Trichoderma hamatum is a filamentous fungus that serves as a biological control agent for multiple phytopathogens and as an important resource promising for fungicides. However, the lack of adequate knockout technologies has hindered gene function and biocontrol mechanism research of this species. This study obtained a genome assembly of T. hamatum T21, with a 41.4 Mb genome sequence comprising 8170 genes. Based on genomic information, we established a CRISPR/Cas9 system with dual sgRNAs targets and dual screening markers. CRISPR/Cas9 plasmid and donor DNA recombinant plasmid were constructed for disruption of the Thpyr4 and Thpks1 genes. The result indicates the consistency between phenotypic characterization and molecular identification of the knockout strains. The knockout efficiencies of Thpyr4 and Thpks1 were 100% and 89.1%, respectively. Moreover, sequencing revealed fragment deletions between dual sgRNA target sites or GFP gene insertions presented in knockout strains. The situations were caused by different DNA repair mechanisms, nonhomologous end joining (NHEJ), and homologous recombination (HR). Overall, we have successfully constructed an efficient and convenient CRISPR/Cas9 system in T. hamatum for the first time, which has important scientific significance and application value for studies on functional genomics of Trichoderma and other filamentous fungi.

Meloidogyne enterolobii, a highly virulent and broad-host-range plant-parasitic nematode, poses an increasing threat to global agricultural production. By inducing the formation of nutrient-rich giant cells in host roots and deploying a diverse array of effector proteins to modulate plant immune responses, this nematode achieves efficient colonization and invasion, resulting in impaired crop growth and significant economic losses. In recent years, global climate warming combined with the rapid development of protected agriculture has broken the traditional geographical limits of tropical and subtropical regions, thereby increasing the risk of M. enterolobii occurrence in temperate and high-latitude areas. Concurrently, conventional chemical control methods are increasingly limited by environmental pollution and the development of resistance, steering research toward green control strategies. This review systematically summarizes the latest research progress of M. enterolobii in terms of ecological diffusion trends, pathogenic mechanisms, and green control, and explored the feasibility of integrating multidisciplinary technologies to construct an efficient and precise control system. The ultimate aim is to provide theoretical support and technical supports for green and sustainable development of global agriculture.

Anthracnose is recognized as a significant agricultural disease. This study investigates the disease symptoms characterized by black dots on walnut fruits observed in the walnut orchards of Longnan City, Gansu Province, China, in June 2022. These symptoms resemble those of anthracnose reported in previous studies. A strain designated Ht-10 was initially isolated and identified as belonging to the Colletotrichum species based on its morphological features. Pathogenicity tests confirmed that this strain induced pronounced anthracnose symptoms in walnuts, consistent with those originally observed in the field. Subsequently, multilocus phylogenetic analysis, which included partial sequences of the internal transcribed spacer (ITS), actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), β-tubulin2 (TUB2), and chitin synthase (CHS-1) genes of Ht-10, indicated that it most likely clustered with Colletotrichum fioriniae. The determination of biological characteristics revealed that the optimal temperature for the growth of Ht-10 was 25 °C in full light at a pH of 6, with soluble starch and yeast paste serving as the optimal carbon and nitrogen sources, respectively. To our knowledge, this is the first report of C. fioriniae as a causal agent of anthracnose in walnut fruits in China.

Cyst nematodes are obligate parasitic nematodes found in the fields of many cultivated crops. These nematodes, which have great economic importance, pose a threat to food security, though they are frequently ignored or misdiagnosed as pests because of covert parasitism. A cyst nematode population parasitizing on Artemisia lavandulaefolia DC., one of the traditional Chinese medicines was collected in Gansu Province. The species was diagnosed using integrative taxonomy and molecular approaches. The cyst population is spherical or lemon-shaped, light brown or dark brown in color, with a long neck and a protruding vulval cone. The stylet of the second-stage juvenile is strong, and the front end of the ball at the base of the stylet is concave; the median bulb and excretory pore are prominent; the tail is blunt and circular, and the transparent tail is usually shorter than the stylet. A phylogenetic analysis was carried out using the internal transcribed spacer (ITS) and 28S genes of ribosomal DNA, which further confirmed the presence of Cactodera chenopodiae. According to our literature review, this is the first report on C. chenopodiae in Compositae. By following this research, we can better understand the challenges posed by A. lavandulaefolia DC. and develop effective strategies for managing its spread and impacts. This will help to protect vulnerable ecosystems and ensure the sustainability of agricultural and forestry activities in affected areas.

Trichoderma longibrachiatum is a filamentous fungus that functions as a biocontrol agent against multiple plant diseases. However, the lack of a genetic transformation system has hindered studies on gene function and the underlying biocontrol mechanisms in this species. In this study, protoplast preparation conditions were optimized using single-factor experiments and response surface methodology, yielding 2.05 × 10⁷ protoplasts/g. PEG-mediated transformation yielded a GFP-labeled strain (GFP-40418). This strain enhanced resistance against Meloidogyne incognita, with root galling decreased by 47.6%. Treatment with GFP-40418 enhanced the activities of antioxidant enzymes, including PAL, POX, SOD, PPO, APX, and CAT. Plants treated with GFP-40418 alone showed the highest expression value, with relative transcriptional levels of PR2, Pal1, LOX, MYC2, ETR1, and ACO1 that were 29.62, 5.76, 7.30, 3.64, 4.79, 9.99-fold higher in comparison with control plants, respectively. These findings provide a genetic platform for exploring the gene functions of T. longibrachiatum 40418 and validate its biocontrol prospect against root-knot nematodes.