The Tripartite Symbiosis Formed by Indigenous Arbuscular Mycorrhizal Fungi, Bradyrhizobium japonicum and Soya Bean Under Field Conditions

The tripartite symbiosis formed by indigenous arbuscular mycorrhizal fungi (AMF), Bradyrhizobium japonicum (Kirchner) Jordan and soya bean (Glycine max L. Merr. cv. Evans) was investigated under field conditions to test the hypotheses that: (i) the tripartite symbiosis enhances nodulation and nodule activity; and (ii) its establishment does not rely on improved phosphorus (P) uptake through the fungal partner. Soil tillage was used to produce treatments with contrasting AMF colonization potentials while the amount of B. japonicum inoculum was kept constant. Nodulation, AMF colonization and the P and nitrogen (N) nutrition of plants were evaluated at 10 and 51 (full‐bloom) days after emergence. N₂ fixation was estimated by the difference method and by the isotopic dilution method. At the early stage of plant growth, AMF hyphal colonization and nodulation were, respectively, 16 % and 33 % greater in plants from untilled than from rototilled soil. The establishment of the tripartite symbiosis was observed under field conditions, and factors other than P nutrition were critical to its formation. However, the tripartite symbiosis did not promote N₂ fixation under the high soil P conditions of this study.     

Mutations affecting nodulation in grain legumes and their potential in sustainable cropping systems

Many spontaneous and a large number of induced mutants that show altered nodulation pattern have been isolated in pea, soybean, common bean, faba bean, chickpea, groundnut and pigeonpea. Available information on nodulation mutants in these crops is summarised. The importance of nodulation mutants in basic studies on plant-microbe symbiotic interactions, nitrogen fixation and breeding of cultivars with higher yield and nitrogen fixation rate are examined. The nodulation mutants, after inoculation with specific bacterial strains or a number of different strains, show either: no nodulation (nod-), few nodules (nod+/-), ineffective nodulation (fix-), hyper nodulation (nod++) or hypernodulation even in the presence of otherwise inhibitory nitrate levels (nts). No spontaneous hypernodulation or nts mutants have been found, all have been induced in independent experiments using different cultivars of pea, soybean and common bean after mutagenising seeds. Most nodulation mutants show monogenic recessive inheritance, though semi-dominant and dominant inheritance is also reported. Nodule number is controlled by a process known as autoregulation; hypernodulating mutants show relaxed autoregulation. By grafting shoots of hypernodulating soybean mutant on normal nodulating soybean, mungbean and hyacinth bean, presence of a common, translocatable signal has been shown. Nodulation mutants have contributed to the understanding of the genetic regulation of host-symbiont interactions, nodule development and N fixation. Initially, the hypernodulating mutants were found to be poor in yield. Using the induced hypernodulating mutant, a new soybean cultivar ‘Nitrobean 60’, has been released in Australia. This cultivar is reported to have given 15% higher yield over cv. ‘Bragg,’ and contributed a higher amount of fixed N to the following cereal crop in rotation. Prospects of using the nodulation mutants in developing grain legume cultivars that combine high yield with high residual N, within the bioenergetic constraints, for developing sustainable cropping systems are examined. This revised version was published online in August 2006 with corrections to the Cover Date.     

水稻根际微生物肠杆菌E-25和肺炎克雷伯氏菌K-12固氮作用的一些生理特性

我们研究了从京郊地区水稻根际分离出的一株肠杆菌(Enterob acter sp)E-25和一株肺炎克雷伯氏菌(Klebsiella pneumoniae)K-12的固氮特性.两种菌均具有较强的固氮活力.用乙炔还原法测定,E-25的比活为552毫微克分子C_2H_4/小时.毫克菌体蛋白,K-12的比活为676毫微克分子C_2H_4/小时.毫克菌体蛋白.在对数生长期菌体蛋白迅速增长,固氮活性也随之增高.对数生长期后,菌体蛋白停止增长,固氮活性开始下降.E-25和K-12均为兼性好氧细菌,在无氧和微氧条件下固氮活力显著高于常氧条件下.E-25比K-12对氧更加敏感.这两种菌可能是在北方碱性土壤中占据优势的根际固氮细菌.由于长期适应生存条件的结果,这两种菌固氮的最适pH值均偏碱性.E-25对碳源的利用也不同于南方水稻根际分离出的肠杆菌.E-25只能利用果糖、葡萄糖和蔗糖,而不能利用琥珀酸、柠檬酸等有机酸做固氮的能源.我们推测E-25在固氮过程中所需能量主要来自糖酵解途经.     

湖南旱地土壤对铵离子的矿物固定与固定态铵释放的动力学研究

采用振荡平衡法和好气培养-间歇淋洗法研究了湖南省主要旱地土壤对添加铵离子的矿物学固定和土壤固定态铵释放的动力学特性。结果表明,供试土壤对添加铵离子的矿物固定速度很快,反应12小时后基本上达到了最大固定量;而土壤固定态铵的释放则非常缓慢,反应42天后仍未达到最大释放量。这一动力学特征有利于土壤对溶液中NH4+的保持,防止NH4+的环境污染,有效地为作物供氮和提高氮肥利用率。一级动力学方程能较好地拟合供试土壤对铵离子的矿物学固定的动力学资料,一级动力学方程和抛物扩散率方程均能较好地拟合供试土壤固定态铵释放的动力学资料,表现为相关系数r值较高,标准误SE值较低。     

氯化钠影响下蓝藻Anabaena 7120的固氮活性和去铵阻抑

正常培养液中NaCl浓度增高时,蓝藻固氮活性明显下降,去铵阻抑速率也减慢。NaCl浓度越高,阻抑程度越大。如果NaCl浓度过高,固氮酶活性即完全丧失,铵阻抑效应也不能消除。低光照度、光合抑制剂、O_2、N_2以及N_2+CO_2都加剧高浓度NaCl对蓝藻固氮酶活性和去铵阻抑的阻抑,而外源蔗糖、H_2和H_2+O_2则有一定程度的促进。     

小麦/蚕豆间作体系中的氮节约效应及产量优势

通过田间小区试验,研究了小麦/蚕豆间作条件下作物的产量优势及不同施氮水平和种植方式中土壤硝酸盐累积。研究表明,间作可以提高作物单位面积复合产量,增产幅度在6%3～3%之间,不施氮处理间作小麦产量比单作增加达84%;间作和施氮对蚕豆产量没有显著影响。不同种植方式下土壤剖面中硝酸盐累积量趋势表现为蚕豆单作>小麦、蚕豆间作>小麦单作。不施氮、施氮量为20、406、0.kg/hm2条件下,种蚕豆的土壤硝酸盐累积量分别比种小麦的土壤增加了25.4、63.5、50.9、93.4.kg/hm2,间作降低了土壤中硝酸盐累积。小麦、蚕豆间作体系中的产量优势主要是种间氮营养生态位发生了分化,蚕豆通过固定空气氮而减少对土壤有效氮的吸收,把土壤中的有效氮节约供给与之相伴的作物小麦利用。     

Relative efficiency,ureide transport and harvest index in soybeans inoculated with isogenic HUP mutants of Bradyrhizobium japonicum

Summary Differences between isogenic uptake hydrogenase (HUP) mutants of Bradyrhizobium japonicum in terms of nodule efficiency, N₂ fixation and N incorporation into various plant parts were studied in a monoxenic greenhouse experiment in order to confirm previous results with soybeans and beans inoculated with various HUP⁺ and HUP^– strains. The HUP+ revertant PJ17-1 of a HUP^– mutant (PJ17) of strain USDA DES 122 showed a completely restored relative efficiency (100% versus 78±2% for the HUP^– mutant), higher nodule efficiency (N₂ fixed per g nodules), higher ureide-N transport rates, higher N contents in pods and higher N harvest indices. All these observations confirm previous experiments with HUP⁺ and HUP^– strains.     

Competitive ability of selected Cyclopia Vent. rhizobia under glasshouse and field conditions

This study tested the competitive ability of three locally isolated Cyclopia rhizobia and strain PPRICI3, the strain currently recommended for the cultivation of Cyclopia, a tea-producing legume. Under sterile glasshouse conditions, the three locally isolated strains were equally competitive with strain PPRICI3. In field soils, the inoculant strains were largely outcompeted by native rhizobia present in the soil, although nodule occupancy was higher in nodules growing close to the root crown (the original inoculation area). In glasshouse experiments using field soil, the test strains again performed poorly, gaining less than 6% nodule occupancy in the one soil type. The presence of Cyclopia-compatible rhizobia in field soils, together with the poor competitive ability of inoculant strains, resulted in inoculation having no effect on Cyclopia yield, nodule number or nodule mass. The native rhizobial population did not only effectively nodulate uninoculated control plants, they also out-competed introduced strains for nodule occupancy in inoculated plants. Nonetheless, the Cyclopia produced high crop yields, possibly due to an adequate supply of soil N.     

氨固定对土壤微生物生物量氮矿化测定的影响

Two soils with relatively high(Soil 1)and low(Soil 2) ammonium fixation capacities were used in this study to examiune the effect of ammonium fixation on the determination of N mineralized from soil microbial biomass.Organism suspension was quantitatively introduced to Soil 1 at various rates.Both fumigation-incubation (FI) and fumigation-extraction (FE) methods were used to treat the soil.The amount of fixed NH4^ increased with increasing rate of organism-N addition.A close correlation was found between the amount of fixed ammonium and th rate of organism-m addition.The net increases of fixed NH4^ -N were equivalent to 38% and 12% of the added organism-N for FI and FE treatments,rspectively,in this specific soil.To provide isotopic evidence,^15N-labelled organism-N was added to Soils 1 and 2 at 121.4 mg N kg^-1.In FI treatment,22 and 3mg N kg^-1 of labelled N were found in the fraction of fixed NH4^ -N in Soile 1 and 2 respectively;while in FE treatment,9 mg N kg^-1 of labelled N was found in the fraction of fixed NH4^ -N in Soil 1 only.There was no labelled N in the fraction of fixed NH4^ -N in Soil 2.In all of the unfumigated (check) soils,there was little or no labelled N in the fixed fractions,probably because the organism-N added was easily mineralized and nitrified.A mean of 0.64 for KN value,the fraction of N mineralized in the killed microbial biomass,as obtained with inclusion of the net increase of fixed NH4^ -N,The corresponding value calculated with exclusion of the net increase of ficed NH4^ -N was 0.46 ,It was concluded that ammonium fixation was a problem in deterination of KN,particularly for soils with a high ammonium fixation capacity, Results also showed that microbial biomass N measurement by FFE method was less affected by ammonium process than that by FI method.     

Nitrogen fixation by Azospirillium brasilense in soil and the rhizosphere under controlled environmental conditions

Summary Acetylene reduction activity by Azospirillum brasilense, either free-living in soils or associated with wheat roots, was determined in a sterilised root environment at controlled levels of O₂ tension and with different concentrations of mineral N. In an unplanted, inoculated soil nitrogenase activity remained low, at approximately 40 nmol C₂H₄ h^-1 per 2kg fresh soil, increasing to 300 nmol C₂H₄ h^-1 when malic acid was added as a C source via a dialyse tubing system. The N₂ fixation by A. brasilense in the rhizosphere of an actively growing plant was much less sensitive to the repressing influence of free O₂ than the free-living bacteria were. An optimum nitrogenase activity was observed at 10 kPa O₂, with a relatively high level of activity remaining even at an O₂ concentration of 20 kPa. Both NO _inf3 ^sup- and NH _inf4 ^sup+ repressed nitrogenase activity, which was less pronounced in the presence than in the absence of plants. The highest survival rates of inoculated A. brasilense and the highest rates of acetylene reduction were found in plants treated with azospirilli immediately after seedling emergence. Plants inoculated at a later stage of growth showed a lower bacterial density in the rhizosphere and, as a consequence, a lower N₂-fixing potential. Subsequent inoculations with A. brasilense during plant development did not increase root colonisation and did not stimulate the associated acetylene reduction. By using the ¹⁵N dilution method, the affect of inoculation with A. brasilense in terms of plant N was calculated as 0.067 mg N₂ fixed per plant, i.e., 3.3% of the N in the root and 1.6% in the plant shoot were of atmospheric origin. This ¹⁵N dilution was comparable to that seen in plants inoculated with non-N₂-fixing Psudomonas fluorescens.