Nitrogen Fixation and Allantoin Formation in Soybean Plants

The activity of nitrogenase and the concentration of ammonia and allantoin (+ allantoic acid) in root nodules were measured throughout the growth period of soybean plants. Nitrogenase activity measured by acetylene reduction increased with plant growth and reached a maximum level at the flowering period. The level of ammonia and allantoin concentration in nodules was parallel with increased nitrogenase activity. At the late reproductive stage (pod-forming period), nitrogenase activity showed a marked decrease, but the ammonia and allantoin in the nodules remained at a constant level. Detached nodules from 56 day-old soybean plants were exposed to ¹⁵N₂ gas, and the distribution of ¹⁵N among nitrogen compounds was investigated. Enrichment of ¹⁵N in allantoin and allantoic acid reached a fairly high level after 90 min of nitrogen fixation; ca. 22% of ¹⁵N in acid-soluble nitrogen compounds was incorporated into allantoin + allantoic acid. In contrast, enrichment of ¹⁵N in amide nitrogen was relatively low. No significant ¹⁵N was detected in the RNA fraction. The data suggested that ureide formation in nitrogen-fixing root nodules did not take place through the breakdown of nucleic acids, but directly associated with the assimilating system of biologically fixed nitrogen.     

Movements of Organic Nitrogen and Carbohydrates in Pelargonium Plants

Evidence has been obtained suggesting that the movement of organicnitrogen in the phloem is independent of carbohydrate movementin Pelargonium plants. The results obtained cannot readily beexplained by the mass-flow theory of Munch.     

Growth and Nitrogen Accumulation in Young Tomato Plants Treated with Gibberellic Acid

The effects of foliar sprays of gibberellic acid (GA) on thegrowth of tomato plants cv. Potentate were studied in growthrooms and a glasshouse. Four sprays of GA (5 ppm) increasedleaf area and whole plant weight relative to water controlsgrown at constant temperatures (7, 17, 22, and 27 °C) for12 days, the largest plants being obtained with 5 ppm. Experimentsmade at four photoperiods (5, 10, 15, and 20 h) and at two lightintensities (7000 and 10 750 lx) showed that GA increased leafand whole plant weight at 15 h, leaf area at 10 and 15 h andstem height at all photoperiods; area, height, and weight increaseswere obtained at both light intensities, leaf growth being increasedmore by GA at 7000 lx and stem growth more at 10 750 lx. Four foliar sprays of GA (5 ppm) were combined with N supplementsapplied via leaf and/or root to plants in sand culture. Withlow supply to the roots (20 ppm N) GA failed to increase growth,but increased it at higher levels. Total N in leaf and stemwas increased by GA or by NH₄NO₃ (10 sprays 280 ppm N) at alllevels of N supplied to roots, but when applied together theeffect on total leaf N was more than additive except at thehighest level (540 ppm) GA increased the concentration of N(as per cent dry matter) in leaf and stem at all levels of Nsupplied to roots. GA and NH₄NO₃ together resulted in a greateramount and a higher concentration of N in the shoots (and usuallyalso in roots) than did NH₄NO₃ alone. Leaf thickness (as freshweight/unit area) could only be increased appreciably by sprayingwith a complete nutrient solution which reduced leaf area butnot dry weight. Growth increases induced by GA were detectable 43 days afterthe first of four sprays in the glasshouse and after 30 daysin the growth room. The persistence of GA effects was comparedwith those induced by sprays of NH₄NO₃.     

Nitrogen Transfer from Four Nitrogen-Fixer Associations to Plants and Soils

Nitrogen (N) fixation is the main source of ‘new’ N for N-limited ecosystems like subarctic and arctic tundra. This crucial ecosystem function is performed by a wide range of N₂ fixer (diazotroph) associations that could differ fundamentally in their timing and amount of N release to the soil. To assess the importance of different associative N₂ fixers for ecosystem N cycling, we tracked ¹⁵N-N₂ into four N₂-fixer associations (with a legume, lichen, free-living, moss) and into soil, microbial biomass and non-diazotroph-associated plants 3 days and 5 weeks after in situ labelling. In addition, we tracked ¹³C from ¹³CO₂ labelling to assess if N and C fixation are linked. Three days after labelling, half of the fixed ¹⁵N was recovered in the legume soils, indicating a fast release of fixed N₂. Within 5 weeks, the free-living N₂ fixers released two-thirds of the fixed ¹⁵N into the soil, whereas the lichen and moss retained the fixed ¹⁵N. Carbon and N₂ fixation were linked in the lichen shortly after labelling, in free-living N₂ fixers 5 weeks after labelling, and in the moss at both sampling times. The four investigated N₂-fixer associations released fixed N₂ at different rates into the soil, and non-diazotroph-associated plants have no access to ‘new’ N within several weeks after N₂ fixation. Although legumes and free-living N₂ fixers are immediate sources of ‘new’ N for N-limited tundra ecosystems, lichens and especially mosses, do not contribute to increase the N pool via N₂ fixation in the short term.     

Effect of Salinity and Potassium on the Uptake of Nitrogen and on Nitrogen Metabolism in Young Barley Plants

In a solution culture experiment with 4-week-old barley plants (variety Villa) the influence of NaCl salinization and of KCl additions on the uptake and turnover of labelled N (¹⁵NH₄¹⁵NO₃) was studied. Labelled N was applied for 24 h at the end of the growth period. Salinization impaired growth and uptake of labelled N. The incorporation of labelled N into the protein fraction, however, was improved by NaCl salinization. Additions of KCl to the nutrient solution diminished the negative effect of NaCl salinization on growth. At both NaCl salinization levels (60 and 120 mM) K additions favoured the uptake of labelled N and particularly its incorporation into the protein fraction. It is suggested that the negative influence of the NaCl stress is not primarily due to an impaired protein synthesis, but is possibly caused by a deterimental effect of Na on other metabolic processes.     

Cyclic Variations in Nitrogen Uptake Rate in Soybean Plants: Uptake During Reproductive Growth

Net uptake of by non-nodulated soybean plants [Glycme max(L ) Merr cv Ransom] growing in flowing hydroponicculture was measured daily during a 63 d period of reproductivedevelopment between the first florally inductive photopenodand late seed growth Removal of from a replenished solution containing 10 mol m^– was determined by ion chromatography Uptake of continued throughout reproductive development The net uptakerate of cycled between maxima and minima with a periodicity of oscillation of 3 to 7 d during the floralstage and about 6 d during the fruiting stage. Coupled withincreasing concentrations of carbon and C:N ratios in tissues,the oscillations in net uptake rates of are evidence that the demand for carbohydrate by reproductiveorgans is contingent on the availability of nitrogen in theshoot pool rather than that the demand for nitrogen followsthe flux of carbohydrate into reproductive tissues. Key words: Nitrate uptake rate, carbon-nitrogen partitioning, Glycme max (L ) Merrill     

去叶对不同生长习性大豆固氮作用的影响

生殖生长期开始去叶后 ,(1)有限型、亚有限型和无限型 3种类型的大豆根瘤固氮酶活性都降低 ,而根瘤中酰脲含量则不同程度地增加 ;(2 )有限型大豆幼茎中酰脲含量明显增加 ,但亚有限型和无限型大豆变化不大 ;(3) 3种类型大豆幼茎中硝态氮含量增加明显     

The Movement of Organic Nitrogen Compounds in Plants: With two Figures in the Text

The hypothesis that the movement of organic nitrogen, like thatof carbohydrate, is governed by the prevailing concentrationgradients for the mobile compounds in the phloem has been testedby determining the effect on nitrogen distribution when normaltranslocation was interrupted by ringing in Pelargonium spp.and Vicia faba plants. Nitrogen compounds in the plant tissue were fractionated intoprotein, soluble protein, amide, amino-acid, peptide, nitrate,and ammonia nitrogen. The presence of short-chain peptides inthe extracts was confirmed by paper chromatography. Results showed that accumulation above the ringed zone was confinedto the soluble nitrogen fractions and that amino-acids and possiblypeptides were involved in translocation. The direction of movementmay be determined by the prevailing concentration gradientsof these compounds in the phloem tissue.     

Nitrogen Transfer Within and Between Plants Through Common Mycorrhizal Networks (CMNs)

Mycorrhizae play a critical role in nutrient capture from soils. Arbuscular mycorrhizae (AM) and ectomycorrhizae (EM) are the most important mycorrhizae in agricultural and natural ecosystems. AM and EM fungi use inorganic NH₄ ⁺ and NO₃ ^?, and most EM fungi are capable of using organic nitrogen. The heavier stable isotope ¹⁵N is discriminated against during biogeochemical and biochemical processes. Differences in ¹⁵N (atom%) or δ¹⁵N (‰) provide nitrogen movement information in an experimental system. A range of 20 to 50% of one-way N-transfer has been observed from legumes to nonlegumes. Mycorrhizal fungal mycelia can extend from one plant's roots to another plant's roots to form common mycorrhizal networks (CMNs). Individual species, genera, even families of plants can be interconnected by CMNs. They are capable of facilitating nutrient uptake and flux. Nutrients such as carbon, nitrogen and phosphorus and other elements may then move via either AM or EM networks from plant to plant. Both ¹⁵N labeling and ¹⁵N natural abundance techniques have been employed to trace N movement between plants interconnected by AM or EM networks. Fine mesh (25～45 μm) has been used to separate root systems and allow only hyphal penetration and linkages but no root contact between plants. In many studies, nitrogen from N₂-fixing mycorrhizal plants transferred to non-N₂–fixing mycorrhizal plants (one-way N-transfer). In a few studies, N is also transferred from non-N₂–fixing mycorrhizal plants to N₂-fixing mycorrhizal plants (two-way N-transfer). There is controversy about whether N-transfer is direct through CMNs, or indirect through the soil. The lack of convincing data underlines the need for creative, careful experimental manipulations. Nitrogen is crucial to productivity in most terrestrial ecosystems, and there are potential benefits of management in soil-plant systems to enhance N-transfer. Thus, two-way N-transfer warrants further investigation with many species and under field conditions.     

Ubiquity of Insect-Derived Nitrogen Transfer to Plants by Endophytic Insect-Pathogenic Fungi: an Additional Branch of the Soil Nitrogen Cycle

The study of symbiotic nitrogen transfer in soil has largely focused on nitrogen-fixing bacteria. Vascular plants can lose a substantial amount of their nitrogen through insect herbivory. Previously, we showed that plants were able to reacquire nitrogen from insects through a partnership with the endophytic, insect-pathogenic fungus Metarhizium robertsii. That is, the endophytic capability and insect pathogenicity of M. robertsii are coupled so that the fungus acts as a conduit to provide insect-derived nitrogen to plant hosts. Here, we assess the ubiquity of this nitrogen transfer in five Metarhizium species representing those with broad (M. robertsii, M. brunneum, and M. guizhouense) and narrower insect host ranges (M. acridum and M. flavoviride), as well as the insect-pathogenic fungi Beauveria bassiana and Lecanicillium lecanii. Insects were injected with ¹⁵N-labeled nitrogen, and we tracked the incorporation of ¹⁵N into two dicots, haricot bean (Phaseolus vulgaris) and soybean (Glycine max), and two monocots, switchgrass (Panicum virgatum) and wheat (Triticum aestivum), in the presence of these fungi in soil microcosms. All Metarhizium species and B. bassiana but not L. lecanii showed the capacity to transfer nitrogen to plants, although to various degrees. Endophytic association by these fungi increased overall plant productivity. We also showed that in the field, where microbial competition is potentially high, M. robertsii was able to transfer insect-derived nitrogen to plants. Metarhizium spp. and B. bassiana have a worldwide distribution with high soil abundance and may play an important role in the ecological cycling of insect nitrogen back to plant communities.     

植物脂质转移蛋白

脂质转移蛋白(lipid transfer proteins,LTPs)是植物生命活动中一类重要的活性蛋白质,在体外能够可逆地结合和转运多种脂质分子.目前已从多种植物中分离到LTPs基因.随着研究的深入,其不同水平的转录本在不同植物的不同发育阶段和生理条件下的不同组织中被发现,但LTPs体内确切的生理、生化功能和作用机制尚不明确.现介绍目前这一领域细胞与分子生物学方面的研究进展,并结合本课题组的研究结果进行了相关探讨,为进一步研究LTPs在植物生殖发育、抗性和防御反应及信号转导中的作用机制提供了新的线索.     

Inhibition of Nitrogen Fixation in Soybean Plants Supplied with Nitrate II. Accumulation and Properties of Nitrosylleghemoglobin in Nodules

The accumulation of nitrosylleghemoglobin (LbNO) in nodulesand the properties of LbNO in vitro were investigated in connectionwith the inhibition of nitrogen fixation in soybean nodulesby nitrate. The leghemoglobin extracted under argon gas from nodules ofplants supplied with nitrate consisted mainly of LbNO, as judgedfrom the spectrum which corresponded to that of LbNO formedin vitro by the reaction of leghemoglobin with nitrite in thepresence of dithionite or by the combination of ferrous leghemoglobin(Lb²⁺) with nitric oxide. Further, LbNO formed in vivo was easilydissociated by visible light, as was LbNO formed in vitro. Thus,authentic LbNO does actually accumulate in nodules. Most of the leghemoglobin was of the ferrous type in nodulesof plants supplied with nitrate. Some LbNO appeared to be derivedfrom LbO₂ which was deoxygenated by nitrite. The increase inlevels of LbNO in nodules paralleled the decrease in acetylenereducing activity. These results indicate that the decrease in nitrogenase activityin nodules of soybean plants supplemented with nitrate is causedby the decrease in levels of LbO₂ that carries oxygen into bacteroids,which results from the formation of LbNO (Received August 22, 1989; Accepted December 4, 1989)     

Inhibition of Nitrogen Fixation in Soybean Plants Supplied with Nitrate I. Nitrite Accumulation and Formation of Nitrosylleghemoglobin in Nodules

The accumulation of nitrite in nodules was investigated to elucidatethe mechanism of inhibition of nitrogen fixation in nodulesof soybean (Glycine max. [L.] Merr.) plants supplied with nitrate.Acetylene-reducing activity (ARA) in nodules fell within 24h as a result of the supply of exogenous nitrate, accompaniedby an increase in the accumulation of nitrite in the cytosolbut not in the bacteroids of nodules. Nitrate reductase (NR)activity in the nodule cytosol remained high, irrespective ofthe supply of nitrate. Nitrosylleghemoglobin (LbNO) was detectedspectrophotometrically in the extract from nodules in whichnitrogen fixation was inhibited by nitrate. In experiments invitro, it was found that LbNO was easily formed from leghemoglobinin the presence of nitrite and dithionite. Thus, it is suggested that nitrogen fixation was inhibited primarilyby a decrease in the function of leghemoglobin, attributableto the formation of LbNO, which was caused by the accumulationof nitrite generated from nitrate by NR in the nodule cytosol. (Received August 22, 1989; Accepted January 24, 1990)     

Nitrogen Transfer Between Plants: A 15N Natural Abundance Study with Crop and Weed Species

An increasing amount of evidence indicates that N can be transferred between plants. Nonetheless, a number of fundamental questions remain. A series of experiments was initiated in the field to examine N transfer between N₂-fixing soybean (Glycine max [L.] Merr.) varieties and a non-nodulating soybean, and between N₂-fixing peanut (Arachis hypogaea L.) or soybean and neighboring weed species. The experiments were conducted in soils with low N fertilities and used differences in N accumulation and/or ¹⁵N natural abundance to estimate N transfer. Mixtures of N₂-fixing and non-nod soybean indicated that substantial inter-plant N transfer occurred. Amounts were variable, ranging from negligible levels to 48% of the N found in the non-nod at maturity. Transfer did not appear to strongly penalize the N₂-fixing donor plants. But, in cases where high amounts of N were transferred, N content of donors was noticeably lowered. Differences were evident in the amount of N transferred from different N₂-fixing donor genotypes. Results of experiments with N₂-fixing crops and the weed species prickly sida (Sida spinosa L.) and sicklepod (Senna obtusifolia [L.] Irwin & Barneby) also indicated substantial N transfer occurred over a 60-day period, with amounts accounting for 30–80% of the N present in the weeds. Transfer of N, however, was generally very low in weed species that are known to be non-hosts for arbuscular mycorrhizae (yellow nutsedge, Cyperus esculentus L. and Palmer amaranth, Amaranthus palmeri [S.] Watson). The results are consistent with the view that N transfer occurs primarily through mycorrhizal hyphal networks, and they reveal that N transfer may be a contributing factor to weed problems in N₂-fixing crops in low N fertility conditions.     

Studies on Nitrogen Metabolism in Tobacco Plants

Δ¹-Pyrroline-5-carboxylate reductase has been considerably purified from tobacco leaves. This enzyme uses NADPH or NADH for the formation of proline, although the former is better used. This enzyme was found in washed chloroplast extract as well as in cytoplasmic fluid and utilized NADPH, formed by the photosynthetic NADP reduction, for the sythesis of proline in the light.     

Nickel in Plants: II. Distribution and Chemical Form in Soybean Plants

The gross tissue distribution, intracellular fate, and chemical behavior of Ni²⁺ in soybean plants (Glycine max cv. Williams) were investigated. Following root absorption, Ni was highly mobile in the plant, with leaves being the major sink in the shoots for Ni during vegetative growth. A senescence >70% of the Ni present in the shoot was remobilized to seeds. Fractionation of root and leaf tissues showed >90% of the Ni to be associated with the soluble fraction of tissues; ultrafiltration of the solubles showed >77% of the Ni to be associated with the 10,000 to 500 molecular weight components of both roots and leaves. Chemical characterization of the soluble components (10,000 to 500 and >500 molecular weight) by thin layer chromatography and electrophoresis resolved a number of Ni-containing organic complexes. Major Ni-containing components formed in the root are transported in the xylem stream, and undergo partial modification on deposition in leaves. Nickel accumulated in seeds is primarily associated with the cotyledons. Chemical fractionation of cotyledon components showed 80% of the Ni to be associated with the soluble whey fraction, while 70% of this fraction was composed of Ni-containing components with molecular weight <10,000.