Role of extracellular polysaccharide in the colonization of wheat (Triticum vulgare L.) roots by N2-fixing cyanobacteria

The characteristics of the mucilaginous sheaths of the cyanobacteria Nostoc 2S9B and Anabaena C5 and their role in the formation of associations with the roots of wheat plants grown in liquid culture have been assessed. Light and scanning electron microscopy revealed that the filaments of Nostoc 2S9B that formed a tight association with the root surface were contained in a firm mucilaginous shell. In contrast, filaments of Anabaena C5 formed a loose association and were easily detached from the mucilage that had a sheet-like appearance and tended to disintegrate as the culture aged. Similarly, there was a tight attachment of the isolated polysaccharide from Nostoc 2S9B to the root surface and a loose attachment of the Anabaena C5 polysaccharide. When the crude polysaccharide from Nostoc 2S9B was freed from proteins by phenol or pronase treatment, its ability to adhere to the root surface was lost or considerably reduced, suggesting that a protein component contributes to the tight attachment of Nostoc 2S9B. The crude polysaccharide preparation from Nostoc 2S9B contained 2.8% (w/w) protein while that from Anabaena C5 was only 0.6% (w/w) protein. The purified exopolysaccharide from Nostoc 2S9B contained three neutral sugars and glucuronic acid, whereas fucose and a uronic acid were the main components of that from Anabaena C5. Washing the roots or treating them with different sugars did not alter the ability of Nostoc 2S9B to colonize the root surface, indicating that cyanobacterial attachment may not be specific.     

Mechanical damage of roots provides enhanced colonization of the wheat endorhizosphere by the dinitrogen-fixing cyanobacterium Nostoc sp. strain 2S9B

 When wheat (Triticum aestivum L.) seedlings are co-cultivated with Nostoc sp. strain 2S9B in hydroponics, the cyanobacterium colonizes the endorhizosphere at low frequency. Mild sonication of the roots dramatically increased the number of cyanobacteria within the root tissue. The cyanobacteria penetrated the roots in the form of motile filaments (hormogonia), and once inside, they divided and transformed into aseriate packages, which showed nitrogenase activity. Cyanobacteria were found in the intercellular spaces as well as inside the epidermal and cortex cells. By prolonging the sonication time the number of cyanobacterial packages inside the roots increased. However, after reaching a certain abundance the total cyanobacterial biomass within the roots did not change regardless of the sonication time. Besides sonication, other methods of root surface damage also brought about enhanced internal root colonization. Sonication of roots proved to be an efficient way of amplifying the internal root colonization by cyanobacteria and can be suggested as a method for creating a model system in non-legume plant-microbe studies. Received: 12 August 1999     

N2-fixation by ammonium-excreting Azospirillum brasilense in auxine-induced root tumours of wheat (Triticum aestivum L.)

Summary Wheat seedlings, treated with the auxine 2,4-dichlor-phenoxy acetic acid (2,4-D) during germination developed only a residual root system. Root elongation was extremely restricted and root tips were deformed to thick club-shaped tumours. When 2,4-D was added in a later stage of plant growth the plants developed additional nodule-like knots along primary roots. Root and shoot dry-matter production was slightly repressed in all 2,4-D treatments and N translocation from roots to shoots was repressed as well. When transferred to an auxine-free growth medium, the 2,4-D-affected roots were not capable of complete recovery. In plants inoculated gnotobiotically with Azospirillum brasilense, either with the wild type or with the NH ₄ ⁺ -excreting mutant strain C3, a 2,4-D addition increased rhizosphere acetylene-reduction activity at pO₂ 1.5 kPa. The O₂ sensitivity of root-associated nitrogenase activity tended to be reduced. The number of root-colonizing bacteria, at approximately 10⁸ colony-forming units (cfu) per g dry root, was similar in the 2,4-D treatments and untreated controls. Plant treatment with high concentrations of the chemical isomer 3,5-dichlor-phenoxy acetic acid (3,5-D) did not have comparable effects, either on plant development or on rhizosphere-associated nitrogenase activity. Root-tumour tissue inhabited by A. brasilense showed purple staining when subjected to a tetrazolium chloride solution, which may indicate intensive local nitrogenase activity in this tissue. Exposed to an ¹⁵N₂-enriched atmosphere, plants treated with 2,4-D and with A. brasilense incorporated significantly higher amounts of ¹⁵N than untreated controls. In all cases the highest values of ¹⁵N enrichment were found following inoculation with the NH ₄ ⁺ -excreting mutant strain C3. Present address: F. A. Janssens Laboratory of Genetics, Catholic University of Leuven, Willem de Croylan 42, B-3001 Heverlee, Belgium     

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.     

Acetylene reduction,hydrogen evolution and nodule respiration in Phaseolus vulgaris

Summary In an experiment performed under greenhouse conditions, four cultivars of Phaseolus vulgaris L. (Venezuela-350; Aroana; Moruna; Carioca) were inoculated with three Rhizobium leguminosarum biovar phaseoli strains (C-05; C-40 = CIAT 255; C-89 = CIAT 55) and were fertilized with an N-free mineral nutrient solution. The plants were harvested 25, 40, and 55 days after emergence and the following paramenters were evaluated: Nitrogenase activity of nodulated roots, H₂ evolution by the nodules; relative efficiency of nitrogenase; respiration rates of nodulated roots and detached nodules; dry weight and total N of stems, leaves, pods, roots, and nodules. Generally the bean cultivar, Rhizobium strain, had an effect and there was an interaction effect with both symbiotic partners, on all parameters. On average, nodules represented 23% of total root respiration but the best symbiotic combinations showed lower ratios of C respired to N fixed. The maximum N-assimilation rate (between 40 and 55 days after emergence) of 11.93 mg N plant^–1 day^–1 occurred with the symbiotic combination of Carioca × C-05, while the poorest rate of 0.55 mg N plant^–1 day^–1 was recorded with Venezuela-350 × C-89. The best symbiotic combinations always showed the highest relative nitrogenase efficiency, but the differences in N₂-fixation rates cannot be explained solely in terms of conservation of energy by recycling of H₂. This requires further investigation.     

Assessment of the effects of the photosynthesis-inhibiting herbicides Diuron,DCMU, metamitron and metribuzin on growth and nitrogenase activity of Nostoc muscorum and a new cyanobacterial isolate,strain G4

Summary The influence of the photosynthesis-inhibiting herbicides Diuron, DCMU, metamitron, and metribuzin on growth and nitrogenase activity of Nostoc muscorum and a new cyanobacterial isolate, strain G4, was studied. The experiments were performed under N₂-fixing photoautotrophic conditions. Both cyanobacteria showed a high degree of tolerance towards the herbicides tested. In the presence of metamitron (35, 70, and 140 ppm) and metribuzin (7, 14, 35, and 70 ppm) strain G4 proliferated as well as the control culture. Metamitron and metribuzin had no influence on the nitrogenase synthesis of strain G4 and N. muscorum. When treated with 1 M DCMU, strain G4 and N. muscorum showed partial inhibition for the first few days, but entirely recovered during succeeding incubation. In the presence of 10 ppm Diuron, N. muscorum and strain G4, compared to the control, showed 30% and 80% nitrogenase activity, respectively.     

Free-living nitrogen-fixing bacteria in temperate cropping systems: Influence of nitrogen source

Sustainable cropping systems rely on a minimum of external inputs. In these systems N is largely acquired in animal manures and leguminous green manures. Little is known of how these organic forms of N fertilizer influence the presence and activity of free-living N₂-fixing bacteria. High concentrations of inorganic N in soil inhibit N₂-fixation in cyanobacteria and Azotobacter spp. It is likely that manure and fertilizer applications would result in concentrations of inorganic N capable of inhibiting N₂ fixation and, ultimately, the presence of these organisms. We investigated the effect of synthetic and organic N fertilizer sources on the populations and N₂-fixation potential of free-living N₂-fixing bacteria in the Farming Systems Trial at the Rodale Research Institute. Field plots received the following N treatments prior to corn (Zea mays L.) production: (1) Legume rotations and green manures supplying about 165 kg N ha^-1; (2) beef cattle manure applied at a rate of 220 kg N ha^-1 (plus 60 kg N ha^-1 from 1994 hay plow-down); or (3) fertilizer N (urea and NH₄NO₃) applied at a rate of 145 kg N ha^-1. Soil samples were collected at two depths from corn plots four times during the growing season, and analyzed for soil moisture, soil pH, numbers of N₂-fixing cyanobacteria and Azotobacter spp., extractable NH _inf4 ^sup+ and NO _inf3 ^sup- , and potentially mineralizable N. Soil samples collected in mid-July were analyzed for nitrogenase activity (by C₂H₂ reduction) and total C and N. Populations of Azotobacter spp. and cyanobacteria were influenced only slightly by treatment; however, cyanobacteria species composition was notably influenced by treatment. Nitrogenase activity in surface soils was greatest in legume-N plots and in subsurface plots levels were greatest in fertilizer-N plots. Populations and activity of free-living N-fixing bacteria appeared to be somewhat reduced in all plots as a result of low soil pH levels and high concentrations of inorganic N across all treatments. Annual applications of N to all plots resulted in high levels of potentially mineralizable N that in turn may have reduced non-symbiotic N₂-fixation in all plots.     

Dinitrogen fixation by biological soil crusts in an Inner Mongolian steppe

Eurasian steppe ecosystems are nitrogen-limited and suffer additionally from high grazing intensities in many areas. Soil surface-bound cyanobacteria are able to fix nitrogen and can be the major source of plant available nitrogen in such ecosystems. In this study, the abundance and dinitrogen fixation capacity of the most common soil surface-bound microbial and lichen species were determined at an ungrazed, a winter-grazed, and a heavily grazed steppe site in the Xilin River catchment, Inner Mongolia, People’s Republic of China. The microorganisms were identified as Nostoc spec. and the lichen species as Xanthoparmelia camtschadalis (Ach.) Hale by a combination of classical light microscopy, confocal laser scanning microscopy and molecular analysis of the internal transcribed spacer (ITS1) region of ribosomal RNA. Both species were found exclusively at grazed steppe sites, with a clear difference in abundance depending on the grazing intensity. At the winter-grazed site, Nostoc was more abundant than Xanthoparmelia; for the heavily grazed site, the opposite was found. N₂ fixation was quantified with both the acetylene reduction method and ¹⁵N₂ incubation. Cyanobacterial colonies of Nostoc fixed N₂ vigorously, whereas X. camtschadalis did not at all. The fraction of nitrogen derived from the fixation of molecular nitrogen in Nostoc was 73%, calculated from ¹⁵N natural abundance measurements of Nostoc with X. camtschadalis as reference. The conservatively calculated N₂ uptake by Nostoc was 0.030–0.033 kg N ha⁻¹ for the heavily grazed site and 0.080–0.087 kg N ha⁻¹ for the winter-grazed site for the growing seasons of 2004 and 2005, respectively. Together with previous findings, this study demonstrates that N₂ fixation by Nostoc can potentially replace significant amounts, if not all, of the nitrogen lost in the form of N₂O and NO soil emissions in this steppe ecosystem.     

Effects of inoculation with N2-fixing cyanobacteria on the nitrogenase activity in soil and rhizosphere of wetland rice (Oryza sativa L.)

Summary A greenhouse experiment was conducted with wetland rice (Oryza sativa cv. IR-50) in a clay-loam soil (Fluventic Eutrochrept) to study the effect of cyanobacterial inoculation a mixed culture of Aulosira fertilissima, Nostoc muscorum, Nostoc spp., and Anabaena spp., applied at the rate of 0.15 g (dry weight pot^-1 or 43 kg ha^-1) on acetylene reduction activity in soil and the root system (excised root), and the grain and straw yield. The effects of applying P (40 kg ha^-1), N (60 kg ha^-1), and P+N to the soil were also evaluated. Cyanbacterial inoculation significantly increased (more than 200% on average) photo-dependent acetylene reduction activity in soils, particularly where the indigenous activity was considerably low, i.e. under unfertilized and N-fertilized conditions. The effect of inoculation was prominent at the maximum tillering and grain formation stages of the crop. This inoculation benefit was, however, marginal in P-applied soils (P and P+N), where the indigenous activity was stimulated more than threefold. The inoculation led to a remarkable increase in root-associative acetylene reduction activity after the maximum tillering stage of the crop, particularly with applied N but for other treatments this inoculation effect was not significant. Cyanobacterial inoculation also increased the grain and straw yield of the crop when N was not applied. The grain and straw yield was significantly correlated with the acetylene reduction activity in flooded soils and in the root system during the tillering and maximum tillering stages of rice growth, respectively.     

A bradyrhizobial-<Emphasis Type="Italic">Penicillium</Emphasis> spp. biofilm with nitrogenase activity improves N<Subscript>2</Subscript> fixing symbiosis of soybean

A bradyrhizobial-fungal biofilm (i.e. Bradyrhizobium elkanii SEMIA 5019-Penicillium spp.) developed in vitro was assayed for its nitrogenase activity and was evaluated for N₂-fixing symbiosis with soybean under greenhouse conditions. The biofilm showed nitrogenase activity, but the bradyrhizobial strain alone did not. Shoot and root growth, nodulation and N accumulation of soybean increased significantly with an inoculum developed from the biofilm. This study concludes that such biofilmed inoculants can improve N₂-fixing symbiosis in legumes, and can also directly contribute to soil N fertility in the long term. Further studies should be conducted to investigate the performance of these inoculants under field conditions.     

Infectivity and acetylene reduction of diazotrophic rhizosphere bacteria in wheat (Triticum aestivum) seedlings under gnotobiotic conditions

Summary Wheat seedlings were inoculated with rhizosphere nitrogen-fixing bacteria and grown gnotobiotically for 15 days. The growth medium consisted of semisolid agar with or without plant nutrients. The bacteria, isolated from roots of field-grown wheat, were three unidentified Gram-negative rods (A1, A2, E1), one Enterobacter agglomerans (C1) and two Bacillus polymyxa (B1, B2). A strain of Azospirillum brasilense (USA 10) was included for comparison.Nitrogenase activity (acetylene reduction activity, ARA) was tested on intact plants after 8 and 15 days of growth. In semisolid agar without plant nutrients, five isolates showed ARA of 0.01–0.9 nmol C₂H₄ plant^–1 h^–1, while the two strains of B. polymyxa had higher ARA of 3.3–10.6 nmol C₂H₄ plant^–1 h^–1.Plant development was not affected by inoculation with bacteria, except that inoculation with B. polymyxa resulted in shorter shoots and lower root weight.Transmission electronmicroscopy of roots revealed different degrees of infection. A. brasilense, A1 and A2, occurred mainly in the mucilage on the root surface and between outer epidermal cells (low infectivity). B. polymyxa strains and E1 were found in and between epidermal cells (intermediate infectivity) while E. agglomerans invaded the cortex and was occasionally found within the stele (high infectivity).     

Characterization of free-living cyanobacterial strains and their competence to colonize rice roots

A total of 45 cyanobacterial strains isolated from rice fields near Loktak Lake in Manipur, India were tested for their rice root colonization capacity under light and under darkness. Twenty-one of these strains showed significant colonization of rice roots. The average colonization values were 637 and 381?μg chl a g^?1 root dry wt in N₂ medium and 792 and 451?μg chl a g^?1 root dry wt in NO ₃ ^? medium under light and darkness, respectively. Thus, while the colonization was higher under light and in NO ₃ ^? medium, there was significant level of colonization under darkness in N₂ medium (381?μg chl a g^?1 root dry wt). A 16S rRNA gene fragment-based denaturing gradient gel electrophoresis analysis revealed difference in the competence of individual strains to colonize rice roots exposed to individual or mixed population. The colonization pattern of seven strains used in competition experiments was found to be biphasic. A 16S rRNA gene-based phylogenetic analysis revealed high level of molecular similarity among strains of Nostoc and Anabaena.     

Sesbania rostrata as a green manure for lowland rice: Growth,N2 fixation,Azorhizobium sp. inoculation,and effects on succeeding crop yields and nitrogen balance

Summary Root and stem nodulation, nitrogen fixation (acetylene-reducing activity), growth and N accumulation bySesbania rostrata as affected by season and inoculation were studied in a pot experiment. The effects ofS. rostrata as a green manure on succeeding wet-season and dry-season rice yields and total N balance were also studied.S. rostrata grown during the wet season showed better growth, nodulation, and greater acetylene-reducing activity than that grown during the dry season. Inoculation withAzorhizobium caulinodans ORS 571 StrSpc® (resistant to streptomycin and spectinomycin) on the stem alone or on both root and stem significantly increased N₂ fixation by the plants. Soil and seed inoculation yielded active root nodules under flooded conditions. Plants that were not inoculated on the stem did not develop stem nodules. The nitrogenase activity of the root nodules was greater than that of the stem nodules in about 50-day-oldS. rostrata. S. rostrata incorporation, irrespective of inoculation, significantly increased the grain yield and N uptake of the succeeding wet season and dry season rice crops. The inoculated treatments produced a significantly greater N gain (873 mg N pot^–1) than the noinoculation (712 mg N pot^–1) treatment. About 80% of the N gained was transferred to the succeeding rice crops and about 20% remained in the soil. The soil N in the flooded fallow-rice treatment significantly declined (–140 mg N pot^–1) but significantly increased in bothS. rostrata-rice treatments (159 and 151 mg N pot^–1 in uninoculated and inoculated treatments respectively). The N-balance data gave extrapolated values of N₂ fixed per hectare at about 303 kg N ha^–1 per two crops forS. rostrata (uninoculated)-rice and 383 forS. rostrata (inoculated)-rice.     

西红柿对聚烯烃控释肥这种新的施肥方法的反应

The response of tomato （Lycopersicon esculentum） plants basically fertilized with 0.3 g N per plant of compound fertilizer with a N：P2O5：K2O ratio of 20：10：20 to sticks of polyolefin-coated fertilizer （POCF） （LongT0 with a N：P2Os：K2O ratio of 14：12：14） applied 23 d after transplanting was investigated using rooting boxes in the greenhouse. The results at 26 and 40 d after stick fertilizer treatment showed that the use of the stick fertilizer greatly increased the production of many new fine roots from the tomato plants. Compared to the unfertilized control, root length and root length density in the stick fertilizer treatment increased by 3.6-6.7 fold. In the soil zones near the stick fertilizer, root weight and root mass density were also significantly higher for the stick fertilizer treatment. Additionally, the use of the stick fertilizer increased the N, P and K concentrations in the leaves and stems of the tomato plants. The new fine roots growing near the stick fertilizer not only absorbed more nutrients and translocated them to the shoots, but also contained more nutrients within themselves. The soil ammonium and nitrate N data showed that N released from the stick fertilizer played a major role in inducing the production of new fine roots. These results indicated that stick fertilizer could be used as an alternative to the co-situs application technique to change and control the root distribution of crops as well as to increase the potential capacity of roots for water and nutrient absorption.     

Short- and long-term effects of ammonium on photodependent nitrogen fixation in wetland rice fields of Spain

Short- and long-term experiments were conducted in the rice fields of Valencia, Spain, to determine the ecological significance of ammonium on nitrogen fixation. A significant inhibition of nitrogenase activity by ammonium, at concentrations higher than 0.5mM, was observed after 8h of incubation in short-term experiments done with a bloom of the N₂-fixing cyanobacterium Anabaena sp. In a second set of short-term experiments for in situ assays of nitrogenase activity in the field, a significant correlation between nitrogenase activity and the number of N₂-fixing cyanobacteria in soil was found. No significant inhibition of nitrogenase activity by ammonium at concentrations up to 2mM was observed in these assays after 24h of incubation. This lack of inhibition was probably due to the rapid decrease in ammonium content in the flood water. Only 5% of the ammonium initially added remained in the water 24h later. In the long-term experiments, nitrogenase activity was assayed in plots fertilized with 0, 70 and 140kgNha^–1, over the cultivation cycle, for 5 years. A partial inhibition of nitrogenase activity by deep-placed N fertilizers was observed. Differences were only significant in 2 years. Mean results from 5 years only showed significant differences between plots fertilized with 0 and 140kgNha^–1. The partial inhibition of nitrogenase activity by ammonium increased over the cultivation cycle. Inhibition was only significant in September, at the end of the cultivation cycle. Received: 28 January 1996     

受固氮活性制约的蓝藻固氮去铵阻抑及其与生理条件的关系

在不同气体环境中培养的蓝藻Ａｎａｂａｅｎａ７１２０固氮活性各异，Ａｒ＋ＣＯ２中最大，空气中次之，Ａｒ中最小。固氮活性高者去铵阻抑速率大，反之则小。它们对各种生理条件的反应不一样，固氮活性高者，其去铵阻抑速率受氧和氮的抑制小。在氢和氧加合条件下，三种活性的蓝藻去铵阻抑均加快，但活性低者慢而小些。光强减弱或加光合抑制剂时，固氮活性低者去铵阻抑速率显著小于活性高者，而添加外源的蔗糖或丙酮酸时，也是固氮活     

Paddy System with a Hybrid Rice Enhances Cyanobacteria Nostoc and Increases N2 Fixation

Biological nitrogen(N) fixation(BNF) plays a significant role in maintaining soil fertility in paddy field ecosystems. Rice variety influences BNF, but how different rice varieties regulate BNF and associated diazotroph communities has not been quantified. Airtight,field-based ~(15)N_2-labelling growth chamber experiments were used to assess the BNF capacity of different rice varieties. In addition,both the 16 S rRNA and nifH genes were sequenced to assess the influence of different rice varieties on bacterial and diazotrophic communities in paddy soils. After subjecting a rice-soil system to 74 d of continuous airtight, field-based ~(15)N_2 labelling in pots in a growth chamber, the amounts of fixed N were 22.3 and 38.9 kg ha~(-1) in inbred japonica(W23) and hybrid indica(IIY) rice cultivars planted in the rice-soil systems, respectively, and only 1%–2.5% of the fixed N was allocated to the rice plants and weeds. A greater abundance of diazotrophs was found in the surface soil(0–1 cm) under IIY than under W23. Sequencing of the 16 S rRNA gene showed significantly greater abundances of the cyanobacterial genera Nostoc, Anabaena, and Cylindrospermum under IIY than under W23.Sequencing of the nifH gene also showed a significantly greater abundance of Nostoc under IIY than under W23. These results indicate that the hybrid rice cultivar(IIY) promoted BNF to a greater extent than the inbred rice cultivar(W23) and that the increase in BNF might have been due to the enhanced heterocystous cyanobacteria Nostoc.     

Role of Azospirillum brasilense nitrate reductase in nitrate assimilation by wheat plants

Summary The nitrogen metabolism of wheat plants inoculated with various Azospirillum brasilense strains and nitrate reductase negative (NR^–) mutants was studied in two monoxenic test tube experiments. The spontaneous mutants selected with chlorate under anaerobic conditions with nitrite as terminal electron acceptor fixed N₂ in the presence of 10 mM NO₃ ^– and were stable after the plant passage. One strain (Sp 245) isolated from surface-sterilized wheat roots produced significant increases in plant weight at both NO₃ levels (1 and 10 mM) which were not observed with the NR^– mutants or with the two other strains. Similar effects were observed in a pot experiment with soil on dry weight and total N incorporation but only at the higher N fertilizer level. In the monoxenic test tube experiments plants inoculated with the mutants showed lower nitrogenase activities than NR⁺ strains at the low NO₃ ^– level (1 = mM) but maintained the same level of activity with 10 mM NO₃ ^– where the activity of all NR⁺ strains was completely repressed. The nitrate reductase activity of roots increased with the inoculation of the homologous strains and with the mutants at both NO₃ ^– levels. At the low NO₃ ^– level this also resulted in increased activity in the shoots, but at the high NO₃ ^– level the two homologous strains produced significantly lower nitrate reductase activity in shoots while the mutants more than doubled it. The possible role of the bacterial nitrate reductase in NO₃ ^– assimilation by the wheat plant is discussed.     

Effect of successive cuttings on uptake and partitioning of 15N among plant parts of Leucaena leucocephala

Summary We studied the effect of three successive cuttings on N uptake and fixation and N distribution in Leucaena leucocephala. Two isolines, uninoculated or inoculated with three different Rhizobium strains, were grown for 36 weeks and cut every 12 weeks. The soil was labelled with 50 ppm KNO₃ enriched with 10 atom % ¹⁵N excess soon after the first cutting. Except for the atom % ¹⁵N excess in branches of K28 at the second cutting, both the L. leucocephala isolines showed similar patterns of total N, fixed N₂, and N from fertilizer distribution in different parts of the plant at each cutting. The Rhizobium strain did not influence the partitioning of ¹⁵N among the different plant parts. Significant differences in ¹⁵N enrichment occurred in different parts. Live nodules of both isolines showed the lowest atom % ¹⁵N excess values (0.087), followed by leaves (0.492), branches (0.552), stems (0.591), and roots (0.857). The roots contained about 60% of the total plant N and about 70% of the total N derived from fertilizer over the successive cuttings. The total N₂ fixed in the roots was about 60% of that fixed in the whole plant, while the shoots contained only 20% of the fixed N₂. We conclude that N reserves in roots and nodules constitute another N source that must be taken into account when estimating fixed N₂ or the N balance after pruning or cutting plants. ¹⁵N enrichment declined up to about fivefold in the reference and the N₂-fixing plants over 24 weeks following the ¹⁵N application. The proportion and the amounts of N derived from fertilizer decreased, while the amount derived from N₂ fixation increased with time although its proportion remained constant.     

干旱与Cd双重胁迫对土壤-小麦-蚜虫系统Cd转移规律影响的研究

为探究干旱和重金属双重胁迫对土壤-小麦-蚜虫系统内Cd转移规律的影响,为小麦蚜虫的生态调节提供理论依据,本研究以麦长管蚜[Sitobionavenae(Fabricius)]为研究对象,用原子吸收分光光度法分别测定不同土壤Cd含量(100 mg?kg-1、200 mg?kg-1)及不同程度干旱胁迫(无胁迫、中度胁迫、重度胁迫)处理下小麦根茎叶及蚜虫体内的Cd含量。结果表明:土壤Cd含量及干旱单一胁迫均对小麦及蚜虫体内的Cd含量造成了显著影响(P0.05)。两者交互作用对小麦根部及叶部的Cd含量影响显著,而对小麦茎部及蚜虫体内Cd含量影响不显著。在相同胁迫条件下, Cd在小麦中的积累分布为根茎叶。随着干旱胁迫程度增大,小麦根部Cd含量及土壤-根转移系数降低,茎部Cd含量及根-茎转移系数升高,麦长管蚜Cd含量在土壤Cd含量100mg?kg-1下高于土壤Cd含量200 mg?kg-1;中度干旱胁迫增加了麦长管蚜体内Cd累积量,而重度干旱胁迫则降低了其体内Cd累积量。叶-蚜虫的Cd转移系数明显大于土壤-根、根-茎和茎-叶转移系数且大于1,说明Cd在麦长管蚜体内产生了生物富集作用。综上所述,干旱胁迫促进了Cd从土壤向小麦茎部转移和根部Cd累积,但抑制了Cd从根部到茎部转移和茎部Cd累积;中度干旱胁迫促进了麦长管蚜体内Cd的积累,而重度干旱胁迫抑制了麦长管蚜体内Cd的积累。