Hordeum Murinum Research Articles

The Integrated Effect Between Low Doses of Herbicide Glyphosate and Plant Extracts in Management of Carrot Crop Daucus carota L. var. sativa Weeds

This study was carried out at Abu Jarash Farm and Weed Laboratory, Faculty of Agriculture, Damascus University, during 2017 winter to investigate the integrated effect of the non-selective herbicide Glyphosate (360 g/l) (Lancer®) and plant extracts of some weeds: sorghum (Sorghum bicolor L.), alfalfa (Medicago sativa L.) and bermuda grass (Cynodon dactylon L.) by using one third and one fourth of the recommended doses of the herbicide mixed with plant extracts for weed management of plots planted with carrot. The integration between herbicide and plant extracts led to 100% weed control when the herbicide was used at 25% of the recommended dose, and was equally effective to that of the recommended dose. The benefits are cost reduction and decrease of harmful effects of synthetic herbicides to the environment. On the other hand, Sorghum bicolor extract was significantly effective in controlling all tested weed species (82.1% effectiveness on Chenopodium album (most sensitive), plant height of wild barley Hordeum murinum L. was 3.8 cm, dry matter content of 4.96% for purple nutsedge, Cyperus rotundus L. and 6.58% for hairy amaranth, Amaranthus retroflexus L. Keywords: Integration, herbicides, plant extracts, reduced doses, weeds

First records of a potentially new plant community from the ruderal vegetation in the Black Sea Coast, Bulgaria

The paper presents the first data of an unreported plant community dominated by Hordeum leporinum Link and Bituminaria bituminosa (L.) Stirt. (class Stellarietea mediae R. Tx., Lohmeyer et Preising in R.Tx. 1950). It is currently established in four localities in the region of the Black Sea coast, between the town of Ravda and the town of Nessebar, Bulgaria. Among the characteristic features of the community are: (i) it occupies places with an active human presence near the beach; (ii) preferences for shallow san-dy or rocky soils; (iii) Bituminaria bituminosa (L.) Stirt. as a dominant, diagnostic and constant species for the community. Other diagnostic species for the association are Anchusa officinalis L., Malva sylves-tris L., Sonchus arvensis L. and Avena fatua L. (iv) annual and perennial species have an equal participa-tion in the community; (v) increased presence of diagnostic species for the class Artemisietea vulgaris Lohmeyer et al. ex von Rochow 1951. Phytogeographically, the species composition of the community has a strong Mediterranean influence. In close proximity to its reléves there are always communities of xerophytic shrubs or low trees (in many cases alien species, such as Amorpha fruticosa L., Ziziphus jujuba Mill. and Robinia pseudoacacia L.) that participate in the association without forming a full second layer (their cover abundance is less than 30%).

Epigenetic responses of hare barley (Hordeum murinum subsp. leporinum) to climate change: an experimental, trait-based approach

The impact of reduced rainfall and increased temperatures forecasted by climate change models on plant communities will depend on the capacity of plant species to acclimate and adapt to new environmental conditions. The acclimation process is mainly driven by epigenetic regulation, including structural and chemical modifications on the genome that do not affect the nucleotide sequence. In plants, one of the best-known epigenetic mechanisms is cytosine-methylation. We evaluated the impact of 30% reduced rainfall (hereafter “drought” treatment; D), 3 °C increased air temperature (“warming”; W), and the combination of D and W (WD) on the phenotypic and epigenetic variability of Hordeum murinum subsp. leporinum L., a grass species of high relevance in Mediterranean agroforestry systems. A full factorial experiment was set up in a savannah-like ecosystem located in southwestern Spain. H. murinum exhibited a large phenotypic plasticity in response to climatic conditions. Plants subjected to warmer conditions (i.e., W and WD treatments) flowered earlier, and those subjected to combined stress (WD) showed a higher investment in leaf area per unit of leaf mass (i.e., higher SLA) and produced heavier seeds. Our results also indicated that both the level and patterns of methylation varied substantially with the climatic treatments, with the combination of D and W inducing a clearly different epigenetic response compared to that promoted by D and W separately. The main conclusion achieved in this work suggests a potential role of epigenetic regulation of gene expression for the maintenance of homoeostasis and functional stability under future climate change scenarios.

Mumetopia interfeles sp. nov., a new species of Anthomyzidae (Diptera) occurring en masse in an urban grassy habitat in Chile: its taxonomy, phylogeny and biology

A new species of Anthomyzidae (Diptera), viz., Mumetopia interfeles Roháček sp. nov. (both sexes), is described from Chile, based on a large series collected from a small grassy area (consisting of a non-native species, Hordeum murinum L.) among houses in the city of Valparaíso. Its systematic affiliation, phylogenetic relationships, and biology are discussed. Cases of mass occurrence of adult Anthomyzidae, particularly those in anthropogenic habitats, are reviewed and general features of them discussed. A phylogenetic hypothesis is proposed for the relationships of the new species and the Chamaebosca group of genera within Anthomyzidae (64 species in dataset), based on Bayesian inference and maximum likelihood analyses of seven DNA markers (12S, 16S, 28S, COI, COII, CytB, ITS2). Taxonomic limits of the genus Mumetopia Melander, 1913 are discussed. Based on a new molecular hypothesis and a previous cladistic analysis of morphological data (Roháček & Barber 2009), a broadened, better-supported concept of Mumetopia (s. str.) is proposed to include M. interfeles sp. nov. (and its unnamed relatives) and externally aberrant, undescribed species endemic to the Juan Fernández Islands. Apart from Anthomyzidae (M. interfeles sp. nov. only), the community of Diptera associated with H. murinum was dominated by phytophagous Chloropidae: Ceratobarys sacculicornis (Enderlein, 1911) occurred in extremely high numbers similar to those of M. interfeles sp. nov., while the less abundant Opetiophora sp. represents the first record of the genus from South America.

Hordeum murinum in The Right Lower Lobe Bronchus in a 2-year-old Patient Presenting with Hemoptysis

Hordeum murinum in The Right Lower Lobe Bronchus in a 2-year-old Patient Presenting with Hemoptysis

Indicative Value of the Dominant Plant Species for a Rapid Evaluation of the Nutritional Value of Soils

A study was conducted on 14 grassland communities located in the south of the Iberian Peninsula and their edaphology, which is identified as specific plant associations. The edaphic study of each association allows a rapid evaluation of the nutrient content in the soil without the need for laboratory edaphic analysis. For each phytosociological relevé and soil, samplings were carried out. The field data were subjected to various statistical analysis—canonical correspondence analysis (CCA), Bayesian networks, and decision trees—to establish nutrient content. When the abundance value of the species is 9 in the Van der Maarel scale, there is an increase in the values of several soil parameters. In the case of Hordeum leporinum, when the Van der Maarel index is 9, the Kc (exchangeable potassium in cmol/kg) undergoes the greatest variation, to a value of up to 0.729 cmol/kg. The application of the decision tree to this species reveals that the soil attributes with the greatest influence in the classification are conductivity, %_si (silt texture), pH, and pF 15 atm (pressure at 15 atmospheres (water retention capacity) in %). Indeed, this interlaced edaphic and phytosociological study provides us with a high-value tool to obtain quick information on the content of nutrients in the soil.

Resistance Evolution to EPSPS Inhibiting Herbicides in False Barley (Hordeum murinum) Harvested in Southern Spain

A failure of the EPSPS-inhibiting herbicide glyphosate to control several populations of Hordeum murinum subsp. leporinum (or H. murinum) occurred in southern Spain after more than fifteen applications in both crop (olive, orchards, and citrus) and non-crop (dry areas, roadsides and ditches) areas. Eight out of 18 populations studied were resistant (R) to glyphosate with R factors higher than four based on GR50. These populations also had the highest values of LD50 and the lowest levels of shikimic acid accumulation. Two adjuvants tested increased glyphosate efficacy in both susceptible (S) and R populations thanks to better spray foliar retention. Moreover, PS I-, PS II-, and ACCase-inhibiting herbicides, in pre- or post-emergence, proved to be the best chemical alternatives with different sites of action (SoA) to control both S and glyphosate-R populations. This study represents the first report worldwide of glyphosate resistance in H. murinum found in very different crop and non-crop areas from southern Spain. To design chemical strategies to implement integrated weed management programs for glyphosate-R H. murinum, both adjuvants and herbicides with alternative SoA as well as application timings should be considered.

Elevated CO2 , temperature and nitrogen levels impact growth and development of invasive weeds in the Mediterranean region.

Invasive plant species present a serious threat to the environment, as well as human and animal health. An interaction may exist between the climatic changes and invasive plant species. In this 2-year study, we investigated the effects of warming, CO2 and nitrogen application on the biomass, growth and leaf tissue nitrogen concentration of three invasive weed species. Treatments were: (i) simulated (elevated) CO2 (approximately 800-900 ppm); (ii) warming or high temperature (day/night 25/15 °C); (iii) simulated (elevated) CO2 combined with high temperature (CO2 = approximately 800-900 ppm; temperature day/night 25/15 °C); and (iv) control conditions (CO2 = approximately 400-450 ppm; temperature day/night 20/10 °C). The doses of nitrogen were: (i) 0 kg ha-1 (control; low); (ii) 60 kg ha-1 (medium); and (iii) 120 kg ha-1 (high). Elevated CO2 and elevated CO2 combined with high temperature improved biomass and the growth of the tested invasive weed species: Lactuca serriola L., Hordeum murinum L. and Bromus tectorum L. Nitrogen application had little effect on grasses, whereas the broadleaved weed mostly had a positive response to nitrogen application. Invasive weed species were generally negatively or neutrally affected by warming. The results of the present study demonstrate that nitrogen fertilization under different climatic conditions improved few of the parameters, whereas elevated CO2 promoted most of the growth parameters of invasive weeds. Overall, is it concluded that these weeds will be more invasive under climate change conditions. © 2020 Society of Chemical Industry.

Meteorological factors driving airborne grass pollen concentration in central Iberian Peninsula

Grasses are a taxonomic group of considerable environmental importance, playing a major ecological role as well as making a significant contribution to the world’s economy, since they include valuable crop and pasture species. Grass pollen allergens are also among the main causes of respiratory allergies worldwide. The interpretation of airborne grass pollen concentrations is a particularly complex task, given the marked diversity of pollen-emitting species and the influence of weather-related variables. This paper examines the influence of meteorological variables on flowering in the species contributing most to airborne grass pollen concentrations. This study was carried out in the surroundings of the city of Toledo (Spain), a Mediterranean city located in the center of the Iberian Peninsula. Temperature was the variable most influencing flowering onset, which was also affected by relative humidity, rainfall and hours of sunlight. The flowering period of grass species that flower at the start of spring (Bromus rubens and Hordeum leporinum) began earlier in years with higher mean winter temperatures, while the species that flower from mid to late spring (Trisetaria panicea and Dactylis glomerata subsp. hispanica), the flowering period began earlier in years with higher cumulative rainfall in winter and spring, which were also the years with the highest mean temperatures. Research into the influence of weather-related variables on grass phenology can shed important light on variations in airborne pollen concentrations, which determine the potential period of exposure for allergy sufferers.

The Impact of Herbicide Application and Defoliation on Barley Grass (Hordeum murinum subsp. glaucum) Management in Mixed Pasture Legumes

Barley grass (Hordeum murinum subsp. glaucum.) is an annual weed associated with grain revenue loss and sheep carcass damage in southern Australia. Increasing herbicide resistance led to a recent investigation into effective integrated weed management strategies for barley grass in southern Australia. Field studies in Wagga Wagga, New South Wales (NSW) during 2016 and 2017 examined the effect of post-emergent herbicide applications and strategic defoliation by mowing on barley grass survival and seed production in a mixed legume pasture. Statistically significant differences between herbicide-only treatments in both years showed propaquizafop to be more than 98% effective in reducing barley grass survival and seed production. Paraquat was not effective in controlling barley grass (58% efficacy), but led to a 36% and 63.5% decrease in clover and other weed biomass, respectively, after 12 months and increased lucerne biomass by over three-fold after 24 months. A single repeated mowing treatment resulted in a 46% decline in barley grass seedling emergence after 12 months and, when integrated with herbicide applications, reduced other weed biomass after 24 months by 95%. Resistance to acetyl-CoA carboxylase (ACCase)-inhibiting herbicides observed in local barley grass populations led to additional and more focused investigation comparing the efficacy of other pre- and post-emergent herbicides for barley grass management in legume pastures. Haloxyfop-R + simazine or paraquat, applied at early tillering stage, were most efficacious in reducing barley grass survival and fecundity. Impact of defoliation timing and frequency on barley grass seedlings was also evaluated at various population densities, highlighting the efficacy of repeated post-inflorescence defoliations in reducing plant survival and seed production. Results highlight the importance of optimal environmental conditions and application timing in achieving efficacious control of barley grass and improving pasture growth and biomass accumulation.

Kinetic and Thermodynamic Studies for the Efficient Removal of Methylene Blue Using Hordeum Murinum as a New Biosorbent

In this work, Hordeum murinum as a sustainable and cost-effective biosorbent material is utilized without any further treatment to remove Methylene blue dye from aqueous solutions. The effects of various parameters including pH, biosorbent amount, reaction time, initial concentration, and temperature are investigated. The results show that 94% of Methylene blue is removed using the optimal adsorption conditions of pH 5.6, biosorbent dose of 0.1 gm, 60 min equilibrium contact time, 20 mg/L of initial dye concentration at a temperature of 25 °C. Experimental results reveal that the adsorption kinetics can have the best description by employing the pseudo-second order model. The spontaneous and exothermic nature of the adsorption process is indicated by the negative values of the Gibbs free energy change and the enthalpy change, respectively. These results reveal that Hordeum murinum exhibited excellent performance for the removal of Methylene blue from aqueous solutions, which can make it very interesting as an alternative and effective low-cost biosorbent material.

Screening diversity and distribution of Copia retrotransposons reveals a specific amplification of BARE1 elements in genomes of the polyploid Hordeum murinum complex.

We explored diversity, distribution and evolutionary dynamics of Ty1-Copia retrotransposons in the genomes of the Hordeum murinum polyploid complex and related taxa. Phylogenetic and fluorescent in situ hybridization (FISH) analyses of reverse transcriptase sequences identified four Copia families in these genomes: the predominant BARE1 (including three groups or subfamilies, A, B and C), and the less represented RIRE1, IKYA and TAR-1. Within the BARE1 family, BARE1-A elements and a subgroup of BARE1-B elements (named B1) have proliferated in the allopolyploid members of the H. murinum complex (H. murinum and H. leporinum), and in their extant diploid progenitor, subsp. glaucum. Moreover, we found a specific amplification of BARE1-B elements within each Hordeum species surveyed. The low occurrence of RIRE1, IKYA and TAR-1 elements in the allopolyploid cytotypes suggests that they are either weakly represented or highly degenerated in their diploid progenitors. The results demonstrate that BARE1-A and BARE1-B1 Copia elements are particularly well represented in the genomes of the H. murinum complex and constitute its genomic hallmark. No BARE1-A and -B1 homologs were detected in the reference barley genome. The similar distribution of RT-Copia probes across chromosomes of diploid, tetraploid and hexaploid taxa of the murinum complex shows no evidence of proliferation following polyploidization.

Scald on gramineous hosts in Iran and their potential threat to cultivated barley

There are five described Rhynchosporium species, Rhynchosporium commune, R. secalis, R. agropyri, R. orthosporum and R. lolii, that cause scald diseases on Poaceae. This study used morphological (conidial shape and size) and phylogenetic analyses of two loci (the internal transcribed spacer region (ITS) and β-tubulin (TUBB)) to identify Rhynchosporium species and their host ranges in Iran. Despite the large variation observed for Rhynchosporium conidial dimensions, the phylogenetic analyses of the ITS region and concatenated ITS and TUBB loci revealed that all isolates from wild grasses in Iran belong to R. commune. R. commune was isolated from Hordeum murinum ssp. glaucum, Hordeum vulgare ssp. spontaneum, Lolium multiflorum and Avena sativa in Iran. A. sativa has only been reported from Iran as a host for R. commune. After cross inoculation, A. sativa was considered as the most resistant host showing the lowest susceptibility to R. commune isolates. Of the grass hosts tested, H. vulgare ssp. spontaneum was the most susceptible. The most aggressive isolate across all tested hosts was isolated from Hordeum murinum ssp. glaucum. Cross-infection of the R. commune isolates from all hosts onto uncultivated grasses and cultivated barley suggests the potential of the uncultivated grasses as inoculum sources for cultivated barley epidemics and pathogen evolution. Thus, management of uncultivated grasses in the vicinity of barley fields should assist in managing the disease on cultivated barley.

Interspecific molecular variation of Tunisian complex Lolium perenne L. and Festuca arundinacea Schreb. based on the internal transcribed spacer locus (ITS).

To establish phylogenetic relationships and estimate the intra and interspecific divergence, the amplification and the sequencing of the internal transcribed spacers of ribosomal DNA (ITS = ITS1 + 5.8S + ITS2) were analyzed in Tunisian complex Lolium-Festuca DNA. These spacer regions have evolved mainly by point mutations. Results revealed a high level of polymorphism within studied species. Significant similarities were observed between these two species and showed the existence of an important phylogenetic relationship. Besides, this molecular approach has revealed two new clusterings, with a homologous ITS gene namely: Bromus hordeaceus and Hordeum murinum subsp. This could be explained by the conservation of an ancestral ITS gene in some fescue plants. Thus, Tunisian tall fescue and perennial ryegrass may derive from Bromus hordeaceus and Hordeum murinum subsp. Considerable morphology and bioclimatic distribution similarities were discovered in ITS sequences within the same species. This study can be of great help to identify suitable accessions that could be used in local fescue and ryegrass improvement program.

The Fate of Deleterious Variants in a Barley Genomic Prediction Population.

Targeted identification and purging of deleterious genetic variants has been proposed as a novel approach to animal and plant breeding. This strategy is motivated, in part, by the observation that demographic events and strong selection associated with cultivated species pose a "cost of domestication." This includes an increase in the proportion of genetic variants that are likely to reduce fitness. Recent advances in DNA resequencing and sequence constraint-based approaches to predict the functional impact of a mutation permit the identification of putatively deleterious SNPs (dSNPs) on a genome-wide scale. Using exome capture resequencing of 21 barley lines, we identified 3855 dSNPs among 497,754 total SNPs. We generated whole-genome resequencing data of Hordeum murinum ssp. glaucum as a phylogenetic outgroup to polarize SNPs as ancestral vs. derived. We also observed a higher proportion of dSNPs per synonymous SNPs (sSNPs) in low-recombination regions of the genome. Using 5215 progeny from a genomic prediction experiment, we examined the fate of dSNPs over three breeding cycles. Adjusting for initial frequency, derived alleles at dSNPs reduced in frequency or were lost more often than other classes of SNPs. The highest-yielding lines in the experiment, as chosen by standard genomic prediction approaches, carried fewer homozygous dSNPs than randomly sampled lines from the same progeny cycle. In the final cycle of the experiment, progeny selected by genomic prediction had a mean of 5.6% fewer homozygous dSNPs relative to randomly chosen progeny from the same cycle.

Ecological Study on some Grasses Growing Naturally in the Deltaic Mediterranean Coast of Egypt

The present study aims to investigate the floristic composition and vegetation analysis of the floraassociated with some grasses growing naturally on the sand formations in the Deltaic Mediterranean coastof Egypt. The total number of the recorded plant species was 79 species (44 annuals and 35 perennials)belonging to 70 genera and 26 families. The life –form spectra of the recorded species comprised 44therophytes, 13 cryptophytes, eight hemicryptophytes, seven chamaephytes and phanerphytes for each.Chorologically, the Saharo-Sindian element was represented by relatively high number of species (34),followed by Irano-Turanian (32 species), then Euro-Siberian (10 species) and Sudano-Zambezian (9species). Based on the importance alue of each species, the classification of the recorded species in 36sampled stands led to recognition of four vegetation groups (A-D). The characteristics dominant andcodominant species of vegetation groups were Elymus farcatus and Calligonum polygonoides (A),Hordeum murinum (B), Stipagrostis lanata and Echinops spinosus (C) and Lycium schweinfurthii andEchinops spinosus (D). DCA diagram showed overlapping between the identified groups. CCA-biplotexhibited that sulfates, calcium, potassium, magnesium and calcium carbonate were the most effectivesoil variables controlling the abundance of identified vegetation groups.

Occurrence of new case-bearer on cereals, Coleophora perplexella Toll, 1960 (Lepidoptera: Coleophoridae)

Coleophora perplexella Toll, 1960 it is native of Spain and Portugal, but it has detected out of its origin area, and was found in Sardinia, Morocco and Algeria without its host plant being identified. Recently has been observed in France (Toulon) on Hordeum murinum L. and Bromus sterilis L. (Poaceae). However, it was never cited among the Tunisian entomofauna. In addition, C. perplexella was never known as agricultural pest in its area or elsewhere. Moreover, no deep studies diagnosis and biology were conducted. This paper is a first study on C. perplexella, detected as an agricultural pest on oat (Avena sativa L., 1753) and barely (Hordeum vulgare L., 1753) making serious damages on early seedlings. Besides, a description and illustrations of different biological stage was provided. Results highlight a difference between the appearance dates of emerged new adults under both controlled and ambient conditions. According these results we can conclude on adaptation plasticity to the environmental conditions. In fact, C. perplexella can survive and cause damage at temperature between 7-10º C from December to February, and 23-25º C of controlled conditions.

Low genetic diversity of Rhynchosporium commune in Iran, a secondary centre of barley origin

Rhynchosporium commune is a destructive pathogen of barley, causing leaf scald. Previous microsatellite studies used Syria as a representative of cultivated barley's centre of origin, the Fertile Crescent. These suggested that R. commune and Hordeum vulgare (cultivated barley) did not co‐evolve in the host's centre of origin. The present study compares R. commune populations from Syria with those from Iran, which represents a secondary centre of origin for barley at the eastern edge of the Iranian Plateau. Results from this study also suggest that R. commune and barley did not co‐evolve in the centre of origin of cultivated barley. This was evidenced by the low pathogen genetic diversity in Iran, which was even lower than in Syria, indicating that the pathogen may have been introduced recently into Iran, perhaps through infected barley seed. Hierarchical analyses of molecular variance revealed that most genetic diversity in Iran and Syria is distributed within populations, with only 14% among populations. Analyses of multilocus association, genotype diversity and mating type frequency suggest that Iranian populations reproduce predominantly asexually. The presence of both mating types on barley and uncultivated grasses suggest a potential for sexual reproduction. Rhynchosporium commune was also found on Hordeum murinum subsp. glaucum, H. vulgare subsp. spontaneum, Lolium multiflorum and, for the first time, on Avena sativa. The variety of wild grasses that can be infected with R. commune in Iran raises concerns of these grasses acting as evolutionary breeding grounds and sources of inoculum.

Germination ecology of winter annual grasses in Mediterranean climates: Applications for soil cover in olive groves

The sustainability of Mediterranean agroecosystems is threatened by several factors, soil erosion being the most important one due to poor management practices. Seeding native grasses for ground cover is an effective practice to protect soil and enhance ecosystem services, but the species to be used should be adapted to the climatic conditions and the particular requirements of each system. Here, we studied seed germination timing of six winter annual grasses: Aegilops triuncialis, Anisantha madritensis, Anisantha rubens, Bromus hordeaceus, Hordeum murinum and Trachynia distachya; with potential for ground cover in olive groves of the Iberian Peninsula. We conducted germination experiments under eight treatments of constant and alternate temperatures and seven treatments of water potential, and fitted hydro- and thermal-time models to assess possible responses of seed-based populations to regional climate. In all species, recently-harvested seeds showed high germination rates across a wide range of temperatures, while winter-stored seeds were highly tolerant to moisture stress. Our results suggest that environmental conditions rather than dormancy prevent germination of the studied species after dispersal. This germination pattern contrasts with the deep physiological dormancy described for winter annuals in temperate climates and desert regions, indicating a special adaptation of the studied populations for the long dry season in Mediterranean climates. We conclude that the regeneration ecology of these widely-distributed grasses makes them an excellent source of seeds for ground cover in Mediterranean woody crops.

From Concept to Commerce: Developing a Successful Fungal Endophyte Inoculant for Agricultural Crops.

The development of endophyte inoculants for agricultural crops has been bedevilled by the twin problems of a lack of reliability and consistency, with a consequent lack of belief among end users in the efficacy of such treatments. We have developed a successful research pipeline for the production of a reliable, consistent and environmentally targeted fungal endophyte seed-delivered inoculant for barley cultivars. Our approach was developed de novo from an initial concept to source candidate endophyte inoculants from a wild relative of barley, Hordeum murinum (wall barley). A careful screening and selection procedure and extensive controlled environment testing of fungal endophyte strains, followed by multi-year field trials has resulted in the validation of an endophyte consortium suitable for barley crops grown on relatively dry sites. Our approach can be adapted for any crop or environment, provided that the set of first principles we have developed is followed. Here, we report how we developed the successful pipeline for the production of an economically viable fungal endophyte inoculant for barley cultivars.