Carbohydrate accumulation in leaves of plants treated with the herbicide chlorsulfuron or imazethapyr is due to a decrease in sink strength

Herbicides that inhibit branched chain amino acid biosynthesis produce a rapid carbohydrate increase in leaves of treated plants. The relationship between these processes is not known nor is the importance of carbohydrate accumulation in the growth inhibition caused by these herbicides. This work analyzes carbohydrate concentration in sources and sinks after herbicide treatments in pea (Pisum sativum L.), as well as photosynthetic carbon assimilation, using two classes of chemicals, chlorsulfuron and imazethapyr, applied to roots or leaves. The most remarkable result was that, in addition to carbohydrate accumulation in leaves, accumulation of sucrose and/or starch in roots was detected. This pattern of carbohydrate accumulation was similar for both herbicides and independent of whether the herbicides were applied to leaves or roots. This indicates that root growth inhibition was not caused by sugar starvation in sinks. Nevertheless, the results are consistent with a decrease in sink strength, leading to the inhibition of photoassimilate translocation.     

The effect of amino acids and amides on the regulation of nitrate uptake by wheat seedlings

Wheat plants grown hydroponically increased their nitrate uptake rate more than two‐fold after three days of N starvation. Exogenously supplied amino acids and amides had no effect on the nitrate uptake rate of plants well nourished in N. After three days of N starvation, however, some of the amino acids and amides supplied to plants inhibited up to 50% of the nitrate uptake rate. The most effective inhibitor was aspartic acid. Asparagine, glutamine or phenylalanine did not show any inhibitory effect. The percentage of inhibition was not increased by increasing the amino acid concentration, nor did the addition of mixed amino acids and amides increase the inhibition exerted by one amino acid alone. During the three days of N starvation, there was a decrease in the concentration of endogenous amino acids in the roots, but not all amino acids decreased their concentration at the same rate.

It is suggested that the endogenous levels of some amino acids may repress the nitrate uptake system in plants well supplied with N. During the development of the N deficiency, the concentration of these amino acid decreases, de‐repressing the nitrate uptake system.     

Biochemical aspects of nitrogen use efficiency: An overview

Crop productivity relies heavily on nitrogen (N) fertilization. N is an essential macronutrient limiting the growth and development of plants in agriculture. Both organic and inorganic forms of N are metabolized in plants; nitrate and ammonia are common forms of inorganic N that can be metabolized in all plants. In the last 40 years the amount of synthetic N applied to crops has risen dramatically, resulting in significant increases in yield but with considerable impacts on the environment. Production and application of N fertilizers consume huge amounts of energy, and excess is detrimental to the environment. A requirement for crops that require decreased N fertilizer levels has been recognized in the call for a ‘Second Green Revolution’ and research in the field of nitrogen-use efficiency (NUE) has continued to grow. Nitrogen-use efficiency is inherently a complex trait, as each step-including N uptake, translocation, assimilation, and remobilization-is governed by multiple interacting genetic and environmental factors. The limiting factors in plant metabolism for maximizing NUE are different at high and low N supplies, indicating great potential for improving the NUE of present cultivars. Decreasing environmental losses and increasing the productivity of crop-acquired N requires the coordination of carbohydrate and N metabolism to give high yields. This has prompted a search to identify genes that improve the NUE of crop plants, with candidate NUE genes existing in pathways relating to N uptake, assimilation, amino acid biosynthesis, carbon (C)/N storage and metabolism, signaling and regulation of N metabolism and translocation, remobilization and senescence. In this review, we present the over view of N metabolism, relation of C/N metabolism and future prospects of improving NUE in crops using various complementary approaches.     

Fermentative metabolism is induced by inhibiting different enzymes of the branched-chain amino acid biosynthesis pathway in pea plants

The inhibition of branched-chain amino acid (BCAA) biosynthesis was evaluated in pea plants in relation to the ability for induction of fermentative metabolism under aerobic conditions. Chlorsulfuron and imazethapyr (inhibitors of acetolactate synthase, ALS, EC 4.1.3.18) produced a strong induction of pyruvate decarboxylase (PDC, EC 4.1.1.1) and alcohol dehydrogenase (ADH, EC 1.1.1.1) activities and a lesser induction of lactate dehydrogenase (LDH, EC 1.1.1.27) and alanine aminotransferase (AlaAT, EC 2.6.1.2) activities in roots. Inhibition of the second enzyme of the BCAA biosynthesis (ketol-acid reductoisomerase, KARI, EC 1.1.1.86) by Hoe 704 (2-dimethylphosphinoyl-2-hydroxyacetic acid) and CPCA (1,1-cyclopropanedicarboxylic acid) enhanced fermentative enzyme activities including PDC, ADH, and AlaAT. Fermentative metabolism induction occurring with ALS- and KARI-inhibitors was related to a higher expression of PDC. In the case of KARI inhibition, it is proposed that fermentation induction is due to an inhibition of ALS activity resulted from an increase in acetolactate concentration. Fermentative metabolism induction in roots, or at least ethanolic fermentation, appeared to be a general physiological response to the BCAA biosynthesis inhibition.     

Stickstoff-Assimilation in der Wurzel und Transport von N-Verbindungen mit dem Blutungssaft der Wurzeln in Abhängigkeit von der Mn-Versorgung

Nitrogen Assimilation in Roots and the Transport of Nitrogen Compounds in the Bleeding Sap of Roots in relation to Manganese Nutrition. The assimilation of nitrogen in the roots of 27 days old pumpkin plants was examined in relation to manganese nutrition. The transport of nitrogen compounds in the xylem was determined in roots and in the bleeding sap of roots using nitrate as the N-source. The maximum NO₃ content in the roots was observed in the Mn treatment which resulted in the highest shoot yields (0.05 ppm Mn). The bleeding sap of this treatment was lowest in nitrate concentration, but showed the highest rate of transport of organic nitrogen compounds. In experiments with ¹⁵N in the nutrient solution the isotope was found in the roots in organic and in inorganic compounds. The composition of the fraction of free amino acids differed between roots and xylem sap. In the bleeding sap glutamine was especially dominant. In the roots the amino acid composition depended on the extent of Mn-supply. Lowest glutamine concentrations were found in the xylem sap from the treatment with maximum shoot yields. A numerical difference was found in the xylem sap between organic N (N_(org)) and the amino acid nitrogen. This difference which account for more than 50 % of the organically bound nitrogen is suggested to be made up in part by low molecular weight peptides, amino sugars and other compounds. In Mn deficiency a general reduction in the intensity of nitrogen metabolism was found. With Mn toxicity the N assimilation activity was more intensive than for the low Mn supply. Simultaneously, however, the transport of organic N compounds from the root was lower.     

Nitrogen metabolism and seed composition as influenced by glyphosate application in glyphosate-resistant soybean

Previous research has demonstrated that glyphosate can affect nitrogen fixation or nitrogen assimilation in soybean. This 2-year field study investigated the effects of glyphosate application of 1.12 and 3.36 kg of ae ha(-1) on nitrogen metabolism and seed composition in glyphosate-resistant (GR) soybean. There was no effect of glyphosate application on nitrogen fixation as measured by acetylene reduction assay, soybean yield, or seed nitrogen content. However, there were significant effects of glyphosate application on nitrogen assimilation, as measured by in vivo nitrate reductase activity (NRA) in leaves, roots, and nodules, especially at high rate. Transiently lower leaf nitrogen or (15)N natural abundance in high glyphosate application soybean supports the inhibition of NRA. With the higher glyphosate application level protein was significantly higher (10.3%) in treated soybean compared to untreated soybean. Inversely, total oil and linolenic acid were lowest at the high glyphosate application rate, but oleic acid was greatest (22%) in treated soybean. These results suggest that nitrate assimilation in GR soybean was more affected than nitrogen fixation by glyphosate application and that glyphosate application may alter nitrogen and carbon metabolism.     

Why does temperature affect relative uptake rates of nitrate, ammonium and glycine: A test with Eucalyptus pauciflora

Few studies have examined how temperature affects uptake of nitrate, ammonium and amino acids from soil. This study tests the hypothesis that cool temperatures favour uptake of the amino acid glycine while warm temperatures favour uptake of inorganic forms of N such as nitrate. We used glasshouse-grown ectomycorrhizal seedlings of the sub-alpine tree species Eucalyptus pauciflora Sieber ex Spreng. Seedlings were grown in soil (humic umbrosol, from species' habitat) that was dominated by amino acids and ammonium with only small amounts of nitrate. To examine if root physiology affects temperature responses of N uptake, we measured uptake from ¹⁵N-labelled hydrosolutions containing equimolar 100 μmol L⁻¹ mixtures of ammonium, nitrate and glycine at temperatures from 5 to 35 °C. We also examined if the effect of temperature on uptake of N forms was due to plant-microbe competition by following the fate of equimolar amounts of labelled ammonium, nitrate and glycine injected into the soil at temperatures of 5 °C and 25 °C. Hydrosolution experiments showed that uptake of glycine was favoured by warm temperatures and inorganic N by cool temperatures. In contrast, when ¹⁵N was injected into soil the uptake of glycine was favoured by low temperatures and nitrate by warm temperatures. At 25 °C, glycine was 17% of the N taken up from soil and nitrate was 51%; whereas at 5 °C glycine was 30% of the N taken up from soil and nitrate was 23%. Microbes were better competitors than seedlings for all forms of N, but temperature did not affect microbial preference for the different N forms. Hence, while microbes limit N available for plant uptake, they do not seem to be the cause of the greater plant uptake of glycine at cool temperatures and nitrate at warm temperatures. Intact uptake of glycine by plants was suggested by the positive relationship between uptake of ¹³C and ¹⁵N and detection by GC-MS of intact , ¹⁵N glycine molecules in roots. In conclusion, uptake of glycine is favoured by cool temperatures and nitrate by warm temperatures, but this is apparently not a function of root physiology or competition with soil microbes.     

钙螯合剂对小麦幼苗氮代谢和干物重的影响

营养液中添加钙螯合剂EGTA[Glycol bis ( aminoethyl)etherN ,N ,N ,N tetraaceticacid ,乙二醇双 (-氨基乙基 )醚四乙酸 ]导致小麦幼苗干物质累积下降 ,降低地上部和地下部全钙含量 ,抑制小麦幼苗对外源硝态氮的吸收及其向地上部的运输 ,改变其在不同器官中的分配比例和影响其向氨基氮或可溶性蛋白质的转化。叶片和根中硝酸还原酶活性与相应器官中的NO3--N含量相关 ,叶片中谷氨酰胺合成酶 (GS)活性在EGTA处理后明显下降 ,根中GS活性变化不显著。     

Nitrogen assimilation in sunflower leaves and upward and downward transport of nitrogen

The assimilation of ammonium and nitrate nitrogen into amino acids of mature sunflower leaves and their transport to the other plant parts were investigated using nitrogen-15 as a tracer. In the leaf, to which ¹⁵N-labelled ammonium was vacuum-infiltrated, the ¹⁵N content of glutamine was always the highest of the amino acids tested and that of alanine was higher than that of glutamic acid and aspartic acid at 15 min after the infiltration. On the other hand in the leaf to which ¹⁵N-labelled nitrate was vacuum-infiltrated, the ¹⁵N content of glutamic acid and aspartic acid was superior to that of glutamine. The incorporation of ¹⁵N into serine was not active in the case of either ¹⁵N-labelled ammonium or nitrate. In the internodes above and below the treated leaf, through which photosynthates were transported into other parts, the ¹⁵N content of γ-aminobutyric acid and glutamine was markedly high when both nitrogen sources were supplied. There were no differences in the labelling patterns of amino acids between the upper and lower internodes. From these results it may be concluded that glutamine, glutamic acid, and aspartic acid play an important role in the assimilation of ammonium and nitrate nitrogen in leaves and that nitrogen is transported mainly in the forms of γ-aminobutyric acid and glutamine from the leaves to the other plant parts,     

绿茶施用铵态氮和硝态氮后木质部导管中N运移模拟

An experiment was carried out to study the transport process of nitrogen （N） assimilation from tea roots by monitoring the dynamic composition of N compounds in xylem sap after 15^N-NO3 and 15^N-NH4 were fed to the root of tea plants （Camellia sinensis L.）. Results showed that the main amino acids were glutamine, theanine, axginine, asparic acid and glutamic acid, which accounted for 49%, 17%, 8%, 7%, and 4%, respectively, of the total amino acids in the xylem sap. After the tea plants were fed with 15^N-NO3 and 15^N-NH4 for 48 h, the amount of total amino acids in xylem sap significantly increased and those fed with 15^N-NH4 had higher increment than those with 15^N-NOa. Two hours after 15^N- NO3 and 15^N-NH4 were fed, 15N abundance in glutamine, asparagine, glutamic acid, alanine, and arginine were detected and increased quickly over time. This indicated that it took less than 2 h for NO3-N and NH4-N to be absorbed by tea roots, incorporated into the above amino acids and transported to the xylem sap. Rapid increase in 15^N-NO3 in the xylem sap of tea plants fed with 15^N-NO3 indicated that nitrate could be directly transported to the xylem sap. Glutamine, theanine, and alanine were the main amino acids transported in xylem sap of tea plants fed with both 15^N-NO3 and 15^N-NH4.     

Uptake and translocation of nitrogen in patumma (Curcuma alismatifolia) by leaves or root

An adequate supply of nitrogen (N) is important for patumma growth and flower quality. This study aimed to compare the uptake and translocation of N by foliar and root application. Fertilization with ¹⁵ nitrate (NO₃)-N via roots or leaves was carried out at four stages, at the 1st to 4th fully expanded leaf (FEL) stages, and the plants were sampled at each successive stage. The uptake and translocation of ¹⁵N from foliar or root applications showed relatively similar patterns at all stages. Although the N fertilizer utilization rate by roots was higher than that via leaves, the foliar application stimulated reproductive growth by earlier flowering. The N supplied at the 1st FEL and the 2nd FEL was utilized mainly in leaves, whereas supplying N at the 3rd and 4th FEL promoted flower quality. Fertilizer application method and stage of application influence the utilization rate and translocation of N to the sink organs.     

Effects of atmospheric nitrogen dioxide on uptake and assimilation of ammonium in soybean plants

Twelve‐day‐old soybean plants were supplied with 1 mM ammonium chloride (NH₄C1) to roots and exposed to 0.2–0.25 μL.L^‐1 nitrogen dioxide (NO₂) for seven days. Amount and rate of ammonium uptake were decreased by NO₂ exposure. However, the ammonium concentration in leaves and leaf pH of exposed plants were increased by NO₂ exposure. These results suggest that the decrease in ammonium uptake may be due to the decline in hydrogen (H⁺) ion concentration in exposed plants which resulted from the reduction of the nitrate and nitrite from NO₂ absorption. They also suggest that the decrease in ammonium uptake and the competition for energy between nitrate reduction and ammonium assimilation may limit ammonium assimilation to organic nitrogen (N) which would further inhibit acidity increase in exposed plants and ammonium uptake by roots.     

Effect of alternative anions (Cl– vs. SO$ _4^{2-} $) on concentrations of free amino acids in young tea plants

The quality of green tea is highly dependent on the concentration of free amino acids, whose profile is dominated by the unique amino acid theanine (N⁵‐ethyl‐glutamine). A high quality is associated with a high amino acid–to–catechin ratio, but previous results indicated that excessive chloride (Cl^–) supply is detrimental for amino acid accumulation. Several experiments were conducted to investigate the effect of chloride on growth and concentrations of free amino acids in young tea plants. Soil‐grown tea plants supplied with different levels of potassium (K) as K₂SO₄ or KCl exhibited increased concentrations of free amino acid in young shoots only when supplied with K₂SO₄, and the negative effect of KCl supply was mainly due to a reduced concentration of theanine. Concentrations of other nutrients in plant tissues were not influenced. The uptake of Cl^– and its interaction with nitrogen (N) uptake were further investigated in a second experiment, in which soil‐cultivated tea plants were supplied with varying amounts of Cl^–. Chloride application reduced yield of young shoots, and severity of leaf damage was related to the concentration of Cl^– in leaves. Nitrogen uptake was reduced by Cl^– addition. To verify whether the decrease of free amino acids was simply a result of inhibited NO assimilation, a third experiment was conducted, in which tea plants were NH ‐fed in the absence or presence (equivalent to the NH concentration) of Cl^–. Again, concentrations of theanine and total free amino acids in young shoots were reduced by Cl^– supply, but changes of the free–amino acid pool did not contribute to the maintenance of charge balance. However, concentration of theanine in roots, where it is synthesized, was not influenced by Cl^–. Total N concentrations of roots and mature leaves, uptake rate of NH , and activity of glutamine synthetase in fibrous roots and young leaves were all unaffected by Cl^– as well. It is suggested that translocation of theanine from root to shoot and its catabolism in young shoots might be influenced by Cl^–.     

Root uptake of N-containing and N-free low molecular weight organic substances by maize: A C/N tracer study

Most studies showing potential organic nitrogen uptake were conducted with amino acids. They conclude that, in some ecosystems, amino acids significantly contribute to the N demand of plants and that roots have special transporters to re-uptake amino acids released into the rhizosphere. However, the relevance of the uptake of organic N compounds can only be evaluated by comparing the uptake of N-containing and N-free organic substances. We compared the uptake of alanine, glucose and acetate labelled with ¹⁴C by maize. Additionally, the N uptake was estimated by ¹⁵N labelled alanine and KNO₃. We found a similar uptake of ¹⁴C from alanine, glucose and acetate, amounting for the whole plant less than 1% of ¹⁴C input. These results show that maize did not prefer N-containing to N-free organic substances. The uptake of ¹⁵N by maize exceeded that of ¹⁴C (10- to 50-fold), irrespective of the ¹⁵N source. However, plant uptake of nitrate (23.6–35.2% of ¹⁵N input) always exceeded the uptake of N from alanine (9.6–28.8%). The uptake of organically bound N by maize growing in soil occurred mainly by transpiration flow – as dissolved organics. The contribution of specific amino acid transporters was minor.     

Aluminum effects on uptake and translocation of nitrogen in sorghum (Sorghum bicolor,L. Moench)

The effects of aluminum on the uptake and translocation of N in two hybrid cultivars of sorghum with differential tolerance to aluminum were studied.

Aluminum decreased the amount of N accumulated and the % of N in the aerial parts of the plants. In the roots the amount of N accumulated also decreased but the % of N increased, in both cultivars. Besides an effect on dry matter yield, Al probably reduces the uptake of N and its translocation to the aerial parts of the plant. Apparently, this impairment on N translocation resulted from Al effects on the root pressure.

Aluminum not only reduced the amount of N translocated but also changed the sap composition. The % of NO₃ ‐N decreased while the % of amino acid‐N increased suggesting an Al effect on N uptake and also on protein degradation. Asparagine and glutamine contributed about 80% of the free amino acid fraction; however, their proportions changed in presence of Al. Therefore, Al also interfered with the synthesis and/or interconversion of these amino acids.     

氮饥饿水稻利用不同形态氮素的差异及其生理机制

通过水培试验，研究了氮饥饿7d后，恢复供应不同形态氮源对水稻氮吸收和积累及氮同化中关键酶活性和光合色素的影响。结果表明，缺氮促进根系生长，增加根冠比。恢复供氮4d显著增加地上部生物量。铵硝混合营养促进了水稻对氮的吸收和转运，叶片和根系中全氮及叶片中铵态氮的含量以硝酸铵处理最高。与单一铵或硝营养相比，铵硝混合营养增强了根系的谷氨酰胺合成酶和叶片中硝酸还原酶的活性，提高了水稻同化和利用氮的能力。另外，与纯硝营养相比，供应铵态氮显著增加了叶片中总叶绿素，尤其是叶绿素a的含量。因此，改善水肥管理、平衡对水稻供氮的铵硝配比将提高水稻氮素的吸收和利用效率。     

Comparison of uptake of different N forms by soil microorganisms and two wet-grassland plants: A pot study

Two common plant species of temperate wet grasslands, Carex acuta and Glyceria maxima, were tested for their preferences in the uptake of different nitrogen (N) sources (amino acid, ammonium, nitrate) and their ability to compete for these sources with soil microorganisms. The experiment was a one-day incubation study with plants growing in soil obtained from the field, which was supplied with a solution containing the three N sources, one at a time labeled with ¹⁵N. The bulk of the N demand of both species was covered by nitrate-N, which was the dominant N form in the soil at the time of the experiment. Ammonium-N was taken up less strongly, and organic N formed only a negligible part of their nutrition. The assimilated inorganic N was preferentially transported to apical meristem of the youngest leaf, while organic N remained mostly in the roots. The fast-growing Glyceria took up more N and was a better competitor vis-à-vis soil microbes for rarer N forms than Carex. However, both plants were poor competitors for N vis-à-vis soil microbes, irrespective of the N form. Microbes took up nitrate ca. five times faster and organic N more than a hundred times faster than plants. Correspondingly, the calculated turnover time of microbial N was 17 days, compared to 40 days for N in plant roots. A significant amount of added ¹⁵N was found at non-exchangeable sites in the soil, which points to the importance of microbial N transformation and abiotic N fixation for N retention in soil. In summary, the preferential assimilation of inorganic N by the wetland plants studied here and their poor ability to compete for N with soil microbes over the short term agree with the results of studies carried out with other species from temperate grasslands.     

Ammonium assimilation in rice based on the occurrence of 15N and inhibition of glutamine synthetase activity

Assimilation of ammonium (NH₄) into free amino acids and total reduced nitrogen (N) was monitored in both roots and shoots of two‐week old rice seedlings supplied with 5 mM 99% (¹⁵NH₄)₂SO₄ in aerated hydroponic culture with or without a 2 h preincubation with 1 mM methionine sulfoximine (MSX), an inhibitor of glutamine synthetase (GS) activity. ¹⁵NH₄ was not assimilated into amino acids when the GS/GOGAT (glutamate synthase) cycle was inhibited by MSX. Inhibition of glutamine synthetase (GS) activity in roots with MSX increased both the amount of NH₄ and the abundance of ¹⁵N labeled NH₄. In contrast, the amount of Gln and Glu, and their proportions as ¹⁵N, decreased in roots when GS activity was inhibited. This research confirms the importance of GS/GOGAT in NH₄ assimilation in rice roots.

¹⁵N‐labeled studies indicate that NH₄ ions incorporated by roots of rice are transformed primarily into glutamine (Gln) and glutamic acid (Glu) before being converted to other amino acids through transamination (15). The formation of amino acids such as aspartic acid (Asp) and alanine (Ala) directly from free NH₄ in roots also has been reported (4,15). Translocation of free NH₄ to plant shoots, based on the concentration of free NH₄ in xylem exudate, has been reported in tomato (13), although NH₄ in shoots primarily originates from nitrate reduction in the shoot. Photorespiration also can contribute to the accumulation of NH₄ in leaves (7).

The GS/GOGAT cycle appears to be primarily responsible for the assimilation of exogenously supplied NH₄ and NH₄ derived from nitrate reduction in leaves, as well as NH₄ derived from photorespiration (2,3,6,8). Genetic evidence cited to support this conclusion includes the lethal effect of photorespiratory conditions on plant mutants deficient in chloroplast‐localized GS and GOGAT activities (2,3,9), and the rapid accumulation of free NH₄ in GS‐deficient mutants under photorespiratory conditions (2,3,5).

The present study was initiated to quantify the in vivo amino acid synthesis in rice roots and shoots by analysis of ¹⁵N labeling, and should provide a more complete understanding of this important system for NH₄ utilization.