The Usage of Arbuscular Mycorrhizal Fungi (Amf) as a Biofertilizer

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S. Songachan This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2559546/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Arbuscular Mycorrhizal Fungi (AMF) are a group of soil-borne fungi that form symbiotic relationships with the roots of most plants, including crops. In this relationship, the fungus provides the plant with nutrients, such as phosphorous, in exchange for carbohydrates produced by the plant through photosynthesis. The use of AMF as a biofertilizer involves the application of these fungi to soil to enhance plant growth and improve nutrient uptake. Studies have shown that AMF can increase plant growth, drought tolerance, and nutrient uptake, leading to improved crop yields. The fungi form a network of hyphae in the soil, which helps to increase the soil's water-holding capacity, as well as its ability to retain nutrients. This can lead to improved plant growth and health, even in nutrient-poor soils. In addition, the use of AMF as a biofertilizer can help to reduce the dependence on synthetic fertilizers, which can have negative environmental impacts. AMF can help to improve soil fertility, increase plant nutrient uptake, and reduce soil erosion, leading to more sustainable agriculture practices. However, it is important to note that the effectiveness of AMF as a biofertilizer can vary depending on several factors, including the species of AMF used, the type of crop being grown, and the conditions of the soil. Additionally, the proper application and management of AMF is important to ensure its effectiveness. In conclusion, the use of AMF as a biofertilizer has the potential to enhance plant growth, improve nutrient uptake, and promote sustainable agriculture practices. Arbuscular mycorrhizal fungi Biofertilizers Inoculum Sustainable agriculture Microbiome Figures Figure 1 Figure 2 Figure 3 Figure 4 Introduction Mycorrhizae are a type of symbiotic relationship that occur between fungi and plant roots. There are two main types of mycorrhiza: ectomycorrhizae and endomycorrhizae. Ectomycorrhizae form a sheath around the roots of certain plants, such as pines, oaks, and eucalyptus. The fungi involved in ectomycorrhizae do not invade the cells of the plant roots, but rather create a layer around the root cells known as the Hartig net. This allows for the exchange of nutrients and water between the plant and the fungus. Ectomycorrhizae are important for the health and growth of trees in forests, as they help to improve the uptake of nutrients and water from the soil. Endomycorrhizae, on the other hand, form a symbiotic relationship with the plant by actually invading the cells of the root. There are two main types of endomycorrhizae: arbuscular mycorrhizae (AM) and ericoid mycorrhizae (EM). Various kinds of mycorrhizal connections are described below in Fig. 1 . Soil micro-organisms such as AMFs reflect a crucial connection between plants and mineral soil nutrients. They are also gaining rising attention as natural fertilisers. AMFs are mandatory symbiotics of phylum Glomeromycota [ 56 ], establishing mutualistic symbiosis, with about 80 percent of the land plant organisms, including many food crops. In return for photosynthetic materials, they provide the host plant with mineral nutrients and water [ 59 ]. The AMF mycelium from the root system is able to extract nutrients from soil volumes that are inaccessible to the roots [ 59 ]. In comparison to plant roots fungal hyphae are far thinner and can reach narrower pores [ 1 ]. Carbohydrates and mineral nutrients are then spread by the plant and fungi within the roots. AM fungal hyphae colonise root cortex predominantly by forming profusely branchy structures inside the cells, i.e. arbuscules, which are known as the functional nutrient exchange site [ 7 ]. AMF therefore eliminates plant growth restrictions imposed by an insufficient supply of nutrients [ 43 ]. In recent times a non-mycorrhizal state can be regarded as rare in natural habitats for most organisms [ 61 ], even though the AM fungal populations below the ground are significantly different depending on the species composition, soil and seasonal form or variation of these factors [ 61 ]. AM experiences provide plants with additional advantages, in addition to an increased food source, such as enhanced drought and salinity resistance [ 6 , 46 ]. While several studies have been performed on the impact of AM symbiosis on plant reaction to abiotic stress such as drought, salinity and flooding in the last few years, the processes that have contributed to an improved plant stress resistance still remains somewhat elusive [ 6 , 52 , 53 , 9 , 10 ].Metals such as Iron(Fe),Copper(Cu) and Zinc (Zn) perform important functions in a variety of sub-cellular compartments, but they constitute a highly reactive community of elements that are toxic at large concentrations [ 65 ]. AM fungi have been documented to minimise the toxicity of heavy metals in host plant and to withstand high metal concentrations in soil. [ 22 , 36 , 18 , 65 , 38 ]. Metal transporters play a vital function in homeostasis of heavy metals. A Zn transporter was identified in Glomus intraradices (GintZnT1) [ 23 ] and more recently multiple putative genes coding Cu, Fe & Zn transporters were identified in a genome-wide study of the recently published Rhizophagus irregularis (formally, Glomus intraradices) genome [ 66 ]. The next steps would be characterisation of these carriers and discerning their role in the symbiosis. AM fungi may also have a significant influence on the environment, as they promote soil structure and aggregation [ 49 , 35 , 50 ] and control the development and production of plant populations [ 67 ].The impact of AM symbiosis have recently been also studied on greenhouse gas (GHG) emission [ 12 , 32 ]. Evidence presented by Bender et al. (2014) suggests that AM fungus may have a role to play in climate change mitigation due to their ability to significantly reduce emissions of N2O, a key greenhouse gas. By enhancing plant nitrogen (N) absorption and assimilation, AM fungi may be able to reduce N2O emissions by decreasing soluble N in soil and, in turn, denitrification [ 12 ]. Correlations between AM fungal abundance and genes involved in primary N2O production (nirK) and consumption (nosZ) suggest that AM fungi promote shifts in soil microbial biomass and community composition that lead to reduced N2O emissions. According to [ 32 ], AM symbiosis aids in N2O emission management at high soil moisture levels, and it was suggested that AM plant N2O emission control may be mediated by higher soil water use rather than increased N absorption. Therefore, AM fungi are primary biotic soil elements, which, when absent or degraded, for instance, by anthropic input, will contribute to a less effective functioning of the ecosystem. The process of re-establishing AMF may be a promising solution to industrial fertilisation methods in order to achieve organic cultivation, a significant goal for farmers in the midst of a global recession and an environmentally friendly consumer. The key technique for achieving this aim is to directly reintroduce AMF (inoculum) propagules in the target soil. However, in the application of these fungi, awareness of how AMF adapts and reacts to the objective of soil management and ecosystem management and the events which result in a functional symbiosis, including the mechanisms involved in the transfer of nutrients is important. After a brief discussion of the latest studies on the nutritional aspects of AM symbiosis and a short description of the challenges of development of AMF inoculum, descriptions of the application of AM fungi are mentioned and addressed, both under regulated and open field conditions, with specific emphasis on identifying factors contributing to success of the biofertilizer. Essential Features Of Amf Symbiosis The symbiosis of AMF with plants was originally discovered 400 million years ago [ 57 ]. Such links are a series of biological processes that have beneficial impacts on the ecosystems of both wild and cultivated biota [ 67 ]. An example of a reciprocal interaction that may regulate the growth and advancement of a plant is the symbiotic relationship between AMF and its partners. The plant's roots may absorb nutrients that would not otherwise be available thanks to the mycelial fungal network that has grown throughout them [ 28 ]. The fungal mycelium colonises the roots of several plants, even if they are from different species, and creates a shared mycorrhizal network (CMN). According to [ 47 ] Pringle et al. (2009), this CMN is regarded as the main element of the terrestrial ecosystem for many plant populations, including invasive species, and it facilitates the transfer of phosphorus (P) and nitrogen (N) to plants via fungi [ 60 ]. They may improve soil qualities and encourage plant development in both natural and challenging environments [ 42 , 2 , 3 ]. Plant resilience to harsh environments is increased by AMF colonisation, which results in several improvements in morpho-physiological traits [ 2 , 3 25 ]. Researchers encourage the use of AMF as influential bio-fertilizers in sustainable agricultural production, having been employed as bioinoculants [ 8 ]. In comparison to untreated soils, AMF-inoculated soil often forms more consistent masses and much more extraradical hyphal mycelium [64 ]. Glomalin-related soil protein (GRSP) is thought to keep soils with complex abiotic stressors moist [ 71 ], which then controls water levels between soil and plants and naturally promotes plant development. Glomalin contains 30–40% carbon (C) and related chemicals to protect soil from desiccation by enhancing its ability to retain soil-water [ 58 ]. Growth-related processes that affect AMF inoculation include stomatal conductance, leaf water capacity, relative water content (RWC), PSII quality, and CO2 assimilation [ 26 , 17 ]. By altering the biochemistry of the organ and tissues above ground, AMF also contributes to improving water stress tolerance [ 9 ]. Additionally, AMF inoculation promotes the accumulation of dry matter and improves moisture uptake, boosting plant tolerance to stressors like salt and drought. AMF extraction for plant growth in various biological conditions will significantly boost organic farming's ability to promote growth and raise production. Here are the key characteristics of AM fungus symbiosis: A.) Establishment: The AM fungus forms a symbiotic relationship with plant roots through the process of colonization, in which the fungus penetrates the root cells and forms structures known as arbuscules. B.) Nutrient exchange: The AM fungus enhances the plant's nutrient uptake by extending the absorptive surface area of the root system and solubilizing soil nutrients. In return, the plant provides the AM fungus with carbon. C.) Soil structure improvement: AM fungi can contribute to soil structure improvement through the production of extraradical mycelium, which helps bind soil particles and improve soil stability.D.) Enhanced plant growth: The symbiotic relationship between AM fungi and plants results in improved plant growth and health, with higher root and shoot biomass, enhanced root system architecture, and increased stress tolerance. E.) Wider host range: AM fungi have a wide host range, forming symbiotic relationships with many different plant species, including some of the most important crops such as maize, wheat, and soybean. F.) Environmental importance: AM fungi play a crucial role in the ecosystem, by improving soil fertility and plant productivity. They also help to maintain soil health and reduce soil degradation by contributing to soil aggregation and water retention. Overall, the AM fungus symbiosis is a vital component of the terrestrial ecosystem, providing many benefits to plants and the environment. General Life-cycle Of An Amf Fungi Germination: The spores of AM fungi or ascospores germinate and form mycelial colonies. Colonization: The hyphae of the AM fungus colonize the root system of a host plant. The hyphae penetrate the root cells and form arbuscules (small, branched structures) within the root cells. Nutrient exchange: The hyphae of the AM fungus extend into the soil and increase the absorptive surface area of the root system, allowing the plant to access nutrients such as phosphorus and nitrogen. In return, the AM fungus receives photosynthetically derived carbohydrates from the plant. Establishment: The relationship between the AM fungus and the host plant becomes established, and the fungus continues to grow and spread throughout the root system. Dispersal: When the host plant dies, the AM fungus begins to decompose the plant's organic matter and return nutrients to the soil. The spores of the AM fungus are then free to infect other plants and continue the cycle. Methods And Source Of Inoculum Propagation In the reviews studied, the most successful form of AMF propagation before inoculation was found to be the use of trap plants almost 75% and interestingly, only marginalised usage of other approaches was found. There are in fact several other solutions to the usage of potted trap plants. Soil cultivation technologies, such as aeroponics and hydroponics, are contributing to the development of pure clean spores and the maximisation of the host plant growth conditions [27 ] and may soon be massively exploited for mass processing. The monoxenic culture of the root organs is another approach that permits the effective large-scale dissemination of AMF directly into the inoculum. Unfortunately, only a reduced number of AM fungal organisms have so far been introduced under the procedure. The system consists of culture of inoculated excised roots (the so-called hairy roots), which have acquired the capacity to proliferate without developing any epigeous portion, after processing with the soil-borne plasmid Ri (root-inducing) Agrobacterium rhizogenes [ 11 ]. In only a few months, a significant number of spores, mycelium and colonised roots have been derived from one Petri dish [19 ]. Since, AMF may use a variety of propagules in order to expand and colonise new roots with different levels of efficiency [ 31 ], the selection of the source of the inoculum (described above) is a factor of primary importance for a successful colonisation. Spores, mycelium fragments fragmented from the lower hyphal network and other complexes inside both living and dead root fragments are all components of the extraradical and intraradical systems of AM fungi. The main cause of regrowth for some AM fungal organisms was in specific intraradical vesicles [14 ]. Different AM fungal taxonomic ranks vary in their capacity to disperse from a given propagule. The propagation of mycelial fragmentation appears to be of greater importance for organisms of the Glomeraceae family, whereas for representatives of other groups such as Gigasporaceae, Acaulosporaceae and Scutellosporaceae spore germination would be preferential method of propagation [ 15 ]. The most effective and user-friendly method to apply a multi species inoculum, as propagation through trap culture is the most widely utilised strategy, is to sieve the substrate and finely cut the root of the trap culture plant so that all the various kinds of fungal propagules (crude inoculum) can be retrieved. This solution was used in almost 68% of the reviews studied. Inoculum Structure In most of the reviews studied, the latest general tendency is to pursue one or more types of AM fungi for individual inoculation (monospecies inoculum). In the case of shoot biomass, single species inoculation experiments are more effective than inoculation experiments with more than one species concurrently used. [ 24 ] have therefore submitted that, after examining the impact on plant growth after inoculating diverse AM fungal populations with functionally distinct characteristics, fewer fungal species that are able to mitigate stress are likely to be of utmost benefit to the hosts when a host plant is subjected to one cause, such as greenhouse experiments. Another greenhouse research has shown that species composition instead of variety may be more critical in deciding how the species works [ 70 ], Most studies were restricted to the single inoculation of following three species: - Rhizophagus intraradices , Funneliformis mosseae and Rhizophagus irregularis . These are extremely versatile symbionts that can colonise a wide range of host plants, maintain long-term storage, disperse widely across the world and quickly and massively reproduce them [ 44 ]. These organisms have been ideal for premium inoculum components due to their above described characteristics. “Several experiments have found that various isolates within the same species can induce broader differences in plant reaction rather than varying species [ 41 , 20 , 4 ]. This indicates that widespread use of single AM fungal organisms, such as R. intraradices , R. irregularis , and F. mosseae should not be considered a defect in inoculation trials because these organisms may possess substantial functional heterogeneity. In this context, in the presence of R. irregularis reference genome [ 66, 37 ] the partial genome re-sequencing of several isolates from various geographical backgrounds can open up the door to exploring the roles of genetic variation in AMF communities so that it is possible to develop and choose more successful AMF for crop plants [ 51 ]”. Another factor that needs to be addressed is that the receptivity of plant organisms, including seeds, to AMF inoculation differs greatly [ 29 , 62 , 63 ]. The plants' reaction to AM fungi can be used as a selection function in modern farming, resulting in varieties or cultivars with different genetic differences. Amf As A Bio-fertilizer Bio-fertilizers are a combination of natural compounds used to increase soil fertility. These fertilisers are very useful for soil health and plant growth [ 54 ]. Various scientific experiments on AMF in the last two decades have demonstrated their innumerable benefits in terms of soil quality and crop productivity. Therefore, it is commonly assumed that AMF may in the foreseeable future be seen as a substitute for inorganic fertilisers, since mycorrhizal applications will effectively reduce the quantitative use of the chemical fertiliser input, especially phosphorus [ 45 ]. Owing to the adverse influence on food safety, crop health, and air and water systems by inorganic fertilisers, herbicides and fungicides, the continued usage of these have triggered numerous land, plant and human health concerns [ 72 ].AMF may be able to minimise chemical fertiliser usage up to 50 percent for optimal agricultural output, although this calculation depends on the variety of plant species and the prevalence of stressful environments. In order for sustainable agriculture to be accomplished, AMF as a biofertilizer becomes more significant because the proper treatment of these symbiotic fungi might significantly minimise the usage of agrochemicals. Inoculation of AMF propagules (inoculum) into a target soil is the key technique embraced for this aim. Sadly, AMF are compulsory symbionts and cannot be produced without the host plants in pure cultures. The large-scale development of AMF inocula is very difficult and complex because of this constraining feature. There are three major forms of AMF inocula. First, AMF soil may be used as an inoculum from the root zone of a plant since it usually includes colonised root parts, AMF spores, and hyphae. However, without adequate knowledge regarding propagule quantity, variety and infectivity, soil inocula cannot be effective and may be at risk of transmitting weed seeds and pathogenic agents. Spores removed from soil can be used instead as starters for the development of crude inoculum. Crude inoculum can be collected from the inert medium adapted for AMF propagation after the known AMF isolate and the host trap plant (i.e. plant which can be colonised with a lot of AMF species) have been cultivated together. This is the most common form of inoculum used for large-scale inoculation, since it normally includes a more condensed collection of propagules of the same sort present in the soil inoculum. Finally, contaminated root fragments alone from an established AMF host that are isolated from a trap crop may even be used as an inoculum source. AMF's large-scale development of crude inoculum remains very demanding even while new mass processing methods [ 27 ] and the technology of seed coating [ 69 ] are in recent years being developed [ 68 ]. The biggest challenge to an AMF inoculum is the unavoidable symbiotic conduct of the AMF, i.e., its requirement for a host facility to expand and complete its life cycles. Which implies that the propagation stage must involve a time and space-demanding process of cultivation with the host plant. “As a result, the establishment of AMF reference collections involves methodologies which are very different and more binding than those for other microbial collections. In addition, the lack of a prompt way to determine when and how often the host plant is colonised by AMF often contributes in threatening AMF 's agricultural usability. The management of the high inoculum needed for large-scale usage is also a challenging operation. However, AMF inoculation for plant manufacturing systems with a transplant stage is simpler since smaller quantities of inoculum are required. A systematic inoculation therapy may at first sight seem theoretically inefficient and financially prohibitive”. Once AMF biodiversity has been preserved and well developed, and an AMF-friendly management such as fall cover [ 34 ] and conservation tillage [ 55 ] is introduced, the AMF population will prosper and if no damage is done before and after cultivation, it is understood that in the future the network of mycorrhizal hyperbiotics of biodiversity will remain unaltered and contagious. Few of the experiments done to study the effectiveness of AMF as a biofertilizer either alone or in combination of other soil microorganisms are as follows: -The maximum grain production of Triticum aestivum at phosphorus levels was recorded on being inoculated with Pseudomonas striata , followed by Glomus fasciculatum [ 40 ]. [ 48 ] have examined the impact of biofertilizers such as Rhizobium, AM fungi etc. for Acacia nilotica and also demonstrated a substantial improvement in the length of seedlings and biomass compared with control. In the combination of Rhizobium + AM + Azospirillum maximum growth and biomass were reported among the treatments and it was 156.8% above the control. Microbial bioagents have been explored in chickpea for the prevention of collar rot disease [ 5 ]. In combination of Rhizobium and AM, optimum decrease in mortality was observed as compared to control (100% seedling mortality). Different yield parameters have been effectively improved in each of the adjustments by disease control and seedling mortality reduction. “The impact of single and dual Vesicular Arbuscular Mycorrhizal (VAM) fungi of Gigaspora rosea , Glomus intraradi ces + Gigaspora rosea , and Glomus etunicatum + Glomus intraradices on Medicago sativa growth and nutrient absorption (NPK) was studied, with a significant increase in dry shoot and root weight [ 30 ]. The effects of phosphate-solubilizing bacteria ( Glomus intraradices , Pseudomonas putida , Pseudomonas alcaligenes , Aspergillus awamori ) and Rhizobium sp . on chickpea growth, nodulation yield, and root-rot disease complex in field conditions were investigated [ 39 ]”. The number of nodules per root system was substantially larger in plants treated with Rhizobium sp. than in control plants. Under temperate circumstances, [ 13 ] investigated the impact of Rhizobium and Vesicular Arbuscular Mycorrhizae fungi on Green gramme ( Vigna radiata L.Wilczek). Rhizobium and VAM had a substantial influence on nitrogen levels during nodulation, yield metrics, NPK content in grain and straw, and crude protein content in grains. The inoculation of VAM is promising because it is inexpensive, simple to manage and promotes the growth of plants and the seed quality. Chickpea ( Cicer arietinum )'s growth and return reaction to inoculation with Rhizobium sp. and VAM has been studied [ 21 ]. These findings showed an increase of 10.83% of the total weight and 9.0% of the germination over control in the pot experiments. Conclusion According to some estimates, the global population will have reached 9 billion by the year 2050 [ 51 ]. In order to preserve both human and environmental health, the world's agricultural sector is now faced with the problem of almost tripling the amount of food that is produced while simultaneously lowering farmers' dependency on agricultural chemicals. The increase in yield that is anticipated is more than the current capacity for the production of food throughout the world [ 51 ], which highlights the need for the development or revitalization of environmentally friendly technologies such as AMF-based biofertilization. Despite the enormous potential it has, agriculturalists have not yet entirely accepted the use of AMF. According to the findings of this study, overall AMF inoculation has beneficial effects on plant growth in both controlled and open-field conditions. This is mostly due to the many nutritional advantages that this class of soil fungus symbionts provides to the host plant. In point of fact, it has been shown that AMF inoculation in the field is just as effective as inoculation in the greenhouse, where, in contrast to open field conditions, non-inoculated controls are often free of AMF. Because of this, the next significant step for the consistent use of AMF in agriculture is to conduct large-scale field experiments and a cost-benefit study, such as the one suggested in [ 16 ], to enable future end-users to be better aware of the benefits of AMF inoculant. Farmers are urged to create their own AMF inoculant from their local soils since research has shown that the indigenous AMF is as potent as or stronger than commercial or cultural isolates. Even farmers in developing nations like India, who are in desperate need of a technique of crop production that is sustainable, would have an easier time gaining access to biofertilization technology as a result of this. Declarations Acknowledgements The authors are thankful to Prof. R.K.Asthana; Head, Department of Botany, Banaras Hindu University for providing his constant guidance throughout and also for providing an international standard laboratory for carrying out this work. We are also very thankful to University Grants Commission-Council of scientific and industrial research for providing financial assistance required to carry out this work. Credit authorship contribution statement Subhesh Saurabh Jha :- Data curation, Visualization, Conceptualization, Writing original draft, Investigation. L.S.Songachan :- Supervision, Methedology, Writing-review & editing, Validation. Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. References Allen, M. F. (2011). 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(2010). “Modulation of aquaporin genes by the arbuscular mycorrhizal symbiosis in relation to osmotic stress tolerance,” in Symbioses and Stress Cellular Origin, Life in Extreme Habitats and Astrobiology, eds J. Seckbach and M. Grube (Springer), 357–374. Sadhana, B. (2014). Arbuscular mycorrhizal fungi (AMF) as a biofertilizers—a review. Int. J. Curr. Microbiol. App. Sci. 3 (4), 384–400. Säle, V., Aguilera, P., Laczko, E., Mäder, P., Berner, A., Zihlmann, U., et al. (2015). Impact of conservation tillage and organic farming on the diversity of arbuscular mycorrhizal fungi. Soil Biol. Biochem. 84, 38–52. Schüßler, A., Schwarzott, D., and Walker, C. (2001). A new fungal phylum, the Glomeromycota: phylogeny and evolution. Mycol. Res. 105, 1413–1421. Selosse, M. A., Strullu-Derrien, C., Martin, F. M., Kamoun, S., and Kenrick, P.(2015). Plants, fungi and oomycetes: a 400-million years affair that shapes the biosphere. New Phytol. 206, 501–506. Sharma, S., Prasad, R., Varma, A., and Sharma, A. K. (2017). Glycoprotein associated with Funneliformis coronatum, Gigaspora margarita and Acaulospora scrobiculata suppress the plant pathogens in vitro. Asian J. Plant Pathol. 11 (4),192–202. Smith, F. A., Jakobsen, I., and Smith, S. E. (2000). Spatial differences in acquisition of soil phosphate between two arbuscular mycorrhizal fungi in symbiosis with Medicago truncatula. New Phytol. 147, 357–366. Smith, S. E., and Read, D. J. (2008). Mycorrhizal Symbiosis, 3rd Edn. London:Academic. J. Arid Land 3, 155–163. Smith, S. E., and Smith, F. A. (2012). Fresh perspectives on the roles of arbuscular mycorrhizal fungi in plant nutrition and growth. Mycologia 104, 1–13. Smith, S. E., and Smith, F. A. (2011). Roles of arbuscular mycorrhizas in plant nutrition and growth: new paradigms from cellular to ecosystem scales. Annu.Rev. Plant Biol. 62, 227–250. Smith, S. E., Jakobsen, I., Grønlund, M., and Smith, F. A. (2011). Roles of arbuscular mycorrhizas in plant phosphorus nutrition: interactions between pathways of phosphorus uptake in arbuscular mycorrhizal roots have important implications for understanding and manipulating plant phosphorus acquisition. Plant Physiol. 156, 1050–1057. Syamsiyah, J., Herawati, A., and Mujiyo. (2018). The potential of arbuscular mycorrhizal fungi application on aggregrate stability in alfisol soil. IOP Conf. Series: Earth Environ. Sci. 142, 012045. Tamayo, E., Gómez-Gallego, T., Azcón-Aguilar, C., and Ferrol, N. (2014). Genome-wide analysis of copper, iron and zinc transporters in the arbuscular mycorrhizal fungus Rhizophagus irregularis. Plant Traffic Transp. 5:547. Tisserant, E., Malbreil, M., Kuo, A., Kohler, A., Symeonidi, A., Balestrini, R., et al. (2013). Genome of an arbuscular mycorrhizal fungus provides insight into the oldest plant symbiosis. Proc. Natl. Acad. Sci. U.S.A. 110, 20117–20122. Van der Heijden, M. G. A., Klironomos, J. N., Ursic, M., Moutoglis, P., StreitwolfEngel, R., Boller, T., et al. (1998). Mycorrhizal fungal diversity determines plantbiodiversity, ecosystem variability and productivity. Nature 396, 69–72. Van der Heijden, M. G.A., Martin, F. M., Selosse, M. A., and Sanders, I. R. (2015). Mycorrhizal ecology and evolution: the past, the present, and the future. New Phytol. 205, 1406–1423. Vosátka, M., Látr, A., Gianinazzi, S., and Albrechtová, J. (2013). Development of arbuscular mycorrhizal biotechnology and industry: current achievements and bottlenecks. Symbiosis 58, 29–37. Wagg, C., Barendregt, C., Jansa, J., and van der Heijden, M. G. A. (2015).Complementarity in both plant and mycorrhizal fungal communities are not necessarily increased by diversity in the other. J. Ecol. 103, 1233–1244. Wu, Z., McGrouther, K., Huang, J., Wu, P., Wu, W., and Wang, H. (2014). Decomposition and the contribution of glomalin-related soil protein (GRSP) in heavy metal sequestration: field experiment. Soil Biol. Biochem. 68, 283–290. Yang, S., Li, F., Malhi, S. S., Wang, P., Dongrang, S., and Wang, J. (2004). Long term fertilization effects on crop yield and nitrate nitrogen accumulation in soil in North western China. Agron. J. 96, 1039–1049. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-2559546","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":174054639,"identity":"21622650-36b6-4fc6-9ab8-342a799fa81a","order_by":0,"name":"Subhesh saurabh jha","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA/UlEQVRIie2OMWsCMRSA33FwXe5wfdLBvxA5OOjkX7lQqEuVgouDQ6ZzsrNC6W9oF+dI4FxSZ6E3tMuNJZM4ZPCJOhrtVmg+eCE88pEPwOP5k8QRHZJGA5ihzA9bdqUSTPXvlBjCpDgpDlrTXvllbNWB4GNhkteqz2T4beCpOquwdb/bnhU1F7C6x+a8HjAZpQisPq/gY3abCEU9mmF7rvibhIz2yhFGirWqQ0q65S975WbjVGBNCkQqEKAzXIi9Ert/YfrnoTkpFC9IuRNlzWcqHmDuChv3StxSWAN1+mlHFX9ejt+NsY6wExHK4y2kyS8LRENc9czj8Xj+ITuDp19y+MkSTQAAAABJRU5ErkJggg==","orcid":"https://orcid.org/0000-0002-5438-8958","institution":"Banaras Hindu University Faculty of Science","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Subhesh","middleName":"saurabh","lastName":"jha","suffix":""},{"id":174054640,"identity":"673fed97-c078-4a4e-b430-c08caca0a14e","order_by":1,"name":"L. S. Songachan","email":"","orcid":"","institution":"Banaras Hindu University Faculty of Science","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"L.","middleName":"S.","lastName":"Songachan","suffix":""}],"badges":[],"createdAt":"2023-02-07 09:51:41","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2559546/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2559546/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":32668317,"identity":"b7b6857b-e68e-4594-860a-4f9e64363a79","added_by":"auto","created_at":"2023-02-08 20:19:11","extension":"jpg","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":190298,"visible":true,"origin":"","legend":"\u003cp\u003eDifferent types of Mycorrhizal symbiosis in nature. \u003cstrong\u003eEctomycorrhizae\u003c/strong\u003e form a sheath around the roots of certain plants, such as pines, oaks, and eucalyptus. The fungi involved in ectomycorrhizae do not invade the cells of the plant roots, but rather create a layer around the root cells known as the Hartig net. \u003cstrong\u003eMonotropoid mycorrhiza\u003c/strong\u003e is a type of mycorrhizal association that forms between certain plant species and fungi. Monotropoid mycorrhizae are characterized by the fact that the fungus is unable to form a visible sheath around the roots of the host plant, and instead forms a diffuse, intraradical mycelium that is intermingled with the plant's own root tissue. \u003cstrong\u003eEricoid mycorrhiza\u003c/strong\u003e (EM) is a symbiotic relationship between certain species of ericaceous plants (such as blueberries, rhododendrons, and heaths) and a group of fungi known as ericoid mycorrhizal fungi (EMF). In this relationship, the EMF colonize the roots of the host plant and form a dense network of fungal structures called hyphae, which increase the plant's ability to absorb water and nutrients, particularly phosphorous. EM is considered a \"mycoheterotrophic\" symbiosis, meaning that the host plant is dependent on the EMF for carbon, which is obtained by the fungus through its association with other plants. \u003cstrong\u003eOrchidaceous mycorrhizae\u003c/strong\u003e are mutualistic associations between orchid plants and fungi. These relationships are formed in the roots of orchids, where the fungus colonizes the root cells and provides the plant with essential nutrients, such as phosphorous. In return, the orchid provides the fungus with organic carbon through photosynthesis. Orchid mycorrhizae are considered to be obligate, meaning that the orchid cannot survive without the fungus and vice versa. This is because orchids do not have the ability to form nodules on their roots like other plants, which allows them to fix nitrogen from the air. Instead, they rely on the fungus to provide them with this essential nutrient. \u003cstrong\u003eArbuscular mycorrhizal (AM\u003c/strong\u003e) fungi are a group of soil-borne, obligate symbionts that form mutualistic associations with the majority of land plants. These fungi colonize plant roots, forming unique structures called arbuscules within the root cells. In exchange for sugars produced by the plant through photosynthesis, the fungi provide the plant with phosphorous and other essential nutrients that are otherwise difficult for the plant to acquire from the soil. AM fungi are ancient symbionts that have evolved alongside plants for over 400 million years, and they play important roles in ecosystem functioning and plant productivity.\u003c/p\u003e","description":"","filename":"1.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2559546/v1/1f38477b087cc4ff5278d4ed.jpg"},{"id":32668316,"identity":"82ba81d6-dee2-444f-a4d0-9bee0eb01ff7","added_by":"auto","created_at":"2023-02-08 20:19:11","extension":"jpg","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":322660,"visible":true,"origin":"","legend":"\u003cp\u003eA pictorial representation of few of the benefits provided by AMF in plant growth and yield of plants. Increased resistance to foliar pathogens, increased drought tolerance, increased salt tolerance, increase defense mechanism, help in uptake of essential nutrients, bioremediation is few of the many benefits that AMF provides in the growth and development of a plant.\u003c/p\u003e","description":"","filename":"2.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2559546/v1/fbf690e480882404c13bd017.jpg"},{"id":32668318,"identity":"4a292f3f-4bd1-4aea-910c-6c9fa6d46209","added_by":"auto","created_at":"2023-02-08 20:19:11","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":136398,"visible":true,"origin":"","legend":"\u003cp\u003eLife cycle of AMF fungi. The life-cycle of AM Fungi is basically divided in two stages pre-symbiosis and symbiosis. Pre-symbiosis is characterized by the following steps:-a.) Plant root exudating certain chemicals b.) spore germination and hyphal growth c.) attachment of appressorium to the root. This step is followed by the symbiosis stage in the life of AM Fungi which largely consists of two steps which are:-d.) Formation of arbuscules, vesicles inside the root e.) formation of spores and mycelium e.) spores getting attached with the roots.\u003c/p\u003e","description":"","filename":"3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2559546/v1/4bb6903eb57a177712ea6bf9.jpg"},{"id":32668319,"identity":"5a5f24c8-8fa0-49d5-963a-a18dc5d8195a","added_by":"auto","created_at":"2023-02-08 20:19:11","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":89003,"visible":true,"origin":"","legend":"\u003cp\u003eGraphical representation of AMF infection in the root. AMF colonisation is characterised by the formation of arbuscules, vesicles, hyphae and spores.\u003c/p\u003e","description":"","filename":"4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-2559546/v1/c177a5ae27be2435d012d797.jpg"},{"id":33666656,"identity":"9d4176fa-a000-439c-a0a4-fbed165076a8","added_by":"auto","created_at":"2023-03-02 06:29:52","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":694588,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2559546/v1/c3078c0a-eff4-4b4b-9f13-1752e9ae661e.pdf"}],"financialInterests":"","formattedTitle":"\u003cp\u003eThe Usage of Arbuscular Mycorrhizal Fungi (Amf) as a Biofertilizer\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eMycorrhizae are a type of symbiotic relationship that occur between fungi and plant roots. There are two main types of mycorrhiza: ectomycorrhizae and endomycorrhizae. Ectomycorrhizae form a sheath around the roots of certain plants, such as pines, oaks, and eucalyptus. The fungi involved in ectomycorrhizae do not invade the cells of the plant roots, but rather create a layer around the root cells known as the Hartig net. This allows for the exchange of nutrients and water between the plant and the fungus. Ectomycorrhizae are important for the health and growth of trees in forests, as they help to improve the uptake of nutrients and water from the soil. Endomycorrhizae, on the other hand, form a symbiotic relationship with the plant by actually invading the cells of the root. There are two main types of endomycorrhizae: arbuscular mycorrhizae (AM) and ericoid mycorrhizae (EM). Various kinds of mycorrhizal connections are described below in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. Soil micro-organisms such as AMFs reflect a crucial connection between plants and mineral soil nutrients. They are also gaining rising attention as natural fertilisers. AMFs are mandatory symbiotics of phylum Glomeromycota [\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e], establishing mutualistic symbiosis, with about 80 percent of the land plant organisms, including many food crops. In return for photosynthetic materials, they provide the host plant with mineral nutrients and water [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e]. The AMF mycelium from the root system is able to extract nutrients from soil volumes that are inaccessible to the roots [\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e]. In comparison to plant roots fungal hyphae are far thinner and can reach narrower pores [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Carbohydrates and mineral nutrients are then spread by the plant and fungi within the roots. AM fungal hyphae colonise root cortex predominantly by forming profusely branchy structures inside the cells, i.e. arbuscules, which are known as the functional nutrient exchange site [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. AMF therefore eliminates plant growth restrictions imposed by an insufficient supply of nutrients [\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]. In recent times a non-mycorrhizal state can be regarded as rare in natural habitats for most organisms [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e], even though the AM fungal populations below the ground are significantly different depending on the species composition, soil and seasonal form or variation of these factors [\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e]. AM experiences provide plants with additional advantages, in addition to an increased food source, such as enhanced drought and salinity resistance [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]. While several studies have been performed on the impact of AM symbiosis on plant reaction to abiotic stress such as drought, salinity and flooding in the last few years, the processes that have contributed to an improved plant stress resistance still remains somewhat elusive [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e, \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].Metals such as Iron(Fe),Copper(Cu) and Zinc (Zn) perform important functions in a variety of sub-cellular compartments, but they constitute a highly reactive community of elements that are toxic at large concentrations [\u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e]. AM fungi have been documented to minimise the toxicity of heavy metals in host plant and to withstand high metal concentrations in soil. [\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e65\u003c/span\u003e, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e38\u003c/span\u003e]. Metal transporters play a vital function in homeostasis of heavy metals. A Zn transporter was identified in \u003cem\u003eGlomus intraradices\u003c/em\u003e (GintZnT1) [\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e] and more recently multiple putative genes coding Cu, Fe \u0026amp; Zn transporters were identified in a genome-wide study of the recently published \u003cem\u003eRhizophagus irregularis\u003c/em\u003e (formally, \u003cem\u003eGlomus intraradices)\u003c/em\u003e genome [\u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e66\u003c/span\u003e]. The next steps would be characterisation of these carriers and discerning their role in the symbiosis. AM fungi may also have a significant influence on the environment, as they promote soil structure and aggregation [\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e, \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e, \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e] and control the development and production of plant populations [\u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e].The impact of AM symbiosis have recently been also studied on greenhouse gas (GHG) emission [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e]. Evidence presented by Bender et al. (2014) suggests that AM fungus may have a role to play in climate change mitigation due to their ability to significantly reduce emissions of N2O, a key greenhouse gas. By enhancing plant nitrogen (N) absorption and assimilation, AM fungi may be able to reduce N2O emissions by decreasing soluble N in soil and, in turn, denitrification [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. Correlations between AM fungal abundance and genes involved in primary N2O production (nirK) and consumption (nosZ) suggest that AM fungi promote shifts in soil microbial biomass and community composition that lead to reduced N2O emissions. According to [\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e], AM symbiosis aids in N2O emission management at high soil moisture levels, and it was suggested that AM plant N2O emission control may be mediated by higher soil water use rather than increased N absorption. Therefore, AM fungi are primary biotic soil elements, which, when absent or degraded, for instance, by anthropic input, will contribute to a less effective functioning of the ecosystem. The process of re-establishing AMF may be a promising solution to industrial fertilisation methods in order to achieve organic cultivation, a significant goal for farmers in the midst of a global recession and an environmentally friendly consumer. The key technique for achieving this aim is to directly reintroduce AMF (inoculum) propagules in the target soil. However, in the application of these fungi, awareness of how AMF adapts and reacts to the objective of soil management and ecosystem management and the events which result in a functional symbiosis, including the mechanisms involved in the transfer of nutrients is important. After a brief discussion of the latest studies on the nutritional aspects of AM symbiosis and a short description of the challenges of development of AMF inoculum, descriptions of the application of AM fungi are mentioned and addressed, both under regulated and open field conditions, with specific emphasis on identifying factors contributing to success of the biofertilizer.\u003c/p\u003e "},{"header":"Essential Features Of Amf Symbiosis","content":"\u003cp\u003eThe symbiosis of AMF with plants was originally discovered 400\u0026nbsp;million years ago [\u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e]. Such links are a series of biological processes that have beneficial impacts on the ecosystems of both wild and cultivated biota [\u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e67\u003c/span\u003e]. An example of a reciprocal interaction that may regulate the growth and advancement of a plant is the symbiotic relationship between AMF and its partners. The plant's roots may absorb nutrients that would not otherwise be available thanks to the mycelial fungal network that has grown throughout them [\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]. The fungal mycelium colonises the roots of several plants, even if they are from different species, and creates a shared mycorrhizal network (CMN). According to [\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e] Pringle et al. (2009), this CMN is regarded as the main element of the terrestrial ecosystem for many plant populations, including invasive species, and it facilitates the transfer of phosphorus (P) and nitrogen (N) to plants via fungi [\u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e]. They may improve soil qualities and encourage plant development in both natural and challenging environments [\u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e42\u003c/span\u003e, \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Plant resilience to harsh environments is increased by AMF colonisation, which results in several improvements in morpho-physiological traits [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]. Researchers encourage the use of AMF as influential bio-fertilizers in sustainable agricultural production, having been employed as bioinoculants [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. In comparison to untreated soils, AMF-inoculated soil often forms more consistent masses and much more extraradical hyphal mycelium [64 ]. Glomalin-related soil protein (GRSP) is thought to keep soils with complex abiotic stressors moist [\u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e71\u003c/span\u003e], which then controls water levels between soil and plants and naturally promotes plant development. Glomalin contains 30\u0026ndash;40% carbon (C) and related chemicals to protect soil from desiccation by enhancing its ability to retain soil-water [ 58 ]. Growth-related processes that affect AMF inoculation include stomatal conductance, leaf water capacity, relative water content (RWC), PSII quality, and CO2 assimilation [\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. By altering the biochemistry of the organ and tissues above ground, AMF also contributes to improving water stress tolerance [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]. Additionally, AMF inoculation promotes the accumulation of dry matter and improves moisture uptake, boosting plant tolerance to stressors like salt and drought. AMF extraction for plant growth in various biological conditions will significantly boost organic farming's ability to promote growth and raise production. Here are the key characteristics of AM fungus symbiosis: A.) Establishment: The AM fungus forms a symbiotic relationship with plant roots through the process of colonization, in which the fungus penetrates the root cells and forms structures known as arbuscules. B.) Nutrient exchange: The AM fungus enhances the plant's nutrient uptake by extending the absorptive surface area of the root system and solubilizing soil nutrients. In return, the plant provides the AM fungus with carbon. C.) Soil structure improvement: AM fungi can contribute to soil structure improvement through the production of extraradical mycelium, which helps bind soil particles and improve soil stability.D.) Enhanced plant growth: The symbiotic relationship between AM fungi and plants results in improved plant growth and health, with higher root and shoot biomass, enhanced root system architecture, and increased stress tolerance. E.) Wider host range: AM fungi have a wide host range, forming symbiotic relationships with many different plant species, including some of the most important crops such as maize, wheat, and soybean. F.) Environmental importance: AM fungi play a crucial role in the ecosystem, by improving soil fertility and plant productivity. They also help to maintain soil health and reduce soil degradation by contributing to soil aggregation and water retention. Overall, the AM fungus symbiosis is a vital component of the terrestrial ecosystem, providing many benefits to plants and the environment.\u003c/p\u003e "},{"header":"General Life-cycle Of An Amf Fungi","content":"\u003cp\u003e \u003cdiv class=\"BlockQuote\"\u003e \u003cp\u003eGermination: The spores of AM fungi or ascospores germinate and form mycelial colonies. Colonization: The hyphae of the AM fungus colonize the root system of a host plant. The hyphae penetrate the root cells and form arbuscules (small, branched structures) within the root cells. Nutrient exchange: The hyphae of the AM fungus extend into the soil and increase the absorptive surface area of the root system, allowing the plant to access nutrients such as phosphorus and nitrogen. In return, the AM fungus receives photosynthetically derived carbohydrates from the plant. Establishment: The relationship between the AM fungus and the host plant becomes established, and the fungus continues to grow and spread throughout the root system. Dispersal: When the host plant dies, the AM fungus begins to decompose the plant's organic matter and return nutrients to the soil. The spores of the AM fungus are then free to infect other plants and continue the cycle.\u003c/p\u003e \u003c/div\u003e \u003c/p\u003e "},{"header":"Methods And Source Of Inoculum Propagation","content":"\u003cp\u003eIn the reviews studied, the most successful form of AMF propagation before inoculation was found to be the use of trap plants almost 75% and interestingly, only marginalised usage of other approaches was found. There are in fact several other solutions to the usage of potted trap plants. Soil cultivation technologies, such as aeroponics and hydroponics, are contributing to the development of pure clean spores and the maximisation of the host plant growth conditions [27 ] and may soon be massively exploited for mass processing. The monoxenic culture of the root organs is another approach that permits the effective large-scale dissemination of AMF directly into the inoculum. Unfortunately, only a reduced number of AM fungal organisms have so far been introduced under the procedure. The system consists of culture of inoculated excised roots (the so-called hairy roots), which have acquired the capacity to proliferate without developing any epigeous portion, after processing with the soil-borne plasmid Ri (root-inducing) \u003cem\u003eAgrobacterium rhizogenes\u003c/em\u003e [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. In only a few months, a significant number of spores, mycelium and colonised roots have been derived from one Petri dish [19 ]. Since, AMF may use a variety of propagules in order to expand and colonise new roots with different levels of efficiency [\u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e], the selection of the source of the inoculum (described above) is a factor of primary importance for a successful colonisation. Spores, mycelium fragments fragmented from the lower hyphal network and other complexes inside both living and dead root fragments are all components of the extraradical and intraradical systems of AM fungi. The main cause of regrowth for some AM fungal organisms was in specific intraradical vesicles [14 ]. Different AM fungal taxonomic ranks vary in their capacity to disperse from a given propagule. The propagation of mycelial fragmentation appears to be of greater importance for organisms of the Glomeraceae family, whereas for representatives of other groups such as Gigasporaceae, Acaulosporaceae and Scutellosporaceae spore germination would be preferential method of propagation [ 15 ]. The most effective and user-friendly method to apply a multi species inoculum, as propagation through trap culture is the most widely utilised strategy, is to sieve the substrate and finely cut the root of the trap culture plant so that all the various kinds of fungal propagules (crude inoculum) can be retrieved. This solution was used in almost 68% of the reviews studied.\u003c/p\u003e"},{"header":"Inoculum Structure","content":"\u003cp\u003eIn most of the reviews studied, the latest general tendency is to pursue one or more types of AM fungi for individual inoculation (monospecies inoculum). In the case of shoot biomass, single species inoculation experiments are more effective than inoculation experiments with more than one species concurrently used. [\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e] have therefore submitted that, after examining the impact on plant growth after inoculating diverse AM fungal populations with functionally distinct characteristics, fewer fungal species that are able to mitigate stress are likely to be of utmost benefit to the hosts when a host plant is subjected to one cause, such as greenhouse experiments. Another greenhouse research has shown that species composition instead of variety may be more critical in deciding how the species works [\u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e70\u003c/span\u003e], Most studies were restricted to the single inoculation of following three species: - \u003cem\u003eRhizophagus intraradices\u003c/em\u003e, \u003cem\u003eFunneliformis mosseae\u003c/em\u003e and \u003cem\u003eRhizophagus irregularis\u003c/em\u003e. These are extremely versatile symbionts that can colonise a wide range of host plants, maintain long-term storage, disperse widely across the world and quickly and massively reproduce them [\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e]. These organisms have been ideal for premium inoculum components due to their above described characteristics. \u0026ldquo;Several experiments have found that various isolates within the same species can induce broader differences in plant reaction rather than varying species [\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. This indicates that widespread use of single AM fungal organisms, such as \u003cem\u003eR. intraradices\u003c/em\u003e, \u003cem\u003eR. irregularis\u003c/em\u003e, and \u003cem\u003eF. mosseae\u003c/em\u003e should not be considered a defect in inoculation trials because these organisms may possess substantial functional heterogeneity. In this context, in the presence of \u003cem\u003eR. irregularis\u003c/em\u003e reference genome [ 66, 37 ] the partial genome re-sequencing of several isolates from various geographical backgrounds can open up the door to exploring the roles of genetic variation in AMF communities so that it is possible to develop and choose more successful AMF for crop plants [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]\u0026rdquo;. Another factor that needs to be addressed is that the receptivity of plant organisms, including seeds, to AMF inoculation differs greatly [\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e, \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e]. The plants' reaction to AM fungi can be used as a selection function in modern farming, resulting in varieties or cultivars with different genetic differences.\u003c/p\u003e "},{"header":"Amf As A Bio-fertilizer","content":"\u003cp\u003eBio-fertilizers are a combination of natural compounds used to increase soil fertility. These fertilisers are very useful for soil health and plant growth [ 54 ]. Various scientific experiments on AMF in the last two decades have demonstrated their innumerable benefits in terms of soil quality and crop productivity. Therefore, it is commonly assumed that AMF may in the foreseeable future be seen as a substitute for inorganic fertilisers, since mycorrhizal applications will effectively reduce the quantitative use of the chemical fertiliser input, especially phosphorus [\u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]. Owing to the adverse influence on food safety, crop health, and air and water systems by inorganic fertilisers, herbicides and fungicides, the continued usage of these have triggered numerous land, plant and human health concerns [ 72 ].AMF may be able to minimise chemical fertiliser usage up to 50 percent for optimal agricultural output, although this calculation depends on the variety of plant species and the prevalence of stressful environments. In order for sustainable agriculture to be accomplished, AMF as a biofertilizer becomes more significant because the proper treatment of these symbiotic fungi might significantly minimise the usage of agrochemicals. Inoculation of AMF propagules (inoculum) into a target soil is the key technique embraced for this aim. Sadly, AMF are compulsory symbionts and cannot be produced without the host plants in pure cultures. The large-scale development of AMF inocula is very difficult and complex because of this constraining feature. There are three major forms of AMF inocula. First, AMF soil may be used as an inoculum from the root zone of a plant since it usually includes colonised root parts, AMF spores, and hyphae. However, without adequate knowledge regarding propagule quantity, variety and infectivity, soil inocula cannot be effective and may be at risk of transmitting weed seeds and pathogenic agents. Spores removed from soil can be used instead as starters for the development of crude inoculum. Crude inoculum can be collected from the inert medium adapted for AMF propagation after the known AMF isolate and the host trap plant (i.e. plant which can be colonised with a lot of AMF species) have been cultivated together. This is the most common form of inoculum used for large-scale inoculation, since it normally includes a more condensed collection of propagules of the same sort present in the soil inoculum. Finally, contaminated root fragments alone from an established AMF host that are isolated from a trap crop may even be used as an inoculum source. AMF's large-scale development of crude inoculum remains very demanding even while new mass processing methods [\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e] and the technology of seed coating [\u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e69\u003c/span\u003e] are in recent years being developed [\u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e68\u003c/span\u003e]. The biggest challenge to an AMF inoculum is the unavoidable symbiotic conduct of the AMF, i.e., its requirement for a host facility to expand and complete its life cycles. Which implies that the propagation stage must involve a time and space-demanding process of cultivation with the host plant. \u0026ldquo;As a result, the establishment of AMF reference collections involves methodologies which are very different and more binding than those for other microbial collections. In addition, the lack of a prompt way to determine when and how often the host plant is colonised by AMF often contributes in threatening AMF 's agricultural usability. The management of the high inoculum needed for large-scale usage is also a challenging operation. However, AMF inoculation for plant manufacturing systems with a transplant stage is simpler since smaller quantities of inoculum are required. A systematic inoculation therapy may at first sight seem theoretically inefficient and financially prohibitive\u0026rdquo;. Once AMF biodiversity has been preserved and well developed, and an AMF-friendly management such as fall cover [\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e] and conservation tillage [\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e] is introduced, the AMF population will prosper and if no damage is done before and after cultivation, it is understood that in the future the network of mycorrhizal hyperbiotics of biodiversity will remain unaltered and contagious. Few of the experiments done to study the effectiveness of AMF as a biofertilizer either alone or in combination of other soil microorganisms are as follows: -The maximum grain production of \u003cem\u003eTriticum aestivum\u003c/em\u003e at phosphorus levels was recorded on being inoculated with \u003cem\u003ePseudomonas striata\u003c/em\u003e, followed by \u003cem\u003eGlomus fasciculatum\u003c/em\u003e [\u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e40\u003c/span\u003e]. [\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e] have examined the impact of biofertilizers such as Rhizobium, AM fungi etc. for \u003cem\u003eAcacia nilotica\u003c/em\u003e and also demonstrated a substantial improvement in the length of seedlings and biomass compared with control. In the combination of \u003cem\u003eRhizobium\u003c/em\u003e\u0026thinsp;+\u0026thinsp;AM\u0026thinsp;+\u0026thinsp;\u003cem\u003eAzospirillum\u003c/em\u003e maximum growth and biomass were reported among the treatments and it was 156.8% above the control. Microbial bioagents have been explored in chickpea for the prevention of collar rot disease [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. In combination of \u003cem\u003eRhizobium\u003c/em\u003e and AM, optimum decrease in mortality was observed as compared to control (100% seedling mortality). Different yield parameters have been effectively improved in each of the adjustments by disease control and seedling mortality reduction. \u0026ldquo;The impact of single and dual Vesicular Arbuscular Mycorrhizal (VAM) fungi of \u003cem\u003eGigaspora rosea\u003c/em\u003e, \u003cem\u003eGlomus intraradi\u003c/em\u003eces\u0026thinsp;+\u0026thinsp;\u003cem\u003eGigaspora rosea\u003c/em\u003e, and \u003cem\u003eGlomus etunicatum\u003c/em\u003e\u0026thinsp;+\u0026thinsp;\u003cem\u003eGlomus intraradices\u003c/em\u003e on Medicago sativa growth and nutrient absorption (NPK) was studied, with a significant increase in dry shoot and root weight [\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e]. The effects of phosphate-solubilizing bacteria (\u003cem\u003eGlomus intraradices\u003c/em\u003e, \u003cem\u003ePseudomonas putida\u003c/em\u003e, \u003cem\u003ePseudomonas alcaligenes\u003c/em\u003e, \u003cem\u003eAspergillus awamori\u003c/em\u003e) and \u003cem\u003eRhizobium sp\u003c/em\u003e. on chickpea growth, nodulation yield, and root-rot disease complex in field conditions were investigated [\u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e39\u003c/span\u003e]\u0026rdquo;. The number of nodules per root system was substantially larger in plants treated with Rhizobium sp. than in control plants. Under temperate circumstances, [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e] investigated the impact of Rhizobium and Vesicular Arbuscular Mycorrhizae fungi on Green gramme (\u003cem\u003eVigna radiata\u003c/em\u003e L.Wilczek). Rhizobium and VAM had a substantial influence on nitrogen levels during nodulation, yield metrics, NPK content in grain and straw, and crude protein content in grains. The inoculation of VAM is promising because it is inexpensive, simple to manage and promotes the growth of plants and the seed quality. Chickpea (\u003cem\u003eCicer arietinum\u003c/em\u003e)'s growth and return reaction to inoculation with Rhizobium sp. and VAM has been studied [\u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e21\u003c/span\u003e]. These findings showed an increase of 10.83% of the total weight and 9.0% of the germination over control in the pot experiments.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eAccording to some estimates, the global population will have reached 9\u0026nbsp;billion by the year 2050 [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]. In order to preserve both human and environmental health, the world's agricultural sector is now faced with the problem of almost tripling the amount of food that is produced while simultaneously lowering farmers' dependency on agricultural chemicals. The increase in yield that is anticipated is more than the current capacity for the production of food throughout the world [\u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e], which highlights the need for the development or revitalization of environmentally friendly technologies such as AMF-based biofertilization. Despite the enormous potential it has, agriculturalists have not yet entirely accepted the use of AMF. According to the findings of this study, overall AMF inoculation has beneficial effects on plant growth in both controlled and open-field conditions. This is mostly due to the many nutritional advantages that this class of soil fungus symbionts provides to the host plant. In point of fact, it has been shown that AMF inoculation in the field is just as effective as inoculation in the greenhouse, where, in contrast to open field conditions, non-inoculated controls are often free of AMF. Because of this, the next significant step for the consistent use of AMF in agriculture is to conduct large-scale field experiments and a cost-benefit study, such as the one suggested in [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e], to enable future end-users to be better aware of the benefits of AMF inoculant. Farmers are urged to create their own AMF inoculant from their local soils since research has shown that the indigenous AMF is as potent as or stronger than commercial or cultural isolates. Even farmers in developing nations like India, who are in desperate need of a technique of crop production that is sustainable, would have an easier time gaining access to biofertilization technology as a result of this.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors are thankful to Prof. R.K.Asthana; Head, Department of Botany, Banaras Hindu University for providing his constant guidance throughout and also for providing an international standard laboratory for carrying out this work. We are also very thankful to University Grants Commission-Council of scientific and industrial research for providing financial assistance required to carry out this work.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCredit authorship contribution statement\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eSubhesh Saurabh Jha\u003c/strong\u003e :- Data curation, Visualization, Conceptualization, Writing original draft, Investigation. \u003cstrong\u003eL.S.Songachan\u0026nbsp;\u003c/strong\u003e:- Supervision, Methedology, Writing-review \u0026amp; editing, Validation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eDeclaration of Competing Interest\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eAllen, M. F. (2011). Linking water and nutrients through the vadose zone: a fungal interface between the soil and plant systems: linking water and nutrients through the vadose zone: a fungal interface between the soil and plant systems.\u003c/li\u003e\n\u003cli\u003eAlqarawi, A. A., Abd-Allah, E. F., and Hashem, A. (2014a). Alleviation of salt induced adverse impact via mycorrhizal fungi in Ephedra aphylla Forssk.J. Plant. Interact. 9 (1), 802\u0026ndash;810. \u003c/li\u003e\n\u003cli\u003eAlqarawi, A. A., Hashem, A., Abd_Allah, E. F., Alshahrani, T. S., and Huqail, A. A.(2014b). Effect of salinity on moisture content, pigment system, and lipid composition in Ephedra alata Decne. Acta Biol. Hung. 65 (1), 61\u0026ndash;71.\u003c/li\u003e\n\u003cli\u003eAngelard, C., Colard, A., Niculita-Hirzel, H., Croll, D., and Sanders, I. R.(2010). Segregation in a mycorrhizal fungus alters rice growth and symbiosisspecific gene transcription. Curr. 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New Phytol.\u003c/li\u003e\n\u003cli\u003eNavarro, J. M., Perez-Tornero, O., and Morte, A. (2014). Alleviation of salt stress in citrus seedlings inoculated with arbuscular mycorrhizal fungi depends on the root stock salt tolerance. J. Plant Physiol. 171 (1), 76\u0026ndash;85. \u003c/li\u003e\n\u003cli\u003eNouri, E., Breuillin-Sessoms, F., Feller, U., and Reinhardt, D. (2014). Phosphorus and nitrogen regulate arbuscular mycorrhizal symbiosis in Petunia hybrida. PLoS ONE 9:e90841. \u003c/li\u003e\n\u003cli\u003e\u0026Ouml;pik, M., Vanatoa, A., Vanatoa, E., Moora, M., Davison, J., Kalwij, J. M.,et al. (2010). The online database MaarjAM reveals global and ecosystemic distribution patterns in arbuscular mycorrhizal fungi (Glomeromycota). New Phytol. 188, 223\u0026ndash;241. \u003c/li\u003e\n\u003cli\u003eOrtas, I. (2012). The effect of mycorrhizal fungal inoculation on plant yield,nutrient uptake and inoculation effectiveness under long-term field conditions.Field Crops Res. 125, 35\u0026ndash;48. \u003c/li\u003e\n\u003cli\u003ePorcel, R., Aroca, R., and Ruiz-Lozano, J. M. (2011). Salinity stress alleviation using arbuscular mycorrhizal fungi. A review. Agron. Sustain. Dev. 32, 181\u0026ndash;200. \u003c/li\u003e\n\u003cli\u003ePringle, A., Bever, J. D., Gardes, M., Parrent, J. L., Rillig, M. C., and Klironomos,J. N. (2009). Mycorrhizal symbioses and plant invasions. Ann. Rev. Ecol. Evol.Syst. 40, 699\u0026ndash;715. \u003c/li\u003e\n\u003cli\u003eRajendran, K. and Jayasree,R.,2007, Effect of Biofertilizers on quality seedling production of Acacia nilotica. Journal of Non-timber products, 14:5-11.\u003c/li\u003e\n\u003cli\u003eRillig, M. C., and Mummey, D. L. (2006). Mycorrhizas and soil structure. New Phytol. 171, 41\u0026ndash;53. \u003c/li\u003e\n\u003cli\u003eRillig, M. C., Aguilar-Trigueros, C. A., Bergmann, J., Verbruggen, E., Veresoglou,S. D., and Lehmann, A. (2015). Plant root and mycorrhizal fungal traits for understanding soil aggregation. New Phytol. 205, 1385\u0026ndash;1388. \u003c/li\u003e\n\u003cli\u003eRodriguez, A., and Sanders, I. R. (2015). The role of community and population ecology in applying mycorrhizal fungi for improved food security. ISME J. 9, 1053\u0026ndash;1061. \u003c/li\u003e\n\u003cli\u003eRuiz-Lozano, J. M. (2003). Arbuscular mycorrhizal symbiosis and alleviation of osmotic stress. New perspectives for molecular studies. Mycorrhiza 13,309\u0026ndash;317. \u003c/li\u003e\n\u003cli\u003eRuiz-Lozano, J. M., and Aroca, R. (2010). \u0026ldquo;Modulation of aquaporin genes by the arbuscular mycorrhizal symbiosis in relation to osmotic stress tolerance,\u0026rdquo; in Symbioses and Stress Cellular Origin, Life in Extreme Habitats and Astrobiology, eds J. Seckbach and M. Grube (Springer), 357\u0026ndash;374. \u003c/li\u003e\n\u003cli\u003eSadhana, B. 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Development of arbuscular mycorrhizal biotechnology and industry: current achievements and bottlenecks. Symbiosis 58, 29\u0026ndash;37. \u003c/li\u003e\n\u003cli\u003eWagg, C., Barendregt, C., Jansa, J., and van der Heijden, M. G. A. (2015).Complementarity in both plant and mycorrhizal fungal communities are not necessarily increased by diversity in the other. J. Ecol. 103, 1233\u0026ndash;1244. \u003c/li\u003e\n\u003cli\u003eWu, Z., McGrouther, K., Huang, J., Wu, P., Wu, W., and Wang, H. (2014). Decomposition and the contribution of glomalin-related soil protein (GRSP) in heavy metal sequestration: field experiment. Soil Biol. Biochem. 68, 283\u0026ndash;290.\u003c/li\u003e\n\u003cli\u003eYang, S., Li, F., Malhi, S. S., Wang, P., Dongrang, S., and Wang, J. (2004). Long term fertilization effects on crop yield and nitrate nitrogen accumulation in soil in North western China. Agron. J. 96, 1039\u0026ndash;1049. \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Arbuscular mycorrhizal fungi, Biofertilizers, Inoculum, Sustainable agriculture, Microbiome ","lastPublishedDoi":"10.21203/rs.3.rs-2559546/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2559546/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eArbuscular Mycorrhizal Fungi (AMF) are a group of soil-borne fungi that form symbiotic relationships with the roots of most plants, including crops. In this relationship, the fungus provides the plant with nutrients, such as phosphorous, in exchange for carbohydrates produced by the plant through photosynthesis. The use of AMF as a biofertilizer involves the application of these fungi to soil to enhance plant growth and improve nutrient uptake. Studies have shown that AMF can increase plant growth, drought tolerance, and nutrient uptake, leading to improved crop yields. The fungi form a network of hyphae in the soil, which helps to increase the soil's water-holding capacity, as well as its ability to retain nutrients. This can lead to improved plant growth and health, even in nutrient-poor soils. In addition, the use of AMF as a biofertilizer can help to reduce the dependence on synthetic fertilizers, which can have negative environmental impacts. AMF can help to improve soil fertility, increase plant nutrient uptake, and reduce soil erosion, leading to more sustainable agriculture practices. However, it is important to note that the effectiveness of AMF as a biofertilizer can vary depending on several factors, including the species of AMF used, the type of crop being grown, and the conditions of the soil. Additionally, the proper application and management of AMF is important to ensure its effectiveness. In conclusion, the use of AMF as a biofertilizer has the potential to enhance plant growth, improve nutrient uptake, and promote sustainable agriculture practices.\u003c/p\u003e","manuscriptTitle":"The Usage of Arbuscular Mycorrhizal Fungi (Amf) as a Biofertilizer","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-02-08 20:19:06","doi":"10.21203/rs.3.rs-2559546/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"025ec1ac-5a3c-4965-acb1-c38f36951352","owner":[],"postedDate":"February 8th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2023-03-02T06:29:45+00:00","versionOfRecord":[],"versionCreatedAt":"2023-02-08 20:19:06","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-2559546","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2559546","identity":"rs-2559546","version":["v1"]},"buildId":"GqpaHPwrfC8PjnIFayRh5","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}