均可根据不同时期的养分需要随时调整,使水肥在土壤中均匀分布,达到节水节肥的目的。同时可避免传统施肥时挖坑作业对浅土层吸收根的损伤,降低劳动力成本。综合来看,根际注射施肥是提高黄土高原区旱地苹果树肥水利用率、产量和品质的有效方式之一。     

Nitrogen isotope analysis of free glycine in soil using isotope dilution mass spectrometry

To enable the estimation of production and consumption rates of free glycine in soils through ¹⁵N isotope dilution experiments, an isotope dilution mass spectrometric method was developed. The method, which enabled high precision N isotope ratio determination of glycine in soil extracts at δ¹⁵N levels up to 4000‰ and concentrations from approximately 2 μM, is based on the following steps: (i) addition of glycine spike to the soil extract, (ii) removal of humic substances and pre-concentration of glycine using solid phase extraction, (iii) derivatization of amino acids, (iv) separation of the derivatives using gas chromatography (GC), (v) their combustion to yield sample N₂ gas, and (vi) finally the use of N isotope ratio mass spectrometry (IRMS). Judging by uncertainty budget calculations, the precision obtained (SD=0.01-0.06 at% ¹⁵N) is sufficient for detecting differences in N isotopic ratios obtained in ¹⁵N isotope dilution experiments.     

Heterotrophic and autotrophic nitrification in two acid pasture soils

Laboratory incubation experiments, using ¹⁵N-labeling techniques and simple analytical models, were conducted to measure heterotrophic and autotrophic nitrification rates in two acid soils (pH 4.8-5.3; 1/5 in H₂O) with high organic carbon contents (6.2-6.8% in top 5 cm soil). The soils were from pastures located near Maindample and Ruffy in the Northeast Victoria, Australia. Gross rates of N mineralization, nitrification and immobilization were measured. The gross rates of autotrophic nitrification were 0.157 and 0.119 μg N g⁻¹ h⁻¹ and heterotrophic nitrification rates were 0.036 and 0.009 μg N g⁻¹ h⁻¹ for the Maindample and Ruffy soils, respectively. Heterotrophic nitrification accounted for 19% and 7% of the total nitrification in the Maindample and Ruffy soils, respectively. The heterotrophic nitrifiers used organic N compounds and no as the substrate for nitrification.     

Inter‐specific variability in organic nitrogen uptake of three temperate grassland species

We tested the inter‐specific variability in the ability of three dominant grasses of temperate grasslands to take up organic nitrogen (N) in the form of amino acids in soils of differing fertility. Amino acid uptake was determined by injecting dual labeled glycine‐2‐¹³C‐¹⁵N into the soil, and then measuring the enrichment of both ¹³C and ¹⁵N in plant tissue after 50 hours. We found enrichment of both ¹³C and ¹⁵N in root and shoot material of all species in both soils, providing first evidence for direct uptake of glycine. We show that there was considerable inter‐specific variability in amino acid uptake in the low fertility soil. Here, direct uptake of amino acid was greater in the grass Agrostis capillaris, which typically dominates low fertility grassland, than Lolium perenne, which inhabits more fertile sites. Direct uptake of amino acid for Holcus lanatus. was intermediate between the above two species. Unlike in the low fertility soil, there was no difference in uptake of either ¹³C or ¹⁵N by grasses in the high fertility soil, where uptake of mineral N is thought to be the major mechanism of N uptake of these grasses. Overall, our findings may contribute to our understanding of differences in competitive interactions between grasses in soils of different fertility status.