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Pulcherrimin and Beyond: The Multifaceted Role of Metschnikowia pulcherrima in Postharvest Disease Management-A Scoping Review. Pulcherrimin及其他:pulcherrima Metschnikowia pulcherrima在采后病害管理中的多面作用
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2026-04-21 DOI: 10.3390/jof12040298
Juliana Pereira Rodrigues Belas, Caroline Corrêa de Souza Coelho, Leda Maria Fortes Gottschalk, Elisa d'Avila Costa Cavalcanti, Denise Maria Guimarães Freire, Otniel Freitas Silva

Postharvest losses of fruits and vegetables are a global problem that directly affect food security, the economy, and the environment. These losses are mainly associated with fungal diseases during storage. Due to the limitations of synthetic fungicides, including the development of resistance and risks to human health, there is growing interest in sustainable disease control strategies. This scoping review analyzes the potential of the yeast Metschnikowia pulcherrima as a biocontrol agent for postharvest phytopathogens, based on the scientific literature published between 2014 and 2026. The reviewed studies identify several antagonistic mechanisms, including competition for nutrients and space, the production of organic volatile compounds, hydrolytic enzyme activity, biofilm formation, and the induction of resistance in fruits. In vitro and in vivo assays show that M. pulcherrima effectively reduces postharvest disease incidence and severity caused by certain fungi. Furthermore, its synergistic effect when combined with emerging technologies is notable. The results highlight its potential as a sustainable alternative to synthetic fungicides, although further studies are needed for large-scale commercial application.

水果和蔬菜的采后损失是一个全球性问题,直接影响到粮食安全、经济和环境。这些损失主要与储存期间的真菌疾病有关。由于合成杀菌剂的局限性,包括抗药性的发展和对人类健康的风险,人们对可持续的疾病控制战略越来越感兴趣。本文基于2014年至2026年间发表的科学文献,分析了酵母pulcherrima作为采后植物病原体生物防治剂的潜力。综述了几种拮抗机制,包括养分和空间的竞争、有机挥发性化合物的产生、水解酶活性、生物膜的形成和果实抗性的诱导。体外和体内试验表明,白斑分枝杆菌能有效降低某些真菌引起的采后疾病发病率和严重程度。与新兴技术相结合,其协同效应显著。研究结果强调了其作为合成杀菌剂的可持续替代品的潜力,尽管需要进一步的研究以实现大规模的商业应用。
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引用次数: 0
Feasibility of Allogeneic Hematopoietic Stem Cell Transplantation Following Recent Invasive Mold Disease in Pediatric Patients. 近期侵袭性霉菌病患儿异体造血干细胞移植的可行性
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2026-04-21 DOI: 10.3390/jof12040297
Kevin Martínez Pérez, Daniela Torres-Hernández, Nathalia Sanclemente, Oscar Ramirez, Andrés Portilla, Jorge Buitrago, Eduardo López-Medina

A history of invasive mold disease (IMD) often delays or contraindicates allogeneic hematopoietic stem cell transplantation (allo-HSCT) in children. Given the limited data on pediatric patients with pre-allo-HSCT IMD, we aimed to describe the management and clinical outcomes of a cohort of children with IMD prior to allo-HSCT through day +100 post-transplantation. Between 2021 and 2024, ten pediatric patients were identified with proven or probable IMD. Their median age was 8.5 years. The most common pathogens were Aspergillus (n = 5) and Fusarium (n = 4). Infections most frequently involved the lungs followed by paranasal sinuses, bloodstream, liver, and skin. All patients demonstrated clinical improvement before transplantation, and by day +100 post-HSCT, no IMD relapses or infection-related mortality were observed. These findings suggest that complete radiologic or clinical resolution is not a prerequisite for proceeding with transplantation. Recent IMD should not be considered an absolute contraindication to urgent allo-HSCT when clinical improvement is evident, as transplantation facilitates immune reconstitution necessary for definitive infection control.

侵袭性霉菌病(IMD)的病史往往延迟或禁忌症的异体造血干细胞移植(同种异体造血干细胞移植)的儿童。鉴于异基因移植前IMD儿童患者的数据有限,我们旨在描述一组异基因移植前IMD儿童在移植后100天的治疗和临床结果。在2021年至2024年期间,有10名儿科患者被确诊或可能患有IMD。他们的平均年龄为8.5岁。最常见的病原菌为曲霉(n = 5)和镰刀菌(n = 4)。感染最常累及肺部,其次是鼻窦、血液、肝脏和皮肤。所有患者在移植前均表现出临床改善,并且在移植后第100天,未观察到IMD复发或感染相关死亡率。这些发现表明,完全的放射学或临床解决并不是进行移植的先决条件。当临床改善明显时,近期的IMD不应被视为紧急同种异体造血干细胞移植的绝对禁忌症,因为移植促进了最终感染控制所必需的免疫重建。
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引用次数: 0
Ndt80 Orchestrates Copper Stress Responses and Mitochondrial Homeostasis in Candida albicans. Ndt80调控白色念珠菌铜胁迫反应和线粒体稳态。
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2026-04-20 DOI: 10.3390/jof12040294
Hsuan-Yu Chen, Hsiu-Jung Lo, Chi-Jan Lin, Chung-Yu Lan

Copper is a crucial cofactor that sustains multiple cellular electron-transfer reactions, making it an essential element for life. However, cytotoxic levels of copper can cause structural damage and cell death through the production of reactive oxygen species (ROS) and nonspecific attacks on proteins. Moreover, immune cells, including neutrophils and macrophages, accumulate copper to induce oxidative bursts that kill engulfed pathogens. Therefore, a well-regulated copper homeostasis system is required for the human commensal fungus Candida albicans to thrive in extreme host environments. Remarkably, C. albicans exhibits higher copper tolerance than the nonpathogenic model yeast Saccharomyces cerevisiae, suggesting the presence of a specific copper tolerance mechanism that supports its adaptability to copper stress. Ndt80 is a versatile transcription factor that regulates several biological processes in C. albicans, ranging from morphological control to drug resistance. This study further reveals that Ndt80 may contribute to copper tolerance by regulating copper transporters and copper-dependent superoxide dismutases (Sods). Additionally, RNA sequencing and complementary approaches uncovered the involvement of Ndt80 in plasma membrane integrity and mitochondrial respiration under copper stress, further linking Ndt80 to copper tolerance. Together, these results broaden our understanding of Ndt80 functions and provide new insights into copper tolerance in C. albicans.

铜是维持多种细胞电子转移反应的关键辅助因子,使其成为生命的基本元素。然而,铜的细胞毒性水平可以通过产生活性氧(ROS)和对蛋白质的非特异性攻击导致结构损伤和细胞死亡。此外,免疫细胞,包括中性粒细胞和巨噬细胞,积累铜来诱导氧化爆发,杀死被吞噬的病原体。因此,一个良好调节的铜稳态系统是人类共生真菌白色念珠菌在极端宿主环境中茁壮成长所必需的。值得注意的是,白色念珠菌表现出比非致病性模式酵母更高的铜耐受性,这表明存在特定的铜耐受性机制,支持其对铜胁迫的适应性。Ndt80是一种多功能转录因子,调节白色念珠菌的几个生物过程,从形态控制到耐药性。本研究进一步揭示Ndt80可能通过调节铜转运体和铜依赖性超氧化物歧化酶(Sods)来促进铜耐受性。此外,RNA测序和互补方法揭示了Ndt80在铜胁迫下参与质膜完整性和线粒体呼吸,进一步将Ndt80与铜耐受性联系起来。总之,这些结果拓宽了我们对Ndt80功能的理解,并为白色念珠菌的铜耐受性提供了新的见解。
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引用次数: 0
Duration of Antifungal Therapy in Disseminated Coccidioidomycosis Patients: A Real-World Treatment Utilization Study. 播散性球孢子菌病患者抗真菌治疗的持续时间:一项现实世界治疗利用研究。
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2026-04-20 DOI: 10.3390/jof12040293
Craig I Coleman, Belinda Lovelace, Mark Bresnik

Background: Disseminated coccidioidomycosis (DCM) often requires prolonged antifungal therapy (AFT). Real-world data on AFT duration in DCM are limited. We evaluated time to AFT discontinuation among patients with DCM in the United States clinical practice. Methods: This retrospective, longitudinal study used STATinMED data (2016-2024). Patients had ≥1 International Classification of Diseases, Tenth Revision (ICD-10) code for DCM (B38.3, B38.4, B38.7, B38.81) during January 2017-December 2023, ≥1 claim for a triazole or amphotericin B within 21 days of the DCM diagnosis (index date), and continuous medical/pharmacy coverage during the 6-month baseline period. Discontinuation was defined as a ≥21-day gap without AFT. Antifungal agent/formulation switches were not considered discontinuations unless accompanied by a qualifying gap. The Kaplan-Meier methods were used to estimate time to discontinuation. Results: We identified 991 patients with DCM. Median age was 52 years (IQR 36, 65); 60.0% were men. Most resided in California (42.8%) or Arizona (33.6%). Initial AFT consisted predominantly of triazoles (96.8%), primarily fluconazole (83.2%). Discontinuation occurred in 27.6%, 40.0%, 54.2%, and 68.0% of patients by 3, 6, 12, and 36 months. Median AFT duration was 9.9 months. Conclusions: In a large US claims cohort, there was substantial variability in AFT duration in routine practice. Many patients had AFT durations under the lower limit of guideline recommendations for DCM, suggesting potential under-treatment, though appropriate clinical justifications may have existed.

背景:播散性球孢子菌病(DCM)通常需要长时间的抗真菌治疗(AFT)。关于DCM中AFT持续时间的实际数据是有限的。在美国临床实践中,我们评估了DCM患者到停药的时间。方法:回顾性、纵向研究使用STATinMED数据(2016-2024)。患者在2017年1月至2023年12月期间具有≥1个国际疾病分类第十版(ICD-10) DCM代码(B38.3, B38.4, B38.7, B38.81),在DCM诊断(索引日期)后21天内具有≥1个三唑或两性霉素B索赔,并且在6个月基线期间具有连续的医疗/药房覆盖。停药定义为≥21天无AFT间隔。抗真菌药物/制剂切换不视为停药,除非伴有合格间隔。Kaplan-Meier方法用于估计停药时间。结果:我们发现991例DCM患者。中位年龄52岁(IQR 36,65);60.0%为男性。大多数人居住在加利福尼亚州(42.8%)或亚利桑那州(33.6%)。最初的AFT主要由三唑类药物(96.8%)组成,主要是氟康唑(83.2%)。在3、6、12和36个月时停药的患者分别为27.6%、40.0%、54.2%和68.0%。中位AFT持续时间为9.9个月。结论:在一个大型的美国索赔队列中,在常规实践中AFT持续时间有很大的变化。许多患者的AFT持续时间低于DCM指南建议的下限,尽管可能存在适当的临床理由,但这表明潜在的治疗不足。
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引用次数: 0
Novel Antifungal Pyridine Alkaloids from Endophytic Fungus Penicillium citrinum VDL118. 内生真菌柑橘青霉VDL118新型抗真菌吡啶生物碱。
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2026-04-20 DOI: 10.3390/jof12040296
Mei Yang, Shan Hu, Zhi-Yu Zhang, Fa-Zhong Yang, Xiao-Qin Yang, Si-Da Xie, Ying-Jun Zhang, Ping Zhao, Guo-Lei Zhu

Three novel alkaloids, penicitrioids A-C (1-3), and two known compounds (4-5) were isolated from the ethyl acetate (EtOAc) extract of the solid fermentation of Penicillium citrinum VDL118, an endophytic fungus harbored in the leaves of Vaccinium dunalianum Wight (Ericaceae), a perennial evergreen shrub native to the southwestern regions of China, Myanmar, and Vietnam. Compounds 1 and 2 are novel pyridine alkaloids characterized by an unprecedented dihydrofuro[3,4-c]pyridine core, while 3 features a distinct pyrrolo[3,4-c]pyridine framework. Their structures were unambiguously established by comprehensive spectroscopic analysis and electronic circular dichroism (ECD) calculations. In vitro antifungal assays revealed that compounds 1-5 exhibited moderate to potent inhibitory effects against five tested phytopathogenic fungi, with minimum inhibitory concentrations (MICs) ranging from 3.1 to 100 μg/mL. Notably, four of them (1-4) displayed broad-spectrum and potent activity against Gloeophyllum trabeum, Coriolus versicolor, Fusarium solani, and Botrytis cinerea, with MIC values as low as 3.1-12.5 μg/mL. Furthermore, a plausible biosynthetic pathway for compounds 1-3 was proposed.

从柠檬酸青霉VDL118的固体发酵乙酸乙酯(EtOAc)萃取物中分离出3个新的生物碱,penicitriids a - c(1-3)和2个已知化合物(4-5)。柠檬酸青霉VDL118是一种内生真菌,生长在中国西南部、缅甸和越南的多年生常绿灌木滇青霉(Ericaceae)的叶片中。化合物1和2是新型吡啶类生物碱,具有前所未有的二氢呋喃[3,4-c]吡啶核心,而化合物3具有独特的吡罗[3,4-c]吡啶框架。通过综合光谱分析和电子圆二色性(ECD)计算明确了它们的结构。体外抑菌实验表明,化合物1 ~ 5对5种植物病原真菌均有中强抑制作用,最低抑菌浓度(mic)为3.1 ~ 100 μg/mL。其中4种(1 ~ 4)对黄球菌、花斑革菌、茄枯菌和灰霉病菌具有广谱和强效活性,MIC值低至3.1 ~ 12.5 μg/mL。此外,还提出了化合物1 ~ 3的生物合成途径。
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引用次数: 0
Bioremediation Potential of Leaf Endophytic Fungi in Allium ampeloprasum and Brassica oleracea var. capitata. 黑胡椒和甘蓝叶片内生真菌的生物修复潜力。
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2026-04-20 DOI: 10.3390/jof12040295
Dayani Pavalakumar, Sagarika Kannangara, Nadeema Dharmasiri, Chamani Amarasekara, Lanka Undugoda, Kasun M Thambugala, Jayantha Munasinghe, Sukanya Haituk, Ratchadawan Cheewangkoon

Polycyclic aromatic hydrocarbons (PAHs) are toxic air pollutants mainly released through vehicular emissions and can accumulate on edible plants, posing health risks to humans. This study aimed to isolate and identify endophytic fungi from Allium ampeloprasum and Brassica oleracea var. capitata, which are widely cultivated along roadside areas in the upcountry region of Sri Lanka. Sampling sites included Nuwara Eliya town, Nanu Oya, St. Clair's, and Meepilimana (control), where above-ground parts of the selected vegetables were collected in six replicates. Fungal isolates were obtained through surface sterilization, and their ability to degrade PAHs (naphthalene, phenanthrene, anthracene, and pyrene) was evaluated using plate assays, spectrophotometric analysis, and high-performance liquid chromatography (HPLC). Phyllosphere PAH concentrations were also measured using HPLC. It revealed significantly higher concentrations of all four PAHs in the phyllosphere of both vegetables at polluted sites, with the highest levels recorded in A. ampeloprasum from Nuwara Eliya town: naphthalene (145.92 ng/g), phenanthrene (97.67 ng/g), anthracene (88.71 ng/g), and pyrene (63.82 ng/g). Most endophytic fungal strains isolated from both vegetables were able to grow on Bacto Bushnell-Haas (BBH) medium supplemented with PAHs, producing colonies exceeding 20 mm in diameter. Spectrophotometric analysis showed that Fusarium liriodendri SP2 (PV400499.1) and Trichoderma atroviride SP1 (PV400486.1) achieved approximately 75% degradation of selected PAHs. Furthermore, HPLC analysis confirmed that these isolates effectively degraded all tested PAHs, with degradation rates of approximately 70%. F. liriodendri was the most efficient degrader, achieving degradation rates of 68.50 ± 2.34% for naphthalene, 65.26 ± 1.21% for phenanthrene, 69.21 ± 1.45% for pyrene, and 66.89 ± 1.98% for anthracene. The PAH degradation byproducts of the selected fungal isolates were non-toxic to Artemia salina, confirming their environmental safety. These results highlight the bioremediation potential of endophytic fungi isolated from A. ampeloprasum and B. oleracea var. capitata in PAH-contaminated environments.

多环芳烃(PAHs)是一种主要通过车辆排放释放的有毒空气污染物,可在食用植物上积累,对人类健康构成威胁。本研究旨在分离鉴定斯里兰卡内陆地区路边广泛种植的大蒜(Allium ampeloprasum)和甘蓝(Brassica oleracea var. capitata)内生真菌。采样地点包括Nuwara Eliya镇、Nanu Oya、St. Clair's和Meepilimana(对照),其中所选蔬菜的地上部分分为6个重复收集。通过表面灭菌获得真菌分离株,并使用平板测定、分光光度分析和高效液相色谱(HPLC)评估其降解多环芳烃(萘、菲、蒽和芘)的能力。用高效液相色谱法测定层球多环芳烃浓度。结果表明,污染地点两种蔬菜层圈中4种多环芳烃的浓度均显著高于污染地点,其中以Nuwara Eliya镇的ampeloprasum最高,分别为萘(145.92 ng/g)、菲(97.67 ng/g)、蒽(88.71 ng/g)和芘(63.82 ng/g)。从这两种蔬菜中分离的大多数内生真菌菌株都能在添加了多环芳烃的BBH培养基上生长,菌落直径超过20 mm。分光光度分析表明,liriodendri Fusarium SP2 (PV400499.1)和atroviride Trichoderma SP1 (PV400486.1)对所选多环芳烃的降解率约为75%。此外,HPLC分析证实,这些分离物有效降解了所有测试的多环芳烃,降解率约为70%。liriodendri对萘的降解率为68.50±2.34%,对菲的降解率为65.26±1.21%,对芘的降解率为69.21±1.45%,对蒽的降解率为66.89±1.98%。所选真菌菌株的多环芳烃降解副产物对盐渍蒿无毒,证实了其环境安全性。这些结果突出了葡萄球菌和甘蓝芽孢杆菌内生真菌在多环芳烃污染环境中的生物修复潜力。
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引用次数: 0
Dual Biocontrol and Plant Growth-Promoting Effects of Trichoderma nordicum V1 Against Oomycete Plant Pathogens. 北方木霉V1对植物卵菌的双重生防和促生作用
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2026-04-20 DOI: 10.3390/jof12040292
Songrong Li, Xian Wen, Siqiao Chen, Yishen Zhao, Jinhao Chen, Wanrong Li, Yajuan Chen, Mingyue Ding, Siqi Jiang, Wilfred Mabeche Anjago, Dongmei Zhou, Feng M Cai, Irina S Druzhinina, Min Jiu, Lihui Wei, Paul Daly

The potential of Trichoderma nordicum (Hypocreales, Ascomycota), a recently described species, for antagonism and use in the biocontrol of oomycete-caused plant diseases is unknown. Trichoderma is a well-known genus for containing microbial antagonists and biocontrol agents. The T. nordicum in this study was isolated from decomposing wood, and rpb2 and tef1 barcode sequencing demonstrated that the isolates were a match to the reference T. nordicum and T. nigricans strains. Since T. nordicum was described before T. nigricans, the isolates were assigned to T. nordicum, although taxonomic uncertainty between these species requires future clarification. In dual-culture confrontation assays, T. nordicum overgrew five economically important oomycete plant pathogens (Phytophthora capsici, P. sojae, Pythium aphanidermatum, P. myriotylum, and Globisporangium ultimum). The inability to recover viable P. aphanidermatum and P. capsici from the parts of the plate overgrown by T. nordicum, coupled with protease and endo-cellulase activities, correlates with T. nordicum having antagonistic abilities. Inoculation with T. nordicum preventively reduced the levels of cucumber seedling damping-off caused by P. aphanidermatum by up to 70%. The T. nordicum biocontrol effects against pepper blight caused by P. capsici were greater than 80%, compared to an autoclaved T. nordicum spore control. T. nordicum could also significantly promote the growth of pepper, with plant weight increased by up to 40%, compared to an autoclaved-spore control. In contrast, T. nordicum could not be used to control Pythium soft rot of ginger caused by P. myriotylum, even though P. myriotylum was overgrown by T. nordicum, suggesting host- or pathosystem-specific factors influence biocontrol efficacy. In summary, T. nordicum is a promising biocontrol agent for use in the control of pepper blight caused by P. capsici, and also has potential for use in the control of other oomycete-caused plant diseases in vegetable production systems.

北木霉(下creales, Ascomycota)是最近发现的一种植物,其拮抗和生物防治卵菌引起的植物病害的潜力尚不清楚。木霉是一种众所周知的含有微生物拮抗剂和生物防治剂的属。本研究分离到的北方T. nordicum是从木质分解中分离到的,rpb2和tef1条形码测序结果表明,分离到的菌株与参考菌株nordicum和nigricans相匹配。由于黑螺旋体早于黑螺旋体被描述,因此分离物被归为黑螺旋体,尽管这些物种之间的分类不确定性需要进一步澄清。在双培养对抗试验中,北方T. nordicum超过了5种经济上重要的卵菌植物病原体(辣椒疫霉、大豆疫霉、隐皮霉、myriotyum和Globisporangium ultimum)。在培养皿中无法从被褐条霉侵染的部分恢复有活力的葡萄皮霉和辣椒霉,再加上蛋白酶和内切纤维素酶的活性,与褐条霉具有拮抗能力有关。接种北方镰刀菌,可预防黄瓜黄瓜枯苗率达70%。与高压灭菌法相比,辣椒粉对辣椒枯萎病的防效大于80%。与蒸压孢子对照相比,北方赤霉病菌对辣椒的生长也有显著的促进作用,植株重量增加了40%。与此相反,尽管紫霉在姜中被紫霉侵染,但紫霉却不能有效防治姜中霉软腐病,这表明寄主或病理系统特异性因素会影响其生物防治效果。综上所述,nordicum是一种很有前途的生物防治剂,可以用于辣椒疫病的防治,也可以用于蔬菜生产系统中其他由卵霉菌引起的植物病害的防治。
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引用次数: 0
Endophytic Fusarium commune G3-29-Mediated dsRNA Delivery for Efficient Control of Western Flower Thrips. 内生镰刀菌g3 -29介导的dsRNA传递有效防治西花蓟马。
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2026-04-18 DOI: 10.3390/jof12040291
Xueyuan Sheng, Yanfei Wang, Chang Chen, Chao Ma, Shuangchao Wang, Endong Wang, Yan Zhao, Lihua Guo

RNA interference (RNAi) provides a sequence-specific strategy for pest management, but efficient and stable double-stranded RNA (dsRNA) delivery remains a key challenge. Here, we established a plant-probiotic-based gene silencing system using the endophytic fungus Fusarium commune G3-29 as a dsRNA delivery vector against western flower thrips (WFTs, Frankliniella occidentalis). Recombinant G3-29 strains expressing dsRNA targeting the essential WFT genes ACT and SNF were constructed and confirmed to colonize kidney bean leaves without pathogenicity. Bioassays showed that feeding on leaves colonized by dsRNA-expressing G3-29 significantly decreased survival and downregulated target gene expression in both WFT larvae and adults. Within 4 days, survival of both larvae and adults fell below 10%. In larvae, target gene expression decreased by 63% (ACT) and 33% (SNF), while in adults, reductions of 74% (ACT) and 65% (SNF) were observed. In contrast, in vitro-synthesized dsRNA failed to induce significant gene silencing or mortality in larvae, and its control efficacy against adults was also inferior to that of endophytic fungus-mediated dsRNA delivery. Our findings establish endophytic fungus F. commune G3-29 as an effective and sustainable dsRNA delivery vehicle for RNAi-based pest control, offering distinct advantages over existing strategies such as HIGS and SIGS. This approach provides a promising new direction for managing WFTs and other insect pests.

RNA干扰(RNAi)为害虫防治提供了一种序列特异性策略,但有效和稳定的双链RNA (dsRNA)递送仍然是一个关键挑战。本研究以内生真菌镰刀菌G3-29为载体,建立了一种基于植物益生菌的基因沉默系统,以抵御西方花蓟马(Frankliniella occidentalis)。构建了以WFT必需基因ACT和SNF为靶点表达dsRNA的重组G3-29菌株,证实其在芸豆叶片上定植无致病性。生物测定表明,以表达G3-29的dsrna定殖的叶片为食,WFT幼虫和成虫的存活率均显著降低,靶基因表达下调。4 d内,幼虫和成虫存活率均低于10%。幼虫靶基因表达量分别降低63% (ACT)和33% (SNF),成虫靶基因表达量分别降低74% (ACT)和65% (SNF)。相比之下,体外合成的dsRNA未能诱导幼虫明显的基因沉默或死亡,其对成虫的控制效果也不如内生真菌介导的dsRNA递送。我们的研究结果表明,内生真菌F. commune G3-29作为基于rnai的害虫防治的有效和可持续的dsRNA递送载体,与现有的HIGS和SIGS策略相比具有明显的优势。该方法为wft和其他害虫的管理提供了一个有希望的新方向。
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引用次数: 0
Two-Step Engineering of Food-Grade Aspergillus oryzae via Endogenous Signal Peptides and Vesicle Trafficking Proteins to Enhance Carrier-Free Protein Secretion. 利用内源性信号肽和囊泡转运蛋白促进无载体蛋白分泌的食品级米曲霉两步工程
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2026-04-18 DOI: 10.3390/jof12040289
Sarocha Panchanawaporn, Nakul Rattanaphan, Sukanya Jeennor, Jutamas Anantayanon, Weerapong Woraprayote, Laphaslada Pumpuang, Thipphiya Karirat, Nuttamon Prompakdee, Kobkul Laoteng, Chanikul Chutrakul

Heterologous protein secretion in filamentous fungi is often constrained by limitations in signal peptide recognition and intracellular trafficking. Aspergillus oryzae, a food-grade industrial fungus, has a robust native secretory system. However, its capacity for recombinant protein secretion remains suboptimal. Here, we developed a two-step, carrier-free engineering strategy to enhance protein secretion in A. oryzae. We identified endogenous signal peptides among highly secreted proteins using a green fluorescent protein (GFP) reporter. The oryzin signal peptide SPAoalp1 increased GFP secretion 5.50-fold compared with a no-signal-peptide control. We co-overexpressed Aosly1, a Sec1/Munc18 family protein that regulates soluble N-ethylmaleimide-sensitive factor attachment protein receptor-mediated vesicle trafficking, which, in combination with SPAoalp1, increased secretion approximately two-fold compared with SPAlp1 control and ten-fold with no-SP control. Applying the engineered platform for genetic improvement of heterologous bovine κ-casein increased secretion from 0.11 to 0.24 mg/L. Physiological optimization further increased secretion. The developed system provided initial evidence for secretion of a ~12 kDa band consistent with Aopafb transcription, with MIC90 values of 4.56-8.24% (v/v) against two Candida albicans strains and 4.68% (v/v) against Aspergillus niger. The system offers a modular framework for engineering fungal secretion and expands the utility of A. oryzae for recombinant protein production.

丝状真菌的异源蛋白分泌经常受到信号肽识别和细胞内运输的限制。米曲霉是一种食品级工业真菌,具有强大的天然分泌系统。然而,它的重组蛋白分泌能力仍然不够理想。在这里,我们开发了一个两步,无载体的工程策略,以提高a . oryzae的蛋白质分泌。我们使用绿色荧光蛋白(GFP)报告基因在高分泌蛋白中鉴定内源性信号肽。与无信号肽对照相比,米蛋白信号肽SPAoalp1使GFP分泌增加5.50倍。我们共过表达了一种调节可溶性n-乙基丙烯酰亚胺敏感因子附着蛋白受体介导的囊泡运输的Sec1/Munc18家族蛋白,该蛋白与SPAoalp1结合后,与SPAlp1对照相比,其分泌量增加了约两倍,与无sp对照相比增加了十倍。应用该工程平台对异源牛κ-酪蛋白进行遗传改良,使其分泌量由0.11 mg/L提高到0.24 mg/L。生理优化进一步增加了分泌。初步证明,该系统可分泌与Aopafb转录一致的~12 kDa条带,对2株白色念珠菌的MIC90值为4.56 ~ 8.24% (v/v),对黑曲霉的MIC90值为4.68% (v/v)。该系统为工程真菌分泌提供了模块化框架,并扩展了m.o ryzae在重组蛋白生产中的应用。
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引用次数: 0
Genotype-Phenotype Relationships in Azole-Resistant Aspergillus: Two Sides of the Same Coin. 抗唑曲霉的基因型-表型关系:同一硬币的两面。
IF 4.2 2区 生物学 Q2 MICROBIOLOGY Pub Date : 2026-04-18 DOI: 10.3390/jof12040290
Merlijn H I van Haren, Willem J G Melchers, Jianhua Zhang, Sarah Dellière, Christine C Bii, Felicia A Stanford, Michael Voetz, P Lewis White, Paul S Dyer, Suzan D Pas, Paul E Verweij, Jochem B Buil

Aspergillus fumigatus is a leading cause of invasive fungal disease in humans and is classified as a critical priority threat by the World Health Organization. Triazole antifungals remain the cornerstone of therapy, yet their effectiveness is steadily being eroded by the continuous rise in drug resistance. Most resistance mechanisms trace back to mutations in Cyp51A, spawning well-defined genotypes such as TR34/L98H and TR46/Y121F/T289A. However, the Cyp51A genotype-phenotype landscape in A. fumigatus is far from straightforward. Isolates that share an identical TR genotype can display strikingly divergent susceptibility profiles, and mutational hotspots in Cyp51A, such as G54, M220 and G448, are linked to varying resistances, challenging assumptions about predictable resistance behavior. Complicating matters further, an expanding array of resistance mechanisms, independent of Cyp51A, is now being uncovered. This review summarizes the current state of knowledge on azole resistance in A. fumigatus, dissecting the intricate genotype-phenotype relationships, spotlighting emerging non-Cyp51A pathways and outlining future strategies to enhance the detection and clinical management of antifungal resistance.

烟曲霉是人类侵袭性真菌疾病的主要原因,被世界卫生组织列为严重的优先威胁。三唑类抗真菌药物仍然是治疗的基石,但由于耐药性的持续上升,它们的有效性正在稳步受到侵蚀。大多数耐药机制可追溯到Cyp51A突变,产生明确的基因型,如TR34/L98H和TR46/Y121F/T289A。然而,烟曲霉的Cyp51A基因型-表型格局远不是直截了当地的。具有相同TR基因型的分离株可以显示出明显不同的易感性谱,并且Cyp51A的突变热点,如G54, M220和G448,与不同的抗性相关,挑战了可预测抗性行为的假设。使问题进一步复杂化的是,目前正在发现一系列独立于Cyp51A的耐药机制。本文综述了烟曲霉对唑类药物耐药的研究现状,剖析了烟曲霉复杂的基因型-表型关系,重点介绍了新出现的非cyp51a途径,并概述了未来加强抗真菌药物耐药检测和临床管理的策略。
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引用次数: 0
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Journal of Fungi
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