High-throughput shoot phenotyping reveals temporal growth responses to nitrogen and inorganic and organic phosphorus sources in tomato.

The application of nitrogen (N) and phosphorus (P) fertilizers to soils is required to maintain crop yields, so the sufficient and timely delivery of nutrients to match crop demand is important in fertilizer management. We quantified temporal growth responses of tomato plants with different rates of N and P application using high-throughput shoot phenotyping. The tomato plants were grown in soil that had organic, inorganic or a combination of sources of P incorporated. Additional N was added to each pot at low and high rates, 13 days after planting. At the same rate of total P application, the inorganic P source resulted in greater shoot growth at the early time points. Later on, the plants supplied with organic or mixed P sources grew faster than those that received the inorganic P source, resulting in comparable shoot biomass in all treatments at the time of destructive harvest. The shoot phenotyping data demonstrated that readily available soil P was important for early tomato growth while available N was more important in later stages of vegetative growth. These results suggest that a fertilizer formulation of combined inorganic and organic P sources may be able to sustain rapid and great shoot growth in tomato plants, while also reducing additional N input.

The function of the Medicago truncatula ZIP transporter MtZIP14 is linked to arbuscular mycorrhizal fungal colonization.

Soil micronutrient availability, including zinc (Zn), is a limiting factor for crop yield. Arbuscular mycorrhizal (AM) fungi can improve host plant growth and nutrition through the mycorrhizal pathway of nutrient uptake. Although the physiology of Zn uptake through the mycorrhizal pathway is well established, the identity of the related molecular components are unknown. Here, RNA-seq analysis was used to identify genes differentially-regulated by AM colonization and soil Zn concentration in roots of Medicago truncatula. The putative Zn transporter gene MtZIP14 was markedly up-regulated in M. truncatula roots when colonized by Rhizophagus irregularis. MtZIP14 restored yeast growth under low Zn availability. Loss-of-function mutant plants (mtzip14) had reduced shoot biomass compared to the wild-type when colonized by AM fungi and grown under low and sufficient soil Zn concentration; at high soil Zn concentration, there were no genotypic differences in shoot biomass. The vesicular and arbuscular colonization of roots was lower in the mtzip14 plants regardless of soil Zn concentration. We propose that MtZIP14 is linked to AM colonization in M. truncatula plants, with the possibility that MtZIP14 function with AM colonization is linked to plant Zn nutrition.

Establishing a quality management framework for commercial inoculants containing arbuscular mycorrhizal fungi.

Microbial inoculants containing arbuscular mycorrhizal (AM) fungi are potential tools in increasing the sustainability of our food production systems. Given the demand for sustainable agriculture, the production of such inoculants has potential economic value and has resulted in a variety of commercial inoculants currently being advertised. However, their use is limited by inconsistent product efficacy and lack of consumer confidence. Here, we propose a framework that can be used to assess the quality and reliability of AM inoculants. First, we set out a range of basic quality criteria which are required to achieve reliable inoculants. This is followed by a standardized bioassay which can be used to test inoculum viability and efficacy under controlled conditions. Implementation of these measurements would contribute to the adoption of AM inoculants by producers with the potential to increase sustainability in food production systems.

Track and trace: how soil labelling techniques have revealed the secrets of resource transport in the arbuscular mycorrhizal symbiosis.

Arbuscular mycorrhizal (AM) fungi colonise plant roots, and by doing so forge the 'mycorrhizal uptake pathway(s)' (MUP) that provide passageways for the trade of resources across a specialised membrane at the plant-fungus interface. The transport of nutrients such as phosphorus (P), nitrogen and zinc from the fungus, and carbon from the plant, via the MUP have mostly been quantified using stable or radioactive isotope labelling of soil in a specialised hyphae-only compartment. Recent advances in the study of AM fungi have used tracing studies to better understand how the AM association will function in a changing climate, the extent to which the MUP can contribute to P uptake by important crops, and how AM fungi trade resources in interaction with plants, other AM fungi, and friend and foe in the soil microbiome. The existing work together with well-designed future experiments will provide a valuable assessment of the potential for AM fungi to play a role in the sustainability of managed and natural systems in a changing climate.

Arbuscular mycorrhizas increased tomato biomass and nutrition but did not affect local soil P availability or 16S bacterial community in the field.

While interest in arbuscular mycorrhizal (AM) fungal effects on soil phosphorus (P) have recently increased, field experiments on this topic are lacking. While microcosm studies provided valuable insights, the lack of field studies represents a knowledge gap. Here, we present a field study in which we grew a mycorrhiza-defective tomato (Solanum lycopersicum L.) genotype (named rmc) and its mycorrhizal wild-type progenitor (named 76R) with and without additional fertiliser, using a drip-irrigation system to examine the impacts of the AM symbiosis on soil P availability and plant growth and nutrition. AM effects on fruit biomass and nutrients, soil nutrient availability, soil moisture and the soil bacterial community were examined. At the time of harvest, the AM tomato plants without fertiliser had the same early season fruit biomass and fruit nutrient concentrations as plants that received fertiliser. The presence of roots reduced the concentration of available soil P, ammonium and soil moisture in the top 10 cm soil layer. Arbuscular mycorrhizas did not significantly affect soil P availability, soil moisture, or 16S bacterial community composition. These findings suggest an indirect role for AM fungi in tomato production but not necessarily a direct role in determining soil physicochemical traits, during the one season that this experiment was conducted. While longer-term field studies may be required in the future, the present study provides new insights into impacts of AM fungi on P availability and uptake in a field soil system.

Development of an organomineral fertiliser formulation that improves tomato growth and sustains arbuscular mycorrhizal colonisation.

Achieving sustainable agricultural development requires the efficient use of nutrient resources for crop production. Recovering nutrients from animal manures may play a key role in achieving this. Animal manures typically have low nutrient concentrations, and in ratios that are often not ideal for balanced crop nutrition. Here, combinations of organic and inorganic phosphorus (P) were formulated as granular products (organomineral fertilisers) with granule size suitable for transport and spreading. The fertilisers were produced by granulating powdered chicken litter with MAP and urea powders making the following formulations: 0:4, 1:3, 2:2, 3:1, 4:0. The kinetics of NH4+-N and P release from the fertilisers, and the effects on tomato growth and nutrition, as well as arbuscular mycorrhizal formation in roots following fertiliser application, were determined. Cumulative NH4+-N release ceased within 12 h, and was lower in the formulations with higher proportions of chicken litter. The cumulative P released reached approximately 80% of total P in all formulations, and the time to obtain maximum P dissolution was 19 days in the formulation that contained only chicken litter. The organomineral fertilisers increased tomato shoot growth by 15-28% compared to the chicken litter only, MAP only and MAP/urea formulations. Reasonable levels of mycorrhizal colonisation of tomato roots was achieved with the organomineral fertilisers. The results demonstrated that optimum plant growth does not depend solely on immediately available P, and that timing of nutrient supply to match plant demand is important. The combination of chicken litter with MAP sustained nutrient supply and improved plant growth. Taken together, organomineral fertiliser formulations are potential alternatives to inorganic P fertilisers that can improve crop growth and nutrition, while provide a sustainable use for animal production wastes.

Effects of plant roots and arbuscular mycorrhizas on soil phosphorus leaching.

While the impact of arbuscular mycorrhizal fungi (AMF) on phosphorus (P) uptake is well understood, the mechanism(s) of how these fungi affect P leaching from soil is still unclear. Here we present results of a study in which we grew a mycorrhiza-defective tomato (Solanum lycopersicum L.) genotype (named rmc) and its mycorrhizal wild-type progenitor (named 76R) in microcosms containing non-sterile soil, to examine the influence of roots and AMF on P leaching. More P was leached from the planted microcosms as compared to the plant-free controls. Further, although there was more plant biomass and greater P uptake in the mycorrhizal plant treatments, these treatments were associated with the most leaching of total P, reactive P, and dissolved organic carbon (DOC). There was a strong correlation between the total P and DOC leached, suggesting that root and fungal exudates may have affected P leaching. These findings provide new insights into the impact of roots and AMF on nutrient leaching in soils.

Smoothing and extraction of traits in the growth analysis of noninvasive phenotypic data.

BACKGROUND: Non-destructive high-throughput plant phenotyping is becoming increasingly used and various methods for growth analysis have been proposed. Traditional longitudinal or repeated measures analyses that model growth using statistical models are common. However, often the variation in the data is inappropriately modelled, in part because the required models are complicated and difficult to fit. We provide a novel, computationally efficient technique that is based on smoothing and extraction of traits (SET), which we compare with the alternative traditional longitudinal analysis methods. RESULTS: The SET-based and longitudinal analyses were applied to a tomato experiment to investigate the effects on plant growth of zinc (Zn) addition and growing plants in soil inoculated with arbuscular mycorrhizal fungi (AMF). Conclusions from the SET-based and longitudinal analyses are similar, although the former analysis results in more significant differences. They showed that added Zn had little effect on plants grown in inoculated soils, but that growth depended on the amount of added Zn for plants grown in uninoculated soils. The longitudinal analysis of the unsmoothed data fitted a mixed model that involved both fixed and random regression modelling with splines, as well as allowing for unequal variances and autocorrelation between time points. CONCLUSIONS: A SET-based analysis can be used in any situation in which a traditional longitudinal analysis might be applied, especially when there are many observed time points. Two reasons for deploying the SET-based method are (i) biologically relevant growth parameters are required that parsimoniously describe growth, usually focussing on a small number of intervals, and/or (ii) a computationally efficient method is required for which a valid analysis is easier to achieve, while still capturing the essential features of the exhibited growth dynamics. Also discussed are the statistical models that need to be considered for traditional longitudinal analyses and it is demonstrated that the oft-omitted unequal variances and autocorrelation may be required for a valid longitudinal analysis. With respect to the separate issue of the subjective choice of mathematical growth functions or splines to characterize growth, it is recommended that, for both SET-based and longitudinal analyses, an evidence-based procedure is adopted.

The effect of zinc fertilisation and arbuscular mycorrhizal fungi on grain quality and yield of contrasting barley cultivars.

Zinc is essential for the functioning of many enzymes and plant processes and the malting process. Arbuscular mycorrhizal fungi (AMF) can improve zinc (Zn) uptake in the important cereal crop barley (Hordeum vulgare) on Zn-deficient soils. Here we investigated the impacts of Zn fertilisation and AMF on the yield and grain quality of malting barley cultivars. Five barley genotypes were inoculated or not with the AMF Rhizophagus irregularis, and grown in pots either fertilised with Zn or not. Measurements of Zn nutrition and yield were made for all cultivars. Further analyses of grain biochemical composition, including starch, ß-glucan and arabinoxylan contents, and analysis of ATR-MIR spectra were made in two contrasting cultivars. Mycorrhizal colonisation generally resulted in decreased biomass, but increased grain dimensions and mean grain weight. Barley grain yield and biochemical qualities were highly variable between cultivars, and the ATR-MIR spectra revealed grain compositional differences between cultivars and AMF treatments. Mycorrhizal fungi can affect barley grain Zn concentration and starch content, but grain biochemical traits including ß-glucan and arabinoxylan contents were more conserved by the cultivar, and unaffected by AMF inoculation. The ATR-MIR spectra revealed that there are other grain characteristics affected by AMF that remain to be elucidated.

The effects of soil phosphorus and zinc availability on plant responses to mycorrhizal fungi: a physiological and molecular assessment.

The positive effects of arbuscular mycorrhizal fungi (AMF) have been demonstrated for plant biomass, and zinc (Zn) and phosphorus (P) uptake, under soil nutrient deficiency. Additionally, a number of Zn and P transporter genes are affected by mycorrhizal colonisation or implicated in the mycorrhizal pathway of uptake. However, a comprehensive study of plant physiology and gene expression simultaneously, remains to be undertaken. Medicago truncatula was grown at different soil P and Zn availabilities, with or without inoculation of Rhizophagus irregularis. Measures of biomass, shoot elemental concentrations, mycorrhizal colonisation, and expression of Zn transporter (ZIP) and phosphate transporter (PT) genes in the roots, were taken. Mycorrhizal plants had a greater tolerance of both P and Zn soil deficiency; there was also evidence of AMF protecting plants against excessive Zn accumulation at high soil Zn. The expression of all PT genes was interactive with both P availability and mycorrhizal colonisation. MtZIP5 expression was induced both by AMF and soil Zn deficiency, while MtZIP2 was down-regulated in mycorrhizal plants, and up-regulated with increasing soil Zn concentration. These findings provide the first comprehensive physiological and molecular picture of plant-mycorrhizal fungal symbiosis with regard to soil P and Zn availability. Mycorrhizal fungi conferred tolerance to soil Zn and P deficiency and this could be linked to the induction of the ZIP transporter gene MtZIP5, and the PT gene MtPT4.

Arbuscular mycorrhizal fungal inoculation and soil zinc fertilisation affect the productivity and the bioavailability of zinc and iron in durum wheat.

There is a growing recognition of the role of arbuscular mycorrhizal fungi (AMF) in food security, specifically the potential for AMF to enhance the yield and mineral nutrition-including phosphorus, zinc (Zn), and iron (Fe)-of food crops. However, the bioavailability of Zn and Fe for humans in the grain of cereal crops can be overestimated by failing to consider the abundance of phytic acid (PA). This is because PA can chelate the micronutrients, making them difficult to absorb. In order to understand the effect of an AM fungus and soil Zn concentration on the productivity and nutritional quality of food parts, this study examined the growth and nutritional responses of durum wheat, with and without inoculation with Rhizophagus irregularis, at five soil Zn concentrations. Growth and nutrient responses of the plants to soil Zn amendment was stronger than responses to AMF. However, the protective effect of AMF under soil Zn toxicity conditions was observed as reduced Zn concentration in the mycorrhizal durum wheat grain at Zn50. Here, AMF inoculation increased the concentration of PA in durum wheat grain but had no effect on the concentration of Zn and Fe; this consequently reduced the predicted bioavailability of grain Zn and Fe, which could lead to a decrease in nutritional quality of the grain. This research suggests that in soil with low (available) phosphorus and Zn concentrations, AMF may reduce the food quality of durum wheat because of an increase in PA concentration, and thus, a decrease in the bioavailability of Zn and Fe.

Impact of an arbuscular mycorrhizal fungus on the growth and nutrition of fifteen crop and pasture plant species.

The formation of arbuscular mycorrhizas (AM) can result in positive, neutral or negative responses in the growth and mineral nutrition of host plants, particularly that of P, Zn and other micronutrients. This study examined the growth and nutritional responses of 15 agriculturally important plant species, including cereals, legumes and vegetables, with and without inoculation with the AM fungus (AMF) Rhizophagus irregularis. Furthermore, we explored whether the responses differed between different functional groups of plants such as monocots and dicots, C3 and C4 plants, and N-fixing and non-N-fixing plants. We found that that mycorrhizal colonisation of roots, plant growth and plant nutrient responses differed between plant species. Among the species analysed, leek (Allium ampeloprasum L. var. porrum) was the most mycorrhiza-responsive, displaying the highest mycorrhizal colonisation and biomass response, and the greatest increases in most mineral nutrients. In other plant species, the concentration of P, Cu, Zn and S were generally enhanced by inoculation with AMF. Furthermore, ionomes differed more greatly between plant species than in response to inoculation with AMF. This research further improves our understanding of the responses of different and diverse plant species to the formation of AM in terms of growth and ionomics under standardised growth conditions. The results of this study may be used in further studies and to inform agricultural practices.

The mycorrhizal pathway of zinc uptake contributes to zinc accumulation in barley and wheat grain.

BACKGROUND: Increasing zinc (Zn) concentrations in crops is important for alleviation of human Zn deficiency. Arbuscular mycorrhizal fungi (AMF) contribute to plant Zn uptake, but their contribution to Zn in the edible portion of crops has not yet been investigated. This study aimed to quantify the mycorrhizal pathway of Zn uptake into grain of wheat and barley under varying soil Zn availabilities. Bread wheat (Triticum aestivum) and barley (Hordeum vulgare) were grown in pots with a hyphal compartment containing 65Zn. Plants were inoculated with Rhizophagus irregularis and grown at three soil Zn concentrations. Radioactive Zn in grain and straw was measured and the contribution of AMF to Zn uptake was calculated. RESULTS: The mycorrhizal pathway of Zn uptake contributed up to 24.3% of total above-ground Zn in wheat, and up to 12.7% of that Zn in barley. The greatest contribution by the mycorrhizal pathway was observed in barley at the lowest Zn addition, and in wheat at the highest one. In addition, grain yield of bread wheat was increased by AMF. CONCLUSIONS: These results suggest that AMF have a substantial role in uptake of Zn into cereals, and the proportional contribution by the MPU is dependent on plant species, as well as available soil Zn.

Variable effects of arbuscular mycorrhizal fungal inoculation on physiological and molecular measures of root and stomatal conductance of diverse Medicago truncatula accessions.

Association with arbuscular mycorrhizal fungi (AMF) can impact on plant water relations; mycorrhizal plants can exhibit increased stomatal conductance (gs ) and root hydraulic conductance (normalized to root dry weight, Lo ), and altered expression of aquaporins (AQP). Many factors regulate such responses; however, plant intraspecific diversity effects have yet to be explored. Twenty geographically diverse accessions of Medicago truncatula were inoculated with the AMF Funneliformis mosseae or mock-inoculated, and grown under well-watered conditions. Biomass, gs , shoot nutrient concentrations and mycorrhizal colonization were measured in all accessions, and Lo and gene expression in five accessions. The diverse accessions varied in physiology and gene expression; some accessions were also larger or had higher gs when colonized by F. mosseae. In the five accessions, Lo was higher in two accessions when colonized by AMF and also maintained within a much smaller range than the mock-inoculated plants. Expression of MtPIP1 correlated with both gs and Lo , and when plants were more than 3% colonized, mycorrhizal colonization correlated with Lo . Accession and AMF treatments had profound effects on M. truncatula, including several measures of plant water relations. Correlations between response variables, especially between molecular and physiological variables, across genotypes, highlight the findings of this study.

Diverse Sorghum bicolor accessions show marked variation in growth and transcriptional responses to arbuscular mycorrhizal fungi.

Sorghum is an important crop grown worldwide for feed and fibre. Like most plants, it has the capacity to benefit from symbioses with arbuscular mycorrhizal (AM) fungi, and its diverse genotypes likely vary in their responses. Currently, the genetic basis of mycorrhiza-responsiveness is largely unknown. Here, we investigated transcriptional and physiological responses of sorghum accessions, founders of a bioenergy nested association mapping panel, for their responses to four species of AM fungi. Transcriptome comparisons across four accessions identified mycorrhiza-inducible genes; stringent filtering criteria revealed 278 genes that show mycorrhiza-inducible expression independent of genotype and 55 genes whose expression varies with genotype. The latter suggests variation in phosphate transport and defence across these accessions. The mycorrhiza growth and nutrient responses of 18 sorghum accessions varied tremendously, ranging from mycorrhiza-dependent to negatively mycorrhiza-responsive. Additionally, accessions varied in the number of AM fungi to which they showed positive responses, from one to several fungal species. Mycorrhiza growth and phosphorus responses were positively correlated, whereas expression of two mycorrhiza-inducible phosphate transporters, SbPT8 and SbPT9, correlated negatively with mycorrhizal growth responses. AM fungi improve growth and mineral nutrition of sorghum, and the substantial variation between lines provides the potential to map loci influencing mycorrhiza responses.

Arbuscular mycorrhizal fungi increase grain zinc concentration and modify the expression of root ZIP transporter genes in a modern barley (Hordeum vulgare) cultivar.

The positive effects of arbuscular mycorrhizal fungi (AMF) on the zinc (Zn) nutrition of a number of cereal species has been demonstrated, but for Hordeum vulgare (barley), this has been scarcely investigated. Zn is taken up by ZIP transporters in the roots, and several barley ZIP transporter genes are up-regulated under Zn deficient conditions. We grew a modern cultivar of barley (cv. Compass) at five different soil Zn concentrations ranging from no addition through to a toxic concentration. The plants were either inoculated with the AMF Rhizophagus irregularis, or mock-inoculated. At harvest, measurements of biomass, tissue Zn concentration, and expression of ZIP transporter genes were taken. Inoculation of barley with AMF resulted in improved grain and straw Zn concentrations, especially at low soil Zn concentrations, but did not increase the biomass of the plants. Of the five HvZIP genes tested that are up-regulated under low Zn conditions, one gene (HvZIP13) was significantly up-regulated by mycorrhizal colonisation at the lowest Zn treatment. Two other ZIP genes were down-regulated in mycorrhizal plants under low soil Zn. Inoculation with AMF has an effect on ZIP transporter genes in the roots of barley plants. Furthermore, AMF may be more useful for improving quality of barley grain in terms of Zn concentrations, rather than improving yield.

Suppression of the activity of arbuscular mycorrhizal fungi by the soil microbiota.

Arbuscular mycorrhizal fungi (AMF) colonise roots of most plants; their extra-radical mycelium (ERM) extends into the soil and acquires nutrients for the plant. The ERM coexists with soil microbial communities and it is unresolved whether these communities stimulate or suppress the ERM activity. This work studied the prevalence of suppressed ERM activity and identified main components behind the suppression. ERM activity was determined by quantifying ERM-mediated P uptake from radioisotope-labelled unsterile soil into plants, and compared to soil physicochemical characteristics and soil microbiome composition. ERM activity varied considerably and was greatly suppressed in 4 of 21 soils. Suppression was mitigated by soil pasteurisation and had a dominating biotic component. AMF-suppressive soils had high abundances of Acidobacteria, and other bacterial taxa being putative fungal antagonists. Suppression was also associated with low soil pH, but this effect was likely indirect, as the relative abundance of, e.g., Acidobacteria decreased after liming. Suppression could not be transferred by adding small amounts of suppressive soil to conducive soil, and thus appeared to involve the common action of several taxa. The presence of AMF antagonists resembles the phenomenon of disease-suppressive soils and implies that ecosystem services of AMF will depend strongly on the specific soil microbiome.