Background: Elaeocarpus spp. stem blight, caused by Pseudocryphonectria elaeocarpicola, is a destructive disease, which will significantly reduce the productivity and longevity of Elaeocarpus spp. plants, especially in the Guangdong Province of China. However, few information is available for P. elaeocarpicola. To unravel the potential adaptation mechanism of stem adaptation, the whole genome of P. elaeocarpicola was sequenced by using the DNBSEQ and PacBio platforms. Results: P. elaeocarpicola harbors 44.49 Mb genome with 10,894 predicted coding genes. Genome analysis revealed that the P. elaeocarpicola genome encodes a plethora of pathogenicity-related genes. Analysis of carbohydrate-active enzymes (CAZymes) revealed a rich variety of enzymes participated in plant cell wall degradation, which could effectively degrade cellulose, hemicellulose and xyloglucans in the plant cell wall and promote the invasion of the host plant. There are 213 CAZyme families found in P. elaeocarpicola, among which glycoside hydrolase (GH) family has the largest number, far exceeding other tested fungi by 53%. Besides, P. elaeocarpicola has twice as many genes encoding chitin and cellulose degradation as Cryphonectria parasitica, which belong to the same family. The predicted typical secreted proteins of P. elaeocarpicola are numerous and functional, including many known virulence effector factors, indicating that P. elaeocarpicola has great potential to secrete virulence effectors to promote pathogenicity on host plants. AntiSMASH revealed that the genome encoded 61 secondary metabolic gene clusters including 86 secondary metabolic core genes which was much higher than C. parasitica (49). Among them, two gene cluster of P. elaeocarpicola, cluster12 and cluster52 showed 100% similarity with the mycotoxins synthesis clusters from Aspergillus steynii and Alternaria alternata, respectively. In addition, we annotated cytochrome P450 related enzymes, transporters, and transcription factors in P. elaeocarpicola, which are important virulence determinants of pathogenic fungi. Conclusions: Taken together, our study represents the first genome assembly for P. elaeocarpicola and reveals the key virulence factors in the pathogenic process of P. elaeocarpicola, which will promote our understanding of its pathogenic mechanism. The acquired knowledge lays a foundation for further exploration of molecular interactions with the host and provide target for management strategies in future research.

Sirex noctilio and Amylostereum areolatum form a highly specific mutualistic symbiosis. The growth and host-degrading activities of the symbiotic fungus are critical to the woodwasps, which directly influence the larval survival rate and adult body size of the woodwasps. Gene expression analysis has been extensively employed to decrypt the intricate growth patterns of symbiotic fungi and identify the associated functional genes underpinning their degradation pathways. Appropriate reference genes are crucial for enhancing the accuracy of studies on gene expression. In an effort to refine gene expression analysis in A. areolatum, our study cultivated the symbiotic fungi on the wood powder medium of Pinus sylvestris var. mongolica, aligning closer to its natural growth conditions. Thirteen reference genes underwent meticulous evaluation via algorithms such as delta Ct, geNorm, BestKeeper, RefFinder, and NormFinder, depending on their stability amidst diverse growth and developmental epochs of A. areolatum. α-TUB, P450, and the combination (α-TUB + P450) were distinguished as the most stable candidates for RT-qPCR analysis, confirmed through AaLac1 expression validation. These findings contribute significantly to the investigation of gene expression in A. areolatum and facilitate a deeper understanding of its symbiotic relationship with S. noctilio.

The stem blight disease on Elaeocarpus spp. was newly identified in 2022 on E. rugosus and E. hain in Guangzhou, representing a serious stem disease that causes branch rot and cankers in Elaeocarpus species and can lead to whole-plant death in severe cases. Strain ZX-13 was isolated from Elaeocarpus rugosus stems and showed excellent antagonism against the Elaeocarpus stem blight pathogen Pseudocryphonectria elaeocarpicola. Based on morphological characteristics, physiological and biochemical results, and molecular biology analyses, the strain was identified as Erwinia phyllosphaerae. Whole-genome sequencing and annotation indicate that strain ZX-13 has a high-quality draft assembly of 4,686,433 bp, with a GC content of 53.85%, encoding 4189 genes, 86 tRNA, 22 rRNA, and 110 ncRNA. Eight antifungal enzymes from the GH family may be crucial factors to its antagonistic activity. Using antiSMASH 7.1.0, the ZX-13 genome predicted eight BGCs (RiPPs, NRPs, terpenes, etc.), with one showing 100% similarity to carotenoid biosynthesis. Novel candidates include an O-antigen, hassallidin C, and lankacidin C. The study identified Erwinia phyllosphaerae ZX-13 as an antagonistic bacterial strain for the first time, indicating substantial potential for biocontrol applications.