Arbuscular mycorrhizal (AM) fungi enhance plant stress tolerance, but it is unclear whether AM fungi affect heat tolerance in cucumbers. This study aimed to analyze how an AM fungus, Diversispora versiformis, affected growth, chlorophyll, five osmolytes, and plasma membrane intrinsic proteins (PIPs) and heat shock protein 70 (Hsp70) gene expression in cucumber leaves after a short-term (80 h) heat stress. Heat treatment significantly reduced root AM fungal colonization rate (0.26 folds). Heat treatment also distinctly suppressed plant height, stem diameter, and biomass, whereas AM fungal inoculation improved these growth variables as well as the chlorophyll index, with the benefit being more obvious under heat than under no-heat stress conditions. Heat treatment triggered differential changes in osmolytes (sucrose, fructose, and betaine) of inoculated and uninoculated cucumbers, whereas inoculation with AM fungus significantly raised leaf sucrose, fructose, glucose, betaine, and proline levels when compared to non-AM fungal inoculation. Heat treatment increased the expression of two (CsPIP1;6 and CsPIP2;1) of eight CsPIPs in inoculated and uninoculated plants, whereas AM fungal inoculation up-regulated the expression of CsPIP1;6, CsPIP2;1, and CsPIP2;6 under heat stress conditions. Hsp70s expressed differently in inoculated and uninoculated plants under heat versus no-heat stress, with 6 of 11 CsHsp70s down-regulated in inoculated plants. Under heat stress conditions, AM fungus only up-regulated CsHsp70-8 expression in 11 Hsp70s, while another eight CsHsp70s were down-regulated. Heat treatment and AM fungal inoculation both increased the expression of CsHsp70-8 and CsPIP1;6. It was concluded that AM fungus-inoculated cucumbers have high levels of growth, chlorophyll, and osmolytes under heat stress and do not require high CsPIPs and CsHsp70s expression to tolerate a short-term heat treatment.

Glomalin-related soil protein (GRSP) is a specific glycoprotein secreted into the soil by hyphae and spores of arbuscular mycorrhizal fungi that have many potential functions. It is not clear whether exogenous GRSP has an effect on plant growth and soil properties or whether the effects are related to the type of GRSP used. In this study, trifoliate orange (Poncirus trifoliata L. Raf.) seedlings were used to analyze the effects of easily extractable GRSP (EE-GRSP) and difficultly extractable GRSP (DE-GRSP) at a quarter-, half-, and full-strength concentration on shoot and root biomass as well as soil properties The results showed that, at different strengths, exogenous EE-GRSR significantly increased shoot and root biomass compared to the control, which displayed the most significant effects from the half-strength EE-GRSP. In contrast, DE-GRSP, at various strengths, significantly reduced shoot and root biomass. Furthermore, the application of exogenous EE-GRSP stimulated soil water-stable aggregate (WSA) content at 2–4 mm and 0.5–1 mm sizes, while DE-GRSP strongly reduced WSA content at the 2–4 mm, 1–2 mm, 0.5–1 mm, and 0.25–0.5 mm sizes, consequently leading to an increase or decrease in the WSA stability, according to the mean weight diameter. However, exogenous EE-GRSP decreased soil pH and DE-GRSP increased it, which was related to WSA stability. Exogenous EE-GRSP almost significantly increased soil acidic, neutral, and alkaline phosphatase activity at different strengths, while exogenous DE-GRSP, also at different strengths, significantly inhibited soil acidic phosphatase activity. The application of both exogenous EE-GRSP and DE-GRSP increased the organic carbon content of the soil. This study concluded that exogenous GRSP exerted differential effects on plant biomass and soil properties, and EE-GRSP can be considered as a soil stimulant for use in citrus plants. To our knowledge, this is the first report on the negative effects of exogenous DE-GRSP on plant biomass and soil properties.

Arbuscular mycorrhizal fungi (AMF) confer positive and negative effects on many plants, but it is unclear whether AMF has an effect on soil fertility, aggregate distribution, and stability. The aim of this study was to analyze the effects of Rhizoglomus intraradices on plant growth, root morphology, leaf chlorophyll and gas exchange, sugar concentrations, and soil nutrients, aggregate distribution, and stability in marigold (Tagetes erecta L.), maize (Zea mays L.), white clover (Trifolium repens L.), and vetch (Vicia villosa Roth.) plants. Twelve weeks after R. intraradices inoculation, maize presented the highest mycorrhizal development, while mycorrhizal dependence was shown to be the decreasing trend in marigold > white clover > vetch > maize. AMF inoculation significantly increased the chlorophyll index of marigold and white clover, the net photosynthetic rate of white clover, the stomatal conductance of maize and white clover, and the transpiration rate of maize. Fructose, glucose, and sucrose in the four plants were differentially affected by R. intraradices. R. intraradices significantly increased the soil organic carbon (SOC) of marigold, maize, and white clover, the Olsen-P of white clover, the available K content of marigold, the easily extractable glomalin-related soil protein (GRSP) of maize, and the difficultly extractable and total GRSP levels of marigold and vetch. In addition, R. intraradices significantly increased the stability of soil water-stable aggregates (WSAs) in all four plants, plus it increased WSA at 0.5–4 mm sizes. Root AMF colonization was significantly positively correlated with WSA stability, SOC, difficultly extractable GRSP, and total GRSP. It is concluded that AMF-triggered changes in plant growth, physiological activities, and soil fertility depended on plant species, but AMF-improved WSA distribution and stability were not dependent on plant species.

The role of arbuscular mycorrhizal fungi in sweet oranges is well known, but the function of their secondary metabolite, especially the easily extractable glomalin-related soil protein (EE-GRSP), an active fraction of glomalin, is still unclear. The proposed study aimed to analyze the field response of foliar application of exogenous EE-GRSP on tree mycorrhizal development and fruit quality of two sweet orange (Citrus sinensis L. Osbeck) varieties viz., Lane Late Navel (LLN) and Rohde Red Valencia (RRV). Application of EE-GRSP significantly increased the root mycorrhizal colonization and soil mycorrhizal hyphal length in both the sweet orange varieties. The external quality of fruits (fruit weight, polar diameter, and equatorial diameter) also improved in response to foliar application of EE-GRSP in both sweet orange varieties. However, EE-GRSP treatment showed no change in fruit soluble solid content, while it increased the Vc content, solids-acid ratio, fructose, glucose, and sucrose content of sarcocarp in the two sweet oranges varieties. The LLN variety treated with EE-GRSP recorded significantly higher N, P, K, Fe, and Si content of sarcocarp as a mark of nutritional quality, while the RRV variety treated with EE-GRSP displayed a higher concentration of nutrients like Cu, Fe, Si, and Zn in the sarcocarp as compared with the corresponding non-treated control. To the best of our knowledge, this is the first report regarding the improvement in fruit quality of late-ripening sweet oranges (especially LLN) in response to foliar application of EE-GRSP as another potential biostimulant.

Endophytes have the ability to improve plant nutrition alongside their agronomic performance, among which arbuscular mycorrhizal fungi provide the most benefits to their host. Previously, we reported for the first time that an arbuscular mycorrhizal-like fungus Piriformospora indica had the ability to colonize roots of trifoliate orange (Poncirus trifoliata) and conferred positive effects on nutrient acquisition. Present study showed the changes in fatty acids and sugars to unravel the physiological and symbiotic association of trifoliate orange with P. indica and an arbuscular mycorrhizal fungus, Funneliformis mosseae singly or in combination. All the endophytic fungi collectively increased fructose, glucose, and sucrose content in leaves and roots, along with a relatively higher increase with P. indica inoculation than with F. mosseae alone or dual inoculation. Treatment with P. indica increased the concentration of part unsaturated fatty acids such as C18:3N6, C20:2, C20:3N6, C20:4N6, C20:3N3, C20:5N3, C22:1N9, and C24:1. Additionally, P. indica induced the increase in the concentration of part saturated fatty acids such as C6:0, C8:0, C13:0, C14:0, and C24:0. F. mosseae hardly changed the content of fatty acids, except for increase in C14:0 and C20:5N3. Double inoculation only reduced the C21:0, C10:0, C12:0, C18:3N3, and C18:1 content and increased the C20:5N3 content. These endophytic fungi up-regulated the root PtFAD2, PtFAD6, PtΔ9, and PtΔ15 gene expression level, coupled with a higher expression of PtFAD2 and PtΔ9 by P. indica than by F. mosseae. It was concluded that P. indica exhibited a stronger response, for sugars and fatty acids, than F. mosseae on trifoliate orange. Such results also reveal the Pi (an in vitro culturable fungus) as a bio-stimulator applying to citriculture.

Many terrestrial plants form reciprocal symbioses with beneficial fungi in roots; however, it is not clear whether Vicia villosa, an important forage and green manure crop, can co-exist with these fungi and how such symbiosis affects plant growth and soil properties. The aim of this study is to analyze the effects of inoculation with three arbuscular mycorrhizal fungi (AMF) such as Diversisporaspurca, Funneliformismosseae, and Rhizophagusintraradices and an endophytic fungus Serendipitaindica on plant growth, root morphology, chlorophyll and sugar levels, soil nutrients, and aggregate size distribution and stability in V. villosa plants. After 63 days of inoculation, the beneficial fungi colonized the roots with colonization rates of 12% to 92%, and also improved plant growth performance and root morphology to varying degrees, accompanied by the most significant promoted effects after R.intraradices inoculation. All AMF significantly raised chlorophylls a and b, carotenoids and total chlorophyll concentrations, along with a significant increase in leaf sucrose, which consequently formed a significantly higher accumulation of glucose and fructose in roots providing carbon sources for the symbionts. Root fungal colonization was significantly (p < 0.01) positively correlated with chlorophyll compositions, leaf sucrose, and root glucose. In addition, inoculation with symbiotic fungi appeared to trigger a significant decrease in soil Olsen-P and available K and a significant increase in NH₄-N, NO₃-N, and glomalin-related soil protein levels, plus a significant increase in the proportion of water-stable aggregates at the size of 0.5–4 mm as well as aggregate stability. This improvement in soil aggregates was significantly (p < 0.01) positively correlated with root fungal colonization rate and glomalin-related soil protein concentrations. The study concludes that symbiotic fungi, especially R. intraradices, improve the growth of V. villosa, which is associated with fungal modulation of sugars, soil fertility and root structural improvement.