Aug 2026· Journal of Integrative Plant Biology· 0 citations· 64 references
Medicine
TL;DR
This integrated approach combines the advantages of microbial biocontrol with RNAi precision, establishing Y. lipolytica as a versatile chassis for sustainable crop protection and offers a scalable, eco-friendly alternative to chemical fungicides for gray mold management.
Abstract
Plant gray mold disease, caused by the necrotrophic fungus Botrytis cinerea, threatens global food security by infecting over 1,400 species. The resistance of B. cinerea to fungicides poses a major challenge for its control. RNA interference (RNAi) shows promise for pathogen management, but high production costs, poor stability, and inefficient delivery of double-stranded RNA (dsRNA) restrict its practical use. In this study, we developed Yarrowia lipolytica strain MP181-2 as a novel RNAi platform, leveraging its ability to colonize the phyllosphere of crop plants (tomato and alfalfa) to enable simultaneous dsRNA production and pathogen targeting via microbe-induced gene silencing (MIGS). Through target screening, we identified three essential fungal genes, BcRpd3, BcNat1, and BcArd1, whose silencing significantly reduced the pathogenicity of B. cinerea. We further constructed a chimeric dsRNA molecule (BcANR-dsRNA) targeting these three genes, which demonstrated superior disease suppression compared to single-gene targeting. The engineered Y. lipolytica strains that produce dsRNA could effectively interfere with the expression of target genes in B. cinerea, and the strain expressing BcANR-dsRNA (designated Yl-dsANR) acquired the ability to suppress the growth of B. cinerea. In planta tests confirmed that Yl-dsANR successfully interfered with the expression of three target genes, leading to decreased fungal biomass and reduced lesion development on different hosts. Notably, foliar application of Yl-dsANR provided durable protection (over 70% efficacy for 5 d) in potted plants. Our integrated approach combines the advantages of microbial biocontrol with RNAi precision, establishing Y. lipolytica as a versatile chassis for sustainable crop protection. This study also provides an effective solution to current limitations in RNA pesticide development and offers a scalable, eco-friendly alternative to chemical fungicides for gray mold management.
Gray mold caused by Botrytis cinerea poses severe threats to global horticultural production. Conventional chemical fungicides face mounting restrictions due to rising pathogen resistance and stringent food safety regulations, creating an urgent need for sustainable alternatives. RNA pesticides offer an eco-friendly alternative to chemical fungicides. However, the application of RNA pesticides faces challenges including double-stranded RNA (dsRNA) instability, delivery inefficiency, and high production costs. Here, we utilized an endophyte to produce dsRNA, enabling effective control of gray mold. The Beauveria bassiana strain QSE-F1 was proved as tomato endophyte and engineered to express dsRNA targeting BcMucin, an essential gene in B. cinerea and was validated as an effective RNA interference (RNAi) target. The engineered B. bassiana (Bb-dsMucin) produced and delivered dsRNA to achieve significant silencing of BcMucin at transcriptional and translational levels. The Bb-dsMucin strain showed superior antifungal activity and gray mold control efficacy compared with control strains. The combined use of conidia and fermentative broth of Bb-dsMucin attained optimal and lasting control efficacy. Our research establishes phyllosphere-derived Microbe-induced gene silencing (MIGS) as a promising biocontrol strategy for managing gray mold, and promotes the advancement of RNA pesticides technologies.
Mengjie Liu, Fan Yang, Huiwen Zheng et al.· Horticulture Research· 0 citations
Sclerotinia sclerotiorum, the causal agent of white mold, is a globally distributed fungal pathogen responsible for major yield losses in many crops. Spray-induced gene silencing (SIGS) has emerged as a promising alternative for disease control. Building on prior research, this study advanced SIGS by targeting S. sclerotiorum argonaute 2 (SsAgo2), using FF4 (PAZ domain to 5'-PIWI domain) and FF5 (PIWI central domain) fragments which are important regions involved in RNA binding and cleavage. We optimized spray tank mix/dilution in the greenhouse and scaled up dsRNA production using E. coli. The greenhouse results demonstrated that dsRNA with nanosheets significantly inhibited white mold disease on tomato and N. benthamiana compared to empty vector control (p = 0.03 and 0.004, respectively). To evaluate off-target effects, SIGS was tested on Botrytis cinerea, a related necrotrophic pathogen. No significant differences in lesion size were observed between SsAgo2 dsRNA-treated and water-treated Nicotiana benthamiana leaves. Gene ontology analysis of differential expressions in sunflower leaves revealed no significant differences between SsAgo2-FF4 or SsAgo2-FF5 compared to control. Field trials in 2024 demonstrated effectiveness. Parameters assessed included disease severity, wet weight, head width, and seed weight. Area under the disease progress curve (AUDPC) analysis revealed that dsRNA sprays, applied before and after manual inoculation or during natural infection, significantly reduced infection levels with p < 0.1 as the cut off. Sunflower fresh weight increased by 104.4 g (p = 0.04) and 96.7 g (p = 0.07), while head width increased by 2.5 cm (p = 0.03) and 1.5 cm (p = 0.10), respectively. This report demonstrates that the dsRNA mixture has potential to incorporate SIGS into field applications for the management of white mold disease.
Y.-W. Tseng, C. Pedersen, Zhi-Yuan Chen et al.· Plant Disease· 0 citations
It is demonstrated that endochitinase overexpression activates defense signaling and improves broad‐spectrum resistance, representing a promising strategy for developing disease‐resistant tomato cultivars.
Lucas José de Sousa, Ana Carolina Mendes Bezerra, Ivonaldo Reis Santos et al.· Biotechnology Journal· 0 citations
BgtH12 is established as a critical virulence factor that suppresses basal immunity to promote Bgt colonization and stable overexpression of BgtH12 in wheat enhanced susceptibility.
It is indicated that banana DMR6 functions as a negative regulator of plant immunity and is closely associated with susceptibility to Fusarium wilt, providing a molecular basis for future functional validation and support DMR6 as a potential target for precise genome editing to develop resistant banana cultivars.
Márcio Leandro da Silveira Fonseca, L. C. de Souza, Fernanda dos SantosNascimento et al.· Molecular Biology Reports· 0 citations
Chinese chive (Allium tuberosum) suffers severe yield and quality losses from white leaf spot disease caused by Alternaria alternata. Spray-induced gene silencing (SIGS) presents a sustainable alternative to traditional chemical fungicides. To maximize the biocontrol efficacy of this approach, we designed dsRNAs targeting two candidate virulence-associated genes of Alternaria alternata: AaGH10, encoding a cell wall-degrading enzyme critical for host penetration, and AaSOD, an antioxidant enzyme crucial for reactive oxygen species (ROS) scavenging. We comprehensively evaluated the antifungal efficacy by assessing mycelial growth inhibition, spore germination, and lesion development through in vitro and in vivo assays. The results demonstrated that both single and combinatorial treatments effectively inhibited fungal growth and spore germination, thereby reducing disease incidence on detached leaves and intact greenhouse plants. The application of the dual-target dsRNA formulation achieved an 86.7% control efficacy on detached leaves. Furthermore, it significantly alleviated in vivo disease severity, decreasing the average number of necrotic lesions from 15.6 to 1.3 per leaf. These results demonstrate that the dual-target combination exerts an enhanced protective effect compared with individual interventions. This specific dual-target dsRNA formulation establishes a robust foundation for the control of Chinese chive white leaf spot disease.
Lian-Zhe Wang, Ke-Hao Huang, Yi-Xian Gou et al.· Journal of Fungi· 0 citations
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