Simultaneous improvement of grain yield and grain protein concentration in durum wheat by using association tests and weighted GBLUP.

KEY MESSAGE: Simultaneous improvement for GY and GPC by using GWAS and GBLUP suggested a significant application in durum wheat breeding. Despite the importance of grain protein concentration (GPC) in determining wheat quality, its negative correlation with grain yield (GY) is still one of the major challenges for breeders. Here, a durum wheat panel of 200 genotypes was evaluated for GY, GPC, and their derived indices (GPD and GYD), under eight different agronomic conditions. The plant material was genotyped with the Illumina 25 k iSelect array, and a genome-wide association study was performed. Two statistical models revealed dozens of marker-trait associations (MTAs), each explaining up to 30%. phenotypic variance. Two markers on chromosomes 2A and 6B were consistently identified by both models and were found to be significantly associated with GY and GPC. MTAs identified for phenological traits co-mapped to well-known genes (i.e., Ppd-1, Vrn-1). The significance values (p-values) that measure the strength of the association of each single nucleotide polymorphism marker with the target traits were used to perform genomic prediction by using a weighted genomic best linear unbiased prediction model. The trained models were ultimately used to predict the agronomic performances of an independent durum wheat panel, confirming the utility of genomic prediction, although environmental conditions and genetic backgrounds may still be a challenge to overcome. The results generated through our study confirmed the utility of GPD and GYD to mitigate the inverse GY and GPC relationship in wheat, provided novel markers for marker-assisted selection and opened new ways to develop cultivars through genomic prediction approaches.

Raptors bred in captivity: semen characteristics and assisted reproduction outcome in goshawk (Accipiter gentilis).

Three sexually mature goshawks reared in captivity and imprinted on humans to express reproductive behavior according to the cooperative method were studied for three consecutive breeding seasons to assess the quality of their sperm. The following parameters were analyzed: ejaculate volume and sperm concentration, motility, viability, and morphology. Ejaculate volume, sperm concentration and motility fluctuated along the reproductive season, revealing the greatest quality of the reproductive material at full springtime (i.e., April). Motility of the sperm collected in March strongly reduced with age, contrary to samples collected in April or May. Sperm viability was not influenced by either age or month of collection within each season. Ultrastructural investigations provided information on normal sperm morphology for the first time in this species. The morphological categories of sperm defects in fresh semen, present at low percentages, are also described. Functional analyses (perivitelline membrane assay and artificial inseminations) confirmed the good quality of the semen obtained using the cooperative method. The reported data provide the basis for further studies aimed at developing protocols to improve the outcome of artificial insemination and semen cryopreservation in the goshawk as well as other bird of prey species.

Pruning harvesting with modular towed chipper: Little effect of the machine setting and configuration on performance despite strong impact on wood chip quality.

Pruning residues can have a high quality as feedstock for energy purposes and are largely available in Europe. However, it is still an untapped resource. Such scarce use is due to the need to optimize their supply chain in term of collection machines and the associate cost of collection. A modular chipper (prototype PC50) for pruning harvest was developed. Such prototype is adaptable to various harvesting logistics and may produce a higher quality woodchip compared with the one produced by shredders available in the market. In this work, we tested the performance and quality of the product delivered by the prototype PC50 in various conditions and plant species, after a modulation of the machine settings (counter-rotating toothed rollers [CRR] speed), loading systems ([LS], either big bag or container), and knife types ([KT], either discontinuous hoe shaped knives or continuous helicoidal knives). To take into account of the covariates in the experiment (Cropping season and plant species), LSmeans were computed to have an unbiased estimate of the treatments means. The modulation of LS and KT scarcely affected the performance of the machine. In particular, the choice of the KT affected the field efficiency when the LS was a Tilting box but not a Big Bag. Whereas the continuous knife resulted in a 97% higher material capacity compared to hoe shape knives, the last of which the amount of short sized (<16 mm) fractions compared to helicoidal knives. No role of the CCR was found on the machine performance, but increasing CRR speed reduced the chip apparent bulk density and the fraction chips with a size <8 mm.

An Endophytic Fungi-Based Biostimulant Modulates Volatile and Non-Volatile Secondary Metabolites and Yield of Greenhouse Basil (Ocimum basilicum L.) through Variable Mechanisms Dependent on Salinity Stress Level.

Salinity in water and soil is one of the major environmental factors limiting the productivity of agronomic and horticultural crops. In basil (Ocimum basilicum L., Lamiaceae) and other Ocimum species, information on the plant response to mild salinity levels, often induced by the irrigation or fertigation systems, is scarce. In the present work, we tested the effectiveness of a microbial-based biostimulant containing two strains of arbuscular mycorrhiza fungi (AMF) and Trichoderma koningii in sustaining greenhouse basil yield traits, subjected to two mild salinity stresses (25 mM [low] and 50 mM [high] modulated by augmenting the fertigation osmotic potential with NaCl) compared to a non-stressed control. The impact of salinity stress was further appraised in terms of plant physiology, morphological ontogenesis and composition in polyphenols and volatile organic compounds (VOC). As expected, increasing the salinity of the solution strongly depressed the plant yield, nutrient uptake and concentration, reduced photosynthetic activity and leaf water potential, increased the Na and Cl and induced the accumulation of polyphenols. In addition, it decreased the concentration of Eucalyptol and ß-Linalool, two of its main essential oil constituents. Irrespective of the salinity stress level, the multispecies inoculum strongly benefited plant growth, leaf number and area, and the accumulation of Ca, Mg, B, p-coumaric and chicoric acids, while it reduced nitrate and Cl concentrations in the shoots and affected the concentration of some minor VOC constituents. The benefits derived from the inoculum in term of yield and quality harnessed different mechanisms depending on the degree of stress. under low-stress conditions, the inoculum directly stimulated the photosynthetic activity after an increase of the Fe and Mn availability for the plants and induced the accumulation of caffeic and rosmarinic acids. under high stress conditions, the inoculum mostly acted directly on the sequestration of Na and the increase of P availability for the plant, moreover it stimulated the accumulation of polyphenols, especially of ferulic and chicoric acids and quercetin-rutinoside in the shoots. Notably, the inoculum did not affect the VOC composition, thus suggesting that its activity did not interact with the essential oil biosynthesis. These results clearly indicate that beneficial inocula constitute a valuable tool for sustaining yield and improving or sustaining quality under suboptimal water quality conditions imposing low salinity stress on horticultural crops.

Validating the regional estimates of changes in soil organic carbon by using the data from paired-sites: the case study of Mediterranean arable lands.

BACKGROUND: Legacy data are unique occasions for estimating soil organic carbon (SOC) concentration changes and spatial variability, but their use showed limitations due to the sampling schemes adopted and improvements may be needed in the analysis methodologies. When SOC changes is estimated with legacy data, the use of soil samples collected in different plots (i.e., non-paired data) may lead to biased results. In the present work, N = 302 georeferenced soil samples were selected from a regional (Sicily, south of Italy) soil database. An operational sampling approach was developed to spot SOC concentration changes from 1994 to 2017 in the same plots at the 0-30 cm soil depth and tested. RESULTS: The measurements were conducted after computing the minimum number of samples needed to have a reliable estimate of SOC variation after 23 years. By applying an effect size based methodology, 30 out of 302 sites were resampled in 2017 to achieve a power of 80%, and an α = 0.05. A Wilcoxon test applied to the variation of SOC from 1994 to 2017 suggested that there was not a statistical difference in SOC concentration after 23 years (Z = - 0.556; 2-tailed asymptotic significance = 0.578). In particular, only 40% of resampled sites showed a higher SOC concentration than in 2017. CONCLUSIONS: This finding contrasts with a previous SOC concentration increase that was found in 2008 (75.8% increase when estimated as differences of 2 models built with non-paired data), when compared to 1994 observed data (Z = - 9.119; 2-tailed asymptotic significance < 0.001). This suggests that the use of legacy data to estimate SOC concentration dynamics requires soil resampling in the same locations to overcome the stochastic model errors. Further experiment is needed to identify the percentage of the sites to resample in order to align two legacy datasets in the same area.

New pedotransfer approaches to predict soil bulk density using WoSIS soil data and environmental covariates in Mediterranean agro-ecosystems.

For the estimation of the soil organic carbon stocks, bulk density (BD) is a fundamental parameter but measured data are usually not available especially when dealing with legacy soil data. It is possible to estimate BD by applying pedotransfer function (PTF). We applied different estimation methods with the aim to define a suitable PTF for BD of arable land for the Mediterranean Basin, which has peculiar climate features that may influence the soil carbon sequestration. To improve the existing BD estimation methods, we used a set of public climatic and topographic data along with the soil texture and organic carbon data. The present work consisted of the following steps: i) development of three PTFs models separately for top (0-0.4 m) and subsoil (0.4-1.2 m), ii) a 10-fold cross-validation, iii) model transferability using an external dataset derived from published data. The development of the new PTFs was based on the training dataset consisting of World Soil Information Service (WoSIS) soil profile data, climatic data from WorldClim at 1 km spatial resolution and Shuttle Radar Topography Mission (SRTM) digital elevation model at 30 m spatial resolution. The three PTFs models were developed using: Multiple Linear Regression stepwise (MLR-S), Multiple Linear Regression backward stepwise (MLR-BS), and Artificial Neural Network (ANN). The predictions of the newly developed PTFs were compared with the BD calculated using the PTF proposed by Manrique and Jones (MJ) and the modelled BD derived from the global SoilGrids dataset. For the topsoil training dataset (N = 129), MLR-S, MLR-BS and ANN had a R2 0.35, 0.58 and 0.86, respectively. For the model transferability, the three PTFs applied to the external topsoil dataset (N = 59), achieved R2 values of 0.06, 0.03 and 0.41. For the subsoil training dataset (N = 180), MLR-S, MLR-BS and ANN the R2 values were 0.36, 0.46 and 0.83, respectively. When applied to the external subsoil dataset (N = 29), the R2 values were 0.05, 0.06 and 0.41. The cross-validation for both top and subsoil dataset, resulted in an intermediate performance compared to calibration and validation with the external dataset. The new ANN PTF outperformed MLR-S, MLR-BS, MJ and SoilGrids approaches for estimating BD. Further improvements may be achieved by additionally considering the time of sampling, agricultural soil management and cultivation practices in predictive models.

Addition of high C:N crop residues to a P-limited substrate constrains the benefits of arbuscular mycorrhizal symbiosis for wheat P and N nutrition.

Many aspects concerning the role of arbuscular mycorrhizal (AM) fungi in plant nutrient uptake from organic sources remain unclear. Here, we investigated the contribution of AM symbiosis to N and P uptake by durum wheat after the addition of a high C:N biomass to a P-limited soil. Plants were grown in pots in the presence or absence of a multispecies AM inoculum, with (Org) or without (Ctr) the addition of 15N-labelled organic matter (OM). A further treatment, in which 15N was applied in mineral form (Ctr+N) in the same amount as that supplied in the Org treatment, was also included. Inoculation with AM had positive effects on plant growth in both control treatments (Ctr and Ctr+N), mainly linked to an increase in plant P uptake. The addition of OM, increasing the P available in the soil for the plants, resulted in a marked decrease in the contribution of AM symbiosis to plant growth and nutrient uptake, although the percentage of mycorrhization was higher in the Org treatment than in the controls. In addition, mycorrhization drastically reduced the recovery of 15N from the OM added to the soil whereas it slightly increased the N recovery from the mineral fertiliser. This suggests that plants and AM fungi probably exert a differential competition for different sources of N available in the soil. On the whole, our results provide a contribution to a better understanding of the conditions under which AM fungi can play an effective role in mitigating the negative effects of nutritional stresses in plants.

Genome-Wide Association Mapping of Prostrate/Erect Growth Habit in Winter Durum Wheat.

By selecting for prostrate growth habit of the juvenile phase of the cycle, durum wheat cultivars could be developed with improved competitive ability against weeds, and better soil coverage to reduce the soil water lost by evaporation. A panel of 184 durum wheat (Triticum turgidum subsp. durum) genotypes, previously genotyped with DArT-seq markers, was used to perform association mapping analysis of prostrate/erect growth habit trait and to identify candidate genes. Phenotypic data of plant growth habit were recorded during three consecutive growing seasons (2014-2016), two different growth conditions (field trial and greenhouse) and two sowing periods (autumn and spring). Genome-wide association study revealed significant marker-trait associations, twelve of which were specific for a single environment/year, 4 consistent in two environments, and two MTAs for the LSmeans were identified across all environments, on chromosomes 2B and 5A. The co-localization of some MTAs identified in this study with known vernalization and photoperiod genes demonstrated that the sensitivity to vernalization and photoperiod response are actually not only key components of spring/winter growth habit, but they play also an important role in defining the magnitude of the tiller angle during the tillering stage. Many zinc-finger transcription factors, such as C2H2 or CCCH-domain zinc finger proteins, known to be involved in plant growth habit and in leaf angle regulation were found as among the most likely candidate genes. The highest numbers of candidate genes putatively related to the trait were found on chromosomes 3A, 4B, 5A and 6A. Moreover, a bioinformatic approach has been considered to search for functional ortholog genes in wheat by using the sequence of rice and barley tiller angle-related genes. The information generated could be used to improve the understanding of the mechanisms that regulate the prostrate/erect growth habit in wheat and the adaptive potential of durum wheat under resource-limited environmental conditions.

Growth-promoting bacteria and arbuscular mycorrhizal fungi differentially benefit tomato and corn depending upon the supplied form of phosphorus.

The ability of plants to take up phosphorus (P) from soil depends on root morphology and root exudates release and can be modulated by beneficial soil microbes. These microbes can solubilize P, affect root elongation and branching, and lead to a higher uptake of P and other nutrients. However, coordination of these mechanisms is unclear, especially the mechanism for changing the available form of P. We aimed to dissect the effects of two different beneficial microbial taxa (plant growth-promoting bacteria (PGPB) and arbuscular mycorrhizal fungi (AMF)) on root morphological traits, plant nutrient content, and growth in tomato and corn fertilized with either Gafsa rock phosphate (RP) or triple superphosphate (TSP), which have contrasting solubility levels. Tomato and corn were grown in pots and inoculated with one of three PGPB species or a mix of two AMF species or were not inoculated. Root traits, botanical fractions, and the contents of various mineral nutrients were measured. TSP stimulated tomato biomass accumulation compared to RP but did not stimulate corn biomass accumulation. PGPB improved the growth of both plant species under RP, with limited differences among the strains, whereas AMF only improved tomato growth under TSP. These differences between microbial systems were explained by a bacterial effect on the total root length but not on the mean root diameter and by the ability of AMF to improve the mineral nutrient content. The effects of PGPB were less dependent on the plant species and on P form than the effects of AMF.These results have implications for the improvement of the early plant growth through the management of beneficial microbes.

Metabolomics Provides Valuable Insight for the Study of Durum Wheat: A Review.

Metabolomics is increasingly being applied in various fields offering a highly informative tool for high-throughput diagnostics. However, in plant sciences, metabolomics is underused, even though plant studies are relatively easy and cheap when compared to those on humans and animals. Despite their importance for human nutrition, cereals, and especially wheat, remain understudied from a metabolomics point of view. The metabolomics of durum wheat has been essentially neglected, although its genetic structure allows the inference of common mechanisms that can be extended to other wheat and cereal species. This review covers the present achievements in durum wheat metabolomics highlighting the connections with the metabolomics of other cereal species (especially bread wheat). We discuss the metabolomics data from various studies and their relationships to other "-omics" sciences, in terms of wheat genetics, abiotic and biotic stresses, beneficial microbes, and the characterization and use of durum wheat as feed, food, and food ingredient.

Milling overrides cultivar, leavening agent and baking mode on chemical and rheological traits and sensory perception of durum wheat breads.

Smell and aroma are important determinants of consumer acceptance, so gaining deeper insight into bread smell and aroma perception is a research goal. Sixteen combinations of four variables were investigated, to evaluate the contributions of bread chemical and rheological properties and volatile organic compounds (VOCs) towards sensory acceptability of breads: genotypes (landrace vs. modern); types of flour (wholemeal vs. semolina); leavening agents (brewing yeast vs. sourdough starter); and baking modes (gas-fired vs. wood-fired). Milling had the greatest impact over the other treatments for the rheological and chemical properties, including for VOCs, with great impact on the sensory traits of the flours and breads. The processing phases had great impact on smell and aroma, as defined through formation of alcohols, aldehydes, terpenes, and other compounds (e.g., ethylbenzene, 2-pentylfuran, methoxyphenyl oxime). Leavening agent had great impact on sensory perception, although breads from the sourdough starter were perceived as with lower taste and colour than the brewing yeast. Baking mode had no relevant role on sensory perception. These data strongly undermine the belief of a 'better product' that is frequently attributed to old genotypes versus modern cultivars, and indicate that the milling and the bread-making processes determine the quality of the end product.

Spatio-temporal topsoil organic carbon mapping of a semi-arid Mediterranean region: The role of land use, soil texture, topographic indices and the influence of remote sensing data to modelling.

SOC is the most important indicator of soil fertility and monitoring its space-time changes is a prerequisite to establish strategies to reduce soil loss and preserve its quality. Here we modelled the topsoil (0-0.3m) SOC concentration of the cultivated area of Sicily in 1993 and 2008. Sicily is an extremely variable region with a high number of ecosystems, soils, and microclimates. We studied the role of time and land use in the modelling of SOC, and assessed the role of remote sensing (RS) covariates in the boosted regression trees modelling. The models obtained showed a high pseudo-R2 (0.63-0.69) and low uncertainty (s.d.<0.76gCkg-1 with RS, and <1.25gCkg-1 without RS). These outputs allowed depicting a time variation of SOC at 1arcsec. SOC estimation strongly depended on the soil texture, land use, rainfall and topographic indices related to erosion and deposition. RS indices captured one fifth of the total variance explained, slightly changed the ranking of variance explained by the non-RS predictors, and reduced the variability of the model replicates. During the study period, SOC decreased in the areas with relatively high initial SOC, and increased in the area with high temperature and low rainfall, dominated by arables. This was likely due to the compulsory application of some Good Agricultural and Environmental practices. These results confirm that the importance of texture and land use in short-term SOC variation is comparable to climate. The present results call for agronomic and policy intervention at the district level to maintain fertility and yield potential. In addition, the present results suggest that the application of RS covariates enhanced the modelling performance.

An assessment of factors controlling N2O and CO2 emissions from crop residues using different measurement approaches.

Management of plant residues plays an important role in maintaining soil quality and nutrient availability for plants and microbes. However, there is considerable uncertainty regarding the factors controlling residue decomposition and their effects on greenhouse gas (GHG) emissions from the soil. This uncertainty is created both by the complexity of the processes involved and limitations in the methodologies commonly used to quantify GHG emissions. We therefore investigated the addition of two soil residues (durum wheat and faba bean) with similar C/N ratios but contrasting fibres, lignin and cellulose contents on nutrient dynamics and GHG emission from two contrasting soils: a low-soil organic carbon (SOC), high pH clay soil (Chromic Haploxerert) and a high-SOC, low pH sandy-loam soil (Eutric Cambisol). In addition, we compared the effectiveness of the use of an infrared gas analyser (IRGA) and a photoacoustic gas analyser (PGA) to measure GHG emissions with more conventional gas chromatography (GC). There was a strong correlation between the different measurement techniques which strengthens the case for the use of continuous measurement approaches involving IRGA and PGA analyses in studies of this type. The unamended Cambisol released 286% more CO2 and 30% more N2O than the Haploxerert. Addition of plant residues increased CO2 emissions more in the Haploxerert than Cambisol and N2O emission more in the Cambisol than in the Haploxerert. This may have been a consequence of the high N stabilization efficiency of the Haploxerert resulting from its high pH and the effect of the clay on mineralization of native organic matter. These results have implication management of plant residues in different soil types.

Arbuscular mycorrhizal fungi altered the hypericin, pseudohypericin, and hyperforin content in flowers of Hypericum perforatum grown under contrasting P availability in a highly organic substrate.

St. John's Wort (Hypericum perforatum) is a perennial herb able to produce water-soluble active ingredients (a.i.), mostly in flowers, with a wide range of medicinal and biotechnological uses. However, information about the ability of arbuscular mycorrhizal fungi (AMF) to affect its biomass accumulation, flower production, and concentration of a.i. under contrasting nutrient availability is still scarce. In the present experiment, we evaluated the role of AMF on growth, flower production, and concentration of bioactive secondary metabolites (hypericin, pseudohypericin, and hyperforin) of H. perforatum under contrasting P availability. AMF stimulated the production of aboveground biomass under low P conditions and increased the production of root biomass. AMF almost halved the number of flowers per plant by means of a reduction of the number of flower-bearing stems per plant under high P availability and through a lower number of flowers per stem in the low-P treatment. Flower hyperforin concentration was 17.5% lower in mycorrhizal than in non-mycorrhizal plants. On the contrary, pseudohypericin and hypericin concentrations increased by 166.8 and 279.2%, respectively, with AMF under low P availability, whereas no effect of AMF was found under high P availability. These results have implications for modulating the secondary metabolite production of H. perforatum. However, further studies are needed to evaluate the competition for photosynthates between AMF and flowers at different nutrient availabilities for both plant and AM fungus.

Artemisia arborescens Essential Oil Composition, Enantiomeric Distribution, and Antimicrobial Activity from Different Wild Populations from the Mediterranean Area.

Aerial parts of Artemisia arborescens were collected from different sites of the Mediterranean area (southwestern Algeria and southern Italy) and the chemical composition of their essential oil (EO) extracted by hydrodistillation was studied by both gas chromatography (GC) equipped with an enantioselective capillary column and GC/mass spectrometry (GC/MS). The EOs obtained were tested against several Listeria monocytogenes strains. Using GC and GC/MS, 41 compounds were identified, accounting for 96.0 - 98.8% of the total EO. All EOs showed a similar terpene profile, which was rich in chamazulene, ß-thujone, and camphor. However, the concentration of such compounds varied among the EOs. A. arborescens EO inhibited up to 83.3% of the L. monocytogenes strains, but the inhibitory spectrum varied among the EOs, with those from Algeria showing a higher inhibition degree than the Italian EOs. Such effect likely depended on the ketone (ß-thujone + camphor) content of the EO. The differences in the EO composition support the hypothesis that A. arborescens has at least two different chemotypes: a ß-thujone and a chamazulene type. The EO inhibitory spectrum indicates the A. arborescens EO as a valuable option in the control of the food-borne pathogens.

Rhynchophorus ferrugineus attack affects a group of compounds rather than rearranging Phoenix canariensis metabolic pathways.

The red palm weevil (RPW; Rhynchophorus ferrugineus) is spreading worldwide and severely harming many palm species. However, most studies on RPW focused on insect biology, and little information is available about the plant response to the attack. In the present experiment, we used metabolomics to study the alteration of the leaf metabolome of Phoenix canariensis at initial (1st stage) or advanced (2nd stage) attack by RPW compared with healthy (unattacked) plants. The leaf metabolome significantly varied among treatments. At the 1st stage of attack, plants showed a reprogramming of carbohydrate and organic acid metabolism; in contrast, peptides and lipid metabolic pathways underwent more changes during the 2nd than 1st stage of attack. Enrichment metabolomics analysis indicated that RPW attack mostly affected a particular group of compounds rather than rearranging plant metabolic pathways. Some compounds selectively affected during the 1st rather than 2nd stage (e.g. phenylalanine; tryptophan; cellobiose; xylose; quinate; xylonite; idonate; and iso-threonate; cellobiotol and arbutine) are upstream events in the phenylpropanoid, terpenoid and alkaloid biosynthesis. These compounds could be designated as potential markers of initial RPW attack. However, further investigation is needed to determine efficient early screening methods of RPW attack based on the concentrations of these molecules.

Soil inoculation with symbiotic microorganisms promotes plant growth and nutrient transporter genes expression in durum wheat.

In a field experiment conducted in a Mediterranean area of inner Sicily, durum wheat was inoculated with plant growth-promoting rhizobacteria (PGPR), with arbuscular mycorrhizal fungi (AMF), or with both to evaluate their effects on nutrient uptake, plant growth, and the expression of key transporter genes involved in nitrogen (N) and phosphorus (P) uptake. These biotic associations were studied under either low N availability (unfertilized plots) and supplying the soil with an easily mineralizable organic fertilizer. Regardless of N fertilization, at the tillering stage, inoculation with AMF alone or in combination with PGPR increased the aboveground biomass yield compared to the uninoculated control. Inoculation with PGPR enhanced the aboveground biomass yield compared to the control, but only when N fertilizer was added. At the heading stage, inoculation with all microorganisms increased the aboveground biomass and N. Inoculation with PGPR and AMF+PGPR resulted in significantly higher aboveground P compared to the control and inoculation with AMF only when organic N was applied. The role of microbe inoculation in N uptake was elucidated by the expression of nitrate transporter genes. NRT1.1, NRT2, and NAR2.2 were significantly upregulated by inoculation with AMF and AMF+PGPR in the absence of organic N. A significant down-regulation of the same genes was observed when organic N was added. The ammonium (NH4 (+)) transporter genes AMT1.2 showed an expression pattern similar to that of the NO3 (-) transporters. Finally, in the absence of organic N, the transcript abundance of P transporters Pht1 and PT2-1 was increased by inoculation with AMF+PGPR, and inoculation with AMF upregulated Pht2 compared to the uninoculated control. These results indicate the soil inoculation with AMF and PGPR (alone or in combination) as a valuable option for farmers to improve yield, nutrient uptake, and the sustainability of the agro-ecosystem.