Litchi (Litchi chinensis Sonn.) is a commercially important fruit but its production and quality are restricted by litchi downy blight, caused by the oomycete pathogen Peronophythora litchii Chen. Volatile substances produced by a biocontrol antinomycetes Streptomyces fimicarius BWL-H1 could inhibited P. litchii growth and development both in vitro and in detached litchi leaf and fruit infection assay. Transmission Electron Microscopy (TEM) and Scanning Electron Microscopy (SEM) analyses indicated that volatile organic compounds (VOCs) from BWL-H1 resulted in severe damage to the endomembrane system and cell wall of P. litchii cells in vitro and abnormal morphology of appressoria, as well as deformed new hyphae in infection process. VOCs could suppress mycelial growth, sporulation, while with no obvious effect on sporangia germination. Based on gas chromatography-mass spectrophotometric analyses, 32 VOCs were identified from S. fimicarius BWL-H1, the most abundant of which was phenylethyl alcohol. Eight VOCs, including phenylethyl alcohol, ethyl phenylacetate, methyl anthranilate, α-copaene, caryophyllene, humulene, methyl salicylate and 4-ethylphenol, that are commercially available, were purchased and their bioactivity was tested individually. Except for humulene, the other seven tested volatile compounds shown strong inhibitory activity against mycelial growth, sporulation, sporangia germination and germ-tube growth of P. litchii. Especially, 4-ethylphenol showed the highest inhibitory effect on sporulation at a very low concentration of 2 µL/L. Overall, our results provided a better understanding of the mode of action of volatiles from BWL-H1 on P. litchii, and showed that volatiles from BWL-H1 have the potential for control of postharvest litchi downy blight.

MicroRNAs (miRNAs) are small non-coding RNAs that regulate protein-coding gene expression primarily found in plants and animals. Fungi produce microRNA-like RNAs (milRNAs) that are structurally similar to miRNAs and functionally important in various biological processes. The fungus Fusarium oxysporum f. sp. cubense (Foc) is the causal agent of Banana Fusarium vascular wilt that threatens global banana production. It remains uncharacterized about the biosynthesis and functions of milRNAs in Foc. In this study, we investigated the biological function of milRNAs contributing to Foc pathogenesis. Within 24 hours post infecting the host, the Argonaute coding gene FoQDE2, and two Dicer coding genes FoDCL1 and FoDCL2, all of which are involved in milRNA biosynthesis, were significantly induced. FoQDE2 deletion mutant exhibited decreased virulence, suggesting the involvement of milRNA biosynthesis in the Foc pathogenesis. By small RNA sequencing, we identified 364 small RNA-producing loci in the Foc genome, 25 of which were significantly down-regulated in the FoQDE2 deletion mutant, from which milR-87 was verified as a FoQDE2-depedent milRNA based on qRT-PCR and Northern blot analysis. Compared to the wild-type, the deletion mutant of milR-87 was significantly reduced in virulence, while overexpression of milR-87 enhanced disease severity, confirming that milR-87 is crucial for Foc virulence in the infection process. We furthermore identified FOIG_15013 (a glycosyl hydrolase-coding gene) as the direct target of milR-87 based on the expression of FOIG_15013-GFP fusion protein. The FOIG_15013 deletion mutant displayed similar phenotypes as the overexpression of milR-87, with a dramatic increase in the growth, conidiation and virulence. Transient expression of FOIG_15013 in Nicotiana benthamiana leaves activates the host defense responses. Collectively, this study documents the involvement of milRNAs in the manifestation of the devastating fungal disease in banana, and demonstrates the importance of milRNAs in the pathogenesis and other biological processes. Further analyses of the biosynthesis and expression regulation of fungal milRNAs may offer a novel strategy to combat devastating fungal diseases.

Fusarium oxysporum f. sp. cubense, particularly the tropical race 4 (TR4), is the most destructive fungal pathogen of banana and can cause significant economic losses. However, the molecular mechanisms underlying the pathogenicity and virulence of the fungus are poorly understood. In this study, we characterized the mutant W2987, which was derived through T-DNA insertional mutagenesis of a wild-type TR4 isolate. W2987 showed slower growth, produced fewer conidia and was less virulent than the wild-type strain. Thermal asymmetric interlaced-PCR (TAIL-PCR) analysis showed that the T-DNA was inserted in the 3′ untranslated region (UTR) of an open reading frame (ORF) in W2987. BLAST search results indicated that the ORF encodes a hypothetical protein with unknown function. Phylogenetic analysis of the gene sequences from various Fusarium isolates showed that the F. oxysporum f. sp. cubense TR4 isolates were distinct from other F. oxysporum isolates. One pair of primers based on the 3′ UTR sequence of the putative virulence gene amplified a specific PCR product from the F. oxysporum f. sp. cubense TR4 isolates but not from other races and Fusarium species. This result indicates that this genetic locus possibly associated with virulence is unique and reliable for diagnosis and detection of F. oxysporum f. sp. cubense TR4.

This paper proposes PestNet, a lightweight method for classifying crop pests, which improves upon MobileNet-V2 to address the high model complexity and low classification accuracy commonly found in pest classification research. Firstly, the training phase employs the AdamW optimizer and mixup data augmentation techniques to enhance the model’s convergence and generalization capabilities. Secondly, the Adaptive Spatial Group-Wise Enhanced (ASGE) attention mechanism is introduced and integrated into the inverted residual blocks of the MobileNet-V2 model, boosting the model’s ability to extract both local and global pest information. Additionally, a dual-branch feature fusion module is developed using convolutional kernels of varying sizes to enhance classification performance for pests of different scales under real-world conditions. Lastly, the model’s activation function and overall architecture are optimized to reduce complexity. Experimental results on a proprietary pest dataset show that PestNet achieves classification accuracy and an F1 score of 87.62% and 86.90%, respectively, marking improvements of 4.20 percentage points and 5.86 percentage points over the baseline model. Moreover, PestNet’s parameter count and floating-point operations are reduced by 14.10% and 37.50%, respectively, compared to the baseline model. When compared with ResNet-50, MobileNet V3-Large, and EfficientNet-B1, PestNet offers superior parameter efficiency and floating-point operation requirements, as well as improved pest classification accuracy.

During initial stages of microbial invasion, the extracellular space (apoplast) of plant cells is a vital battleground between plants and pathogens. The oomycete plant pathogens secrete an array of apoplastic carbohydrate active enzymes, which are central molecules for understanding the complex plant-oomycete interactions. Among them, pectin acetylesterase (PAE) plays a critical role in the pathogenesis of plant pathogens including bacteria, fungi and oomycetes. Here, we demonstrated that Peronophythora litchii (syn. Phytophthora litchii) PlPAE5 suppresses litchi (Litchi chinensis) plant immunity by interacting with litchi lipid transfer protein 1 (LcLTP1). The LcLTP1-binding activity and virulence function of PlPAE5 depend on its PAE domain but not on its pectin acetylesterase activity. The high expression of LcLTP1 enhances plant resistance to oomycete and fungal pathogens, and this disease resistance depends on BRASSINOSTEROID INSENSITIVE 1-associated receptor kinase 1 (BAK1) and Suppressor of BIR1 (SOBIR1) in Nicotiana benthamiana. LcLTP1 activates the plant salicylic acid (SA) signaling pathway, while PlPAE5 subverts the LcLTP1-mediated SA signaling pathway by destabilizing LcLTP1. Conclusively, this study reports a virulence mechanism of oomycete pectin acetylesterase suppressing plant LTP-mediated SA immune signaling and will be instrumental for boosting plant resistance breeding.