Nitrogen affects cluster root formation and expression of putative peptide transporters

Non-mycorrhizal Hakea actites (Proteaceae) grows in heathland where organic nitrogen (ON) dominates the soil nitrogen (N) pool. Hakea actites uses ON for growth, but the role of cluster roots in ON acquisition is unknown. The aim of the present study was to ascertain how N form and concentration affect cluster root formation and expression of peptide transporters. Hydroponically grown plants produced most biomass with low molecular weight ON>inorganic N>high molecular weight ON, while cluster roots were formed in the order no-N>ON>inorganic N. Intact dipeptide was transported into roots and metabolized, suggesting a role for the peptide transporter (PTR) for uptake and transport of peptides. HaPTR4, a member of subgroup II of the NRT1/PTR transporter family, which contains most characterized di- and tripeptide transporters in plants, facilitated transport of di- and tripeptides when expressed in yeast. No transport activity was demonstrated for HaPTR5 and HaPTR12, most similar to less well characterized transporters in subgroup III. The results provide further evidence that subgroup II of the NRT1/PTR family contains functional di- and tripeptide transporters. Green fluorescent protein fusion proteins of HaPTR4 and HaPTR12 localized to tonoplast, and plasma- and endomembranes, respectively, while HaPTR5 localized to vesicles of unknown identity. Grown in heathland or hydroponic culture with limiting N supply or starved of nutrients, HaPTR genes had the highest expression in cluster roots and non-cluster roots, and leaf expression increased upon re-supply of ON. It is concluded that formation of cluster roots and expression of PTR are regulated in response to N suppl

Addressing the nitrogen problem in sugarcane production to reduce pollution of the Great Barrier Reef

The N pollution footprint of sugarcane cropping is large due to inefficiencies caused by mismatched N supply and crop N demand over sugarcane’s long N accumulation phase. The Great Barrier Reef lagoon receives excessive N loads that contribute to the rapidly declining reef health. Exceeding international average nitrous oxide emission rates several fold, sugarcane soils contribute significantly to Australia’s agricultural emissions. Nitrogen pollution reduction schemes over recent decades have mostly targeted reducing N fertiliser rates in line with expected yields and improving soil quality. Overall, these measures have not resulted in the desired N pollution reduction and further innovation is needed to address this problem. We present research that aims to aid agronomic innovation with (i) next-generation fertilisers that are based on repurposed nutrient-rich wastes and sorbent materials to better match N supply and crop demand and to improve soil function and carbon levels, (ii) understanding of soil N cycling and microbial processes, (iii) legume companion cropping as a source of biologically fixed N, and (iv) genetic improvement of sugarcane that more effectively captures and uses N. We conclude that evidence-based innovation has to support crop growers across climate and soil gradients in the 400,000 hectares of catchments of the Great Barrier Reef. This should include investment into new technologies to support ecologically-sound agriculture and a circular economy without waste and pollution

Legume crop rotation suppressed nitrifying microbial community in a sugarcane cropping soil

Nitrifying microorganisms play an important role in nitrogen (N) cycling in agricultural soils as nitrification leads to accumulation of nitrate (NO3 −) that is readily lost through leaching and denitrification, particularly in high rainfall regions. Legume crop rotation in sugarcane farming systems can suppress soil pathogens and improve soil health, but its effects on soil nitrifying microorganisms are not well understood. Using shotgun metagenomic sequencing, we investigated the impact of two legume break crops, peanut (Arachis hypogaea) and soybean (Glycine max), on the nitrifying communities in a sugarcane cropping soil. Cropping with either legume substantially increased abundances of soil bacteria and archaea and altered the microbial community composition, but did not significantly alter species richness and evenness relative to a bare fallow treatment. The ammonia oxidisers were mostly archaeal rather than bacterial, and were 24–44% less abundant in the legume cropping soils compared to the bare fallow. Furthermore, abundances of the archaeal amoA gene encoding ammonia monooxygenase in the soybean and peanut cropping soils were only 30–35% of that in the bare fallow. These results warrant further investigation into the mechanisms driving responses of ammonia oxidising communities and their nitrification capacity in soil during legume cropping

Soil N availability, rather than N deposition, controls indirect N2O emissions

Ammonia volatilised and re-deposited to the landscape is an indirect N2O emission source. This study established a relationship between N2O emissions, low magnitude NH4 deposition (0–30  kg N ha − 1 ), and soil moisture content in two soils using in-vessel incubations. Emissions from the clay soil peaked ( < 0.002 g N [ g soil ] − 1 min − 1 ) from 85 to 93% WFPS (water filled pore space), increasing to a plateau as remaining mineral-N increased. Peak N2O emissions for the sandy soil were much lower ( < 5 × 10 − 5 μg N [ g soil ] − 1 min − 1 ) and occurred at about 60% WFPS, with an indistinct relationship with increasing resident mineral N due to the low rate of nitrification in that soil. Microbial community and respiration data indicated that the clay soil was dominated by denitrifiers and was more biologically active than the sandy soil. However, the clay soil also had substantial nitrifier communities even under peak emission conditions. A process-based mathematical denitrification model was well suited to the clay soil data where all mineral-N was assumed to be nitrified ( R 2 = 90 % ), providing a substrate for denitrification. This function was not well suited to the sandy soil where nitrification was much less complete. A prototype relationship representing mineral-N pool conversions (NO3− and NH4+) was proposed based on time, pool concentrations, moisture relationships, and soil rate constants (preliminary testing only). A threshold for mineral-N was observed: emission of N2O did not occur from the clay soil for mineral-N <70 mg ( kg of soil ) − 1 , suggesting that soil N availability controls indirect N2O emissions. This laboratory process investigation challenges the IPCC approach which predicts indirect emissions from atmospheric N deposition alone

Effects of externally supplied protein on root morphology and biomass allocation in Arabidopsis

Growth, morphogenesis and function of roots are influenced by the concentration and form of nutrients present in soils, including low molecular mass inorganic N(IN, ammonium, nitrate) and organic N (ON, e.g.amino acids). Proteins, ON of high molecular mass, are prevalent in soils but their possible effects on roots have received little attention. Here, we investigated how externally supplied protein of a size typical of soluble soil proteins influences root development of axenically grown Arabidopsis. Addition of low to intermediate concentrations of protein (bovine serum albumen, BSA) to IN-replete growth medium increased root dry weight, root length and thickness, and root hair length. Supply of higher BSA concentrations inhibited root development. These effects were independent of total N concentrations in the growth medium. The possible involvement of phytohormones was investigated using Arabidopsis with defective auxin (tir1-1 and axr2-1) and ethylene (ein2-1) responses. That no phenotype was observed suggests a signalling pathway is operating independent of auxin and ethylene responses. This study expands the knowledge on N form-explicit responses to demonstrate that ON of high molecular mass elicits specific responses

Combination of Inorganic Nitrogen and Organic Soil Amendment Improves Nitrogen Use Efficiency While Reducing Nitrogen Runoff

Improved nitrogen fertiliser management and increased nitrogen use efficiency (NUE) can be achieved by synchronising nitrogen (N) availability with plant uptake requirements. Organic materials in conjunction with inorganic fertilisers provide a strategy for supplying plant-available N over the growing season and reducing N loss. This study investigated whether a combined application of inorganic N with an organic soil amendment could improve nitrogen use efficiency by reducing N loss in runoff. Nitrogen runoff from a ryegrass (Lolium multiflorum) cover was investigated using a rainfall simulator. Nitrogen was applied at low, medium and high (50, 75 and 100 kg/ha) rates as either (NH4)2SO4 or in combination with a poultry manure-based organic material. We showed that the NUE in the combination (58–75%) was two-fold greater than in (NH4)2SO4 (24–42%). Furthermore, this combination also resulted in a two-fold lower N runoff compared with the inorganic fertiliser alone. This effect was attributed to the slower rate of N release from the organic amendment relative to the inorganic fertiliser. Here, we demonstrated that the combined use of inorganic and organic N substrates can reduce nutrient losses in surface runoff due to a better synchronisation of N availability with plant uptake requirements. View Full-Tex

Combination of Inorganic Nitrogen and Organic Soil Amendment Improves Nitrogen Use Efficiency While Reducing Nitrogen Runoff

Improved nitrogen fertiliser management and increased nitrogen use efficiency (NUE) can be achieved by synchronising nitrogen (N) availability with plant uptake requirements. Organic materials in conjunction with inorganic fertilisers provide a strategy for supplying plant-available N over the growing season and reducing N loss. This study investigated whether a combined application of inorganic N with an organic soil amendment could improve nitrogen use efficiency by reducing N loss in runoff. Nitrogen runoff from a ryegrass (Lolium multiflorum) cover was investigated using a rainfall simulator. Nitrogen was applied at low, medium and high (50, 75 and 100 kg/ha) rates as either (NH4)2SO4 or in combination with a poultry manure-based organic material. We showed that the NUE in the combination (58–75%) was two-fold greater than in (NH4)2SO4 (24–42%). Furthermore, this combination also resulted in a two-fold lower N runoff compared with the inorganic fertiliser alone. This effect was attributed to the slower rate of N release from the organic amendment relative to the inorganic fertiliser. Here, we demonstrated that the combined use of inorganic and organic N substrates can reduce nutrient losses in surface runoff due to a better synchronisation of N availability with plant uptake requirements. View Full-Tex

Nitrate Paradigm Does Not Hold Up for Sugarcane

Modern agriculture is based on the notion that nitrate is the main source of nitrogen (N) for crops, but nitrate is also the most mobile form of N and easily lost from soil. Efficient acquisition of nitrate by crops is therefore a prerequisite for avoiding off-site N pollution. Sugarcane is considered the most suitable tropical crop for biofuel production, but surprisingly high N fertilizer applications in main producer countries raise doubt about the sustainability of production and are at odds with a carbon-based crop. Examining reasons for the inefficient use of N fertilizer, we hypothesized that sugarcane resembles other giant tropical grasses which inhibit the production of nitrate in soil and differ from related grain crops with a confirmed ability to use nitrate. The results of our study support the hypothesis that N-replete sugarcane and ancestral species in the Andropogoneae supertribe strongly prefer ammonium over nitrate. Sugarcane differs from grain crops, sorghum and maize, which acquired both N sources equally well, while giant grass, Erianthus, displayed an intermediate ability to use nitrate. We conclude that discrimination against nitrate and a low capacity to store nitrate in shoots prevents commercial sugarcane varieties from taking advantage of the high nitrate concentrations in fertilized soils in the first three months of the growing season, leaving nitrate vulnerable to loss. Our study addresses a major caveat of sugarcane production and affords a strong basis for improvement through breeding cultivars with enhanced capacity to use nitrate as well as through agronomic measures that reduce nitrification in soil

Crystal structure of plant acetohydroxyacid synthase, the target for several commercial herbicides

Acetohydroxyacid synthase (AHAS, EC 2.2.1.6) is the first enzyme in the branched-chain amino acid biosynthesis pathway. Five of the most widely used commercial herbicides (i.e. sulfonylureas, imidazolinones, triazolopyrimidines, pyrimidinyl-benzoates and sulfonylamino-cabonyl-triazolinones) target this enzyme. Here we have determined the first crystal structure of a plant AHAS in the absence of any inhibitor (2.9 Å resolution) and it shows that the herbicide-binding site adopts a folded state even in the absence of an inhibitor. This is unexpected because the equivalent regions for herbicide binding in uninhibited Saccharomyces cerevisiae AHAS crystal structures are either disordered, or adopt a different fold when the herbicide is not present. In addition, the structure provides an explanation as to why some herbicides are more potent inhibitors of Arabidopsis thaliana AHAS compared to AHASs from other species (e.g. S. cerevisiae). The elucidation of the native structure of plant AHAS provides a new platform for future rational structure-based herbicide design efforts. Database: The coordinates and structure factors for uninhibited AtAHAS have been deposited in the Protein Data Bank (www.pdb.org) with the PDB ID code 5K6Q

Turning the Table: Plants Consume Microbes as a Source of Nutrients

Interactions between plants and microbes in soil, the final frontier of ecology, determine the availability of nutrients to plants and thereby primary production of terrestrial ecosystems. Nutrient cycling in soils is considered a battle between autotrophs and heterotrophs in which the latter usually outcompete the former, although recent studies have questioned the unconditional reign of microbes on nutrient cycles and the plants' dependence on microbes for breakdown of organic matter. Here we present evidence indicative of a more active role of plants in nutrient cycling than currently considered. Using fluorescent-labeled non-pathogenic and non-symbiotic strains of a bacterium and a fungus (Escherichia coli and Saccharomyces cerevisiae, respectively), we demonstrate that microbes enter root cells and are subsequently digested to release nitrogen that is used in shoots. Extensive modifications of root cell walls, as substantiated by cell wall outgrowth and induction of genes encoding cell wall synthesizing, loosening and degrading enzymes, may facilitate the uptake of microbes into root cells. Our study provides further evidence that the autotrophy of plants has a heterotrophic constituent which could explain the presence of root-inhabiting microbes of unknown ecological function. Our discovery has implications for soil ecology and applications including future sustainable agriculture with efficient nutrient cycles