A study was conducted for endophytic antagonistic fungi obtained from the roots of healthy apple trees growing in nine replanted orchards in Shandong Province, China. The fungi were assessed for their ability to inhibit Fusarium proliferatum f. sp. malus domestica MR5, a fungal strain associated with apple replant disease (ARD). An effective endophyte, designated as strain 6S-2, was isolated and identified as Trichoderma asperellum. Strain 6S-2 demonstrated protease, amylase, cellulase, and laccase activities, which are important for the parasitic and antagonistic functions of pathogenic fungi. The inhibition rate of 6S-2 against Fusarium proliferatum f. sp. malus domestica MR5 was 52.41%. Strain 6S-2 also secreted iron carriers, auxin, ammonia and was able to solubilize phosphorus. Its fermentation extract and volatile substances inhibited the growth of MR5, causing its hyphae to twist, shrink, swell, and rupture. The antifungal activity of the 6S-2 fermentation extract increased with increasing concentrations. It promoted the production and elongation of Arabidopsis thaliana lateral roots, and the strongest effects were seen at a concentration of 50 mg/mL. A GC-MS analysis of the 6S-2 fermentation extract and volatile substances showed that they comprised mainly alkanes, alcohols, and furanones, as well as the specific volatile substance 6-PP. The application of 6S-2 spore suspension to replanted apple orchard soils reduced plant oxidative damage and promoted plant growth in a pot experiment. Therefore, the endophytic strain T. asperellum 6S-2 has the potential to serve as an effective biocontrol fungus for the prevention of ARD in China, and appears to promote plant growth.

Trichoderma asperellum strain 6S-2 with biocontrol effects and potential growth-promoting properties was made into a fungal fertilizer for the prevention of apple replant disease (ARD). 6S-2 fertilizer not only promoted the growth of Malus hupehensis Rehd seedlings in greenhouse and pot experiments, but also increased the branch elongation growth of young apple trees. The soil microbial community structure changed significantly after the application of 6S-2 fertilizer: the relative abundance of Trichoderma increased significantly, the relative abundance of Fusarium (especially the gene copy numbers of four Fusarium species) and Cryptococcus decreased, and the relative abundance of Bacillus and Streptomyces increased. The bacteria/fungi and soil enzyme activities increased significantly after the application of 6S-2 fertilizer. The relative contents of alkenes, ethyl ethers, and citrullines increased in root exudates of M. hupehensis Rehd treated with 6S-2 fertilizer and were positively correlated with the abundance of Trichoderma. The relative contents of aldehydes, nitriles, and naphthalenes decreased, and they were positively correlated with the relative abundance of Fusarium. In addition, levels of ammonium nitrogen (NH₄-N), nitrate nitrogen (NO₃-N), available phosphorus (AP), available potassium (AK), organic matter (SOM), and pH in rhizosphere soil were also significantly related to changes in the microbial community structure. In summary, the application of 6S-2 fertilizer was effective in alleviating some aspects of ARD by promoting plant growth and optimizing the soil microbial community structure.

(1) Background: The cultivation of resistant rootstocks is an effective way to prevent ARD. (2) Methods: 12-2 (self-named), T337, and M26 were planted in replanted and sterilized soil. The aboveground physiological indices were determined. (3) Results: The plant heights and the stem thicknesses of T337 and M26 were significantly affected by ARD. Relative chlorophyll content (June–October), P_n (August–September), and G_s (August) of T337 and relative chlorophyll content (June–July, September), P_n (September–October), and C_i (September) of M26 were significantly affected by ARD. ARD had a significant effect on F_v/F_m (June), qP (June–July), and NPQ of T337 (June–October, except August) and F_v/F_m (June) and NPQ (June-October, except July) of M26. Additionally, ARD affected Rfd of M26 and T337 during August. SOD (August and October), POD (August–September), and CAT (July-August, October) activities and MDA (September–October) content of T338 as well as SOD (July–October), POD (June–October), and CAT (July-October) activities and MDA (July, September–October) content of M26 were significantly affected by ARD. ARD significantly reduced nitrogen (October), phosphorus (September–October), and zinc (July) contents of M26 and potassium (June) content of T337. The above physiological indices were not affected by ARD in 12-2. (4) Conclusions: 12-2 could be useful as an important rootstock to relieve ARD due to strong resistance.

Apple replant disease (ARD) is a common soil disease that occurs in apple-growing areas around the world, causing root tip rot and necrosis, plant growth retardation and even plant death. Biofumigation is a promising strategy for controlling ARD due to its advantages of convenient application and being environmentally friendly. Tagetes erecta is an effective biological fumigant, but its effect on ARD is unclear. In the present study, we used Malus hupehensis Rehd. seedlings as the test material to detect the mitigating effects of different forms of T. erecta: air-dried sample (DS), fresh samples (FS) and fresh sample infusion solution (IS) on ARD. The effects of different forms of T. erecta on the growth of apple seedlings, leaf photosynthesis, root antioxidant enzyme, soil enzymatic activity and microbial environment were investigated. Compared with the CK treatments, DS, FS, and IS treatments all significantly increased the biomass of apple seedlings and promoted root growth under replanting conditions. Among them, DS showed the best results. The activity of antioxidant enzyme including superoxide dismutase, peroxidase and catalase were significantly increased in roots grown in soils treated with T. erecta. Moreover, T. erecta treatment also increased the activity of soil urease, phosphatase, sucrase and catalase enzyme, significantly altered the abundance of soil fungal communities and, in particular, reduced the abundance of Fusarium oxysporum, the main causal fungus of ARD. Therefore, our results suggest that biofumigation of different forms of T. erecta enhanced the resistance of ARD by regulating root reactive oxygen levels and improving the soil fungal communities.