Partitioning of Sugar between Growth and Nitrate Reduction in Cotton Roots

The level of endogenous sugars was inversely related to nitrate availability in young cotton (Gossypium hirsutum L.) plants, with high nitrate causing a greater decline in sugar content of roots than of shoots. High nitrate (low sugar) plants also displayed relatively more shoot growth and less root growth than low nitrate (high sugar) plants. These data are consistent with the theory that roots are poor competitors for sugar, and that sugar supply is a major factor limiting root growth in vivo.

The effects of endogenous sugar level on root growth and on nitrate reductase activity in the root were different. When root sugar level was experimentally controlled by varying nitrate concentration in the nutrient solution, root growth was less sensitive than nitrate reductase activity to sugar deficiency. Also, in sterile root tips cultured on media containing a wide range of sucrose concentrations, growth rate was considerably less sensitive to endogenous sugar deficiency than was nitrate assimilation rate. Similarly, in plants which were detopped or girdled, nitrate reductase activity in the roots declined more rapidly than did root sugars, especially glucose and fructose. These results suggest that when sugar is deficient, cotton roots preferentially use it for growth at the expense of nitrate reduction.

     

Interactive regulation of nitrogen and aluminum in rice

Despite many studies on the high aluminum (Al) tolerance of rice (Oryza sativa), its exact mechanisms remain largely unknown. It is also unclear why Al improves growth of some plants. Our research on interactions between nitrogen (N) and Al may help to understand these phenomena. Previously, we found that ammonium-supplemented rice was more Al tolerant than nitrate-supplemented rice. Furthermore, Al-tolerant rice varieties preferred ammonium, while Al-sensitive ones preferred nitrate; in fact, Al tolerance was significantly correlated with the ammonium/nitrate preference among rice varieties. Al even enhanced growth of ammonium-supplemented rice, while it inhibited growth of nitrate-supplemented rice. Based on our own and other reports on N-Al interactions, we propose that intermediate products of N metabolism may play a role in rice Al tolerance. Al-enhanced ammonium utilization may explain why Al promotes growth of some plants, since Al often coexists with higher levels of ammonium than nitrate in acid soils.     

钙对黄瓜子叶愈伤组织中硝酸还原酶(NR)活性的影响

本文采用黄瓜子叶愈伤组织,研究了不同钙培养下,对愈伤组织生长、硝酸盐吸收,NR活性以及组织中钙、镁含量的影响。结果表明,缺钙后,愈伤组织生长、硝酸盐吸收、NR活性等都比正常钙培养下的愈伤组织降低。这与用整株植物所得结果基本一致,对此结果进行了讨论。     

Map-based cloning in crop plants: tomato as a model system II. Isolation and characterization of a set of overlapping yeast artificial chromosomes encompassing the jointless locus

Constructs carrying the entire or part of the tobacco nitrate reductase cDNA (NIA) cloned between the promoter and terminator sequences of the 35S RNA of the cauliflower mosaic virus were introduced into tobacco, in an attempt to improve nitrate assimilation. Several transgenic plants that had elevated NIA mRNA and nitrate reductase (NR) activity were obtained. In addition, a few plants that exhibited a chlorotic phenotype characteristic of NR-deficient mutants were also obtained. One of these plants contained no NIA mRNA, no NR activity and accumulated nitrate. This phenotype was therefore assumed to result from co-suppression of 35S-NIA transgenes and host NIA genes. NR-deficient plants were also found among the progeny of transformants overexpressing NIA mRNA. Genetic analyses indicated that these NR-deficient plants were homozygous for the 35S-NIA transgene, although not all homozygous plants were deficient for NR. The ratio of normal to NR-deficient plants in the progeny of homozygous plants remained constant at each generation, irrespective of the state of expression of the NIA genes (active or inactive) in the previous generation. This ratio also remained unchanged when field trials were performed in two areas of France: Versailles and Bergerac. The analysis of homozygous plants revealed that co-suppression was reversible at some stage of sexual reproduction. Indeed, host genes and transgenes reactivated at each generation, and co-suppression always appeared after a lag period of normal growth, suggesting that the phenomenon is developmentaly regulated. We observed that the triggering of cosuppression was delayed when plants were initially grown under limited light and/or watered with limited nitrate supply (light and nitrate both being required for the expression of the host NIA genes). However, this delay did not affect the final ratio between normal and NR-deficient plants after transfer to nitrate-fertilized fields. Independent transformants exhibited either different co-suppression ratios or no co-suppression at all, irrespective of the transgene copy number, suggesting that genomic sequences surrounding the transgene might play a role in determining co-suppression.     

Diagnosis of nitrogen deficiency and toxicity of Eucalyptus globulus seedlings by foliar analysis

The relationship between shoot growth and foliar nitrogen (N) in E. globulus seedlings was studied in the glasshouse to determine standard values for N deficiency and toxicity diagnosis. Seedlings were grown for 9 weeks in yellow sand, at 10 rates of N, applied as ammonium sulphate, calcium nitrate or ammonium nitrate. Shoot dry weight (DW) increased linearly with N rate for all forms of N in the deficiency range. Seedlings continued to respond to higher rates of ammonium and ammonium nitrate than to nitrate. Maximum shoot DW for nitrate fed plants and ammonium nitrate fed plants were 51% and 84% respectively of ammonium fed plants. Total N concentration in the youngest fully expanded leaf (YFEL) ranged from 1.0% to 3.3% in deficient and adequate plants. The critical N concentration for deficiency diagnosis (corresponding to 90% maximum yield) in the YFEL, determined from these growth response curves averaged over all N forms, was 2.6% N. For ammonium nitrate fed plants, total N concentration in the YFEL for the severely deficient, deficient, adequate, and toxic ranges were <1.4%, 1.4–2.5%, 2.6–3.5%, > 4.3%. High total N concentrations were associated with growth depression and toxicity symptoms, which differed with N form. For nitrate fed plants, a total N concentration above 3.3% in the YFEL was associated with severe growth depression, and leaf tip necrosis. The adequate concentration range for ammonium nitrate was similar to values found on a field trial with 7 month old E. globulus trees grown on an exforest site.     

钙对黄瓜子叶愈伤组织中硝酸还原酶（NR）活性的影响

本文采用黄瓜叶愈伤组织,研究了不同钙培养下,对愈伤组织生长、硝酸盐吸收,NR活性以及组织中钙、镁含量的影响。结果表明,缺钙后,愈伤组织生长、硝酸盐吸收、NR活性等都比正常钙培养下的愈伤组织降低。这与用整株植物所得结果基本一致,对此结果进行了讨论。     

The effect of deep placement and surface application of nitrogen fertilizers at different light intensities on growth and yield of wetland rice

Summary Lowland rice (RD 3) was cultivated in containers of clay soil submerged with 5 cm water under controlled conditions in the phytotron. Deep placement of urea supergranules 5 cm in the soil significantly enhanced both plant growth and fertilizer efficiency when the plants were cultivated under high light intensity (70 Wm^–2). At the highest urea level grain yield increased 119% above the control level, while growth and fertilizer efficiency was not as high when deep placement of calcium nitrate was used.The application of urea prills and calcium nitrate (18.4g Nm^–2) in two split doses on the soil surface increased grain yield as much as 91% above the control level. At the lower nitrogen concentration (9.2 g N m^–2), the urea prills were more efficient than calcium nitrate as indicated by the grain yield. The height of those plants fertilized by surface application was affected by the concentration and not the type of fertilizer. The number of tillers, however, was significantly higher on urea fertilized plants.When the rice plants were cultivated under low light intensity 930 Wm^–2), neither the nitrogen fertilizers nor the method of application had a significant effect on growth and yield.     

Effects of different N fertilizers on the activity of <Emphasis Type="Italic">Glomus mosseae</Emphasis> and on grapevine nutrition and berry composition

Grapevine N fertilization may affect and be affected by arbuscular mycorrhizal (AM) fungal colonization and change berry composition. We studied the effects of different N fertilizers on AM fungal grapevine root colonization and sporulation, and on grapevine growth, nutrition, and berry composition, by conducting a 3.5-year pot study supplying grapevine plants with either urea, calcium nitrate, ammonium sulfate, or ammonium nitrate. We measured the percentage of AM fungal root colonization, AM fungal sporulation, grapevine shoot dry weight and number of leaves, nutrient composition (macro- and micronutrients), and grapevine berry soluble solids (total sugars or °Brix) and total acidity. Urea suppressed AM fungal root colonization and sporulation. Mycorrhizal grapevine plants had higher shoot dry weight and number of leaves than non-mycorrhizal and with a higher growth response with calcium nitrate as the N source. For the macronutrients P and K, and for the micronutrient B, leaf concentration was higher in mycorrhizal plants. Non-mycorrhizal plants had higher concentration of microelements Zn, Mn, Fe, and Cu than mycorrhizal. There were no differences in soluble solids (°Brix) in grapevine berries among mycorrhizal and non-mycorrhizal plants. However, non-mycorrhizal grapevine berries had higher acid content with ammonium nitrate, although they did not have better N nutrition and vegetative growth.     

High supply of NO3 ? mitigates salinity effects through an enhancement in the efficiency of photosystem II and CO2 assimilation in Jatropha curcas plants

This study was performed to determine if a high supply of N-NO₃ ^? is capable of mitigating negative salinity effects on photosynthesis and growth through the stimulation of nitrate assimilation, which could act as an sink from photosynthetic electron transport chain and restrict the over reduction in thylakoid membrane in Jatropha curcas leaves. The experiment was arranged in a factorial design with two nitrate concentrations (1 and 10?mM) and two NaCl levels (0 and 100?mM). Salt-stressed plants supplied with high NO₃ ^? demonstrated a higher nitrate uptake rate, nitrate reductase activity and soluble-protein content when compared with plants that presented low nitrate uptake. High nitrate assimilation was associated with higher leaf growth, CO₂ assimilation and lower membrane damage in salt-stressed plants. The superior performance of salt-stressed plants grown with high NO₃ ^? was indicated by a higher effective quantum yield of PSII and electron transport rate and lower energy excess at the PSII level and non-photochemical quenching. Interestingly, a high NO₃ ^? level in the absence of NaCl did not alter the leaf growth, photochemical activity and gas exchange parameters when compared with plants supplied with low nitrate. The proline and glycinebetaine contents were similarly increased in both low- and high-NO₃ ^? salt-stressed plants. Our data suggest that the favorable effects induced by high nitrate supply were possibly associated with stimulation in the nitrate assimilatory pathway. This process might have acted as a sink of electrons from the thylakoid membranes minimizing photo-damage and stimulating CO₂ assimilation under salinity in J. curcas.     

Nitrate Signaling and the Two Component High Affinity Uptake System in Arabidopsis

Nitrate transporters are important for nitrogen acquisition by plants and in algae some require two gene products, NRT2 and NAR2, for function. The NRT2 family was already described and the recent identification of a family of the NAR2-type genes in higher plants showed that there was a homologue in Arabidopsis, AtNAR2.1. Using heterologous expression in yeast and oocytes we showed that the two Arabidopsis AtNRT2.1 and AtNAR2.1 proteins interacted to give a functional high affinity nitrate transport system (HATS). The gene knock out mutant atnar2.1-1 is deficient specifically for HATS activity and the resulting growth phenotype on low nitrate concentration is more severe than for the atnrt2.1-1 knock out mutant. Physiological characterisation of the plant N status and gene expression revealed a pattern that was characteristic of severe nitrogen deficiency. Consistent with the down regulation of AtNRT2.1 expression, the atnar2.1-1 plants also displayed the same phenotype as atnrt2.1 mutants in lateral root (LR) response to low nitrate supply. Using atnar2.1-1 plants constitutively expressing the NpNRT2.1 gene, we now show a specific role for AtNAR2.1 in LR response to low nitrate supply. AtNAR2.1 is also involved in the repression of LR initiation in response to high ratios of sucrose to nitrogen in the medium. Therefore the two component system itself is likely to be involved in the signaling pathway integrating nutritional cues for LR architecture regulation. Using a green fluorescent protein-NRT2.1 protein fusion we show the essential role of AtNAR2.1 for the presence of AtNRT2.1 to the plasma membrane.Key Words: high affinity nitrate transport, nitrate transporter, nitrate signalling, root growth     

Growth but Not Photosynthesis Response of a Host Plant to Infection by a Holoparasitic Plant Depends on Nitrogen Supply

Parasitic plants can adversely influence the growth of their hosts by removing resources and by affecting photosynthesis. Such negative effects depend on resource availability. However, at varied resource levels, to what extent the negative effects on growth are attributed to the effects on photosynthesis has not been well elucidated. Here, we examined the influence of nitrogen supply on the growth and photosynthesis responses of the host plant Mikania micrantha to infection by the holoparasite Cuscuta campestris by focusing on the interaction of nitrogen and infection. Mikania micrantha plants fertilized at 0.2, 1 and 5 mM nitrate were grown with and without C. campestris infection. We observed that the infection significantly reduced M. micrantha growth at each nitrate fertilization and more severely at low than at high nitrate. Such alleviation at high nitrate was largely attributed to a stronger influence of infection on root biomass at low than at high nitrate fertilization. However, although C. campestris altered allometry and inhibited host photosynthesis, the magnitude of the effects was independent of nitrate fertilizations. The infection reduced light saturation point, net photosynthesis at saturating irradiances, apparent quantum yield, CO₂ saturated rate of photosynthesis, carboxylation efficiency, the maximum carboxylation rate of Rubisco, and maximum light-saturated rate of electron transport_, and increased light compensation point in host leaves similarly across nitrate levels, corresponding to a similar magnitude of negative effects of the parasite on host leaf soluble protein and Rubisco concentrations, photosynthetic nitrogen use efficiency and stomatal conductance across nitrate concentrations. Thus, the more severe inhibition in host growth at low than at high nitrate supplies cannot be attributed to a greater parasite-induced reduction in host photosynthesis, but the result of a higher proportion of host resources transferred to the parasite at low than at high nitrate levels.     

Repression of nitrate reductase in cucumber leaves caused by calcium deficiency

Growth of young cucumber plants was strongly inhibited, whencalcium was removed from the culture solution. The activitiesof nitrate reductase, glutamate dehydrogenase and glutaminesynthetase were investigated after the removal of calcium. Thoughthe activities of glutamine synthetase and glutamate dehydrogenasewere not altered much, nitrate reductase activity, measuredby in vitro and in vivo assays, decreased dramatically. Theloss of nitrate reductase activity coincided with the levelof nitrate in the leaves. When nitrate was supplied to the cucumberswith a nitrate deficiency, the plants induced nitrate reductasetogether with a distinct accumulation of nitrate. However, cucumberstreated for both calcium and nitrate deficiency failed to inducenitrate reductase and to accumulate nitrate on the additionof large amounts of nitrate. Leaf sections that had been treatedfor both calcium and nitrate deficiency could induce nitratereductase when floated on nitrate solution under the light.This indicates that the drastic loss of nitrate reductase causedby the removal of calcium was due mainly to the deficiency ofnitrate as the inducer in leaves. (Received December 19, 1979; )     

Effects of nitrogen dioxide and nitrate nutrition on nodulation, nitrogenase activity, growth, and nitrogen content of bean

The influence of nutrient nitrate level (0-20 millimolar) on the effects of NO₂ (0-0.5 parts per million) on nodulation and in vivo acetylene reduction activity of the roots and on growth and nitrate and Kjeldahl N concentration in shoots was studied in bean (Phaseolus vulgaris L. cv Kinghorn Wax) plants. Exposing 8-day old seedlings for 6 hours each day, for 15 days, to 0.02 to 0.5 parts per million NO₂ decreased total nodule weight at 0 and 1 millimolar nitrate, and nitrogenase (acetylene reduction) activity at all concentrations of nitrate. The pollutant had little effect on root fresh or dry weights. Shoot growth was inhibited by NO₂. The NO₂ exposure increased nitrate concentration in roots only at 20 millimolar nutrient nitrate. Exposure to NO₂ markedly increased Kjeldahl N concentration in roots but generally decreased that in shoots. The experiments demonstrated that nutrient N level and NO₂ concentration act jointly in affecting nodulation and N fixing capability, plant growth and composition, and root/shoot relationships of bean plants.     

The Opposing Actions of Arabidopsis CHROMOSOME TRANSMISSION FIDELITY7 and WINGS APART-LIKE1 and 2 Differ in Mitotic and Meiotic Cells

Sister chromatid cohesion, which is mediated by the cohesin complex, is essential for the proper segregation of chromosomes during mitosis and meiosis. Stable binding of cohesin with chromosomes is regulated in part by the opposing actions of CTF7 (CHROMOSOME TRANSMISSION FIDELITY7) and WAPL (WINGS APART-LIKE). In this study, we characterized the interaction between Arabidopsis thaliana CTF7 and WAPL by conducting a detailed analysis of wapl1-1 wapl2 ctf7 plants. ctf7 plants exhibit major defects in vegetative growth and development and are completely sterile. Inactivation of WAPL restores normal growth, mitosis, and some fertility to ctf7 plants. This shows that the CTF7/WAPL cohesin system is not essential for mitosis in vegetative cells and suggests that plants may contain a second mechanism to regulate mitotic cohesin. WAPL inactivation restores cohesin binding and suppresses ctf7-associated meiotic cohesion defects, demonstrating that WAPL and CTF7 function as antagonists to regulate meiotic sister chromatid cohesion. The ctf7 mutation only had a minor effect on wapl-associated defects in chromosome condensation and centromere association. These results demonstrate that WAPL has additional roles that are independent of its role in regulating chromatin-bound cohesin.     

Root symbioses ofAlnus glutinosa (L.) gaertn. and their possible role in alder decline: A preliminary study

During the last few years alder has declined in South Bohemia. The possible role of mycorrhizal and actinorhizal symbioses is reviewed and some of the preliminary results from experiments testing the influence of these symbioses on alder growth and the influence of eutrophication on the development of these symbioses are reported. Seedlings ofAlnus glutinosa were inoculated with arbuscular mycorrhizal (AM) fungi and the actinomyceteFrankia in experiment 1, and with rhizosphere soil collected from field sites with different degrees of alder damage in experiment 2. In both experiments, a solution containing nitrate, ammonia and phosphorus in concentrations simulating eutrophic waters, was applied. Both symbioses markedly promoted the growth of the seedlings in experiment 1. The plants inoculated with the rhizosphere soil microflora in experiment 2 were larger than the control plants. Response of the seedlings to the inoculation with the soil from the rhizosphere of damaged alder trees from six field sites differs, even though no correlation was found relating growth to the health status of the trees. Nutrient treatment did not have any effect on the growth of seedlings in either experiment. The dry weight ofFrankia was greater in mycorrhizal plants compared to nonmycorrhizal plants and mycorrhizal colonization is reduced inFrankia inoculated plants supplemented with phosphorus in experiment 1. Nitrogen enhanced mycorrhizal colonization in nodulated plants which were not supplemented with phosphorus no effect of nitrogen on actinorhiza was observed.     

The <Emphasis Type="Italic">trans</Emphasis> and <Emphasis Type="Italic">cis</Emphasis> zeatin isomers play different roles in regulating growth inhibition induced by high nitrate concentrations in maize

Abscisic acid (ABA), auxins, and cytokinins (CKs) are known to be closely linked to nitrogen signaling. In particular, CKs control the effects of nitrate availability on plant growth. Our group has shown that treatment with high nitrate concentrations limits root growth and leaf development in maize, and conditions the development of younger roots and leaves. CKs also affect source-sink relationships in plants. Based on these results, we hypothesized that CKs regulate the source-sink relationship in maize via a mechanism involving complex crosstalk with the main auxin indole-3-acetic acid (IAA) and ABA. To evaluate this hypothesis, various CK metabolites, IAA, and ABA were quantified in the roots and in source and sink leaves of maize plants treated with high and normal nitrate concentrations. The data obtained suggest that the cis and trans isomers of zeatin play completely distinct roles in maize growth regulation by a complex crosstalk with IAA and ABA. We demonstrate that while trans-zeatin (tZ) and isopentenyladenine (iP) regulate nitrate uptake and thus control final leaf sizes, cis-zeatin (cZ) regulates source and sink strength, and thus controls leaf development. The implications of these findings relating to the roles of ABA and IAA in plants’ responses to varying nitrate concentrations are also discussed.     

Nitrate Assimilation Contributes to Ralstonia solanacearum Root Attachment,Stem Colonization,and Virulence

Ralstonia solanacearum, an economically important plant pathogen, must attach, grow, and produce virulence factors to colonize plant xylem vessels and cause disease. Little is known about the bacterial metabolism that drives these processes. Nitrate is present in both tomato xylem fluid and agricultural soils, and the bacterium''s gene expression profile suggests that it assimilates nitrate during pathogenesis. A nasA mutant, which lacks the gene encoding the catalytic subunit of R. solanacearum''s sole assimilatory nitrate reductase, did not grow on nitrate as a sole nitrogen source. This nasA mutant exhibited reduced virulence and delayed stem colonization after soil soak inoculation of tomato plants. The nasA virulence defect was more severe following a period of soil survival between hosts. Unexpectedly, once bacteria reached xylem tissue, nitrate assimilation was dispensable for growth, virulence, and competitive fitness. However, nasA-dependent nitrate assimilation was required for normal production of extracellular polysaccharide (EPS), a major virulence factor. Quantitative analyses revealed that EPS production was significantly influenced by nitrate assimilation when nitrate was not required for growth. The plant colonization delay of the nasA mutant was externally complemented by coinoculation with wild-type bacteria but not by coinoculation with an EPS-deficient epsB mutant. The nasA mutant and epsB mutant did not attach to tomato roots as well as wild-type strain UW551. However, adding either wild-type cells or cell-free EPS improved the root attachment of these mutants. These data collectively suggest that nitrate assimilation promotes R. solanacearum virulence by enhancing root attachment, the initial stage of infection, possibly by modulating EPS production.