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.

Tinospora sagittata is a columbin-rich medicinal plant, but its columbin levels are reduced under artificial cultivation conditions. The objective of this study was to analyze the effects of inoculations with Diversispora versiformis (Dv), Funneliformis mosseae (Fm), Rhizophagus intraradices (Ri), and mixed inoculation (Dv + Fm + Ri) (Mix) on growth performance, root morphology, leaf photosynthetic physiology, and root columbin levels in T. sagittata. These arbuscular mycorrhizal fungi (AMF) were able to colonize the roots, as evidenced by a root mycorrhizal colonization rate ranging from 17% to 48% and soil hyphal lengths ranging from 17.51 cm/g to 32.02 cm/g, with the Mix treatment being the greatest. AMF inoculations improved plant height (16–151%), leaf number (119–283%), shoot (37–211%), and root biomass (22–318%) to varying extents, with Ri and Mix treatments being the most prominent. AMF-treated plants presented relatively greater root total length, projected area, surface area, volume, and average diameter, especially those treated with Ri and Mix. AMF inoculations also significantly improved the leaf nitrogen balance index, transpiration rate, and stomatal conductance, while the photosynthesis rate and chlorophyll index varied by AMF species, along with a decrease in intercellular CO₂ levels. Root columbin levels ranged from 0.524 mg/g to 5.389 mg/g, and AMF inoculation significantly increased root columbin levels by 228–928%, with Ri being the most significant. Root columbin levels were significantly positively correlated with soil hyphal length, root total length, root projected area, root surface area, root volume, and root average diameter, but not root AMF colonization rate. This study demonstrates for the first time that AMF, especially Ri, can be employed as a biostimulant to promote growth as well as root columbin levels in T. sagittata, where AMF-triggered improvement in root morphology is an important reason for promoting root columbin levels.

Salt stress poses a substantial challenge to melon cultivation, but grafting techniques have shown promise in enhancing salt tolerance. This study aims to identify key genes involved in salt tolerance within melon rootstocks. The salt tolerance of four melon cultivars was evaluated, revealing that ‘ST2’ exhibited salt sensitivity, whereas ‘XZM17’ demonstrated salt tolerance. Grafting experiments indicated that salt-sensitive melons benefit significantly from being grafted onto salt-tolerant rootstocks. Transcriptome analysis further identified the CmDUF239-1 gene as a critical factor contributing to improved salt tolerance in grafted melons. Functional studies demonstrated that knocking out CmDUF239-1 reduces salt tolerance, reflected in decreased activities of antioxidant enzymes (SOD, POD, CAT) and diminished expression levels of related genes (CmSOD1, CmPRX53-1, CmPRX53-2, CmCAT2). Conversely, overexpression of CmDUF239-1 leads to enhanced enzyme activity and gene expression, along with improved Na+/K+ homeostasis, evidenced by decreased Na+ accumulation and increased K+ absorption. Furthermore, CmDUF239-1 overexpression upregulated Na+/K+ transport-related genes (CmSOS1, CmNHX6, CmKUP3, CmSKOR), whereas CmDUF239-1 knockout had the opposite effect. These findings indicate that CmDUF239-1 plays a dual role in promoting salt tolerance by regulating antioxidant defenses and ion transport, contributing to our understanding of the molecular mechanisms behind grafting-induced salt tolerance and providing insights for the breeding of resilient melon varieties.