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Background Nitrogen (N) is a macronutrient that is essential for optimal plant growth and seed yield. Allotetraploid rapeseed (AnAnCnCn, 2n = 4x = 38) has a higher requirement for N fertilizers whereas exhibiting a lower N use efficiency (NUE) than cereal crops. N limitation adaptation (NLA) is pivotal for enhancing crop NUE and reducing N fertilizer use in yield production. Therefore, revealing the genetic and molecular mechanisms underlying NLA is urgent for the genetic improvement of NUE in rapeseed and other crop species with complex genomes. Results In this study, we integrated physiologic, genomic and transcriptomic analyses to comprehensively characterize the adaptive strategies of oilseed rape to N limitation stresses. Under N limitations, we detected accumulated anthocyanin, reduced nitrate (NO3−) and total N concentrations, and enhanced glutamine synthetase activity in the N-starved rapeseed plants. High-throughput transcriptomics revealed that the pathways associated with N metabolism and carbon fixation were highly over-represented. The expression of the genes that were involved in efficient N uptake, translocation, remobilization and assimilation was significantly altered. Genome-wide identification and molecular characterization of the microR827-NLA1-NRT1.7 regulatory circuit indicated the crucial role of the ubiquitin-mediated post-translational pathway in the regulation of rapeseed NLA. Transcriptional analysis of the module genes revealed their significant functional divergence in response to N limitations between allotetraploid rapeseed and the model Arabidopsis. Association analysis in a rapeseed panel comprising 102 genotypes revealed that BnaC5.NLA1 expression was closely correlated with the rapeseed low-N tolerance. Conclusions We identified the physiologic and genome-wide transcriptional responses of oilseed rape to N limitation stresses, and characterized the global members of the BnamiR827-BnaNLA1s-BnaNRT1.7s regulatory circuit. The transcriptomics-assisted gene co-expression network analysis accelerates the rapid identification of central members within large gene families of plant species with complex genomes. These findings would enhance our comprehensive understanding of the physiologic responses, genomic adaptation and transcriptomic alterations of oilseed rape to N limitations and provide central gene resources for the genetic improvement of crop NLA and NUE. Electronic supplementary material The online version of this article (10.1186/s12870-018-1507-y) contains supplementary material, which is available to authorized users.

A high concentration of ammonium (NH4+) as the sole source of nitrogen in the growth medium often is toxic to plants. The nitrate transporter NRT1.1 is involved in mediating the effects of NH4+ toxicity; however, the mechanism remains undefined. In this study, wild-type Arabidopsis (Arabidopsis thaliana Columbia-0 [Col-0]) and NRT1.1 mutants (chl1-1 and chl1-5) were grown hydroponically in NH4NO3 and (NH4)2SO4 media to assess the function of NRT1.1 in NH4+ stress responses. All the plants grew normally in medium containing mixed nitrogen sources, but Col-0 displayed more chlorosis and lower biomass and photosynthesis than the NRT1.1 mutants in (NH4)2SO4 medium. Grafting experiments between Col-0 and chl1-5 further confirmed that NH4+ toxicity is influenced by NRT1.1. In (NH4)2SO4 medium, NRT1.1 induced the expression of NH4+ transporters, increasing NH4+ uptake. Additionally, the activities of glutamine synthetase and glutamate synthetase in roots of Col-0 plants decreased and soluble sugar accumulated significantly, whereas pyruvate kinase-mediated glycolysis was not affected, all of which contributed to NH4+ accumulation. By contrast, the NRT1.1 mutants showed reduced NH4+ accumulation and enhanced NH4+ assimilation through glutamine synthetase, glutamate synthetase, and glutamate dehydrogenase. Moreover, the up-regulation of genes involved in ethylene synthesis and senescence in Col-0 plants treated with (NH4)2SO4 suggests that ethylene is involved in NH4+ toxicity responses. This study showed that NH4+ toxicity is related to a nitrate-independent signaling function of NRT1.1 in Arabidopsis, characterized by enhanced NH4+ accumulation and altered NH4+ metabolism, which stimulates ethylene synthesis, leading to plant senescence.

The effect of exogenous abscisic acid (ABA) on the nitrogen use efficiency (NUE) of Brassica napus was studied by pot experiments using ABA (1 mg L−1) daubed on the leaves at the early siliquing stage. The results showed that activities of proteases and glutamine synthetase (GS) were significantly promoted by the ABA treatment, which increased the capacity of degradation of foliar proteins and biosynthesis of glutamine (N-transport compounds), both of which promote decreased N in leaves of the ABA treatment. L-glutamine, other free amino acids, and soluble sugar content in the phloem sap of the ABA treatment were significantly higher than those of the control. A 15N label trial showed that more 15N was distributed from leaves to the grain by the ABA treatment, which resulted in a significantly higher NUE in the ABA treatment relative to the control.

The aim of this study was to determine the remediation efficiency of petroleum-contaminated soil from an oilfield using different types of remediation treatments under laboratory conditions. Compared with unamended soil as the control treatment (T1), soil samples were amended with bulrush straw powder (T2), with biochar alone (T3) and in combination with nutrients (nitrogen and phosphorus) (T4). The remediation experiment was carried out for 8 weeks. The extent of hydrocarbon degradation was monitored gravimetrically, and the residual oil fractions were analyzed by gas chromatography. The characteristics of the polluted soil (water-holding capacity and nutrients) were improved significantly by biochar addition (p

Background: Vacuoles are unique compartments in plant cells. Their multiple functions require massive fluxes of molecules across their limiting membrane, the tonoplast. Transport across the tonoplast is energized by its membrane potential and the proton gradient established by two proton pumps, the vacuolar H+-ATPase (V-ATPase), and the vacuolar H+-pyrophosphatase (V-PPase), which play important roles in the growth and development of plants. However, the mechanisms by which they affect plant growth and development, in the absence of tonoplast proton pumps, remain unclear. Results: In this study, we show that the Arabidopsis thaliana double mutant, vha-a2vha-a3, which lacks two tonoplast-localized isoforms of the membrane-integral V-ATPase subunit VHA-a, is viable, but shows a phenotype of inhibited growth and leaf chlorosis. Reduced total nitrogen absorption and increased nitrate assimilation lead to the accumulation of ammonium in the shoot, which results in the growth retardation of vha-a2vha-a3. Concurrently, the distribution and concentration of cations are abnormal in plants when VHA-a2 and VHA-a3 genes are absent, which is another major growth-limiting factor. Interestingly, the growth inhibition of the vha-a2vha-a3 double mutant was alleviated and senescence delayed, while the concentration of total nitrogen increased and that of ammonium decreased, when adding additional potassium. Conclusions: Our results show that plants can be viable without VHA-a2 and VHA-a3 but are affected by multiple factors that result in growth retardation, which can be alleviated by adding additional potassium, which provides a new insight on the relationship between vacuolar H+-ATPase and potassium.

This study examined the performances of Acorus calamus, Pontederia cordata, and Alisma plantagoaquatica in removing nitrogen (N) from farmland wastewater. P. cordata showed the fastest rate of N removal, followed by A. plantagoaquatica, whereas that of A. calamus was slowest. P. cordata and A. plantagoaquatica achieving a greater rate of TN reduction in soil than that by A. calamus. A. plantagoaquatica demonstrated the highest N adsorption capacity, 32.6% and 392.1% higher than that of P. cordata and A. calamus, respectively. The higher potential nitrification and denitrification rate, and abundance of functional genes in the P. cordata microcosm resulted in a stronger process of nitrification–denitrification, which accounted for 65.99% of TN loss. Plant uptake and nitrification–denitrification were responsible for 50.06% and 49.94% of TN removed within the A. plantagoaquatica. Nitrification-denitrification accounted for 86.35% of TN loss in A. calamus. These findings helped to insight into N removal mechanisms in different plants.

Plant vacuoles are unique compartments that play a critical role in plant growth and development. The vacuolar H+-ATPase (V-ATPase), together with the vacuolar H+-pyrophosphatase (V-PPase), generates the proton motive force that regulates multiple cell functions and impacts all aspects of plant life. We investigated the effect of V-ATPase activity in the vacuole on plant growth and development. We used an Arabidopsisthaliana (L.) Heynh. double mutant, vha-a2 vha-a3, which lacks two tonoplast-localized isoforms of the membrane-integral V-ATPase subunit VHA-a. The mutant is viable but exhibits impaired growth and leaf chlorosis. Nitrate assimilation led to excessive ammonium accumulation in the shoot and lower nitrogen uptake, which exacerbated growth retardation of vha-a2 vha-a3. Ion homeostasis was disturbed in plants with missing VHA-a2 and VHA-a3 genes, which might be related to limited growth. The reduced growth and excessive ammonium accumulation of the double mutant was alleviated by potassium supplementation. Our results demonstrate that plants lacking the two tonoplast-localized subunits of V-ATPase can be viable, although with defective growth caused by multiple factors, which can be alleviated by adding potassium. This study provided a new insight into the relationship between V-ATPase, growth, and ammonium accumulation, and revealed the role of potassium in mitigating ammonium toxicity.

Adequate amount of essential plant nutrients and plant density are vital factors influencing the productivity of oilseed rape (Brassica napus L.). Field experiments were conducted in southern China to study the influence of different fertilizing levels and planting densities on the seed yield, chlorophyll content and some growth traits of winter oilseed rape. Five fertilizing levels and six planting densities were set in a randomized block design. The tested planting densities were D1, D2, D3, D4, D5 and D6 with 7.5 × 104, 15 × 104, 22.5 × 104, 30 × 104, 37.5 × 104 and 45 × 104 plants ha−1, respectively. The tested fertilizing levels were F0 (control without fertilization), F1 (N = 60, P2O5 = 30, K2O = 52.5, S = 3.75, B =0.3 kg ha−1), F2 (N = 120, P2O5 = 60, K2O = 105, S = 7.50, B =0.60 kg ha−1), F3 (N = 180, P2O5 = 90, K2O = 157.5, S = 11.25, B =0.90 kg ha−1) and F4 (N = 240, P2O5 = 120, K2O = 210, S = 15, B =1.20 kg ha−1). The results showed that there were close relationships between the seed yield and fertilizing levels × planting densities. Below the fertilization level of F2, the seed yield increased as the planting density increased. Application of F3 caused a seed yield decline when the planting density surpassed 37.5 × 104 plants ha−1. The tested fertilizing levels caused remarkable increases in the seed yield but the plant density had little effects. The highest seed yield (2487 kg ha−1) was recorded in F4D3 followed by F4D1 (2308 kg ha−1), F4D5 (2303 kg ha−1) and F3D6 (2302 kg ha−1); while the minimum yield was obtained from the unfertilized soil treatments. The current research paper highlighted the importance of balanced N, P, K, S and B application under the different plant densities in winter oilseed rape. The individual growth of plants compensated the low plant density, but increasing the fertilization rates caused remarkable increases in the oilseed rape, suggesting that optimum nutrients rates are a critical factor in seed yield production.

The nitrate transporter NRT1.1 is involved in plant NH4+ toxicity; however, its mechanism remains undefined. In this study, wild-type Arabidopsis (Col-0) and NRT1.1 mutants (chl1-1 and chl1-5) were grown hydroponically in NH4NO3 and (NH4)2SO4 media to evaluate NRT1.1 function in NH4+ stress responses. All plants grew normally in mixed N sources, but Col-0 displayed more chlorosis, and lower biomass and photosynthesis than the NRT1.1 mutants in the (NH4)2SO4 condition. Grafting experiments between Col-0 and chl1-5 further confirmed that NH4+ toxicity is NRT1.1-dependent. In (NH4)2SO4 medium, NRT1.1 facilitated the higher expression of NH4+ transporters, increasing NH4+ uptake. Additionally, glutamine synthetase (GS) and glutamate synthetase (GOGAT) in roots of Col-0 plants decreased and soluble sugar accumulated significantly, whereas pyruvate kinase (PK)-mediated glycolysis was not affected, all of which contributed to NH4+ accumulation. In contrast, the NRT1.1 mutants reduced NH4+ accumulation and enhanced NH4+ assimilation through glutamate dehydrogenase (GDH) and glutamate-oxaloacetate transamination (GOT) activity. In addition, the upregulation of genes involved in senescence in Col-0 plants treated with (NH4)2SO4 suggests that ethylene could be involved in NH4+ toxicity responses. Our results indicate that NH4+ toxicity is dependent on NRT1.1 in Arabidopsis, characterized by enhanced NH4+ accumulation and by perturbed NH4+ metabolism, which stimulated ethylene-induced plant senescence.

Oilseed rape (Brassica napus L.) is a crucial source of edible oil and livestock feeding, and is a promising biofuel crop. The increasing demand for oilseed rape requires strategies to increase yield while retaining quality. Field experiments were performed in southern China to evaluate the impacts of fertiliser level and planting density on dry matter accumulation, seed oil and protein content (%) and yield (per ha), oil quality, and the profitability of oilseed rape. Fertiliser treatments contained nitrogen (N), phosphorus (P), potassium (K), sulfur (S) and boron (B) at four increasing rates, compared with nil fertiliser, and there were six planting densities (range 7.5–45 × 104 plants ha–1). Dry matter accumulation significantly (P < 0.05) increased in response to increasing plant population and fertiliser level, whereas harvest index decreased, suggesting that increasing the production of oilseed rape is limited to improvement of population quality. Seed oil and protein contents were significantly affected by fertiliser level rather than planting density. Increasing the fertiliser rate increased seed protein content, simultaneously slightly decreasing oil content. Glucosinolate content of rapeseed slightly decreased with increasing fertiliser but erucic and oleic acid contents were not affected, indicating that increasing the fertiliser rate might not reduce oilseed rape quality. Manipulation of fertiliser level was more effective than altering planting density for increasing seed oil and protein yields. Highest seed oil and protein yields resulted from the highest fertiliser application of 240 kg N, 52.4 kg P, 174.3 kg K, 15 kg S and 1.2 kg B ha–1, under a planting density of 22.5 × 104 plants ha–1. Maximum economic gain occurred with the two highest fertiliser levels, whereas planting density has no significant effect on profitability. A balanced application of NPKSB fertilisers should be employed with direct-sowing cultivation of oilseed rape, aimed at building a suitable population structure that balances plant population density and individual growth.

Nitrate, once taken up by plants, can either be stored in vacuoles or reduced by nitrate reductase in the cytoplasm. High accumulation of NO3 − in the vacuole occurs when assimilation into the cytoplasm is saturated. This study elucidates how proton pumps at the tonoplast (V-ATPase and V-PPase) affect the NO3 − content of Brassica napus by controlling the distribution of NO3 − between the cytoplasm and vacuole. Pot experiments were conducted in a greenhouse under normal N (15.0 mM nitrate) conditions using B. napus genotypes that demonstrated either high (Xiangyou15) or low (814) nitrogen use efficiency (NUE). The NO3 − content of the high NUE genotype was significantly lower than that of the low NUE genotype, whereas the total N per plant of the two genotypes was almost the same, suggesting that the different NUE between the two genotypes is not due to the difference of NO3 − uptake. The relative expression levels of V-ATPase (vha-a3) and V-PPase (avp1) genes in the high NUE genotype were significantly lower than in the low NUE genotype, resulting in lower V-ATPase and V-PPase activities in the high NUE genotype. The transport of NO3 − and protons from the cytoplasm to the vacuole is powered by V-ATPase and V-PPase, so their lower activities increase H+ efflux from and reduce NO3 − influx into the vacuoles of the high NUE genotype. We conclude that the lower activity of proton pumps at the tonoplast is the main reason the high NUE genotype possesses lower NO3 − content and higher N-use efficiency.

Two winter oilseed rape varieties grown in sand culture were labeled with 15Nitrogen (15N) at different growth stages to show the route of nitrogen (N) absorption, distribution, and transfer in oilseed rape (Brassica napus L.). Averaged over the two varieties, 84% of the N absorbed at the seedling stage, and 67% of the N absorbed at the stem elongation stage were distributed into the leaves. Of the N absorbed at flowering stage, 43% was distributed into the leaves and 36% into stems. However, 42.4% of the N absorbed at siliquing stage was directly found into the silique. The proportion of the N redistributed from vegetative organs into reproductive organs was 34%, 44%, 41%, and 32%, at seedling, stem elongation, flowering, and siliquing stages, respectively. The amounts of N transferred were 203, 326, 218, and 82 mg per plant, respectively. This corresponded to 65% of the total in the grain. The proportions of N lost after absorption were 24%, 11%, 12%, and 7% at the four growth stages, respectively. The ...

With hydroponics culture, we monitored the response of the growth and nutrient accumulation of oilseed rape (Brassica napus L.) to five ethanol concentrations: 0 mL•L−1 (control), 0.0125 mL...

The relationship between nitrogen efficiency (NE), defined as seed yield per unit nitrogen (N) application, and seed quality was examined in two oilseed rape (Brassica napus L.) varieties at 5 N application levels, 0.6, 3, 6, 12, 15 mmol L−1, N1, N2, N3, N4 and N5, respectively. Seed yield, oil yield and protein content were increased with the increase in N application level, but NE and oil content were decreased, and the fatty acid composition in seed was hardly changed. Analysis of seven fatty acids revealed aslight decrease in the contents of erucic acid and arachidonic acid with the increase in N application level, but no obvious change in the contents of palmitic acid, stearic acid, oleic acid, linoleic acid and linolenic acid. Compared with the low NE variety H29, the seed yield and contents of erucic acid and arachidonic acid in the high NE variety bin270 were more markedly increased with the increase in N application level, and the oil content was hardly changed. The seed yield, oilcontent and oil yield were higher in the high NE variety than in the low NE variety at all 5 N application levels. There were no significant differences in protein, palmitic acid, stearic acid and oleic acid contents between the varieties at any of the 5 N application levels, but there were slight differences in the linoleic acid and linolenic acid contents between the two varieties. In brief, N application improved oil yield more greatly in the high NE variety than in the low NE variety, but hardly affected the fatty acid composition. Therefore, the seed quality and oil content of oilseed rape may not be decreased by breeding of a high NE variety with a high N absorption efficiency and high N use efficiency.

To determine the relationship between nitrogen (N) redistribution of oilseed rape (Brassica napus L.) varieties and N-use efficiency during later growth stages, the differences in N-use efficiency between two varieties (X-36 and X-50) were studied using the 15N labeling method with sand culture under complete nutrient solution conditions. Results showed that X-36 had greater grain yield not because of greater N uptake but because of greater grain yield per unit N in the plant and N-harvesting index. The average proportion of N in the two varieties that was redistributed from the vegetative organs to the grain was 65.1%. The redistribution amount and the proportion of N absorbed at the stem elongation stage and redistributed into the grain of two varieties were the greatest; the least were amounts absorbed at the siliquing stage. The high-N-use-efficiency variety (X-36), when compared with the low-N-use-efficiency variety (X-50), had slower redistribution speed of N before the siliquing stage, which then b...

Differences of nitrogen efficiency of oilseed rape (Brassica napus L.) cultivars and their physiological properties were studied in a pot experiment, and the ratio of seed yield with no nitrogen supplied to that with normal nitrogen supply was adopted as a nitrogen efficiency coefficient. Results showed that the nitrogen efficiency coefficient determined for eight oilseed rape cultivars varied from 0.37 to 0.69, the ratio of nitrogen uptake amounts per plant, nitrogen transfer velocity from stems and leaves to seeds, and nitrogen physiological efficiency of oilseed rape cultivars under nitrogen stressed condition differed from with normal nitrogen supply. The higher the nitrogen efficiency of a cultivar, the higher the ratio of N uptake in no nitrogen to with N supplied. Under low nitrogen-supplying conditions, high nitrogen efficiency cultivars had longer roots, more lateral roots, higher amounts of reuse of nitrate from stem and leaves, and higher nitrate reductase activities in leaves.

Nitrate (NO3−) can accumulate in high concentrations in plant cell vacuoles if it is not reduced, reutilized or transported into the cytoplasm. Such accumulation of NO3− in the vacuole occurs when mechanisms for NO3− assimilation in the cytoplasm are saturated. Moreover, other processes such as efflux across the plasma membrane might affect NO3− accumulation in the vacuole. These are the main reasons limiting nitrogen use efficiency (NUE) in plants. This study elucidates mechanisms for NO3− transport from the cytoplasm to vacuoles by the V-proton pump (V-ATPase and V-PPase) and their relationship with different NUE in four Brassica napus genotypes. Pot experiments were conducted in a greenhouse under normal (15.0 mmol L−1) and limited N (7.5 mmol L−1) concentrations of nitrate using B. napus genotypes that demonstrated either high (742 and Xiangyou 15) or low (814 and H8) NUE (g g−1). Specific inhibitors of V-ATPase and V-PPase increased nitrate reductase (NR) activity, resulting in greatly decreased NO3− in plant tissues. Nitrate reductase activity and NO3− content correlated more highly to V-PPase activity than they did to V-ATPase activity, and correlation between V-PPase activity and NO3− content was significantly higher than it was to V-ATPase. Genotypes with high NUE had significantly lower activities of V-ATPase and V-PPase than those with low NUE. In the high-NUE plants, lower activities of V-proton pump underlie mechanisms that result in significantly lower NO3− content in plant tissues of the high-NUE genotypes than those found in plant tissues of the low-NUE genotypes. Our results show that the tonoplast proton pumps V-PPase and V-ATPase strongly negatively affect NR activity and positively affect NO3− content. V-PPase contributed more to this regulatory mechanism than did V-ATPase.