Strigolactones: New players in the nitrogen-phosphorus signalling interplay.

Nitrogen (N) and phosphorus (P) are among the most important macronutrients for plant growth and development, and the most widely used as fertilizers. Understanding how plants sense and respond to N and P deficiency is essential to optimize and reduce the use of chemical fertilizers. Strigolactones (SLs) are phytohormones acting as modulators and sensors of plant responses to P deficiency. In the present work, we assess the potential role of SLs in N starvation and in the N-P signalling interplay. Physiological, transcriptional and metabolic responses were analysed in wild-type and SL-deficient tomato plants grown under different P and N regimes, and in plants treated with a short-term pulse of the synthetic SL analogue 2'-epi-GR24. The results evidence that plants prioritize N over P status by affecting SL biosynthesis. We also show that SLs modulate the expression of key regulatory genes of phosphate and nitrate signalling pathways, including the N-P integrators PHO2 and NIGT1/HHO. The results support a key role for SLs as sensors during early plant responses to both N and phosphate starvation and mediating the N-P signalling interplay, indicating that SLs are involved in more physiological processes than so far proposed.

Cytosolic ascorbate peroxidase 1 modulates ascorbic acid metabolism through cooperating with nitrogen regulatory protein P-II in tea plant under nitrogen deficiency stress.

Nitrogen (N) element is essential nutrient, and affect metabolism of secondary metabolites in higher plants. Ascorbate peroxidase (APX) plays an important role in ascorbic acid (AsA) metabolism of tea plant. However, the roles of cytosolic ascorbate peroxidase 1 (CsAPX1) in AsA metabolism under N deficiency stress in tea plant remains unclear in detail. In this work, nitrogen regulatory protein P-II (CsGLB1) and CsAPX1 were identified by isobaric tags for relative and absolute quantitation (iTRAQ) from tea plant. The cell growth rates in transgenic Escherichia coli overexpressing CsAPX1 and CsGLB1 were higher than empty vector under N sufficiency condition. Phenotype of shoots and roots, AsA accumulation, and expression levels of AtAPX1 and AtGLB1 genes were changed in transgenic Arabidopsis hosting CsAPX1 under N deficiency stress. These findings suggested that cytosolic CsAPX1 acted a regulator in AsA accumulation through cooperating with GLB1 under N deficiency stress in tea plant.

Ozone-induced impairment of night-time stomatal closure in O3-sensitive poplar clone is affected by nitrogen but not by phosphorus enrichment.

Nocturnal transpiration may be a key factor influencing water use in plants. Tropospheric ozone (O3) and availability of nutrients such as nitrogen (N) and phosphorus (P) in the soil can affect daytime water use through stomata, but the combined effects of O3, N and P on night-time stomatal conductance (gs) are not known. We investigated the effects of O3 and soil availability of N and P on nocturnal gs and the dynamics of stomatal response after leaf severing in an O3-sensitive poplar clone (Oxford) subjected to combined treatments over a growing season in an O3 free air controlled exposure (FACE) facility. The treatments were two soil N levels (0 and 80 kg N ha-1; N0 and N80), three soil P levels (0, 40 and 80 kg P ha-1; P0, P40 and P80) and three O3 levels (ambient concentration, AA [35.0 ppb as hourly mean]; 1.5 × AA; 2.0 × AA). The analysis of stomatal dynamics after leaf severing suggested that O3 impaired stomatal closure execution. As a result, nocturnal gs was increased by 2.0 × AA O3 in August (+39%) and September (+108%). Night-time gs was correlated with POD0 (phytotoxic O3 dose) and increased exponentially after 40 mmol m-2 POD0. Such increase of nocturnal gs was attributed to the emission of ethylene due to 2.0 × AA O3 exposure, while foliar abscisic acid (ABA) or indole-3-acetic acid (IAA) did not affect gs at night. Interestingly, the O3-induced stomatal opening at night was limited by N treatments in August, but not limited in September. Phosphorus decreased nocturnal gs, although P did not modify the O3-induced stomatal dysfunction. The results suggest that the increased nocturnal gs may be associated with a need to improve N acquisition to cope with O3 stress.

Arbuscular mycorrhizal fungi alleviate phosphorus limitation by reducing plant N:P ratios under warming and nitrogen addition in a temperate meadow ecosystem.

Global change apart from ecosystem processes also influences the community structure of key organisms, such as arbuscular mycorrhizal fungi (AMF). We conducted a 3-year experiment where we suppressed with benomyl mycorrhiza to understand how AMF alter the plant community structure under warming and nitrogen (N) addition. The elemental content and foliar tissue stoichiometry of the dominant species Leymus chinensis and the subordinate species Puccinellia tenuiflora were studied along with soil nutrient stoichiometries. Overall, N addition enhanced plant N: phosphorus (P) ratios at a greater level than experimental warming did. Under global change conditions, AMF symbionts significantly increased soil available P concentrations, promoted plant P absorption and decreased the plant N:P ratios. AMF alleviate P limitation by reducing plant N:P ratios. Our results highlight that the negative influence of global change on plant productivity might cancel each other out through the additive effects of AMF and that global change will increase the dependency of plants on their mycorrhizal symbionts.

The cost of surviving nitrogen excess: energy and protein demand in the lichen Cladonia portentosa as revealed by proteomic analysis.

MAIN CONCLUSION: Different nitrogen forms affect different metabolic pathways in lichens. In particular, the most relevant changes in protein expression were observed in the fungal partner, with NO 3- mostly affecting the energetic metabolism and NH 4+ affecting transport and regulation of proteins and the energetic metabolism much more than NO 3- did. Excess deposition of reactive nitrogen is a well-known agent of stress for lichens, but which symbiont is most affected and how, remains a mystery. Using proteomics can expand our understanding of stress effects on lichens. We investigated the effects of different doses and forms of reactive nitrogen, with and without supplementary phosphorus and potassium, on the proteome of the lichen Cladonia portentosa growing in a 'real-world' simulation of nitrogen deposition. Protein expression changed with the nitrogen treatments but mostly in the fungal partner, with NO3- mainly affecting the energetic metabolism and NH4+ also affecting the protein synthesis machinery. The photobiont mainly responded overexpressing proteins involved in energy production. This suggests that in response to nitrogen stress, the photobiont mainly supports the defensive mechanisms initiated by the mycobiont with an increased energy production. Such surplus energy is then used by the cell to maintain functionality in the presence of NO3-, while a futile cycle of protein production can be hypothesized to be induced by NH4+ excess. External supply of potassium and phosphorus influenced differently the responses of particular enzymes, likely reflecting the many processes in which potassium exerts a regulatory function.

Nutrient enrichment effects on mycorrhizal fungi in an Andean tropical montane Forest.

Nitrogen (N) and phosphorus (P) deposition are increasing worldwide largely due to increased fertilizer use and fossil fuel combustion. Most work with N and P deposition in natural ecosystems has focused on temperate, highly industrialized, regions. Tropical regions are becoming more developed, releasing large amounts of these nutrients into the atmosphere. Nutrient enrichment in nutrient-poor systems such as tropical montane forest can represent a relatively large shift in nutrient availability, especially for sensitive microorganisms such as arbuscular mycorrhizal fungi (AMF). These symbiotic fungi are particularly critical, given their key role in ecosystem processes affecting plant community structure and function.To better understand the consequences of nutrient deposition in plant communities, a long-term nutrient addition experiment was set up in a tropical montane forest in the Andes of southern Ecuador. In this study, we investigated the impacts of 7 years of elevated N and P on AMF root colonization potential (AMF-RCP) through a greenhouse bait plant method in which we quantified root colonization. We also examined the relationship between AMF-RCP and rarefied tree diversity.After 7 years of nutrient addition, AMF-RCP was negatively correlated with soil P, positively correlated with soil N, and positively correlated with rarefied tree diversity. Our results show that AMF in this tropical montane forest are directly affected by soil N and P concentrations, but may also be indirectly impacted by shifts in rarefied tree diversity. Our research also highlights the need to fully understand the benefits and drawbacks of using different sampling methods (e.g., AMF-RCP versus direct root sampling) to robustly examine AMF-plant interactions in the future.

Size evolution in microorganisms masks trade-offs predicted by the growth rate hypothesis.

Adaptation to local resource availability depends on responses in growth rate and nutrient acquisition. The growth rate hypothesis (GRH) suggests that growing fast should impair competitive abilities for phosphorus and nitrogen due to high demand for biosynthesis. However, in microorganisms, size influences both growth and uptake rates, which may mask trade-offs and instead generate a positive relationship between these traits (size hypothesis, SH). Here, we evolved a gradient of maximum growth rate (µmax) from a single bacterium ancestor to test the relationship among µmax, competitive ability for nutrients and cell size, while controlling for evolutionary history. We found a strong positive correlation between µmax and competitive ability for phosphorus, associated with a trade-off between µmax and cell size: strains selected for high µmax were smaller and better competitors for phosphorus. Our results strongly support the SH, while the trade-offs expected under GRH were not apparent. Beyond plasticity, unicellular populations can respond rapidly to selection pressure through joint evolution of their size and maximum growth rate. Our study stresses that physiological links between these traits tightly shape the evolution of competitive strategies.

[Nutrients Recovery on the Growth of Nitrogen and Phosphorus Starved Microcystis aeruginosa].

Microcystis in natural water bodies may frequently go through periods of nutrient limitation and then may recover when the limited nutrient becomes available. We investigated changes in cell physiology and expression of photosynthesis-related genes during the recovery of Microcystis aeruginosa from nitrogen starvation and phosphorus starvation with the method of 14C isotope and fluorescent quantitative PCR. Our results suggested that Microcystis cells relieved from N starvation and P starvation resumed growth within 24 h and displayed significantly higher growth rates than not-starved-cells in the first 48 h. Carbon production rates and the expression levels of photosynthesis-related genes all increased rapidly after relieving from N starvation and P starvation in different degrees, enabling the rapid recovery from nutrient starvation. However, N-starved cells can not resume their cellular activity to full capacity when N became available and the damage of N deficiency to M. aeruginosa was unrecoverable, whereas cellular activity of P-starved cells could recover to normal properties.

Feeding protein supplements in alfalfa hay-based lactation diets improves nutrient utilization, lactational performance, and feed efficiency of dairy cows.

Due to the increasing cost of soybean meal and concerns of excess N being excreted into the environment, new protein supplements have been developed. Two products that have shown potential in increasing N utilization efficiency are slow-release urea (SRU; Optigen; Alltech Inc., Nicholasville, KY) and ruminal-escape protein derived from yeast (YMP; DEMP; Alltech Inc.). The objective of this study was to assess the effects of feeding these 2 supplements in alfalfa hay-based [45.7% of forage dietary dry matter (DM)] dairy diets on nutrient utilization, feed efficiency, and lactational performance of dairy cows. Twelve multiparous dairy cows were used in a triple 4 × 4 Latin square design with one square consisting of ruminally cannulated cows. Treatments included (1) control, (2) SRU-supplemented total mixed ration (SRUT), (3) YMP-supplemented total mixed ration (YMPT), and (4) SRU- and YMP-supplemented total mixed ration (SYT). The control consisted only of a mixture of soybean meal and canola meal in a 50:50 ratio. The SRU and the YMP were supplemented at 0.49 and 1.15% DM, respectively. The experiment consisted of 4 periods lasting 28 d each (21 d of adaptation and 7 d of sampling). Cows fed YMPT and SYT had decreased intake of DM, and all supplemented treatments had lower crude protein intake compared with those fed the control. Milk yield tended to have the greatest increase in YMPT compared with the control (41.1 vs. 39.7 kg/d) as well as a tendency for increased milk fat and protein yields. Feed efficiencies based on yields of milk, 3.5% fat-corrected milk, and energy-corrected milk increased at 10 to 16% due to protein supplementation. Cows fed protein supplements partitioned less energy toward body weight gain, but tended to partition more energy toward milk production. Efficiency of use of feed N to milk N increased by feeding SRUT and YMPT, and milk N-to-manure N ratio increased with YMPT. Overall results from this experiment indicate that replacing the mixture of soybean meal and canola meal with SRU and YMP in alfalfa hay-based dairy diets can be a good approach to improve nutrient utilization efficiencies in lactating dairy cows.

Mapping QTLs for yield and nitrogen-related traits in wheat: influence of nitrogen and phosphorus fertilization on QTL expression.

KEY MESSAGE: The present study identified some new important genomic regions and demonstrated the availability of conditional analysis in dissecting QTLs induced by environmental factors. The high input and low use efficiency of nutrient fertilizers require knowledge of the genetic control of crop reaction to nutrient supplements. In this study, 14 morphological and 8 physiological traits of a set of 182 wheat (Triticum aestivum L.) recombinant inbred lines (Xiaoyan 54 × Jing 411) were investigated in six environments to map quantitative trait loci (QTLs). The influence of nitrogen (N) and phosphorus (P) fertilization on QTL expression was studied by unconditional and conditional analysis. A total of 117 and 30 QTLs were detected by unconditional and conditional analysis, respectively, among which 21 were common for both methods. Thirty-four QTL clusters were identified. Eighteen conserved QTLs (15.4 % of the 117 QTLs) between years, but within nutritional treatment were found. The three major QTLs on chromosomes 2D, 4B and 6A were coincident with Rht8, Rht-B1b and TaGW2, respectively. The other two important intervals on chromosomes 4B and 7A for yield component traits were newly detected QTLs that warrant further study. By conditional analysis, spikelet number per spike was found to be induced by P fertilization mostly, whereas N fertilization had more effects on the expression of the QTLs for nitrogen concentration and utilization efficiency traits. QTLs that respond to N and P interactions were also detected. The results are helpful for understanding the genetic basis of N utilization efficiency in wheat under different N and P supplement environments and provide evidence for the availability of conditional analysis in dissecting QTLs induced by environmental factors.

Bioavailability of methionine hydroxy analog-calcium salt relative to DL-methionine to support nitrogen retention and growth in starter pigs.

This study assessed the efficacy of a calcium salt of methionine hydroxy analog (MHA-Ca, 84%) relative to dl-methionine (dl-Met, 99%) in starter pigs. In Experiment 1 (Exp. 1, N balance trial), 42 individually housed barrows (Genesus, average initial BW 19 kg) were used in two blocks of 21 pigs each with six replicates per treatment. Within each block, pigs were randomly allotted to seven diets: a methionine (Met)-deficient basal diet or the basal diet with three added levels of dl-Met (0.02%, 0.04% and 0.06%) or MHA-Ca (0.024%, 0.048% and 0.071%) on an equimolar basis. After a 7-day adaptation period, feces and urine were collected quantitatively for 5 days. Urinary and total N outputs decreased linearly (P < 0.05) with dl-Met or MHA-Ca supplementation. Nitrogen retention, expressed as g/day and as % of intake increased linearly (P < 0.01) with dl-Met and MHA-Ca supplementations. The relative efficacy of MHA-Ca to dl-Met was estimated to be 71.2% on a product-to-product basis for N retention expressed as % of intake. In Exp. 2 (performance trial), 280, 21-day-old crossbred (Pietrain × (Landrace × Large White)) pigs (eight pigs per pen, seven pens per treatment), were allocated to five diets in a completely randomized block design after a 10-day adaptation period. The Met-deficient basal diet contained 16.5% CP and 0.21% Met. Other diets were basal diet supplemented with two graded levels of dl-Met (0.04% and 0.08%) or MHA-Ca (0.062% and 0.12%) on a product basis at a dl-Met to MHA-Ca ratio of 65 : 100 at the expense of maize. BW and feed disappearance were monitored weekly for 3 weeks to determine performance. Final BW, average daily gain and average daily feed intake increased (P < 0.05) and feed to gain ratio decreased (P < 0.05) with the addition of Met to the basal diet irrespective of the source. Overall, the N retention results of Exp. 1 showed that the average relative bioavailability of MHA-Ca to dl-Met to support N retention (% of N intake) was 71% on a product- to-product basis (85% on an equimolar basis). In Exp. 2, pig performance was not different when Met was supplemented in the Met-deficient diet at a dl-Met to MHA-Ca ratio of 65 : 100 on a product basis.

Nitrogen deficiency as well as phosphorus deficiency in sorghum promotes the production and exudation of 5-deoxystrigol, the host recognition signal for arbuscular mycorrhizal fungi and root parasites.

Strigolactones released from plant roots induce hyphal branching of symbiotic arbuscular mycorrhizal (AM) fungi and germination of root parasitic weeds, Striga and Orobanche spp. We already demonstrated that, in red clover plants (Trifolium pratense L.), a host for both AM fungi and the root holoparasitic plant Orobanche minor Sm., reduced supply of phosphorus (P) but not of other elements examined (N, K, Ca, Mg) in the culture medium significantly promoted the secretion of a strigolactone, orobanchol, by the roots of this plant. Here we show that in the case of sorghum [Sorghum bicolor (L.) Moench], a host of both the root hemiparasitic plant Striga hermonthica and AM fungi, N deficiency as well as P deficiency markedly enhanced the secretion of a strigolactone, 5-deoxystrigol. The 5-deoxystrigol content in sorghum root tissues also increased under both N deficiency and P deficiency, comparable to the increase in the root exudates. These results suggest that strigolactones may be rapidly released after their production in the roots. Unlike the situation in the roots, neither N nor P deficiency affected the low content of 5-deoxystrigol in sorghum shoot tissues.

Cadmium toxicity to two marine phytoplankton under different nutrient conditions.

Cd accumulation and toxicity in two marine phytoplankton (diatom Thalassiosira weissflogii and dinoflagellate Prorocentrum minimum) under different nutrient conditions (nutrient-enriched, N- and P-starved conditions) were examined in this study. Strong interactions between the nutrients and Cd uptake by the two algal species were found. Cd accumulation as well as N and P starvation themselves inhibited the assimilation of N, P, and Si by the phytoplankton. Conversely, N starvation strongly inhibited Cd accumulation but no influence was observed under P starvation. However, the Cd accumulation difference between nutrient-enriched and N-starved cells was smaller when [Cd(2+)] was increased in the medium, indicating that net Cd accumulation was less dependent on the N-containing ligands at high-Cd levels. As for the subcellular distribution of the accumulated Cd, most was distributed in the insoluble fraction of T. weissflogii while it was evenly distributed in the soluble and insoluble fractions of P. minimum at low-Cd levels. A small percentage of cellular Cd (<15%) was adsorbed on the cell surface for both algae at the lowest [Cd(2+)], which increased when the [Cd(2+)] increased. Cd toxicity in phytoplankton was quantified as depression of growth and maximal photosynthetic system II quantum yield, and was correlated with the [Cd(2+)], intracellular Cd concentration, and Cd concentrations in the cell-surface-adsorbed, soluble, and insoluble fractions. According to the estimated median inhibition concentration (IC50) based on the different types of Cd concentration, the toxicity difference among the different nutrient-conditioned cells was the smallest when the Cd concentration in the soluble fraction was used, suggesting that it may be the best predictor of Cd toxicity under different nutrient conditions.

Nutrient status and plant growth effects of forest soils in the Basin of Mexico.

The nutrient status of forest soils in the Mexico City Air Basin was evaluated by observing plant growth responses to fertilization with N, P or both nutrients combined. P deficiency was the most frequent condition for soil from two high pollution sites and N deficiency was greatest at a low N deposition site. Concentrations of Pb and Ni, and to a lesser extent Zn and Co, were higher at the high pollution sites. However, positive plant growth responses to P and sometimes to N, and results of wheat root elongation bioassays, suggest that heavy metal concentrations were not directly phytotoxic. Further studies are needed to determine if heavy metal toxicity to mycorrhizal symbionts of eucalyptus (Eucalyptus camaldulensis Dehnh.) from high pollution sites may explain the P deficiency and stunted growth. P deficiency is expected to limit the capacity for biotic N retention in N saturated forested watersheds in the Basin of Mexico dominated by Andisols.

Miniaturized test system for soil respiration induced by volatile pollutants.

A miniaturized method based on 96-well microtitre plates was developed and used to study respiration in pristine and contaminated soils following addition of volatile substrates. Small soil samples were exposed to fuel components, which were volatilized from spatially separate reservoirs of 2,2,4,4,6,8,8-heptamethylnonane (HMN) as an organic carrier. Respiration was determined as CO(2) production by means of a pH-indicator and bicarbonate-containing agar, or as (14)CO(2) evolution from (14)C-labelled substrates. Substrate concentrations inducing maximum microbial activity or inhibition were determined and CO(2) production profiles examined by multivariate analysis. When high concentrations of fuel components were applied, distinction of hydrocarbon exposed soils from unexposed soil was achieved within 6 h of incubation. With low concentrations, adequate distinction was achieved after 24 h, probably as a result of community adaptation. Nutrient limitation was identified with the (14)C method for toluene, and the optimal N and P amendment determined. Further potential applications of this rapid and inexpensive method are outlined.

Nutrient and temperature limitation of bacterioplankton growth in temperate lakes.

Limitation of bacterioplankton production by nutrients and temperature was investigated in eight temperate lakes in summer. Six of the lakes were resampled in autumn. The lakes differ in nutrient content, water color, and concentration of dissolved organic carbon. Nutrients (phosphorus, nitrogen, and organic carbon) were added alone and in all possible combinations to filtered lake water inoculated with bacteria from the lake. After incubation for 36-40 h at in situ temperatures (ranging from 7 to 20 degrees C), the response in bacterioplankton production was determined. The effect of increased temperature on bacterioplankton growth was also tested. Bacterioplankton production was often limited by phosphorus alone, organic carbon alone, or the two in combination. Phosphorus limitation of bacterioplankton production was more common in the summer, whereas limitation by organic carbon was more frequently observed in the autumn. There was a close balance between limitation by phosphorus and organic carbon in the epilimnion in the summer. In the hypolimnion in the summer, bacterioplankton growth was primarily phosphorus-limited. The effect of phosphorus additions decreased with increasing phosphorus concentrations in the lakes. However, there were no correlations between the effect of added organic carbon and water color, dissolved organic carbon concentration, or phosphorus concentration. When temperature was low (in the hypolimnion in the summer, and throughout the water column in the autumn) temperature also limited bacterioplankton production. Thus, temperature and inorganic nutrients or organic compounds can limit bacterioplankton growth both alone and simultaneously. However, at low temperatures, temperature is the most important factor influencing bacterioplankton growth.

Sex allocation of a cosexual wind-pollinated tree, Quercus dentata, in terms of four currencies.

Sex allocation of a cosexual wind-pollinated species, Quercus dentata (Fagaceae), was analyzed using biomass, carbon, nitrogen and phosphorus as currencies based on data accumulated for 61 individuals from 1997-2004. Strongly female-biased sex allocation was indicated when measured in terms of biomass and carbon, but no significant bias was detected when measured in terms of nitrogen or phosphorus. From an adaptive viewpoint, there is little support for strong female-biased sex allocation, suggesting that sex allocation in terms of nitrogen or phosphorus is closer to the real picture. The relative sex allocation considerably varied from year to year, but the relative femaleness of individuals in the population was rather constant across years. No significant correlation was observed between relative sex allocation and fecundity or tree height, but individuals that showed very low fecundity tended to produce only acorns.

Assessing the generality of global leaf trait relationships.

Global-scale quantification of relationships between plant traits gives insight into the evolution of the world's vegetation, and is crucial for parameterizing vegetation-climate models. A database was compiled, comprising data for hundreds to thousands of species for the core 'leaf economics' traits leaf lifespan, leaf mass per area, photosynthetic capacity, dark respiration, and leaf nitrogen and phosphorus concentrations, as well as leaf potassium, photosynthetic N-use efficiency (PNUE), and leaf N : P ratio. While mean trait values differed between plant functional types, the range found within groups was often larger than differences among them. Future vegetation-climate models could incorporate this knowledge. The core leaf traits were intercorrelated, both globally and within plant functional types, forming a 'leaf economics spectrum'. While these relationships are very general, they are not universal, as significant heterogeneity exists between relationships fitted to individual sites. Much, but not all, heterogeneity can be explained by variation in sample size alone. PNUE can also be considered as part of this trait spectrum, whereas leaf K and N : P ratios are only loosely related.

High nitrogen : phosphorus ratios reduce nutrient retention and second-year growth of wetland sedges.

Shifts from nitrogen (N)- to phosphorus (P)-limited growth due to high N deposition may alter the functioning of wetland vegetation. This experiment tested how N vs P deficiency affects the growth and nutrient use of wetland sedges. Five wetland Carex species were grown at nine N : P supply ratios (0.6-405) with two absolute levels of N and P. Biomass and nutrient concentrations were determined after one and two growing seasons. Shoot biomass was maximal at N : P supply ratios of 15-26 after one season but 5-15 after two seasons. Photosynthesis after the first season, second-year growth, leaf longevity, and the fraction of nutrient supply retained by plants over two seasons were all negatively related to N : P supply ratios, with small effects of absolute supply. The five Carex species responded similarly to N : P ratios but differed in nutrient resorption efficiency and biomass allocation. Plants treated with high N : P ratios appeared to lose nutrients below ground. Such losses may reduce plant performance in P-limited wetlands affected by high N deposition.

The role of alternative oxidase in modulating carbon use efficiency and growth during macronutrient stress in tobacco cells.

When wild-type (wt) tobacco (Nicotiana tabacum cv. Petit Havana SR1) cells are grown under macronutrient (P or N) limitation, they induce large amounts of alternative oxidase (AOX), which constitutes a non-energy-conserving branch of the respiratory electron transport chain. To investigate the significance of AOX induction, wt cells were compared with transgenic (AS8) cells lacking AOX. Under nutrient limitation, growth of wt cell cultures was dramatically reduced and carbon use efficiency (g cell dry weight gain g(-1) sugar consumed) decreased by 42-63%. However, the growth of AS8 was only moderately reduced by the nutrient deficiencies and carbon use efficiency values remained the same as under nutrient-sufficient conditions. As a result, the nutrient limitations more severely compromised the tissue nutrient status (P or N) of AS8 than wt cells. Northern analyses and a comparison of the mitochondrial protein profiles of wt and AS8 cells indicated that the lack of AOX in AS8 under P limitation was associated with increased levels of proteins commonly associated with oxidative stress and/or stress injury. Also, the level of electron transport chain components was consistently reduced in AS8 while tricarboxylic acid cycle enzymes did not show a universal trend in abundance in comparison to the wt. Alternatively, the lack of AOX in AS8 cells under N limitation resulted in enhanced carbohydrate accumulation. It is concluded that AOX respiration provides an important general mechanism by which plant cells can modulate their growth in response to nutrient availability and that AOX also has nutrient-specific roles in maintaining cellular redox and carbon balance.