In the present study, three mitogenomes from the Bipolaris genus (Bipolaris maydis, B. zeicola, and B. oryzae) were assembled and compared with the other two reported Bipolaris mitogenomes (B. oryzae and B. sorokiniana). The five mitogenomes were all circular DNA molecules, with lengths ranging from 106,403 bp to 135,790 bp. The mitogenomes of the five Bipolaris species mainly comprised the same set of 13 core protein-coding genes (PCGs), two rRNAs, and a certain number of tRNAs and unidentified open reading frames (ORFs). The PCG length, AT skew and GC skew showed large variability among the 13 PCGs in the five mitogenomes. Across the 13 core PCGs tested, nad6 had the least genetic distance among the 16 Pleosporales species we investigated, indicating that this gene was highly conserved. In addition, the Ka/Ks values for all 12 core PCGs (excluding rps3) were < 1, suggesting that these genes were subject to purifying selection. Comparative mitogenomic analyses indicate that introns were the main factor contributing to the size variation of Bipolaris mitogenomes. The introns of the cox1 gene experienced frequent gain/loss events in Pleosporales species. The gene arrangement and collinearity in the mitogenomes of the five Bipolaris species were almost highly conserved within the genus. Phylogenetic analysis based on combined mitochondrial gene datasets showed that the five Bipolaris species formed well-supported topologies. This study is the first report on the mitogenomes of B. maydis and B. zeicola, as well as the first comparison of mitogenomes among Bipolaris species. The findings of this study will further advance investigations into the population genetics, evolution, and genomics of Bipolaris species.

In this study, the complete mitogenomes of three Diaporthe species (Diaporthe eres ZM79-3, D. phaseolorum ZM33-4 and Diaporthe sp. ZM41-5) were sequenced, assembled and compared with the other three previously sequenced Diaporthe mitogenomes (D. caulivora VNIIKR SE Dcaul3, D. longicolla MSPL 10-6 and D. sojae VNIIKR SE Dps12). The six Diaporthe mitogenomes were found to be circular DNA molecules, with lengths ranging from 53,646 bp to 108,865 bp. The mitogenomes of the six Diaporthe species mainly comprised the same set of 15 core protein-coding genes (PCGs), two rRNAs, and a certain number of tRNAs and unidentified open reading frames (ORFs). The PCG length, AT skew and GC skew showed large variability among the 15 PCGs in the six mitogenomes. The nad1 gene had the least K2P genetic distance of the 15 core PCGs among the 13 Diaporthales species, indicating that this gene was highly conserved. The Ka/Ks values for all 15 core PCGs were < 1, suggesting that these genes were all subject to purifying selection. Comparative mitogenome analysis showed that introns contributed the most to the size variation of Diaporthe mitogenomes. Frequent intron loss/gain events were detected to have occurred in the cox1 gene during the evolution of the Diaporthales mitogenomes. Although the mitogenomes of 13 species from Diaporthales had undergone large-scale gene rearrangements, six mitogenomes of Diaporthe species had identical gene arrangements. Phylogenetic analysis based on combined mitochondrial gene datasets showed that the six Diaporthe species formed well-supported topologies. To our knowledge, this study is the first report on the mitogenomes of D. phaseolorum ZM33-4 and Diaporthe sp. ZM41-5, as well as the first comparison of mitogenomes among Diaporthe species. Our findings will further promote investigations of the genetics, evolution and phylogeny of the Diaporthe species.

Apple Valsa canker, caused by Valsa mali and Valsa pyri, is a devastating disease of apple trees and poses a severe threat to the sustainable development of apple production. Although the two species’ whole genomes have been sequenced, their mitochondrial genomes are still uncharacterized. In this study, the complete mitochondrial genomes of V. mali and V. pyri were assembled, annotated, and compared by bioinformatic methods. The results indicate that the mitogenomes are both circular DNA molecules with sizes of 213,406 bp and 128,022 bp, respectively. The AT skew values of the two Valsa species’ mitogenomes were positive, while the GC skew values were negative. Comparative mitogenome analysis revealed that the length and base composition of protein-coding genes (PCGs), rRNA genes, and tRNA genes differed between the two Valsa species. It was found that the expansion of V. mali was primarily attributable to the intronic regions. There are large numbers of interspersed repetitive sequences (IRS) in both Valsa mitogenomes; however, the proportion of IRS in V. mali (43.56%) was much higher than that in V. pyri (2.41%). The alignment of large fragments between the mitochondrial and nuclear genomes of both V. mali (1.73 kb) and V. pyri (5.17 kb) indicates that gene transfer between mitochondrial and nuclear genomes occurred during evolution. The ka/ks ratios for 15 core PCGs were below one, suggesting that these genes were subjected to purifying selection pressure. Comparative mitogenomics revealed that the two fungi had significant mitogenomic collinearity and large-scale gene rearrangements. The results of phylogenetic analysis based on Bayesian inference (BI) and maximum likelihood (ML) using a combined mitochondrial gene set confirmed that V. mali and V. pyri were fully independent taxa with a high bootstrap value of 100 (ML) and a high posterior probability of 1.0 (BI). This is the first report on the mitogenomes within the genus Valsa. These results will pave the way to understanding the evolution and differentiation of mitogenomes in the genus Valsa.

Background/Objectives: Wheat root and crown rot, caused by Fusarium pseudograminearum, Fusarium graminearum, and Bipolaris sorokiniana, are devastating soil-borne diseases that cause substantial yield losses worldwide. Endophytic fungi are promising sources of bioactive metabolites for agricultural applications. This study aimed to isolate and characterize an endophytic fungus with antifungal activity against major wheat pathogens, identify its active compound, and investigate the underlying transcriptional response. Methods: An endophytic strain Y2 was isolated from Hedyotis diffusa leaves and identified through morphological and phylogenetic analysis based on TEF-1α and RPB2 sequences. Pathogenicity of strain Y2 was evaluated on wheat stem bases. The bioactive compound was purified by HPLC and identified by HR-ESI-MS and NMR. Antifungal activity was assessed using dual-culture and microbroth dilution assays. Transcriptomic analysis (RNA-seq) was performed on F. pseudograminearum treated with equisetin, with qRT-PCR validation of seven representative differentially expressed genes. Results: Strain Y2 was identified as Fusarium incarnatum or a closely related member of the F. incarnatum–equiseti species complex (FIESC) and confirmed to be non-pathogenic to wheat. The purified bioactive compound was characterized as equisetin, which exhibited significant antifungal activity with MIC values of 16, 32, and 64 μg/mL against F. pseudograminearum, B. sorokiniana, and F. graminearum, respectively. Transcriptomic analysis revealed that equisetin treatment induced a polarized transcriptional response in F. pseudograminearum, characterized by strong upregulation of ribosome and translation-related genes and widespread downregulation of other metabolic pathways, particularly nitrogen metabolism. qRT-PCR validation of seven representative genes confirmed the reliability of the RNA-seq data. Conclusions: Our findings demonstrate that equisetin is the active antifungal metabolite produced by F. incarnatum Y2, with potent in vitro activity against major wheat root and crown rot pathogens. The transcriptomic data provide insights into the potential mechanism of action, while the non-pathogenic nature of strain Y2 supports its biosafety. Although these results highlight equisetin as a promising lead compound for antifungal development, further in planta efficacy and safety studies are required before it can be considered for practical biocontrol.