高肥力土壤条件下不同基因型花生对氮素利用的差异

在桶栽条件下,利用¹⁵N示踪技术,选用20个基因型花生为供试材料,研究了高肥力土壤条件下不同基因型花生对氮素利用的差异.结果表明:高肥力土壤条件下花生氮素营养以土壤氮为主,根瘤固氮次之,肥料氮最低.不同基因型间花生对全氮、肥料氮、土壤氮和根瘤固氮的吸收和积累均存在显著差异,基因型间遗传变异以根瘤固氮最大,肥料氮和土壤氮相当.氮素荚果生产效率和氮肥利用率基因型间差异显著,最高值分别为最低值的3.6和2.1倍.全氮、肥料氮、土壤氮和根瘤固氮的氮素收获指数基因型间均存在显著遗传变异,且以根瘤固氮的氮素收获指数基因型间遗传变异最大.花生荚果产量与不同氮源氮素积累量及氮素收获指数、氮素荚果生产效率和氮肥利用率呈显著或极显著正相关.依据花生对不同氮源氮素吸收积累和荚果产量筛选出全氮高积累高产型、肥料氮高积累高产型、土壤氮高积累高产型和根瘤固氮高积累高产型四大类型花生,其中四大类型特征兼有的有4个花生基因型.     

缺磷条件下低分子量有机酸对大豆氮积累和结瘤固氮的影响

采用砂培方法在温室条件下研究了低分子量有机酸柠檬酸、草酸、苹果酸及3种酸的混合物对大豆植株氮素积累、结瘤和固氮的影响.结果表明：低分子量有机酸对大豆植株氮素积累有显著的抑制作用,使大豆地上部各时期氮素积累量的降低幅度分别为：苗期17.6%～44.9%,花期29.8%～88.4%,鼓粒期9.18%～69.6%,成熟期2.21%～41.7%;低分子量有机酸对大豆根瘤生长和固氮能力也有显著影响,表现为使根瘤数量、根瘤固氮酶活性和豆血红蛋白含量显著降低,降低幅度分别为11.4%～59.6%,80.5%～91.7%和11.9%～59.9%,从而使大豆的固氮效率降低,最终导致大豆的固氮量较对照显著降低(降低幅度9.71%～64.5%).低分子量有机酸对大豆氮积累、根瘤生长和固氮能力的抑制作用随浓度的增加而增加.3种有机酸中,草酸的抑制作用相对大于柠檬酸和苹果酸,3种有机酸混合后,抑制作用加强.     

1株高效花生根瘤菌的筛选与鉴定

从辽宁多地花生种植土壤及其鲜根瘤初步筛选到30株花生根瘤菌,进一步通过回接盆栽花生,测定花生根瘤数、根瘤干重,以及鲜根瘤固氮酶活性、植株全氮量等,筛选出1株结瘤固氮能力较强花生根瘤菌wz-6,通过16S rDNA序列分析,鉴定为圆明慢生根瘤菌(Bradyrhizobium yuanmingense)。为开发优质花生根瘤菌菌剂奠定基础。     

与玉米混作改善花生铁营养对其根瘤形态结构及豆血红蛋白含量的影响

通过土培方法研究了与玉米混作对花生根瘤形态结构及固氮功能的影响。结果表明,玉米与花生混作能够明显地改善花生铁营养、提高根瘤豆血红蛋白的含量。同时,单作花生根瘤细胞液泡化程度较高,正在发育的根瘤细胞内类菌体数量明显地比混作的花生低。成熟根瘤细胞类菌体周膜外空间(细胞壁以内、周膜外的空间)体积变大。说明单作花生固氮酶活性较低的原因是缺铁抑制了豆血红蛋白的合成和改变了根瘤形态结构以及类菌体的超微结构。     

氮素调控花生共生结瘤的机制研究进展

氮是花生生长发育所需的大量元素,共生结瘤固氮是花生获取氮素的主要方式之一。花生共生结瘤固氮涉及复杂的调控机理,揭示氮素对根瘤固氮的调控机制对发挥生物固氮潜力具有重要意义。本文系统总结了花生根瘤形成的“裂隙侵染”机制、花生共生结瘤和数量调控的机制以及氮素影响花生结瘤的调控机制。目前,氮素影响慢生根瘤菌与花生互作进而调控结瘤的分子机理尚不清楚,因此未来的研究重点应该集中在氮素影响花生慢生根瘤菌与花生的信号交流、根瘤数调节和营养交换机制等方面,为提高花生结瘤固氮效率和产量、减少化学氮肥施用提供理论基础。     

一种高效研究大豆根瘤共生固氮的营养液栽培体系

为建立一种既可高效结瘤固氮, 又具有一定产量的大豆(Glycine max)营养液栽培系统, 设计并进行了2个试验。首先在不同供氮条件下, 研究了接种根瘤菌对大豆的结瘤状况、固氮能力、生物量及产量的影响。结果表明, 供氮过高或过低, 均影响大豆生长、产量形成及根瘤固氮; 并且植物生长所需的最适供氮水平远高于生物固氮所需的最适供氮水平。此外, 大豆生物固氮活性最高的时期在生殖期第一期(R1期)之前。由此推断, 大豆R1期前, 供应较低的氮, 有利于根瘤形成及固氮; 而从R1期起, 应提高供氮水平, 以促进植物生长及产量的形成。在此基础上开展第2个试验, 对供氮条件进行了优化处理(即R1期前低氮供应、R1期开始中氮供应)。结果表明, 与持续供应高氮相比, 优化供氮处理不仅可获得较多固氮酶活性较高的大根瘤, 还能保持较好的生长、获得更高的百粒重及维持80%左右的产量。研究结果不仅可为高效研究大豆根瘤共生固氮提供营养液配方, 还可为大豆高产高效栽培提供试验依据。     

玉米/花生混作改善花生铁营养对花生根瘤碳氮代谢及固氮的影响

研究了石灰性土壤上玉米 (Zea mays L.) /花生 (Arachishypogaea L.)混作改善花生铁营养对花生光合速率、光合产物的运输、花生各部位糖类含量、固氮酶活性以及根瘤内碳氮代谢及其有关酶活性的影响。结果表明 ,玉米 /花生混作改善花生铁营养能够明显增强固氮酶活性 ,进而增加了间作花生根瘤氨基酸的含量 ,这主要是由于玉米 /花生混作改善花生铁营养促进了花生光合作用 ,提高光合产物数量 ,增加光合产物由地上部向地下部的运输 ,但是处理间花生根瘤蔗糖和可溶性糖含量变化不大 ,单作花生根瘤还积累较多淀粉 ,说明不是光合产物的供应导致了花生固氮活性的差异。玉米 /花生混作对花生根瘤碳水化合物代谢水平影响较大 ,混作花生根瘤异柠檬酸脱氢酶 (IDH)、苹果酸脱氢酶 (MDH)、琥珀酸脱氢酶活性明显高于单作 ,而磷酸烯醇丙酮酸羧激 (PEPCK)活性低于单作花生 ,表明混作花生根瘤内三羧酸循环代谢水平较高 ,形成类菌体直接吸收利用的能量物质苹果酸和琥珀酸多 ,能够满足类菌体的固氮需求 ,因此 ,玉米 /花生混作改善花生铁营养增强根瘤碳水化合物代谢水平是提高花生固氮作用的重要原因之一     

氮锌复合作用对混播系统白三叶生长与固氮及氮转移的影响

选用南方草地土壤——山地黄棕壤进行盆栽试验,研究了氮锌复合作用对混播系统白三叶生长、根瘤发育、固氮及固氮产物转移的影响。结果表明,氮锌复合作用对白三叶生长、根瘤发育、固氮及氮转移在白三叶不同生长阶段、不同氮锌施用范围表现出不同的效应。生长前期氮锌复合作用对白三叶茎叶及根系产量的协同效应表现在氮0-30mg/kg、锌0-6mg/kg的范围内,而生长后期协同范围扩大到氮0-90mg/kg与锌0-20mg/kg的不同组合处理中;在高于上述氮锌施用范围,氮锌之间表现为拮抗效应。与不施氮相比较,施氮显著降低了白三叶根瘤数量及生长前期固氮百分数,但适量施氮（30mg/kg）显著增加了白三叶根瘤重量及生长后期固氮百分数。在所有施氮水平中,施锌6mg/kg显著增加了根瘤数量、根瘤重量及固氮百分数。与茎叶及根系生长氮锌协同范围相比较,氮锌对根瘤发育与固氮作用的协同范围较小。固氮产物转移与白三叶根瘤衰败有密切相关。试验为我国南方牧场科学施肥提供理论依据.     

NO-3态氮对花生结瘤与固氮作用的影响

NO- 3 - N是环境介质中影响豆科植物共生固氮的重要因素。通过土培方法研究了 NO- 3 - N对花生根瘤生长发育和固氮功能的影响 ,结果表明 ,随着 NO- 3 - N施用量的增加 ,花生根瘤原基、根瘤数和根瘤干重明显降低。当 NO- 3 - N施用量为 5 0、1 0 0 mg·kg- 1时花生单株和单位根瘤固氮酶活性升高 ,而施用 NO- 3 - N为1 5 0、2 0 0 mg·kg- 1时花生的固氮酶活性较低。结果分析表明 ,环境介质中较高浓度的 NO- 3 - N可能是通过影响根瘤菌 -花生间的识别侵染 ,从而抑制根瘤原基形成并导致可见根瘤的数量减少 ;花生根瘤生长发育主要受环境中 NO- 3 - N浓度和其根系含氮量的影响 ,而与植株吸氮量关系不大。     

施氮量对不同类型花生品种衰老特性和产量的影响

为了探讨花生高产的适宜施氮量,在大田高产条件下,以珍珠豆型花生品种白沙1016和普通型花生品种花育17为材料,研究了施氮量对不同类型花生品种衰老特性和产量的影响。结果表明,两花生品种叶片叶绿素含量和光合速率、SOD和CAT活性均随着施氮量的增加而增加,MDA含量随施氮量的增加而降低,只是白沙1016品种在施氮超过135kg/hm2后上述指标增加或降低不显著,说明增施氮肥可以延缓花生叶片的衰老。在一定施氮量范围内,两花生品种有效荚果数随着施氮量的增加而增加,千克果数随着施氮量的增加而降低,导致荚果产量随着施氮量的增加而增加(珍珠豆型花生品种白沙1016施氮在0-90kg/hm2、普通型花生品种花育17在0-135kg/hm2范围内),但是超过此范围后再增加施氮量反而导致有效荚果数下降、千克果数增多、荚果产量下降。     

A functional relationship between leghaemoglobin and nitrogenase based on novel measurements of the two proteins in legume root nodules

A combination of physiological and structural measurements made on nodulated cowpea and soybean plants cultured with roots in different pO(2) permitted the expression of data in various ways. Values of leghemoglobin concentration and nitrogenase activity from the two legumes were expressed conventionally either on a per plant or per gram nodule fresh weight basis, and where microscopy was done, on the basis of nitrogenase-containing, N(2)-fixing units (i.e. per bacteroid, per infected cell, or per gram infected tissue). In both legumes, acetylene reduction, N fixed and ureide content expressed on the basis of whole plants or per nitrogenase-containing units were very significantly correlated with values of leghaemoglobin concentrations expressed in a similar manner. The use of mathematical correlations in this study involving leghaemoglobin concentrations and various indices of N(2) fixation indicated a strong functional relationship between the two proteins in symbiotic legumes. These findings confirm previous suggestions that leghaemoglobin and the nitrogenase complex are two proteins closely associated with N(2)-fixing efficiency in legume root nodules.     

Ontogenetic Changes in Root Nodule Subpopulations of Common Bean (Phaseolus vulgaris L.): II. PROTEIN CONTENT AND CARBOHYDRATE POOLS

Root nodules of common bean (Phaseolus vulgaris L. GN 1140)show different ontogenetic patterns of nitrogen fixation indifferent parts of the root system. The objective of the presentstudy was to search for nodule physiological factors which couldexplain these differences. Plants were grown to maturity undercontrolled environment conditions. Root nodules were regularlyharvested from two zones of the root system, and assayed forsoluble cytosolic content and carbohydrate status. Nodule concentrationsof soluble carbohydrates, glucose, fructose or sucrose, didnot change significantly during the pod-filling decline in nitrogenaseactivity. Nodule concentrations of sucrose appeared more relatedto nodule ontogeny than to the ontogenetic stage of the shoot.Total soluble protein and leghaemoglobin concentrations werehighly correlated to nitrogenase activity. In the top zone ofthe root system proteolytic capacity increased during pod-filling,indicating a role in nodule senescence. Proteolytic capacityalso increased in the mid zone, but to a lesser extent, andwithout correlation to nitrogenase activity. The two nodulesubpopulations showed different nitrogenase activity in relationto their leghaemoglobin and carbohydrate status. We suggestthat physiological differences within the root nodule subpopulations,possibly in combination with the distance from the shoot, makesthe top zone more susceptible to a systemic increase in proteolyticcapacity during rapid pod-filling. Key words: Nitrogen fixation, protein, carbohydrate, ontogeny, Phaseolus vulgaris     

哈茨木霉发酵液中肽类物质对豇豆根瘤结构和功能的影响

通过树脂吸附、离子交换、薄层层析、高效液相色谱系统等分离、纯化方法,从哈茨木霉(Trichoderm aharzianum)T2-16菌株发酵液中分离得到一种对豆科作物生长具促进作用的物质,通过质谱等方法鉴定为肽类物质。为探明该活性物质对豆科作物的促生机制,用该活性物质对豇豆种子浸种处理后,进行盆栽实验,通过对盆栽实验中根瘤结构与固氮活性变化的研究,结果显示,该活性物质可增加根瘤侵染组织的面积和根瘤细胞中类菌体的数量,降低根瘤细胞液泡化程度,促进根瘤中公共细胞周膜较多形成,加快类菌体的发育成熟,提高根瘤豆血红蛋白的含量,从而提高根瘤的固氮活性。     

The effect of potassium deficiency on growth and N2-fixation in Trifolium repens

The effects of potassium (K) deficiency on growth, N₂-fixation and photosynthesis in white clover ( Trifolium repens L.) were investigated using natural occurring gas fluxes on the nodules in real time of plants under three contrasting relative addition rates of K causing mild K deficiency, or following abrupt withdrawal of the K supply causing strong K deficiency of less than 0.65% in dry matter. A steady-state below-optimum K supply rate led to an increase in CO₂-fixation per unit leaf surface area as well as per plant leaf surface. However, nitrogenase activity per unit root weight and per unit nodule weight was maintained, as was the efficiency with which electrons were allocated to the reduction of N₂ in the nodules. Abrupt K removals stimulated nodule growth strongly without delay, but as K concentrations decreased in the plant tissue a significant decline in nitrogenase activity per unit root weight as well as per unit nodule mass occurred. Further, the rate of photosynthesis per unit leaf area was unaffected, while the CO₂ acquisition for the plant as a whole increased due to an expansion of total leaf area whereas the leaf area per unit leaf weight was unaffected. The ratio between CO₂-fixation and N₂-fixation increased, although not statistically significant, under short-term K deprivation as well as under long-term low K supply indicating a downregulation of nodule activity following morphological and growth adjustments. This downregulation took place despite a partly substitution of the K by Na. It is concluded that N₂-fixation does not limit the growth of K-deprived clover plants. K deprivation induces changes in the relative growth of roots, nodules, and shoots rather than changes in N and/or carbon uptake rates per unit mass or area of these organs.     

Effects of exogenous application and stem infusion of ascorbate on soybean (Glycine max) root nodules

Numerous biochemical and physiological studies have demonstrated the importance of ascorbate (ASC) as a reducing agent and antioxidant in higher plant metabolism. Of special note is the capacity of ASC to eliminate damaging activated oxygen species (AOS) including O₂^−· and H₂O₂. N₂-fixing legume nodules are especially vulnerable to oxidative damage because they contain large amounts of leghaemoglobin which produces AOS through spontaneous autoxidation; thus, ASC and other components of the ascorbate–reduced glutathione (ASC–GSH) pathway are critical antioxidants in nodules. In order to establish a meaningful correlation between concentrations of ASC and capacity for N₂ fixation in legume root nodules, soybean ( Glycine max ) plants were treated with excess ASC via exogenous irrigation or continuous intravascular infusion through needles inserted directly into plant stems. Treatment with ASC led to striking increases in nitrogenase activity (acetylene reduction), nodule leghaemoglobin content, and activity of ASC peroxidase, a key antioxidant enzyme. The concentration of lipid peroxides, which are indicators of oxidative damage and onset of senescence, was decreased in ASC-treated nodules. These results support the conclusion that ASC is critical for N₂ fixation and that elevated ASC allows nodules to maintain a greater capacity to fix N₂ over longer periods.     

The effects of shading and defoliation on nodulation and nitrogen fixation by white clover

In a glasshouse experiment, single plants of ten-weeks old white clover (Trifolium repens L.) were subjected to two levels of shading and two levels of defoliation. Nodulation and nitrogen fixation parameters were measured at six sequential harvest over four weeks. Changes in nodule number and hence nodule dry weight per plant were due to nodule decay, sloughing off and non-production and were closely related to losses in root dry weight. Severe defoliation caused degradation of leghaemoglobin, an effect which was seen in less than three days from treatment. It led also to a temporary but marked decrease in the nitrogen fixation capacity of the nodules as measured by the acetylene reduction assay. Recovery of normal activity by the nitrogenase system took about ten days. The effects of shading and defoliation on the pattern of nodulation have been described briefly.     

Leghaemoglobin oxygenation gradients in alfalfa and yellow sweetclover nodules

Respiration in support of N(2) fixation by rhizobia in legume root nodules depends on an adequate supply of O(2), but excessive O(2) can damage nitrogenase, the key enzyme. The movement of O(2) into and within the nodule is driven by gradients in the concentration of O(2) or in the oxygenation of the O(2)-carrier, leghaemoglobin. Steeper gradients may increase flux to the sites of respiration, but gradients also raise the possibility of inadequate O(2) in some nodule zones and excessive O(2) in others. No detailed study of O(2) gradients in the interior of nodules has been published previously. Spectral changes in leghaemoglobin with oxygenation, previously used to measure the average O(2) status of the nodule interior, were used to map longitudinal gradients in O(2) and in respiratory capacity in the elongated nodules of alfalfa (Medicago sativa L.) and sweetclover (Melilotus officinalis L.). Variability among nodules under air in the magnitude and direction of internal O(2) gradients was seen in both species. Despite consistently higher respiratory capacity near the meristematic tip, a majority of nodules had higher O(2) towards the tip than towards the base. These results contrast with a previous report, apparently based on limited data, but they are consistent with anatomical and tracer studies showing higher gas permeability near the tip.