Interactions between nitrate uptake and N2 fixation in white clover

Summary White clover (Trifolium repens L.) plants grown in pots and supplied with the same concentration x days of¹⁵N labelled nitrate, but in contrasting patterns and doses had similar N concentrations but differed in the proportions devived from N₂ fixation and nitrate. N₂-fixation and nodule dry weight responded rapidly (2–3 days) to changes in nitrate availability. Plants exposed frequently to small doses of nitrate took up more nitrate (and hence relied less on N₂-fixation) and had greater dry weights and shoot: root ratios than those exposed to larger doses less often. In mixed ryegrass (Lolium perenne L.)/clover communities clover's ability to either successfully compete for nitrate or fix N₂ gave it consistently higher N concentrations than grass whether they were given high or low nitrate nutrient. This higher N concentration was accompanied by greater dry weights than grass in the low nitrate swards but not where high levels of nitrate were applied.     

Positive effects of continuous low nitrate levels on growth and photosynthesis of Alexandrium tamarense (Gonyaulacales, Dinophyceae)

The growth and photosynthesis of Alexandrium tamarense (Lebour) Balech in different nutrient conditions were investigated. Low nitrate level (0.0882 mmol/L) resulted in the highest average growth rate from day 0 to day 10 (4.58 × 10² cells mL^?1 d^?1), but the lowest cell yield (5420 cells mL^?1) in three nitrate level cultures. High nitrate‐grown cells showed lower levels of chlorophyll a‐specific and cell‐specific light‐saturated photosynthetic rate (P_m^{chl a} and P_m^cell), dark respiration rate (R_d^chla and R_d^cell) and chlorophyll a‐specific apparent photosynthetic efficiency (α^chla) than was seen for low nitrate‐grown cells; whereas the cells became light saturated at higher irradiance at low nitrate condition. When cultures at low nitrate were supplemented with nitrate at 0.7938 mmol/L in late exponential growth phase, or with nitrate at 0.7938 mmol/L and phosphate at 0.072 mmol/L in stationary growth phase, the cell yield was drastically enhanced, a 7–9 times increase compared with non‐supplemented control culture, achieving 43 540 cells mL^?1 and 52 300 cells mL^?1, respectively; however, supplementation with nitrate in the stationary growth phase or with nitrate and phosphate in the late exponential growth phase increased the cell yield by no more than 2 times. The results suggested that continuous low level of nitrate with sufficient supply of phosphate may facilitate the growth of A. tamarense.     

Colonization with arbuscular mycorrhizal fungi improves salinity tolerance of tomato (Solanum lycopersicum L.) plants

The purpose of this study was to investigate the mechanisms underlying alleviation of salt stress by mycorrhization. Solanum lycopersicum L. cultivars Behta and Piazar with different salinity tolerance were cultivated in soil without salt (EC?=?0.63 dSm^?1), with low (EC?=?5 dSm^?1), or high (EC?=?10 dSm^?1) salinity. Plants inoculated with the arbuscular mycorrhizal fungi Glomus intraradices (+AMF) were compared to non-inoculated plants (?AMF). Under salinity, AMF-mediated growth stimulation was higher in more salt tolerant Piazar than in sensitive Behta. Mycorrhization alleviated salt-induced reduction of P, Ca, and K uptake. Ca/Na and K/Na ratios were also better in +AMF. However, growth improvement by AMF was independent from plant P nutrition under high salinity. Mycorrhization improved the net assimilation rates through both elevating stomatal conductance and protecting photochemical processes of PSII against salinity. Higher activity of ROS scavenging enzymes was concomitant with lowering of H₂O_2, less lipid peroxidation, and higher proline in +AMF. Cultivar differences in growth responses to salinity and mycorrhization could be well explained by differences in ion balance, photochemistry, and gas exchange of leaves. Function of antioxidant defenses seemed responsible for different AMF-responsiveness of cultivars under salinity. In conclusion, AMF may protect plants against salinity by alleviating the salt-induced oxidative stress.     

Effect of Rhizobium Sp. Inoculation on N2-Fixing and Photosynthetic Activities of Two Cowpea [Vigna unguiculata (L.) Walp.] Genotypes

Time course of symbiotic N₂-fixing and photosynthetic activities during vegetative growth from 30 d after plantation until pod set was measured in the CB5 and 7964 cowpea [Vigna unguiculata (L.) Walp.] genotypes of contrasting senescence traits. At emergence, seedlings were inoculated with a "non-cowpea miscellany" Rhizobium strain generally used to inoculate Cicer arietinum. Maximum N₂-fixing activity occurred in inoculated CB5 and 7964 plants about 54 and 68 d after plantation, respectively. A similar temporal shift of maximum was found for net photosynthetic rate (P _N), confirming a good coordination between the two processes. A higher P _N was found from the first measurements in inoculated plants of both genotypes as compared with uninoculated plants. Apparently, the maximum activity of both N₂-fixation and P _N was timed to occur at a particular stage of plant ontogeny correlating the high N supply with the high N demand by the plant. Rhizobium inoculation did not significantly affect partitioning coefficients of biomass to various plant organs but extended leaf longevity by about 10 d in the CB5 genotype, retarding thus the monocarpic senescence. This revised version was published online in June 2006 with corrections to the Cover Date.     

硝酸盐对牛粪梭菌甲酸脱氢酶缺失型Wood-Ljungdahl途径固碳的影响

【背景】异于同型产乙酸菌通常利用Wood-Ljungdahl途径将2分子CO₂还原为1分子乙酰辅酶A,Clostridium bovifaecis缺失Wood-Ljungdahl途径甲基支路第1步将CO₂还原为甲酸的甲酸脱氢酶,需甲酸存在时将1分子甲酸和1分子CO₂还原为乙酰辅酶A发生葡萄糖的同型产乙酸型发酵。已有报道显示,硝酸盐也可作为同型产乙酸菌的电子受体,而且对不同同型产乙酸菌的代谢影响有所不同,然而硝酸盐对这种独特的甲酸脱氢酶缺失型Wood-Ljungdahl途径固碳的影响尚不清楚。【目的】探究硝酸盐对C.bovifaecis甲酸脱氢酶缺失型Wood-Ljungdahl途径固碳的影响。【方法】硝酸盐浓度分别为10 mmol/L和30 mmol/L时,以未添加硝酸盐为对照实验,研究C.bovifaecis在葡萄糖+甲酸+CO₂为基质条件下的细菌生长、底物消耗和产物生成情况。【结果】10 mmol/L和30 mmol/L硝酸盐存在时,主要产物乙醇浓度分别为5.80 mmol/L和1.66 mmo...     

Early growth of Brazilian tree Dimorphandra wilsonii is also threatened by African grass Urochloa decumbens

The conversion of the Brazilian savannas for pastures and agricultural use has caused the species Dimorphandra wilsonii (Fabaceae-Caesalpinioideae) to become isolated and restricted to areas occupied by African grasses of Urochloa sp. This highly endangered tree species was cultivated in the presence of Bradyhizobium japonicum (symbiont 1), Glomus etunicatum (symbiont 2), and Urochloa decumbens in low nitrogen (N) availability in order to evaluate its growth in these experimental conditions. Even though the nodulation and mycorrhization was of low occurrence, the inoculated plants with symbionts had the greatest nitrogen and chlorophyll content, photosynthetic radiation use efficiency, and biomass accumulation in relation to the plants which had not been inoculated and/or cultivated in the presence of U. decumbens. The results suggest effective N₂ fixation, independent of the localization of bacteria, whether in the root tissue interior or free in the rhizosphere. Therefore, the presence of N₂-fixing bacteria can benefit the early growth of D. wilsonii, whereas the occurrence and aggressive persistence of U. decumbens can limit this development, increasing the threat of extinction of this species in their habitat.     

Simultaneous use of sodium nitrate and urea as nitrogen sources improves biomass composition of Arthrospira platensis cultivated in a tubular photobioreactor

Arthrospira platensis is widely cultivated in open ponds for industrial purposes. However, high‐protein A. platensis biomass produced in photobioreactors (PBRs) is recommended for pharmaceutical and cosmetic formulations. A. platensis was cultivated in a 3.5 L tubular airlift PBR using both sodium nitrate and urea as nitrogen sources. Sodium nitrate was added from the start of the cultivation using a batch process. Urea was supplied daily at exponentially increasing feeding rate using a fed‐batch process. The simultaneous optimization of the independent variables, namely, total quantity of sodium nitrate (m_T1) and total quantity of urea (m_T2), led to an optimal condition of m_T1 = 15.0 mmol/L and m_T2 = 7.5 mmol/L. Maximum biomass concentration (5183 ± 94 mg/L) corresponding to the highest biomass productivity (683 ± 13 mg/L/day) was obtained under such condition. The addition protocol of both nitrogen sources resulted in high productivities of protein (6.2 ± 0.4 mg/L/day) as well as chlorophyll‐a (372.2 ± 7.7 mg/L/day). Such innovative process could be applied in the large‐scale production of A. platensis using tubular PBR for novel applications.     

Effect of NO3 on N2 fixation and nitrogenous solutes of xylem in two nodulated West African geocarpic legumes,Kersting's bean (Macrotyloma geocarpum L.) and Bambara groundnut (Vigna subterranea L.)

Nodulation, nitrogen (N₂) fixation and xylem sap composition were examined in sand cultured plants of Bambara groundnut (Vigna subterranea L.) and Kersting's bean (Macrotyloma geocarpum L.) inoculated with Bradyrhizobium strain CB756 and supplied via the roots for a 4 week period from the third week onwards with different levels of (¹⁵N)-nitrate (0–15 mM). The separate contributions of nitrate and N₂ to plant nitrogen were measured by isotope dilution. Increasing levels of nitrate inhibited nodule growth (measured as dry matter or nodule N) of both species parallel with decreased dependence on symbiotically-fixed N. Specific nodule activity (N₂ fixed g nodule dry⁻¹ d⁻¹ of nodules) was reduced progressively with time in V. subterranea at higher (5 or 15 mM) levels of NO₃, but this was not so for M. geocarpum. Root xylem bleeding sap of both species showed ureides (allantoin and allantoic acid) as predominant (>90%) solutes of nitrogen when plants were relying solely on atmospheric N. Levels of ureide and glutamine decreased and those of asparagine and nitrate in xylem increased with increasing level of applied nitrate. Relative levels of xylem ureide-N were positively correlated (R²=0.842 for M. geocarpum and 0.556 for V. subterranea), and the ratio of asparagine to glutamine in xylem exudate negatively correlated (R²=0.955 for M. geocarpum and 0.736 for V. subterranea) with plant reliance on nitrogen fixation. The data indicate that xylem sap analyses might be useful for indirect field assays of nitrogen fixation by the species and that Kersting's bean might offer some potential as a symbiosis in which N₂ fixation is relatively tolerant of soil N.     

Intra-specific variation in growth and P accumulation of Leucaena leucocephala and Gliricidia sepium as influenced by soil phosphate status

Twenty-three provenances of Gliricidia sepium and eleven isolines of Leucaena leucocephala were examined at a low and at high phosphate levels (20 and 80 mg P kg^-1 soil) for growth, phosphate (P) uptake and use efficiency. Large differences in growth at the low P level, and in growth response to the higher P rate occurred among L. leucocephala isolines and G. sepium provenances. Shoot dry weight at low P varied from 1.30 to 3.01 g plant^-1 for L. leucocephala and from 1.44 to 3.07 g plant^-1 for G. sepium.Leucaena isolines had only half the root weight of G. sepium provenances yet produced approximately 90% of the shoot weight of the corresponding G. sepium treatments, i.e. more than a 2-fold difference in root/shoot ratios. Total P in shoots of G. sepium was some 85% greater than of the respective L. leucocephala isolines in corresponding treatments. Physiological phosphate use efficiency (g shoot/mg P in shoots) (PPUE) was not a simple reciprocal relation, being markedly lower at higher shoot % P and content. However, for the same shoot P both species produced the same shoot weight. Nevertheless, there were apparent genotypic differences within species in the root development, shoot P and PPUE.In another study, the numbers of rhizobia in the rhizosphere of L. leucocephala, nodulation, N₂ fixation at five different levels of P were determined. The numbers of rhizobia in the rhizosphere of inoculated L. leucocephala during the first two weeks were lower when P was added but later became similar to those without added P. Nodules formed earlier than inoculated plants fertilized with P and in greater numbers (4- to 5-fold) and dry weights than in those without P. However, the percentage of N₂ derived from fixation did not change with increasing levels of P application. These results suggest that the observed P effect did not operate via stimulated growth of rhizobia in the rhizosphere, nor through increased N₂ fixation rate. The major effect appeared to be due to effects via plant growth.     

Delayed inoculation and starter nitrogen for enhancing early growth and nitrogen status ofLespedeza cuneata

Summary Two growth chamber experiments were conducted to determine the response ofLespedeza cuneata (Dumont) G. Don. (sericea lespedeza) to delayed inoculation and low levels of nitrogen fertilization. Nitrogen was supplied either as NH ₄ ⁺ or as NO ₃ ^– in solution. At 0.5 and 5.0 ppm nitrogen early growth and N₂(C₂H₂) fixation was inhibited by NH ₄ ⁺ and promoted by NO ₃ ^– . Inoculation at seeding did not negatively affect growth prior to the onset of N₂(C₂H₂) fixation. Delayed inoculation until the trifoliate stage thus did not increase growth or N₂ fixation during the first 40 days of growth. After 40 days, specific nitrogenase activity was highest for plants inoculated at the first trifoliate stage of growth. In contrast, growth and total shoot nitrogen accumulation were higher in plants inoculated at planting. The experimental results suggest that delaying inoculation is not a useful technique for improving early growth ofL. cuneata for surface mine reclamation.     

Regulation of the High-Affinity Nitrate Transport System in Wheat Roots by Exogenous Abscisic Acid and Glutamine

Nitrate is a major nitrogen （N） source for most crops. Nitrate uptake by root cells is a key step of nitrogen metabolism and has been widely studied at the physiological and molecular levels. Understanding how nitrate uptake is regulated will help us engineer crops with improved nitrate uptake efficiency. The present study investigated the regulation of the high-affinity nitrate transport system （HATS） by exogenous abscisic acid （ABA） and glutamine （Gin） in wheat （Triticum aestivum L.） roots. Wheat seedlings grown in nutrient solution containing 2 mmol/L nitrate as the only nitrogen source for 2weeks were deprived of N for 4d and were then transferred to nutrient solution containing 50 μmol/L ABA, and 1 mmol/L Gin in the presence or absence of 2 mmol/L nitrate for 0, 0.5, 1, 2, 4, and 8 h. Treated wheat plants were then divided into two groups. One group of plants was used to investigate the mRNA levels of the HATS components NRT2 and NAR2 genes in roots through semi-quantitative RT-PCR approach, and the other set of plants were used to measure high-affinity nitrate influx rates in a nutrient solution containing 0.2 mmol/L ^15N-labeled nitrate. The results showed that exogenous ABA induced the expression of the TaNRT2.1, TaNRT2.2, TaNRT2.3, TaNAR2.1, and TaNAR2.2 genes in roots when nitrate was not present in the nutrient solution, but did not further enhance the induction of these genes by nitrate. Glutamine, which has been shown to inhibit the expression of NRT2 genes when nitrate is present in the growth media, did not inhibit this induction. When Gin was supplied to a nitrate-free nutrient solution, the expression of these five genes in roots was induced. These results imply that the inhibition by Gin of NRT2 expression occurs only when nitrate is present in the growth media. Although exogenous ABA and Gin induced HATS genes in the roots of wheat, they did not induce nitrate influx.     

Tomato Growth in Soil Amended with Sugar Mill By-Products Compost

Sugar mill by-products compost may be a good soil amendment to promote tomato (Lycopersicon esculentum L.) growth. In addition, the compost may further promote plant growth by inoculation with N₂-fixing bacteria. Compost from sugar-mill waste was prepared with and without the N₂-fixing bacteria, Azotobacter vinelandii, Beijerinckia derxii and Azospirillum sp. and incubated for 50 days. Each compost type was added to 10 kg of soil in pots at rates of 0, 15, and 45 g with and without fertilizer N at rates of 0, 0.75, and 1.54 g. A blanket application of P and K was applied to all pots. Shoot and root dry weights and N content of the whole plant was measured at 55 days. Dry weight of tomato shoots was increased by 40% by addition of fertilizer N and root weight was increased by 66%. Without fertilizer N the high rate of inoculated compost increased shoot growth 180% and uninoculated compost increased shoot growth 112%. For most treatments with and without fertilizer N, inoculated compost enhanced shoot growth and nitrogen content more than uninoculated compost. Root weights were nearly doubled by addition of either compost in comparison to the 0 N treatment. At the low rate of compost addition without fertilizer N, root weight was the same for uninoculated and inoculated compost but at the high rate of compost addition root weight was 32% higher for inoculated compost. The N₂-fixing bacteria colonized roots when inoculated compost was used. Sugar mill by-products compost proved to be an effective soil amendment for promoting the growth of tomato plants.     

Response ofStylosanthes guianensis to endomycorrhizal fungi inoculation as affected by lime and phosphorus applications

The nutrient concentration in the shoots ofStylosanthes guianensis (Aubl.) Sw. cultivated in a sterilized acid and dystrophic soil (Quartzipsament) amended with 4 levels of lime (0; 0.27; 0.63 and 1.10 meq Ca²⁺/100 g soil, as Ca(OH)₂), 2 levels of P (0 and 20 mg P kg^-1 soil, as KH₂PO₄) and not-inoculated or inoculated with 3 vesicular-arbuscular (VA) mycorrhizal fungi was evaluated under greenhouse conditions. The effectiveness of the different fungal species in increasing the nutrient concentration in the shoots varied with the different edaphic conditions. In general, mycorrhiza formation was associated with increases in the concentrations of most of the nutrients analyzed. Under the experimental conditions, the increments in nutrient concentration were higher overall in plants inoculated withAcaulospora scrobiculata. Inoculation ofS. guianensis with VA mycorrhizal fungi was also associated with alterations in the ratios of nutrients in the shoots, which might be important in understanding and explaining the tolerance of mycorrhizal plants to nutritional stresses such as Al and Mn toxicity.     

Growth and metabolic activity of lemon juice vesicle explants in vitro

The relationship between the relative abundance of ureides ([ureide-N/ureide-N plus nitrate-N] × 100) in the shoot axis (stems plus petioles), nodulated roots and leaflets of “Bragg” soybean (Glycine max [L.] Merrill) and the symbiotic dependence of these plants was examined under glass-house conditions. Plants, inoculated with effective Rhizobium japonicum CB1809, were grown with their roots exposed continuously to a nutrient solution containing either 0, 1.5, 3.0, 6.0 or 12.0 millimolar NO₃-N per liter. Nodulation and N₂-acetylene fixation were correlated inversely with the level of nitrate. Seasonal acetylene reduction profiles for each of the nitrate treatments were integrated and the symbiotic dependence ([N₂ fixed per total plant N] × 100) determined using a conversion ratio of 1.5:1 (acetylene reduced:N₂ fixed), calculated from the zero NO₃ treatment. Examination of the nitrogenous solutes of the shoot axis and nodulated roots showed linear relationships between the relative abundance of ureides and the symbiotic dependence of the plants. Two standard curves, depicting these relationships during vegetative and reproductive growth, were drawn for each plant part. The overriding effect of plant age invalidated any attempt to develop a standard relationship for leaflets. Data from two diurnal studies suggested that relative ureides were insensitive to diurnal fluctuations, thus simplifying sampling procedures. Plant material could be stored at ambient temperatures (20-30°C) for up to 24 h without affecting the relative concentration of ureides and nitrate. It is suggested that the shoot axis provides the most suitable target organ when using this technique as a quantitative assay for N₂ fixation because of ease of sampling of these tissues, especially with field-grown plants.     

不同氮源及其浓度对标志链带藻合成淀粉和油脂的影响北大核心CSCD

【目的】以标志链带藻(Desmodesmus insignis)为实验材料,研究不同氮源及其浓度对该藻生长、总脂和淀粉(碳水化合物)含量的影响,为该藻在生物能源方面的应用提供一定的理论依据。【方法】以硝酸钠、碳酸氢铵或尿素为氮源,5个氮浓度(3、6、9、12和18 mmol/L)的BG-11培养基培养标志链带藻,采用干重法测定生物质浓度、重量法测定总脂、苯酚-硫酸法测定、总碳水化合物和淀粉的含量。【结果】标志链带藻在3种氮源下均能很好的生长。最高油脂含量出现在3 mmol/L硝酸钠实验组,达到32.61%(d.w)。当18 mmol/L碳酸氢铵作为氮源时,总碳水化合物与淀粉的含量以及产率都达到最高,分别为56.54%(d.w)和55.33%(d.w)、0.24和0.23 g/(L·d)。以尿素为氮源时,其生物质浓度和各组分含量与其它氮源实验组差别不大,均有利于该藻的生长及各生化组分含量的积累。【结论】以该藻种生产生物能源的成本等综合考虑,以18 mmol/L碳酸氢铵和尿素为氮源培养标志链带藻最优。     

Growth and nitrate uptake properties of plants grown at different relative rates of nitrogen supply. II. Activity and affinity of the nitrate uptake system in Pisum and Lemna in relation to nitrogen availability and nitrogen demand

Abstract Net nitrate uptake rates were measured and the kinetics calculated in non-nodulated Pisum sativum L. cv. Marma and Lemna gibba L. adapted to constant relative rates of nitrate-N additions (R_A), ranging from 0.03 to 0.27 d^?1 for Pisum and from 0.05 to 0.40 d^?1 for Lemna, V_max of net nitrate uptake (measured in the range 10 to 100 mmol m^?3 nitrate, i.e. ‘system I’) increased with R_A in the growth limiting range but decreased when R_A exceeded the relative growth rate (RGR), K_m was not significantly related to changes in R_A. On the basis of previous ¹³N-flux experiments, it is concluded that the differences in V_max at growth limiting R_A are attributable to differences in influx rates. Linear relationships between V_max and tissue nitrogen concentrations were obtained in the growth limiting range for both species, and extrapolated intercepts relate well with the previously defined minimal nitrogen concentrations for plant growth (Oscarson, Ingemarsson & Larsson, 1989). Analysis of V_max for net nitrate uptake on intact plant basis in relation to nitrogen demand during stable, nitrogen limited, growth shows an increased overcapacity at lower R_A values in both species, which is largely explained by the increased relative root size at low R_A. A balancing nitrate concentration, defined as the steady state concentration needed to sustain the relative rate of increase in plant nitrogen (R_N), predicted by R_A, was calculated for both species. In the growth limiting range, this value ranges from 3.5 mmol m^?3 (R_A 0.03 d^?1) to 44 mmol m^?3 (R_A 0.21 d^?1) for Pisum and from 0.2 mmol m^?3 (R_A 0.05 d^?1) to 5.4 mmol m^?3 (R_A 0.03 d^?1) for Lemna. It is suggested that this value can be used as a unifying measure of the affinity for nitrate, integrating the performance of the nitrate uptake system with nitrate flux and long term growth and demand for nitrogen.     

俄罗斯杨水培苗对硼胁迫的生长和生理响应

该研究采用水培试验,分析了俄罗斯杨(Populus russkii)对短期硼胁迫(1、5、10mmol/L硼酸)的生长和生理响应特征,探明其对硼胁迫的耐受范围,初步揭示其耐硼的生理基础。结果显示:(1)与对照相比,1mmol/L硼酸处理俄罗斯杨水培苗的生长情况更优,叶片光合色素含量和净光合速率显著上升,SOD和APX活性及抗坏血酸含量显著上调。(2)5mmol/L硼酸处理幼苗有轻微胁迫受害症状,叶片褪绿变黄,叶绿素b含量比对照显著上升,而Chla/Chlb比值显著下降,其SOD和APX活性显著上调。(3)10mmol/L硼酸处理幼苗表现出严重毒害症状,叶片发黄、茎尖和根尖生长受抑制;叶片PSⅡ原初光能转换效率(F_v/F_m)和Chla/Chlb比值比对照显著下降;其叶片MDA含量、POD活性、抗坏血酸含量以及游离脯氨酸和可溶性糖含量均比对照显著上升。研究表明,在不同浓度硼酸短期胁迫下,俄罗斯杨水培苗能通过上调抗氧化酶活性和抗氧化物质、渗透调节物质含量来有效清除体内胁迫产生的活性氧,减轻膜脂过氧化伤害,使其在低于5mmol/L硼酸胁迫环境下正常生长,表现出较强的耐硼性。     

RESPONSE OF TROPIC LEAF MOVEMENTS AND PHOTOSYNTHETIC GAS EXCHANGE TO WATER AVAILABILITY IN N2-FIXING AND NO3-FED SOYBEAN

Leaf movements, water status, and gas exchange were measured in soybean inoculated with Bradyrhizobium and grown under high and low soil water and nitrate availabilities. We hypothesized that paraheliotropism in low NO₃-grown plants (which have greater N₂ fixation rates) would differ from that of high NO₃-grown plants (which have lower N₂ fixation rates), such that carbon return on nitrogen investment into photosynthesis would be enhanced. Low NO₃-low water plants had more vertical leaf angles and received lower solar irradiances at midday than high NO₃-low water plants. Under constant, vertical illumination, low NO₃-low water plants had steeper leaf angles, increased rates of leaf movement, lower photosaturated photosynthetic rates, and lower stomatal conductances for a given leaf water potential than high NO₃-low water plants. Leaves of high NO₃ plants had lower photosynthetic nitrogen use efficiencies than did low NO₃ plants. Low water plants had lower leaf osmotic potentials and ratios of intercellular/ambient CO₂ concentration than high water plants, but NO₃ treatment did not affect these parameters. Results provided support for our initial hypothesis, and demonstrated a high degree of correspondence between gas exchange and heliotropic response to soil nitrogen and water availabilities in soybean.     

Most probable numbers of Azospirillum associated with the roots of inoculated pearl millet

Summary Inoculation of pearl millet (Pennisetum americanum (L.) Leeke) with Azospirillum significantly increased the numbers of this organism in the rhizosphere, rhizoplane, washed and crushed roots and surface sterilized and crushed roots. The maximum number of organisms plant^–1 were localized in the rhizosphere. The numbers of Azospirillum on the roots of inoculated plants grown under sterilized conditions were much higher than in the field grown plants. In both cases populations outside the roots were higher than in the surface sterilized roots. The highest numbers per unit root weight were recorded between 60–75 days of growth. N₂-ase activity throughout the growth cycle was very low and was not related to the populations of Azospirillum on the roots. Root exudates and extracts of pearl millet showed a stimulatory effect on the growth of Azospirillum suggesting their possible involvement in the colonization of this organism on the roots of inoculated plants.     

Inhibition of N2 fixation by white clover (Trifolium repens L.) at low concentrations of NO inf3 sup− in flowing solution culture

The impact of sustained low external concentrations of NO ₃ ^– (0, 10, 100 and 1000 mmol m^–3) on plant growth and the relative acquisition of N through N₂ fixation and NO ₃ ^– uptake by established, nodulated white clover (Trifolium repens L. cv. Blanca) was studied over 28 days in flowing solution culture. Nitrogen fixation was measured by N difference and ¹⁵N dilution methods. Plants supplied with NO ₃ ^– achieved higher relative growth rates (% MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGqiVu0Je9sqqrpepC0xbbL8F4rqqrFfpeea0xe9Lq-Jc9% vqaqpepm0xbba9pwe9Q8fs0-yqaqpepae9pg0FirpepeKkFr0xfr-x% fr-xb9adbaqaaeGaciGaaiaabeqaamaabaabaaGcbaGabmiEayaara% aaaa!3702!\[\bar x\]=0.091 d^–1) compared with control plants dependent on N₂ fixation (0.073 d^–1). Nitrate plants showed progressive increases in shoot: root d.w. ratios from 4 to 6.5–7.6 between days 0–28, compared with 5.1 on day 28 for control plants. Increases in both nodule d.w. and numbers per plant were inhibited after day seven at all concentrations of NO ₃ ^– . The severity of inhibition of N₂ fixation increased with increasing NO ₃ ^– concentration and with time. The total amounts of N₂ fixed per plant between days 0–7 after supplying 10, 100 and 1000 mmol m^–3 NO ₃ ^– , respectively, were 37–39, 28–30 and 0–13%, of the total N acquired. Between days 7–28 the proportional contributions of N₂ fixation to total N acquisition declined to 3, 0.5 and 0%, respectively, in these treatments. The corresponding mean specific rates of N₂ fixation between days 0–7 were, respectively, 5.4, 3.2, and 2.0 mmol N d^–1 g^–1 nodule d.w., compared with 7.9 mmol N d^–1 g^–1 nodule d.w. for zero NO ₃ ^– plants. There was no evidence of a transitory increase in N₂ fixation following the addition of NO ₃ ^– , even at the lowest supply concentration.