This study investigated the biodiversity, phylogeny, and antifungal activity of endophytic fungi isolated from Zanthoxylum bungeanum. A total of 940 isolates obtained were grouped into 93 morphotypes, 43 species, and 23 genera, which were authenticated by molecular identification based on rDNA internal transcribed spacer (ITS) sequence analysis. A high diversity of endophytic fungi from Z. bungeanum are observed with high species richness S (43), Margalef index D′ (6.1351), Shannon–Wiener index H′ (3.2743), Simpson diversity index D_s (0.9476), PIE index (0.9486), and evenness Pielou index J (0.8705) but a low dominant index λ (0.0524). Significant tissue specificity of the endophytic fungi was observed in Z. bungeanum, and the highest species richness and diversity indexes were obtained in the stem. Phylogenetic analyses of the 93 endophytic isolates were carried out by the neighbor-joining (NJ) method to demonstrate their evolutionary processes. Antifungal activities of endophytic fungi were assayed and eight endophytic isolates showed strong and long-lasting inhibition against host pathogenic fungi Fusarium sambucinum and Pseudocercospora zanthoxyli. Here, for the first time, we systematically demonstrate the biodiversity, phylogeny, and antifungal activity of endophytic fungi associated with Z. bungeanum and reveal the value of sampling different tissues of a given plant to obtain the greatest endophyte species diversity, which might offer a framework for further investigation and utilization of endophytic fungi as aunique source of interesting and useful bioactive compounds.

Lanzhou lily is a plant native to China with high edible, medicinal, and ornamental value that is relatively susceptible to Fusarium wilt. In this study, the application of Trichoderma longibrachiatum SMF2 (TlSMF2) effectively controlled Lanzhou lily wilt disease caused by Fusarium oxysporum and F. solani. TlSMF2 and the antimicrobial peptaibols trichokonins (TKs) produced by TlSMF2 inhibited the mycelial growth and spore germination of these two pathogens. Transcriptome analysis revealed that the TKs-induced defense responses of Lanzhou lily were mainly related to the production of plant hormones and defense enzymes. In detail, TKs treatment increased the levels of salicylic acid (SA) and jasmonic acid (JA) and the expression of their related genes and upregulated the activities of chitinase and phenylalanine ammonia-lyase (PAL). Moreover, TKs caused the induction of LzWRKY26 and LzWRKY75, which is highly homologous to LrWRKY3 that positively regulates Lilium regale resistance to F. oxysporum. LzWRKY26 expression was also induced by SA and MeJA treatments and F. oxysporum infection, which was consistent with the findings that many cis-acting elements associated with phytohormones and stress responses are present in the promoter region of LzWRKY26. Therefore, the biocontrol mechanisms of TlSMF2 against Lanzhou lily wilt disease involve substrate competition and toxicity against pathogens, as well as the induction of systemic resistance in plants. Our results highlight a promising biological control agent for soil-borne fungal diseases and offer deeper insights into the biocontrol mechanisms of TlSMF2.