用~(15)N天然丰度法估测结瘤作物的共生固N_2量

测定了位于北京西北郊的本所农场耕地及附近几个土样的土壤总Nδ15N值,农场耕地的δ15N值为6.39±0.42‰,本所庭园土壤为5.03±0.26‰,圆明园土壤为1.66±0.49‰。农场耕地土壤总Nδ15N的垂直变化为6.09±0.77～7.71±0.67‰,水平变化为6.52±0.55～7.13±0.73‰。 无N营养液砂培大豆的测定结果表明,根瘤富含15N,δ15N值为8.32±0.16‰和10.54±0.30‰,地上部贫化15N,δ15N为—2.25±0.48～—4.16±0.75‰,说明在固N2过程中和固N2产物输送转化过程中发生了N同位素的分馏,固N2分馏因数β=1.0023～1.0042。 用15N天然丰度法估计不同大豆品种的％Ndfa表明,不同大豆品种的N素自给能力不同。固N2植物的δ15N值明显低于非固N2植物的δ15N值,一般相差3～5δ15N,说明用15N天然丰度法评价这些植物的固N2状况是可能的。然而,不固N02植物也有较低的δ15N值,即便在同一地块上,非固N2植物的δ15N值也很不相同,有待进一步研究。     

大豆根瘤及产量对生物糖氮肥和尿素的响应

为探究生物糖氮肥和尿素2种类型氮肥对大豆根瘤的调控效应,以合丰55为试验材料,设置4个处理,即:生物糖氮肥150 kg·hm-2(N1)、生物糖氮肥225 kg·hm-2(N2)、尿素150 kg·hm-2(N3)、尿素75 kg·hm-2(N4),采用随机区组设计,研究不同类型氮肥对大豆根瘤数量、根瘤干重、根系干重、地上部干物重和有效根瘤的影响。结果表明,不同处理对大豆籽粒产量的影响依次为N1N2N4N3,且各处理间差异达到极显著水平。N1的大豆籽粒产量比N2、N3、N4分别高6.24%、21.57%、9.39%。N1的根瘤数量、根瘤干重、根系干重、地上部干物重以及有效根瘤最高,最终籽粒产量最高。因此,生物糖氮肥对大豆根瘤生长发育起促进作用,进而提高大豆产量。本研究为大豆氮素营养调控和高产施肥提供了理论参考。     

氮肥对花生根系生长和结瘤能力的调控效应

为探明氮肥对花生根系生长和结瘤能力的调控效应,本研究以花育22号为试验材料,设置5个处理(N1:基肥45 kg N·hm~(-2);N2:基肥60 kg N·hm~(-2);N3:基肥24 kg N·hm~(-2)+追肥36 kg N·hm~(-2);N4:基肥75 kg N·hm~(-2);N5:基肥135 kg N·hm~(-2)),在桶栽条件下,探究氮肥水平和氮肥运筹对花生根系、根瘤生长发育的影响以及根系与根瘤性状的量化关系。结果表明,N4花生的根系长度、根系表面积和根系体积在全生育期均最高;氮肥全部作为基肥施用(N2)可促进花生前期根系的生长发育,氮肥分为基肥和追肥施用(N3)可促进花生中后期根系的生长发育;随着生育进程的推进,不同处理下根系长度、根系表面积和根系体积的变异系数逐渐增大,最大变异系数分别为29.3%、25.1%和22.1%。生育前期根瘤数目和根瘤鲜重随氮肥水平的增加逐渐降低,而生育中后期根瘤数目和根瘤鲜重则以N4最高,N3次之。根系长度、根系表面积和根系体积与根瘤数目和根瘤鲜重均呈正相关,相关系数为0.1839~0.5262,此外除根系表面积与根瘤鲜重相关性不显著外,其余均达显著或极显著水平。因此,合理施氮可有效调控花生根系和根瘤性状,协调促进根系生长和根瘤发育。本研究为花生生产合理施用氮肥提供了科学依据。     

不同形态氮添加对长白山阔叶红松林土壤团聚体CO_2和N_2O排放的影响

以长白山阔叶红松林暗棕色森林土为研究对象,研究不同形态氮(N)添加对土壤不同粒级团聚体CO2和N2O排放的影响。采用室内短期培养试验(15 d),研究对照(CK)、氯化铵(NH4Cl,含N 150 mg kg-1)和硝酸钠(Na NO3,含N 150 mg kg-1)添加对全土(bulk soil)、大团聚体(250~1000μm)、微团聚体(53~250μm)、粉粒+黏粒(53μm)土壤组分CO2和N2O排放的影响。结果表明:CO2的排放量为大团聚体微团聚体全土粉粒+黏粒;NH4+-N添加对全土和各粒级团聚体的CO2排放没有显著影响;NO3--N添加对大团聚体和微团聚体的CO2排放有促进作用,并且在微团聚体中影响显著(P0.05),但对全土和粉粒+黏粒的CO2排放影响不显著。不同形态N添加对全土和各粒级团聚体N2O排放影响不同,NO3--N添加显著促进了N2O的排放,NO3--N添加后N2O排放量为全土大团聚体微团聚体粉粒+黏粒;NH4+-N的添加抑制了N2O的排放,NH4+-N添加后的土壤大团聚体、微团聚体和粉粒+黏粒的N2O排放量间无显著差异。由此可见,不同形态N添加影响土壤组分的CO2和N2O排放,且作用效果不一。     

固氮微生物进入小麦类根瘤的特性探索

在小麦上用豆科植物根浸液并接种相庆根瘤菌进行了诱瘤试验，使用ＥＬＩＳＡ技术检测了根瘤菌时入类根瘤的比例，表明了有深入研究的必要性。电镜切片观察到进入小麦细胞间隙。测定了小麦的接种ＯＲＳ５７１后乙炔还原活性随时间的变化情况。     

N2O产生法测定土壤无机态氮15N丰度

用化学方法分别将土壤中微量的铵、硝酸盐和亚硝酸盐转化为N2O气体,然后用带自动预浓缩装置的同位素比值质谱仪测定N2O中的15N丰度.N2O中的15N丰度测量值完全符合铵、硝酸盐和亚硝酸盐的15N参考值.方法快速、简单和准确,不受空气氮的污染.特别是方法的检测限很低,每批次样品中只需含5～ 20μg N.它将有助于土壤氮素的矿化作用、硝化作用和反硝化作用的研究.     

基于15N示踪法的双根大豆系统氮素吸收和分配特性研究

【目的】施氮可以促进大豆生长并提高产量,同时会抑制根瘤生长和固氮。因此研究大豆对不同形态氮素的吸收、分配及再分配特点,可以为解析大豆氮的转运特性及施氮对根瘤的系统性抑制提供参考。【方法】利用嫁接方法,制备具有两个根部和一个地上部的双根大豆植株,在砂培条件下分别以NO₃–和NH₄+为氮源设置两种试验处理。试验Ⅰ,一侧施50 mg/L的15NO₃– 或15NH₄+(A侧),另一侧不施氮 (B侧);试验Ⅱ,一侧施50 mg/L的15NO₃– 或15NH₄+(A侧),另一侧施同形态的50 mg/L的NO₃– 或NH₄+(B侧)。于始花期 (R1) 和始粒期 (R5) 取样两次,将植株分为A根、B根、A侧根瘤、B侧根瘤、茎、叶片、叶柄、荚等部位,用于测定15N丰度、干重和氮含量等指标。【结果】试验Ⅰ和试验Ⅱ结果发现,大豆A和B两侧根瘤的15N丰度均高于自然丰度 (0.365%),说明根瘤的生长发育过程中,所需要的氮不是全部来自自身固氮,还需要从根中吸取氮。与试验Ⅰ相比,试验Ⅱ的根瘤固氮率明显下降,表明大豆植株优先吸收利用肥料氮。NO₃–与NH₄+处理相比,各器官15N丰度均没有显著性差异,说明在50 mg/L的氮浓度下,NO₃–和NH₄+对大豆的氮营养没有显著差异。试验Ⅰ和试验Ⅱ均发现大豆B侧根及根瘤的15N丰度高于自然丰度 (0.365%),且小于施加的肥料氮的15N丰度 (3.63%),表明A侧根吸收的氮会经地上部转移到B侧的根及根瘤中,即根吸收的肥料氮会以一定的比例运输到地上部,随后会再次重新分配回根及根瘤中。本试验将双根大豆系统中地上部和B侧根及根瘤看成一个氮转移系统,利用15N丰度的差异,构建了R1～R5期地上部向根及根瘤转移氮量的计算方法。经计算发现,当施氮浓度为50 mg/L时,在始花期至始粒期,根来自地上部转移的氮占根部氮积累量的28.4%～40.8%,根瘤来自地上部转移的氮占其氮积累量的14.4%～17.2%。【结论】根瘤生长所需要的氮不是全部来源于自身固氮,有一部分来源于根系吸收的氮。在有肥料氮存在时,大豆植株优先吸收肥料氮。根系吸收的肥料氮以及根瘤固氮被运输到地上部后,会再次重新分配回根及根瘤中。在50 mg/L的氮浓度下,氮素形态 (NO₃–和NH₄+) 不会影响大豆植株对氮的吸收及分配。     

利用15N标记研究氮素水平对大豆根瘤生长的影响

为了明确氮肥对大豆根瘤生长的影响,以东农47为试验材料,采用15N 标记和砂培的方法,进行了氮素营养水平对大豆根瘤生长及与根瘤固氮相关性的研究。结果表明,随外源氮水平增加,根瘤干重呈先增加后降低的变化趋势,当营养液氮浓度为50 mg/L时,有利于根瘤的生长;根瘤生长需要一定量启动氮,启动氮的作用维持到第3片复叶完全展开时（V3）对大豆根瘤的生长最有利;在盛荚期 （R4）和鼓粒始期 （R5）补充外源氮的供给抑制根瘤的生长。根瘤干重与植株中根瘤氮积累量、根瘤氮比例呈极显著正相关,根瘤干重与植株中氮积累量呈显著正相关。     

种植大豆对土壤氮素盈亏影响的估算

应用15N示踪方法,研究了种植大豆对土壤氮素盈亏的影响,并对土壤氮素盈亏进行了估算。结果表明:大豆成熟期70.4%~88.6%的氮素转移到籽粒中,大豆氮素收获率很高,导致土壤氮素亏损;秸秆还田时土壤氮素亏损量平均为39.2kg/hm2,秸秆不还田时土壤氮素亏损量平均为49.2kg/hm2;大豆根瘤固氮率与土壤氮素盈亏量呈直线相关,根瘤固氮率越高,土壤氮素亏损量越少;秸秆还田条件下,根瘤固氮率71.5%是土壤氮素盈亏平衡点,秸秆不还田时根瘤固氮率要达到80.9%才能保障土壤氮素盈亏平衡。     

施氮水平对大豆氮素积累与产量影响的研究

以绥农14为材料,利用砂培和15N标记的方法研究了施氮水平对大豆氮素积累及产量的影响。结果表明:随着施氮水平的提高,大豆全株氮素积累量及叶柄、荚皮、籽粒中氮素积累量呈现先增加后下降的趋势;高氮水平增加了叶片和茎中氮素积累量,N150较N0处理叶片的氮素积累量增加了3倍,而茎增加了5倍,但减少了根中氮素积累,降低了大豆全株和籽粒中根瘤固氮量及其所占比例,降低了肥料氮和根瘤氮的收获指数,其中根中氮素积累N150较N50处理降低了60.3%,全株根瘤氮和籽粒中根瘤氮N150较N0分别降低了74.9%、85.7%,肥料氮的收获指数N150较N50降低19.8%,根瘤氮的收获指数N150较N0降低25.5%。随着施氮水平的增加,大豆产量也呈现先增加后下降的趋势,施氮水平的增加促进了大豆植株株高、结荚高度和始荚节位的显著增加,但对节数没有明显影响,N150和N0比较株高增加了55.2%,结荚高度增加了199.7%,始荚节位增加了142.9%。     

Ammonium-excreting Azospirillum sp. become intracellularly established in maize (Zea mays) para-nodules

Maize seedlings develop nodule-like tumour knots (para-nodules) along primary roots when treated with the auxin 2,4-dichlor-phenoxy-acetic acid (2,4-D). Inoculated NH ₄ ⁺ -excreting Azospirillum brasilense cells were shown to colonize these tumours, mostly intracellularly, promoting a high level of N₂ fixation when microaerophilic conditions were imposed. The nitrogenase activity inside the para-nodules was less sensitive to free O₂ than in non-para-nodulating roots. Both light and electron microscopy showed a dense bacterial population inside intact tumour cells, with the major part of the cell infection along a central tumour tissue. The bacteria colonized the cytoplasm with a close attachment to inner cell membranes. In an auxin-free growth medium, young 2,4-D-induced para-nodules grew further to become mature differentiated root organs in which introduced bacteria survived with a stable population. These results provide evidence that gramineous plants are potentially able to create a symbiosis with diazotrophic bacteria in which the NH ₄ ⁺ -excreting symbiont will colonize para-nodule tissue intracellularly, thus becoming well protected.     

果胶细菌诱导油菜结瘤与固氮研究

果胶细菌处理油菜幼根（2天苗龄、根长3cm），同时接种紫云英根瘤菌突变菌株（Rhizobiumhuakuii），诱发类根瘤的结瘤率为96.80％。从这些类根瘤中分离的菌株能回接到原寄主。类根瘤的电镜观察可以明显地见到细胞内充满了细菌。乙炔还原法测定的固氨酶活性（C2H4）为76.80nmol／（planth）。     

外源GA3对高原夏季胡萝卜肉质根发育的影响

为了揭示外源GA3对胡萝卜肉质根发育的影响机理,并为高原夏季胡萝卜安全高效栽培技术研究提供参考理论,本试验以耐先期抽薹的胡萝卜品种为试材,采用酶联免疫吸附法(ELISA),研究了外源GA3对高原夏季胡萝卜肉质根膨大、木质化及其内源激素含量的影响。结果表明,在叶片旺盛生长期和肉质根迅速膨大期分别喷施5.78×10-4 mol·L-1外源GA3 1次,或在苗期、叶片旺盛生长期和肉质根迅速膨大期喷施5.78×10-4 mol·L-1外源GA3 1~3次,能显著降低高原夏季胡萝卜苗期肉质根日鲜重增长速率与库活力,促进肉质根木质化。与对照相比,喷施外源GA3,可显著降低苗期肉质根内源GA3、IAA、GA4、ABA、DHZR与IPA含量,显著提高肉质根内源ZR含量;显著降低叶片旺盛生长期肉质根内源GA3、IAA、GA4、IPA含量,显著提高内源ZR含量;显著提高迅速膨大期肉质根内源IAA、GA4、ZR含量;显著降低肉质根迅速膨大期之前的IAA/ZR和GA4/ZR比值。随着外源GA3喷施次数的增多,IAA/ZR和GA4/ZR比值在肉质根迅速膨大期有逐渐增高的趋势。外源GA3通过改变内源激素含量及其平衡状态来影响高原夏季胡萝卜肉质根的发育。     

Endophytic establishment of Azorhizobium caulinodans through auxin-induced root tumors of rice (Oryza sativa L.)

Rice seedlings developed nodule-like tumors (para-nodules) along primary and secondary roots when treated with the auxin 2,4-dichlor-phenoxy-acetic acid (2,4-D). Histologically, these tumors appeared as cancerous out-grown lateral-root primordes and were thus comparable with stem nodules of the legume Sesbania rostrata. Azorhizobium caulinodans (a diazotroph known as a specific endophyte of Sesbania rostrata) was introduced and became established inside rice para-nodules and in root tissues around tumor bases. The infection with A. caulinodans followed a typical crack-entry invasion at places where paranodule tumors had emerged through the root cortex and epidermis. The bacteria settled with high cell densities in intercellular spaces of the induced tumors and between root cortical cells. Infection of plant cells took place both in the epidermis and in cortical tissue. Intracellularly established A. caulinodans was found inside the cytoplasm, surrounded by membrane-like structures. N₂ fixation by tumor-inhabiting Azorhizobium sp. was increased at low O₂ tensions (1.5–3 kPa) compared with an untreated control. Only a little activity remained at O₂ tensions of 5 kPa and above. The present results confirm that root-tumor induction offers a suitable method of establishing diazotrophs endophytically in the roots of gramineous crops.     

VA菌根真菌对转移Ri T-DNA胡萝卜根器官的侵染

在无菌条件下观察珠状巨孢囊霉（Gigaspora margarita）对转移ＲｉＴ-ＤＮＡ胡萝卜根器官的侵染过程。采用发根土壤杆菌（Agrobacterium rhizogenes）诱导出细胞中整合有ＲｉＴ-ＤＮＡ胡萝卜的不定根作宿主,比较了孢子的４种表面消毒方法,经过２～３个月的双重培养,看到ＶＡ菌根真菌对根器官的侵染,并获得成熟孢子。形成的孢子无须休眠,只要提供发芽条件,就能立即发芽,接种到根的周围又能侵染。文中对侵染的全过程作了较详细的描述,并对此项技术在菌根研究中的意义进行了讨论     

INFLUENCE OF CHEMICAL FORM AND CONCENTRATION OF NITROGEN ON APOPLASTIC pH OF LEAVES

The objective of this study was to investigate the effects of various forms of nitrogen (NO^? ₃, NH⁺ ₄) supplied to the roots via a nutrient solution on the apoplastic pH in intact leaves determined by fluorescence ratio imaging. In contrast to NH⁺ ₄, higher apoplastic pH values in leaves of Phaseolus vulgaris and Helianthus annuus were measured with NO^? ₃ nutrition. In this context no significant differences were found in leaves of Vicia faba and Zea mays supplied with the various forms of N. Comparative studies on apoplastic pH in leaves of Vicia faba, Zea mays and Helianthus annuus demonstrated that NO^? ₃ reductase activity in roots was responsible for the differences in NO^? ₃ concentration and pH in the leaf apoplast. Light-induced pH changes in the leaf apoplast also occur and may overlap the effects of various forms of N. Increasing concentrations of NO^? ₃ supply to the roots did not significantly affect apoplastic pH in leaves of Helianthus annuus. Depletion of NO^? ₃ in the nutrient solution led to lower apoplastic pH in leaves of Zea mays. Leaf fertilization with NH⁺ ₄ led to a decline in apoplastic pH of leaves whereas NH₃ gas exposure caused a biphasic response in apoplastic pH.     

Nitrate versus chloride nutrition effects in a soil‐plant system on the growth,nitrate accumulation,and nitrogen,potassium, sodium,calcium, and chloride content of carrot

The experiment was conducted to evaluate the effects of various nitrate/ chloride (NO₃/Cl) ratios on growth, nitrate accumulation, and mineral absorption in carrot, Daucus carota L., plants in a controlled environment. The experiment included two Cl sources [potassium chloride (KC1) and calcium chloride (CaCl₂)] and five NO/Clratios at 100/0, 90/10, 80/20, 70/30, and 60/40 with total‐nitrogen (N) concentration of 400 mg NO₃ kg^‐1 soil in 100/0 treatment. Fresh and dry weights of shoots and storage roots, and length and diameter of storage roots increased significantly with mixed NO₃/C1 treatments with both Cl sources as compared to single NO₃ (100/0) treatment. Growth was enhanced up to the 80/20 NO₃/C1 treatments. With Cl present in the treatments, the concentration of total‐N unchanged and NO₃ decreased in plants, and Cl and potassium (K) increased with the Cl sources. In KC1 treatments, Na absorption decreased. Calcium (Ca) content of the plants significantly differed by the treatments. It was concluded that N fertilization provided with combined Cl forms and NO₃/Cl rates can enhance production of better quality carrot and at the same time decrease of the N fertilizer input.     

不同水稻品种根尖吸收NO3-过程中表皮细胞膜电位变化特征

越来越多的结果表明给水稻提供部分硝酸盐营养可促进水稻的氮素总吸收量并明显改善水稻的生长发育。利用微电极技术分别测定了4个水稻品种即农垦57(粳稻)、泗优917(杂粳)、扬稻6号(籼稻)和汕优63(杂籼)幼苗根尖表皮细胞在3种NO3^-浓度(0．1mmol L^-1、1mmol L^-1、10mmol L^-1)处理过程中膜电位的变化特征。结果表明：水稻根系吸收NO3^-引起膜的去极化，去极化到一定程度出现复极化。去极化程度随外界处理液中NO3^-浓度的增加而加强，0．1mmol L^-1 NO3^-处理产生的去极化值平均为7mV，1mmol L^-1 NO3^-处理的平均为11mV，10mmol L^-1 NO3^-处理的平均为21mV；就复极化来说，0．1mmol L^-1 NO3^-处理自动出现复极化，1mmol L^-1 NO3^-处理和处理10mmol L^-1 NO3^-处理只有当除去NO3^-时才出现复极化。就单位时间膜电位变化大小而言，扬稻6号对外界NO3^-较敏感，3种NO3^-浓度引起的去极化值均高于其他3个品种，表现出对NO3^-的吸收能力较强；泗优917和汕优63表现出相似的去极化大小和相似的反应时间，而农垦57对NO3^-相对不敏感，3种NO3^-浓度引起的去极化值均低于其他3个品种，表现出对NO3^-的吸收能力较弱。另外，有部分品种的水稻根在吸收NO3^-以后表现为膜电位先超极化后去极化。上述结果表明用根系对NO3^-响应的细胞膜电位变化来研究水稻的硝酸盐营养是有可行性的。     

Timing Nitrogen Applications for Quality Tops and Healthy Root Production in Carrot

When carrots (Daucus carota L.) are mechanically harvested, sufficient nitrogen (N) must be balanced between the roots and carrot tops; weak tops reduce yield. A 2-year study was conducted in Montcalm County, Michigan, where four replications of four N treatments (45, 90, 135, and 180 kg ha^?1), were arranged in a randomized complete block design. Results showed the importance of determining pre-existing N sources, in as much as the deep taproot of carrot accessed unmeasured N in the subsoil and nitrate concentrations in irrigation water added N. The greatest yield occurred at 153–189 kg ha^?1 available N while tops continued to take up N through 200–232 kg ha^?1. When the last N application was made less than 35 days before harvest, the rate of N uptake exceeded dry-matter accumulation rate.     

Methionine sulfoximine suppressed the stimulation of dark carbon fixation by ammonium nutrition in wheat roots

To evaluate the role of NH₄ ⁺ assimilates in dark carbon fixation in roots in providing carbon skeletons expended for NH₄ ⁺ assimilation, the rate of dark carbon fixation in roots was measured using NaH¹⁴CO₃. The ¹⁴C-metabolites were analyzed in wheat (Triticum aestivum L.) plants grown in NH₄ ⁺ media for various periods of time with or without methionine sulfoximine (MSX) treatment. The dark carbon fixation rate in the roots of wheat plants that had been grown with NH₄ ⁺ for 1 d was approximately 6-fold higher than the rate in control roots. The stimulation of dark carbon fixation in NH₄ ⁺-grown plants, however, was not observed in MSX-treated roots. In the roots of NH₄ ⁺-grown plants, the concentration and ¹⁴C-Iabeling of acidic metabolites such as citrate and malate considerably decreased whereas those of basic metabolites, especially asparagine, increased noticeably. With MSX treatment, the incorporation of ¹⁴C into basic metabolites was negligible. In response to NH₄ ⁺, phosphoenolpyruvate carboxylase (PEPC) activity increased, and PEPC proteins accumulated in wheat roots. Neither activity nor amounts of PEPC in roots increased in the presence of MSX. These findings suggest that primary assimilation of NH₄ ⁺ in roots is essential for the stimulation of dark carbon fixation, which coincides with the increased activity of root PEPC, to sufficiently replenish carbon skeletons necessary for NH₄ ⁺ assimilation.