Commercial Inoculants Research Articles

The Biosynthesis and Functions of Polyamines in the Interaction of Plant Growth-Promoting Rhizobacteria with Plants.

Plant growth-promoting rhizobacteria (PGPR) are members of the plant rhizomicrobiome that enhance plant growth and stress resistance by increasing nutrient availability to the plant, producing phytohormones or other secondary metabolites, stimulating plant defense responses against abiotic stresses and pathogens, or fixing nitrogen. The use of PGPR to increase crop yield with minimal environmental impact is a sustainable and readily applicable replacement for a portion of chemical fertilizer and pesticides required for the growth of high-yielding varieties. Increased plant health and productivity have long been gained by applying PGPR as commercial inoculants to crops, although with uneven results. The establishment of plant-PGPR relationships requires the exchange of chemical signals and nutrients between the partners, and polyamines (PAs) are an important class of compounds that act as physiological effectors and signal molecules in plant-microbe interactions. In this review, we focus on the role of PAs in interactions between PGPR and plants. We describe the basic ecology of PGPR and the production and function of PAs in them and the plants with which they interact. We examine the metabolism and the roles of PAs in PGPR and plants individually and during their interaction with one another. Lastly, we describe some directions for future research.

Screening Digitaria eriantha cv. Suvernola Endophytic Bacteria for Maize Growth Promotion.

The search for sustainable agriculture has increased interest in using endophytic bacteria to reduce fertilizer use and increase stress resilience. Stress-adapted plants are a potential source of these bacteria. Some species of these plants have not yet been evaluated for this, such as pangolão grass, from which we considered endophytic bacteria as potential plant growth promoters. Bacteria from the root, colm, leaves, and rhizospheric soil were isolated, and 132 strains were evaluated for their in vitro biological nitrogen fixation, IAA and siderophores production, and phosphate solubilization. Each mechanism was also assessed under low N availability, water stress, and low-solubility Fe and P sources in maize greenhouse experiments. All strains synthesized IAA; 63 grew on N-free media, 114 synthesized siderophores, and 46 solubilized P, while 19 presented all four mechanisms. Overall, these strains had better performance than commercial inoculant in all experiments. Still, in vitro responses were not good predictors of in vivo effects, which indicates that the former should not be used for strain selection, since this could lead to not testing strains with good plant growth promotion potential. Their heterologous growth promotion in maize reinforces the potential of stress-adapted plant species as potential sources of strains for inoculants.

Meta-omics integration approach reveals the effect of soil native microbiome diversity in the performance of inoculant Azospirillum brasilense.

Plant growth promoting bacteria (PGPB) have been used as integrative inputs to minimize the use of chemical fertilizers. However, a holistic comprehension about PGPB-plant-microbiome interactions is still incipient. Furthermore, the interaction among PGPB and the holobiont (host-microbiome association) represent a new frontier to plant breeding programs. We aimed to characterize maize bulk soil and rhizosphere microbiomes in irradiated soil (IS) and a native soil (NS) microbial community gradient (dilution-to-extinction) with Azospirillum brasilense Ab-V5, a PGPB commercial inoculant. Our hypothesis was that plant growth promotion efficiency is a result of PGPB niche occupation and persistence according to the holobiont conditions. The effects of Ab-V5 and NS microbial communities were evaluated in microcosms by a combined approach of microbiomics (species-specific qPCR, 16S rRNA metataxonomics and metagenomics) and plant phenomics (conventional and high-throughput methods). Our results revealed a weak maize growth promoting effect of Ab-V5 inoculation in undiluted NS, contrasting the positive effects of NS dilutions 10-3, 10-6, 10-9 and IS with Ab-V5. Alpha diversity in NS + Ab-V5 soil samples was higher than in all other treatments in a time course of 25 days after sowing (DAS). At 15 DAS, alpha diversity indexes were different between NS and IS, but similar in all NS dilutions in rhizospheric samples. These differences were not persistent at 25 DAS, demonstrating a stabilization process in the rhizobiomes. In NS 10-3 +Ab-V5 and NS 10-6 Ab-V5, Ab-V5 persisted in the maize rhizosphere until 15 DAS in higher abundances compared to NS. In NS + Ab-V5, abundance of six taxa were positively correlated with response to (a)biotic stresses in plant-soil interface. Genes involved in bacterial metabolism of riboses and amino acids, and cresol degradation were abundant on NS 10-3 + Ab-V5, indicating that these pathways can contribute to plant growth promotion and might be a result of Ab-V5 performance as a microbial recruiter of beneficial functions to the plant. Our results demonstrated the effects of holobiont on Ab-V5 performance. The meta-omics integration supported by plant phenomics opens new perspectives to better understanding of inoculants-holobiont interaction and for developing better strategies for optimization in the use of microbial products.

Biological nitrogen fixation by soybean (Glycine max [L.] Merr.), a novel, high protein crop in Scotland, requires inoculation with non-native bradyrhizobia

It is currently not recommended to grow soybean (Glycine max [L.] Merr.) further than 54° North, but climate change and the development of new high latitude-adapted varieties raises the possibility that it could be introduced into Scotland as a novel high protein crop deriving most of its nitrogen (N) requirements through biological N fixation (BNF). This was evaluated via field trials in 2017 and 2018 near Dundee (56.48°N). As there are no native soybean-nodulating bacteria (SNB) in UK soils, soybean requires inoculation to exploit its BNF potential. In 2017, three commercial inoculants containing elite Bradyrhizobium strains significantly increased plant biomass in plot trials with a soybean 000 maturity group variety (ES Comandor). Rhizobia were isolated from the nodules and identified as the original inoculant species, B. diazoefficiens and B. japonicum. One inoculant (Rizoliq Top) was used for larger-scale trials in 2018 with two varieties (ES Comandor, ES Navigator); inoculation doubled the grain yield to 1 t ha-1 compared to the uninoculated crop. The inoculated soybean obtained most of its N through BNF in both years regardless of plant genotype i.e. >73%Ndfa, with BNF contributions to aerial biomass exceeding 250 kg N ha-1 yr-1 in 2017 and that to grain 50 kg N ha-1 yr-1 in 2018. These data suggest that N-fixing soybean could be grown in Scotland without mineral N-fertiliser, either for forage as animal feed, or as green pods for human consumption (“edamame”), and potentially, even as dry grain. The potential for survival of the Bradyrhizobium inoculant strains in soils was also demonstrated through the detection of the inoculant strain B. diazoefficiens SEMIA 5080 at relatively high populations (104 g-1 dry soil) using a qRT-PCR method with SNB-specific nodZ primers. Microbiome data obtained from soil using 16S rRNA primers demonstrated that the diversity of bacteria belonging to the genus Bradyrhizobium increased in soybean-cropped soils compared to bulk soil regardless of inoculation status. The economic and practical implications of residual inoculum, as well as those arising from introducing a non-native plant and alien bacteria into Scottish soils in terms of their impact on the native soil microbiota are discussed.

Can mycorrhizal fungi fix farming? Benefits and limitations of applying them to agroecosystems

The intensification of agriculture over the past decades has helped increase crop yields but has also led to various environmental issues, potentially undermining future productivity. One of the options considered for increasing the sustainability of agriculture is the stimulation or enhancement of microorganisms that associate with plants and help them acquire more nutrients from the soil leading to benefits such as increased growth. One key player considered are arbuscular mycorrhizal fungi (AMF), a group of below-ground fungi that form symbiotic relationships with the vast majority of land plants. However, evidence from lab and field trials suggests that not all plants respond equally to colonization by these fungi, and research is on-going to better understand the context-dependency of the symbiosis. Moreover, introducing AMF to agricultural fields through the application of commercial inoculants that contain fungal spores and hyphae has been inconsistent in their desired effects of boosting crop yield and quality. Recently, a quality framework has been put forward to try and increase the reliability of these inocula so farmers can make better use of them. Although further research on the fundamental and applied aspects of plant–arbuscular mycorrhizal associations is required, agricultural practices that favour these fungi should be encouraged as they are likely to lead to wider benefits and ultimately contribute to a more sustainable agricultural system.

Effect of adding lactic acid bacteria to maize silage on nutritive quality, fermentation properties and in vitro digestibility

This study aimed to determine the effects of adding lactic acid bacteria to maize silage on the nutritionalf quality, its fermentation properties and its in vitro organic matter digestion (IVOMD). In the study, pre-fermented juices (PFJ) prepared from different water-soluble carbohydrate (WSC) sources at the rate of 5% and commercial homofermentative and heterofermentative lactic acid bacteria (LAB) were added to silages. Groups were designed as (I) control, (II) Glucose-PFJ, (III) Fructose-PFJ, (IV) Sucrose-PFJ, (V) Molasses-PFJ, (VI) Homofermentative LAB (HoLAB) and (VII) Heterofermentative LAB (HetLAB). When the lactic acid bacteria (LAB) count, lactic acid (LA), acetic acid (AA), LA/AA ratio, pH and yeast values of the natural fermented lactic acid bacteria liquids prepared by adding 5% different easily soluble carbohydrate sources to meadow grass fermentation values of the groups were examined, the differences between them were statistically significant. 
 The differences between the groups in the CA, ADF, IVOMD and methane (CH4) values of the silage groups prepared by adding PFJ were found to be statistically significant. The differences in the fermentation characteristics of the silages (pH, ammonia-nitrogen (NH3-N), LA, AA, LA/AA, CO2 and total yeast mold after aerobic stability) were statistically significant. When all parameters were examined, it was concluded that the addition of PFJ, which is prepared by adding 5% fructose to the meadow grass plant, to the maize silage has positive effects on IVOMD, ME, CH4, LA and yeast-mold and can be used instead of commercial inoculants.

A Commercial Arbuscular Mycorrhizal Inoculum Alleviated the Effects of Acid Water on Lupinus angustifolius Grown in a Sterilized Mining Dump.

Lupinus species have been sporadically reported to be colonized by arbuscular mycorrhizal fungi (AMF). The interactions between AMF and lupine plants could also be non-symbiotic, from positive to negative, as controlled by the stress conditions of the plant. The goal of the study was to reveal the existence of such positive interactions and provide preliminary data for a myco-phytoremediation technology of mining dumps using L. angustifolius as a first crop. The objective was to test the hypothesis that the AMF inoculation of an acidified dump material contaminated with heavy metals would improve the growth of L. angustifolius and decrease oxidative stress. The design consisted of a one-month bivariate pot experiment with plants grown in a mining dump soil inoculated and not inoculated with a commercial AMF inoculum sequestered in expanded clay and watered with acidic and neutral water. There was no AMF root colonization under the experimental conditions, but under neutral and acidic water conditions, the phosphorus concentrations in roots and leaves increased, and the superoxide dismutase and peroxidase activities significantly decreased due to AMF inoculation. The increase in leaf phosphorus concentration was correlated with the decrease in peroxidase activity. The fresh weight of shoots and leaves significantly increased due to the commercial inoculum (under acidic water conditions). At the end of the experiment, the ammonium concentration in the substrate was higher in the inoculated treatments than in the not inoculated ones, and the concentrations of many elements in the dump material decreased compared to the start of the experiment. A comprehensive discussion of the potential mechanisms underlying the effects of the commercial AMF inoculum on the non-host L. angustifolius is completed.

Response of Sagebrush Steppe Grass Species to AMF Inoculum Sources and Biochar.

The sagebrush steppe has presented increasing levels of degradation. The addition of arbuscular mycorrhizal fungi (AMF) and biochar have been suggested to restore ecosystems. However, little is known about their effects on sagebrush steppe plants. We tested three sources of AMF inoculum: soil from a disturbed site (Inoculum A), soil from an undisturbed site (Inoculum B), and commercial inoculum (Inoculum C), all with and without biochar, to test if they could mediate growth of Pseudoroegneria spicata (native perennial), Taeniatherum caput-medusae (early seral; exotic annual) and Ventenata dubia (early seral; exotic annual) under greenhouse conditions. We measured AMF colonization and biomass. We hypothesized that the plant species would be differently affected by the inoculum types. The colonization of T. caput-medusae and V. dubia was greatest when inoculated with Inoculum A (38.8% and 19.6%). In contrast, the colonization of P. spicata was greatest with Inoculum B and Inoculum C (32.1% and 32.2). Biochar decreased biomass production but increased colonization with Inoculum A for P. spicata and V. dubia and with Inoculum C for T. caput-medusae. This study reveals the response of early and late seral sagebrush steppe grass species to contrasting sources of AMF and suggests that late seral plant species respond better to late seral inocula.

Restoration on magnetite mine waste substrates using Western Australian native plants only marginally benefited from a commercial inoculant

Post-mining landscapes often lack self-sustaining plant communities and functional belowground microbial communities. Inappropriate management of soil can hinder ecological restoration of mine sites. However, the potential role of microbial inoculants and plant nutrient-acquisition strategies in improving mined substrates and facilitating mine-site restoration remains relatively unexplored. An eight-month glasshouse experiment was conducted to test: (1) whether a commercially-available microbial inoculant was effective in restoring biological properties of mined substrates, and (2) the effect of plant nutrient-acquisition strategies on improving the chemical properties of mined substrates for the growth of native plants. There was no significant improvement in plant growth by adding a commercially-sourced microbial inoculant. Soil microbial biomass carbon and phosphorus increased significantly after plant growth (main effect of species * substrates * inoculation interaction). The non-mycorrhizal disturbance-specialist plant species Maireana georgei was effective in improving hostile conditions of the mine waste. Our results highlight that re-vegetating stockpiled topsoil using local keystone species is a desirable practice that can improve soil biological properties and benefit mine-site restoration. Commercially-available agricultural-based microbial inoculants may not be compatible for mine-site restoration using native plants, but future research using indigenous soil microbes is warranted.

Inoculum source dependent effects of ericoid, mycorrhizal fungi on flowering and reproductive success in highbush blueberry (Vaccinium corymbosum).

Most terrestrial angiosperms form mutualisms with both mycorrhizal fungi and animal pollinators. Yet, the effects of mycorrhizae on pollinator behavior and plant reproduction are unknown for most species, and whether the source or type of mycorrhizal fungi affects reproductive success has rarely been examined. We examined whether inoculating highbush blueberry (Vaccinium corymbosum; Ericaceae) with ericoid mycorrhizal fungi enhanced investment in flowering and attractiveness to pollinators, and thus reduced their levels of pollen limitation over that of non-inoculated plants. We also examined the degree to which pollen limitation was dependent on inoculation source and the surrounding pollinator community context. Three-year-old saplings of Vaccinium corymbosum 'Bluecrop' or highbush blueberry (Ericaceae) were inoculated with a) ericoid mycorrhizal fungi within soil of the rhizosphere of plants growing at a local blueberry farm, b) a commercially available ericoid inoculant, c) both the local soils and commercial inoculum, or d) were not inoculated and served as controls. They were grown for one year in pots in a common garden and, in the following year, were moved to six farms in central Vermont that were known from prior studies to differ in pollinator abundance and diversity. We conducted a hand pollination experiment at each farm to examine if inoculation or pollinator abundance (i.e., farm context) affected reproductive success. Plants treated with all types of inoculums were more likely to flower, and produced more inflorescence buds than non-inoculated plants in 2018. However, in 2019, plants in the combination inoculum treatment, alone, produced more inflorescence buds than those in the other treatments. Neither the source of inoculum nor hand pollination affected fruit set (the proportion of flowers setting fruit), or fruit sugar content. Hand pollination, but not inoculation, increased berry mass and the average number of seeds produced/berry. Our results add to the growing body of evidence that mycorrhizal fungi can affect reproductive traits of their hosts but that the effects of mycorrhizal fungi depend on the mycorrhizal symbionts.

Bradyrhizobium inoculants impact on promiscuous nodulating soybeans cultivars in Ghana's farming systems

AbstractSoybean is rarely inoculated with commercial Bradyrhizobium inoculants in Ghana's cropping systems. A 2‐year field study was conducted at Council for Scientific and Industrial Research–Savanna Agricultural Research Institute's experimental field at Nyankpala, Ghana to assess how commercial inoculants affect growth, nodulation, nitrogen fixation, and grain yield of promiscuous nodulating soybean cultivars. The experiment was a split‐plot design with the main plot factor as soybean [tropical Glycine max (L.) Merr. crosses (TGX) cultivars]; Jenguma (TGX1448‐2E), Afayak (TGX1834‐5E), and Songda (TGX 1445‐3E). The subplot factor consisted of three commercial Bradyrhizobium japonicum inoculants, namely, Biofix (strain USDA 110), NoduMax (strain USDA 110), and Legumefix (strain 532c) plus uninoculated control. Shoot biomass and nodulation were assessed at vegetative (V8, 8‐leaf stage), full bloom (R2), beginning to pod (R3), full pod (R4), and full seed (R6) stages, respectively, and N fixation and grain yield at maturity. Shoot dry matter increased with growth stage and peaked at the R6 stage with significant effect associated with Biofix and NoduMax. Nodulation on the upper root and the whole root also peaked at the R4 stage, with pronounced effects associated with Biofix and NoduMax. Enhanced grain yields of up to 30% were achieved with Biofix and NoduMax compared to the uninoculated control in both 2016 and 2017. Regarding the cultivars, shoot biomass increased with growth stage and peaked at R6 stage and Afayak yielded greater shoot biomass. On average (2 years), Afayak produced greater grain yields than Jenguma and Songda. In conclusion, Biofix and NoduMax were the most promising commercial inoculants for soybean production in Northern Ghana.

Inoculation with Azospirillum argentinense Az19 improves the yield of maize subjected to water deficit at key stages of plant development

Water deficit constitutes a severe limitation to agricultural productivity. In the context of sustainable crop production, the potential of microbial biotechnology to increase plant drought tolerance and improve crop yields under adverse conditions is gaining relevance. This work aimed to compare the performance of Azospirillumargentinense strain Az19 to that of strain Az39, the most widely used for commercial inoculants, when inoculated in maize plants exposed to water deficit. For this purpose, greenhouse and field assays were conducted. In the greenhouse experiment, strain Az19 prevented the adverse effect of water deficit at V2 stage on maize growth. Moreover, the percentage of fertile plants and the ear weight decreased significantly under water deficits imposed at V2 and flowering in Az39-inoculated plants but not in Az19-inoculated plants. In the first field trial with the commercial maize hybrid DOW DS 515 PW, Az19-inoculated plants were those which better tolerated the water deficit imposed. In the second field trial, two maize genotypes with differential drought sensitivity (LP 29×LP 2542, sensitive; LP 882 (923)×LP 4703, tolerant) were tested. Higher tolerance to water deficit was detected in plants inoculated with A. argentinense Az19, with a noticeable effect on grain yield components in the sensitive genotype. Based on these results, we propose the use of A. argentinense Az19 for the formulation of more targeted Azospirillum-based inoculants, suitable for agroecological areas subjected to seasonal water deficits.

Microbiome analysis revealed distinct microbial communities occupying different sized nodules in field-grown peanut.

Legume nodulation is the powerhouse of biological nitrogen fixation (BNF) where host-specific rhizobia dominate the nodule microbiome. However, other rhizobial or non-rhizobial inhabitants can also colonize legume nodules, and it is unclear how these bacteria interact, compete, or combinedly function in the nodule microbiome. Under such context, to test this hypothesis, we conducted 16S-rRNA based nodule microbiome sequencing to characterize microbial communities in two distinct sized nodules from field-grown peanuts inoculated with a commercial inoculum. We found that microbial communities diverged drastically in the two types of peanut nodules (big and small). Core microbial analysis revealed that the big nodules were inhabited by Bradyrhizobium, which dominated composition (>99%) throughout the plant life cycle. Surprisingly, we observed that in addition to Bradyrhizobium, the small nodules harbored a diverse set of bacteria (~31%) that were not present in big nodules. Notably, these initially less dominant bacteria gradually dominated in small nodules during the later plant growth phases, which suggested that native microbial communities competed with the commercial inoculum in the small nodules only. Conversely, negligible or no competition was observed in the big nodules. Based on the prediction of KEGG pathway analysis for N and P cycling genes and the presence of diverse genera in the small nodules, we foresee great potential of future studies of these microbial communities which may be crucial for peanut growth and development and/or protecting host plants from various biotic and abiotic stresses.

Factors Impacting Rhizobium-legume Symbiotic Nitrogen Fixation with the Physiological and Genetic Responses to Overcome the Adverse Conditions: A Review

Symbiotic N2 fixation is essential for the plant’s growth because it can fix reactive nitrogen compounds in soil. However, all steps of this process can be hampered by several biotic and abiotic environmental factors. This study mainly focused on discussing the impacts of 12 major factors on this process by reviewing the significant numbers of research works. According to the information from these works, we found some significant physiological and genetic impacts caused by these factors like plasmid deletion, genomic mismanagement, abnormal molecular signals, toxicity, deficiency of minerals, deformation of rhizobial cells, protein denaturation, nucleic acid damage, acetylene reduction and nod factors limitation. Furthermore, expression of heat or acid shock proteins, internal buffering, genes spanning, extracellular immobilization, periplasmic allocation, change of lipopolysaccharides composition, intracellular accumulation of inorganic and organic solutes (Osmolyte) and activation of hydrogenase expression are shown by both micro and macro symbionts as a natural response to adapt to these stress conditions. Though the stress-tolerant strains like HR-3, HR-6, HR-10, HR-12, acta, actP, exoR, lpiA, actR, actS and phrR can be used to sense the external environment and make signals to change gene transcription during the adverse condition, the application of genetic engineering should be expanded more to promote the commercial inoculation by the production of novel stress-tolerant strains or modified genes of rhizobia and legumes.

Diversity and enzymatic activity of the microbiota isolated from compost based on restaurant waste and yard trimmings

IntroductionThe bad management of organic waste negatively affects environmental quality and composting has been a viable recycling alternative. Microorganisms are responsible for waste degradation during the composting process and, consequently, for transforming this waste into natural fertilizer. This work aimed to analyze and identify the biodiversity of yeasts and filamentous fungi throughout a composting process based on organic residues under different treatments (commercial inoculum, non-commercial inoculum, and control treatment) and to investigate the enzymatic activity of these microorganisms.MethodsMicroorganisms were isolated and identified from samples at 0, 5, 10, 20, 40, 60, and 120 days. Filamentous fungi were identified according to their macroscopic and microscopic characteristics, and yeasts were identified by sequencing the 18S rDNA region. All identified strains were evaluated for ligninolytic, cellulolytic, hemicellulolytic, amylolytic, pectinolytic, proteolytic, lipolytic, and ammonification. During the composting phases, the filamentous fungi were higher than the yeast population.Results and discussionAt the beginning of the process, a higher species diversity was observed, and the population of yeasts and filamentous fungi was, on average, 6.50 log CFU g−1. The microbial communities were similar throughout the process in the two inoculated treatments, which showed more significant microbial activity, diversity, and efficiency in the transformation of organic matter, and consequently, advantages in terms of the final product quality compared to the control treatment. The yeasts Pichia kudriavzevii, Pichia farinosa, Issatchenkia orientalis, and the filamentous fungi of the genus Aspergillus spp. proved to have high biotechnological value and could be used as starter cultures to accelerate the composting process.

Arbuscular mycorrhizal fungi associated with maize plants during hydric deficit

The objective of this study was to verify the physiological behavior and development of maize plants under hydric deficit inoculated with the AMF Rhizophagus clarus and Claroideoglomus etunicatum and the commercial inoculant ROOTELLA BR in nonsterilized soil as a strategy to mitigate the effects of drought in the crop. Corn seeds were grown and inoculated with R. clarus, C. etunicatum and the commercial inoculant ROOTELLA BR separately at sowing. The plants were grown in a greenhouse and submitted to water deficit in stage V3, keeping the pots at 20% field capacity for 10 days. The first analyses were performed, followed by reirrigation for 2 days, and the analyses were performed again. The experiment was a double factorial, with 2 water treatments (irrigated and water deficit) × 4 inoculation treatments (control, ROOTELLA BR, R. clarus, C. etunicatum) × 5 replicates per treatment, totaling 40 vessels. The results indicate that the plants were able to recover favorably according to the physiological data presented. It is noted that in inoculated plants, there was no damage to the photosynthetic apparatus. These data demonstrate that AMF contribute greatly to better plant recovery after a dry period and a new irrigation period. Inoculation with AMF favors postwater stress recovery in plants.

The Effects of Short-Time Delayed Sealing on Fermentation, Aerobic Stability and Chemical Composition on Maize Silages

Despite the efforts to improve the methodological standards of silage trials, many factors that can influence the results of lab-scale studies need to be better understood. This study aimed to determine the effects of short-time delayed sealing and inoculation with a blend of Lentilactobacillus buchneri and Lactiplantibacillus plantarum on fermentation, aerobic stability, and chemical composition of silages. Whole-crop maize was treated with or without a commercial inoculant and ensiled (29.3% dry matter) for 55 days in 8.8 L PVC silos that were sealed immediately (up to 30 min delay) or after a delay (90, 150, or 210 min between chopping and sealing) with five replicates each. The increasing air exposure before sealing increased fermentation losses and reduced silage nutritional value. Crude protein and ash were significantly affected by inoculation, with control treatments showing higher ash and lower protein values. Lignin, neutral detergent fiber, and acid detergent fiber were only affected by the delay period. The longer the sealing delay, the higher the gas production, and the lower the starch values and lactic acid content observed in samples. Inoculation was inefficient in reducing total dry matter losses, but it increased aerobic stability, acetic acid, and ethanol contents of silages and reduced effluent loss. Control silages had higher total dry matter loss during the aerobic exposure than inoculated silages. The results confirmed that the delay periods tested were long enough to negatively interfere with the chemical composition of silages, especially the fibrous fraction content.

Native Arbuscular Mycorrhizal Inoculum Modulates Growth, Oxidative Metabolism and Alleviates Salinity Stresses in Legume Species.

Soil salinity constitutes a major abiotic stress that contributes to soil degradation and crop yield reduction. Using arbuscular mycorrhizal fungi (AMF) inoculation can help to alleviate these deleterious effects. Most researches on AMF application are dealing with ecological restoration, whereas little consideration has been given to agriculture and legume production. The comparison of the efficacy of two AMF inoculums, one native originating from Algerian semiarid saline soils and one commercial inoculum, was carried out regarding their effects on the growth and the mineral nutrition of several legumes species, Medicago sativa, Medicago falcata, Trifolium repens and Trifolium alexandrinum, cultivated in semiarid Algerian saline soil under greenhouse conditions. Our results showed that native mycorrhizal inoculum enhanced shoot biomasses by 20%, mycorrhizal rate by 30%, shoot phosphorus content by 25% and K+/Na+ ratio by 45% for studied plants when compared with commercial inoculum. The best efficiency of the native AMF inoculum is probably due to the complementarity between the AMF strains which composed the inoculum. Funneliformis geosporum was the most abundant species recorded at the end of the experience in all plant roots especially with native inoculum. Our findings pointed out the effectiveness of native AMF inoculum application to promote agricultural production in semiarid saline soils.

Flavonoids promote Rhizophagus irregularis spore germination and tomato root colonization: A target for sustainable agriculture.

The use of arbuscular mycorrhizal (AM) fungi has great potential, being used as biostimulants, biofertilizers and bioprotection agents in agricultural and natural ecosystems. However, the application of AM fungal inoculants is still challenging due to the variability of results when applied in production systems. This variability is partly due to differences in symbiosis establishment. Reducing such variability and promoting symbiosis establishment is essential to improve the efficiency of the inoculants. In addition to strigolactones, flavonoids have been proposed to participate in the pre-symbiotic plant-AM fungus communication in the rhizosphere, although their role is still unclear. Here, we studied the specific function of flavonoids as signaling molecules in AM symbiosis. For that, both in vitro and in planta approaches were used to test the stimulatory effect of an array of different subclasses of flavonoids on Rhizophagus irregularis spore germination and symbiosis establishment, using physiological doses of the compounds. We show that the flavone chrysin and the flavonols quercetin and rutin were able to promote spore germination and root colonization at low doses, confirming their role as pre-symbiotic signaling molecules in AM symbiosis. The results pave the way to use these flavonoids in the formulation of AM fungal-based products to promote the symbiosis. This can improve the efficiency of commercial inoculants, and therefore, help to implement their use in sustainable agriculture.

Screening and field evaluation of white clover rhizobia for New Zealand pastures

Context Biological nitrogen (N) fixation through the rhizobia–legume symbiosis is a sustainable and cost-effective source of N for agriculture. In New Zealand (NZ), white clover (Trifolium repens) is a key component of pastures and rhizobial inoculation of clover is widely used. The current commercial inoculant for white clover, TA1, was isolated in Australia in the 1950s and may not be the best partner for modern white clover cultivars. Aims To identify Rhizobium leguminosarum bv. trifolii (Rlt) isolates suitable for use in NZ pastures. Methods The symbiotic potential of >230 isolates collected from throughout NZ was evaluated in plant bioassays. Selected isolates were further evaluated in pot and field trials. Key results Approximately 40% of NZ isolates supported better clover growth than TA1 under N-limited conditions in vitro. Of 24 Rlt isolates evaluated in a glasshouse trial, five produced significantly higher clover biomass than TA1. Three (S11N9, S20N7, S4N6) of nine isolates evaluated in two field trials in 2018–2019 significantly increased clover growth (12–38%) compared with paired uninoculated clover at several harvests, whereas inoculation with TA1 did not improve yield. In a third trial in 2020, S11N9 and S20N7 increased clover growth compared with the uninoculated control at two of three harvests; S4N6 performed better than TA1 at one harvest. When tested with four white clover cultivars, five Rlt isolates had higher symbiotic potential than TA1. Conclusions Inoculating white clover would be beneficial if improved inoculant isolates were available. Implications We recommend some NZ Rlt isolates could be developed into commercial inoculants to improve white clover performance in NZ.