NH3 Volatilization,N2O Emission and Microbial Biomass Turnover from 15N-Labeled Manure Under Laboratory Conditions

On irrigated agricultural soils from semi-arid and arid regions, ammonia (NH₃) volatilization and nitrous oxide (N₂O) emission can be a considerable source of N losses. This study was designed to test the capture of ¹⁵N loss as NH₃ and N₂O from previous and recent manure application using a sandy, calcareous soil from Oman amended one or two times with ¹⁵N labeled manure to elucidate microbial turnover processes under laboratory conditions. The system allowed to detect ¹⁵N enrichments in evolved N₂O-N and NH₃-N of up to 17% and 9%, respectively, and total N, K₂SO₄ extractable N and microbial N pools from previous and recent ¹⁵N labeled manure applications of up to 7%, 8%, and 15%. One time manured soil had higher cumulative N₂O-N emissions (141 µg kg^?1) than repeatedly manured soil with 43 µg kg^?1 of which only 22% derived from recent manure application indicating a priming effect.     

开垦年限对黑土氮初级转化速率和净转化速率的影响

以东北黑土区开垦2 a和开垦30 a的典型旱作土壤为研究对象,采用15N同位素成对标记技术开展室内培养试验,利用数值计算模型(FLUAZ)计算不同开垦年限土壤的氮初级转化速率,以比较不同开垦年限黑土氮初级转化速率和净转化速率的差异,明确开垦年限对黑土氮转化过程的影响。结果表明,与开垦2a土壤相比,开垦30a土壤的有机碳和水溶性有机碳含量显著降低,导致土壤氮初级矿化速率和初级固定速率也显著降低。但开垦30a土壤的初级硝化速率、净硝化速率和净氮矿化速率却显著高于开垦2a土壤。两个开垦年限土壤的初级硝化速率分别为净硝化速率的1.15倍和1.02倍,说明土壤微生物对硝态氮的固定很少。开垦30a土壤的m/i值(氮初级矿化速率与初级固定速率之比)和n/ia值(初级硝化速率与初级铵态氮固定速率之比)均显著大于1,而开垦2 a土壤的m/i值和n/ia值均接近1。表明开垦2 a土壤的氮矿化与固定过程紧密偶联,氮素损失的风险较小,而开垦30 a土壤中氮矿化量超过了固定量,这为硝化作用的进行提供了底物,增加了硝酸盐反硝化和淋溶风险。     

Promoter identification and effect on activation of NF-κB of porcine ISG58

Interferon (IFN)-stimulated gene (ISG) 56 family (composed of ISG54, ISG56, ISG58, and ISG60) plays important roles in defense against viral infection in mammalian cells. Numerous studies have been conducted on ISG54, ISG56, and ISG60; however, little is known on ISG58. In the present study, the upstream sequence of porcine ISG58 gene was first characterized as functional promoter by luciferase reporter assay, and then two directly adjacent IFN-stimulated response elements (ISREs), one at ?206 to ?194 (ISRE-I) and a second one, directly upstream of this element at ?219 to ?207?bp (ISRE-II), were identified using the bioinformatics method. The subsequent site-directed deletion and transient transfection experiments showed the candidate ISREs are functional. ISRE-I works better than ISRE-II and synergistic cooperation exists between two ISREs. Additionally, the effect of porcine ISG58 on activation of NF-κB was analyzed using the dual-luciferase reporter assay. The results will contribute to revealing the role of ISG58 in immune response.     

稳定碳、氮同位素在牛肉加工过程中的分馏效应

为了探究牛肉及副产物中稳定碳、氮同位素在加工过程中的变化规律,确证稳定碳、氮同位素在牛肉加工制品产地溯源中的稳定性和有效性。本试验通过对牛肉进行不同时间的水煮、烤制和油炸3种处理,其中水煮和烤制加工时间分别为5、10、15、20、25和30 min,油炸加工时间分别为1、2、3、4和5 min;采用元素分析仪-稳定同位素比率质谱仪(EA-IRMS)测定脱脂牛肉、粗脂肪及副产物中δ¹³C和δ¹⁵N值。结果表明,脱脂牛肉中δ¹³C值在水煮、烤制和油炸3种加工方式不同加工时间之间均无显著差异,水煮和烤制后粗脂肪中δ¹³C值无显著变化,油炸后的牛肉粗脂肪δ¹³C值主要受植物油的影响,加工方式及加工时间对其无显著影响;水煮脱脂牛肉δ¹⁵N值在加工25、30 min时与对照组牛肉存在显著差异,但平均差值仅为0.3‰~0.9‰。牛肉稳定碳、氮同位素在不同加工过程中分馏效应较小,可有效用于牛肉加工半成品及成品的原产地溯源。     

基于15N示踪技术的干旱区滴灌葡萄氮素利用分析

为研究水氮调控对干旱区滴灌葡萄氮素吸收利用的影响,以新疆鲜食葡萄弗雷为试验材料,利用¹⁵N 示踪技术,设置2种灌水处理(灌水量为4 950、5 400 m³·hm^-2,分别记作W1、W2),3种施氮处理(施氮量为177、235、292 kg·hm^-2,分别记作F1、F2、F3)进行大田试验。结果表明,各处理土壤全氮含量和¹⁵N丰度差异极显著(P<0.01),并且在0~20 cm土层出现富集现象。各器官征调氮素能力随水氮投入加大极显著增强(P<0.01)。根系、茎秆、叶片器官的生物量随着吸氮量的增加极显著提高(P<0.01),而较高施氮量(F3)不利于果实器官生物量的积累。果树吸收的肥料氮量随水氮投入的加大逐渐增加,受水氮调控影响极显著(P<0.01),果树吸收的土壤氮量大于肥料氮量。W2F1的¹⁵N标记氮肥利用率和¹⁵N标记氮肥偏生产力最高,分别为38.36%和114.20 kg·kg^-1,W2F2的产量最高,为20 253 kg·hm^-2,但与W2F1差异不显著。表明水氮投入过多会引起茎秆和叶片器官徒长且不利于提高¹⁵N标记氮肥利用率。综上所述,灌水量5 400 m³·hm^-2、施氮量177 kg·hm^-2(W2F1)是兼顾经济效益与生态效应的可行水氮运筹模式。本研究结果为干旱区滴灌葡萄高产高效种植提供了参考。     

Dynamics of nitrogen translocation from mature leaves to new shoots and related gene expression during spring shoots development in tea plants (Camellia sinensis L.)

The contribution of N remobilization is crucial for new shoots growth and quality formation during spring tea shoots development. However, the translocation mechanism of N from source leaves to sink young shoots is not well understood. In the present study, ¹⁵N urea was applied to mature tea leaves one week before bud break to track N remobilization in a field experiment. The dynamic changes in plant ¹⁵N abundance, contents of amino acids, and the expression levels of genes related to N metabolism and translocation were followed during the 18‐d development of new spring shoots until three expanding young leaves. The results showed that during the growth of new shoots the amount of ¹⁵N in the shoots increased, whereas the N_dff (N derived from ¹⁵N‐urea) in mature leaves decreased, showing that the foliar‐applied N in mature leaves was readily exported to new shoots. This process was found to be accompanied by decline of chlorophylls. In the mature leaves, expression CsATG18a and CsSAG12 involved in autophagy was dramatically induced (> 4‐fold) at approximately nine days after the bud breaking. The genes involved in the transformation of amino acids, including primarily CsGDH2, CsGDH4, CsGLT3, CsGS1;3, and CsASN2 were upregulated by > 3‐fold after bud breaking. The expression levels of CsATG8A, CsATG9, CsSAG12, CsGS1;1, CsGDH1, and CsAAP6 correlated negatively with the N_dff in mature leaves, but positively with ¹⁵N amount and total N amount in new shoots, suggesting these genes played important roles in N export from mature leaves. In the new shoots, the expression of most genes showed two defined peaks, one on six days and one on 12 days after bud breaking. The expression of CsGS2, CsASN3, CsGLT1, and CsAAP4 positively correlated with the ¹⁵N amount and total N amount in new shoots. These genes might be involved in the transport and re‐assimilation of N from mature leaves. The overall results demonstrated that the translocation of ¹⁵N from mature leaves to new spring shoots was regulated by the genes involved in autophagy, protein degradation, amino acid transformation and transport.     

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%。     

不同营养水平日粮对陕北白绒山羊生长性能和小肠组织SLC7A7、SLC3A1及SLC15A1 mRNA表达的影响

本试验旨在研究日粮营养水平对断奶后2~6月龄陕北白绒山羊生长性能及小肠组织中与氨基酸转运吸收相关的SLC7A7、SLC3A1和SLC15A1 mRNA表达的影响。选取健康、日龄（（60±1.60）d）和体重（（10.73±1.03）kg）相近的雌性陕北白绒山羊羔羊36只，随机分为4组，分别饲喂4种试验日粮，其消化能和粗蛋白质水平分别为标准日粮的85%、100%、115%和130%。标准日粮营养水平参考肉羊饲养标准NY/T816-2004，依据生长阶段（10~19 kg、15~27 kg）和目标增重设置。试验期间，分别于120和180日龄称重，于180日龄，每个重复屠宰1只试验羊，采集十二指肠中上部、空肠中段、回肠末端组织样品，通过实时荧光定量PCR检测SLC7A7、SLC3A1和SLC15A1表达水平。结果表明：1）第一阶段（60~120 d）115%水平组平均日增重显著高于其他三组（P<0.05），第二阶段（121~180 d）115%水平组平均日增重显著高于85%和100%水平组（P<0.05），与130%水平组无明显差异。两阶段115%水平组羔羊干物质采食量极显著高于其他3组（P<0.01）。两阶段料重比115%水平组显著低于85%、100%水平组（P<0.05）。2）相同营养水平下，SLC7A7和SLC15A1 mRNA的表达丰度顺序均为回肠>空肠>十二指肠；3）随日粮营养水平的增加，SLC7A7、SLC3A1和SLC15A1 mRNA在小肠各段的相对表达量呈先上升后下降的趋势，且115%水平组表达量最高。115%水平组SLC7A7和SLC15A1 mRNA相对表达量显著高于其他3组（P<0.05）；115%水平组SLC3A1 mRNA的相对表达量显著高于85%和130%水平组组（P<0.05）。本试验条件下，与其他营养水平组相比，115%水平组陕北白绒山羊羔羊在2~6月龄生长性能最佳，SLC7A7、SLC3A1和SLC15A1 mRNA在小肠各段的表达量最高。     

3种介导猪NR2F2基因转染PK15细胞方法的比较研究

试验旨在研究3种转染方法对猪肾上皮细胞（PK15）的转染效率，为以PK15细胞为模型研究外源基因的功能提供参考。本研究以PK15细胞为研究对象，用脂质体、电穿孔、慢病毒3种转染方法转染后，在荧光显微镜下观察绿色荧光蛋白的表达，采用实时荧光定量PCR检测EGFP和NR2F2基因的表达情况，采用CCK-8检测细胞存活率，进而比较3种方法的转染效果。结果显示，转染PK15细胞后，电穿孔和慢病毒方法转染效率极显著高于脂质体（P<0.01），但电穿孔方法和慢病毒方法之间差异不显著（P>0.05）；EGFP和NR2F2基因的实时荧光定量PCR结果显示慢病毒方法效果最好，脂质体方法较差，与细胞转染效率基本一致；CCK-8结果显示，电穿孔转染后细胞存活率最低，极显著低于对照组（P<0.01），脂质体方法显著低于对照组（P<0.05），慢病毒方法与对照组间差异不显著（P>0.05）。综合考虑转染效率及细胞活性，本研究认为慢病毒转染方法最适合转染PK15细胞，为今后高效转染PK15细胞提供了理想的方法。