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学者姓名：汤蔚
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Abstract ：
Rice (Oryza sativa L.) is one of the most important staple foods for human population worldwide. However, rice production continues to be severely threatened by rice blast disease caused by an ascomycete fungus Magnaporthe oryzae. Tail-anchored (TA) proteins are conserved across diverse organisms and belong to a class of polypeptides that are inserted into the membrane by a hydrophobic sequence located at the C-terminal region. The Guided Entry of Tail-anchored (GET) complex is responsible for the post-translational insertion of nascent TA proteins into the Saccharomyces cerevisiae ER lipid bilayer. In S. cerevisiae, the GET pathway comprises six known associated components Get1, Get2, Get3, Get4, Get5, Sgt2 and Ssa1 that have been identified and extensively studied. However, the role of the GET complex in rice blast fungus has not been elucidated. Here, we identified five proteins of the GET Complex in M. oryzae, namely MoGet1, MoGet2, MoGet3, MoGet4 and MoSgt2 and generated the gene knock-out mutants. Deletion of MoGET1 and MoGET2 revealed that they are required for vegetative growth, asexual reproduction, pathogenesis, and right localization of TA protein, MoYsy6, while MoGet3 negatively regulates hyphal growth, asexual development and pathogenesis of M. oryzae. In contrast, loss of MoGet4 and MoSgt2 had no effect on the normal development of the rice blast fungus. We demonstrated that the MoGet2 is important in osmotic stress response and positively regulates cell wall integrity. The MoGet1 and MoGet2 were ER-localized and indispensable for DTT-induced ER stress response. In vitro and in vivo interaction assay revealed MoGet3 has physical interaction with both MoGet1 and MoGet2, indicating the existence of a possible synergistic function amongst the Get components in rice blast fungus. In summary, this finding provides valuable insight into the biological functions of the GET components in plant fungal pathogens.
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| GB/T 7714 | Abah, Felix , Zheng, Qiaojia , Chen, Xinru et al. Characterisation of guided entry of tail-anchored proteins in Magnaporthe oryzae [J]. | PLOS PATHOGENS , 2025 , 21 (7) . |
| MLA | Abah, Felix et al. "Characterisation of guided entry of tail-anchored proteins in Magnaporthe oryzae" . | PLOS PATHOGENS 21 . 7 (2025) . |
| APA | Abah, Felix , Zheng, Qiaojia , Chen, Xinru , Huang, Linwan , Chen, Xiaomin , Biregeya, Jules et al. Characterisation of guided entry of tail-anchored proteins in Magnaporthe oryzae . | PLOS PATHOGENS , 2025 , 21 (7) . |
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由稻瘟病菌(Magnaporthe oryzae)侵染引起的稻瘟病是全球水稻生产中极具破坏性的病害,严重威胁粮食安全。效应蛋白作为稻瘟病菌的关键致病因子,通过干扰宿主免疫反应促进侵染。但是,目前大部分已鉴定的植物病原真菌效应蛋白都具有N-端信号肽,对无N-端分泌信号肽的效应蛋白鲜有报道。本研究聚焦稻瘟病菌无N端分泌信号肽的非典型效应蛋白MoLEPA,发现其缺失突变体的致病性显著性降低。进一步分析表明MoLEPA基因缺失影响了稻瘟病菌附着胞萌发、膨压积累以及脂质代谢,从而影响稻瘟病菌的致病性;亚细胞定位观察发现, MoLepa在烟草和水稻原生质体中均可定位于细胞核内;此外,异源表达MoLEPA则抑制病原相关分子模式(PAMP)激发的免疫反应,增强水稻感病性。通过酵母双杂交文库筛选,鉴定到水稻靶标蛋白OsLIP1,其与MoLEPA在细胞核内互作,后续将进一步阐明MoLepa与OsLIP1的互作特点。相关结果有望揭示稻瘟病菌非典型效应蛋白的致病新机制,为抗病育种及靶向效应蛋白的绿色防控技术开发提供理论依据。
Keyword ：
植物与真菌互作 植物与真菌互作 水稻免疫 水稻免疫 稻瘟病菌 稻瘟病菌 非典型效应蛋白 非典型效应蛋白
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| GB/T 7714 | 高大明 , 包瑛 , 蔡世国 et al. 稻瘟病菌无N-端信号肽效应蛋白MoLepa靶向水稻细胞核调控其免疫机制的研究 [C] //中国植物病理学会2025年学术年会 . 2025 . |
| MLA | 高大明 et al. "稻瘟病菌无N-端信号肽效应蛋白MoLepa靶向水稻细胞核调控其免疫机制的研究" 中国植物病理学会2025年学术年会 . (2025) . |
| APA | 高大明 , 包瑛 , 蔡世国 , 陈小敏 , 汤蔚 , 王宗华 . 稻瘟病菌无N-端信号肽效应蛋白MoLepa靶向水稻细胞核调控其免疫机制的研究 中国植物病理学会2025年学术年会 . (2025) . |
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Rice blast caused by Magnaporthe oryzae pathotype is the worst disease that leads to serious food insecurity globally. Understanding rice blast disease pathogenesis is therefore essential for the development of a blast disease mitigation strategy. Reverse phosphorylation mediated by phosphatases performs a vital function in the activation of diverse biological mechanisms within eukaryotic. However, little has been reported on the roles of PP2Cs in the virulence of blast fungus. In this current work, we deployed functional genomics and biochemical approaches to characterize type 2C protein phosphatase MoPtc6 in blast fungus. Deletion of MoPTC6 led to a drastic reduction in conidiophore development, conidia production, hyphal growth, and stress tolerance. Western blotting assay demonstrated that the phosphorylation level of MoOsm1 was decreased while MoMps1 was increased in the MoPtc6 deletion mutant, and comparative transcriptome assay revealed a higher number of expressed genes between mutant and wild type. Localization assay confirmed that MoPtc6 is sub-localized in the cytoplasm of mycelia, spores, and in the appressoria of M. oryzae. Furthermore, disruption of MoPTC6 impaired appressoria turgor pressure and glycogen utilization; more findings revealed attenuation of hyphal penetration and virulence upon deletion of MoPTC6. Generally, present findings suggested the role of MoPtc6 in the growth and virulence of M. oryzae.
Keyword ：
Magnaporthe oryzae Magnaporthe oryzae phosphorylation phosphorylation protein phosphatases protein phosphatases stress tolerance stress tolerance virulence virulence
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| GB/T 7714 | Jagero, Frankline Otieno , Felix, Abah , Abubakar, Yakubu Saddeeq et al. Type 2C Protein Phosphatase MoPtc6 Plays Critical Roles in the Development and Virulence of Magnaporthe oryzae [J]. | JOURNAL OF FUNGI , 2025 , 11 (5) . |
| MLA | Jagero, Frankline Otieno et al. "Type 2C Protein Phosphatase MoPtc6 Plays Critical Roles in the Development and Virulence of Magnaporthe oryzae" . | JOURNAL OF FUNGI 11 . 5 (2025) . |
| APA | Jagero, Frankline Otieno , Felix, Abah , Abubakar, Yakubu Saddeeq , Chen, Meilian , Anjago, Wilfred M. , Mediatrice, Hatungimana et al. Type 2C Protein Phosphatase MoPtc6 Plays Critical Roles in the Development and Virulence of Magnaporthe oryzae . | JOURNAL OF FUNGI , 2025 , 11 (5) . |
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Protein phosphatases are crucial enzymes that regulate key cellular processes such as the cell cycle, gene transcription, and translation in eukaryotes. Seven PP2C protein phosphatases have been identified in Magnaporthe oryzae. However, their synergistic roles in the pathology and physiology of M. oryzae remain poorly investigated. By qRT-PCR analysis, we found that PTC1 and PTC2 are significantly upregulated in the PTC5 deletion mutant. The double deletion of the MoPTC5/MoPTC1 and MoPTC5/MoPTC2 genes significantly reduced hyphal growth, conidiophore formation, sporulation, and virulence in M. oryzae. In addition, the double-knockout mutants were increasingly sensitive to different osmotic, oxidative, and cell wall stresses. Western blot analysis revealed that MoPtc5 plays a synergistic function with MoPtc1 and MoPtc2 in the regulation of MoMps1 and MoOsm1 phosphorylation levels. Lastly, appressorium formation and turgor generation were remarkably affected in the Delta Moptc5 Delta Moptc1 and Delta Moptc5 Delta Moptc2 double-deletion mutants. These findings demonstrate the overlapping roles of PP2c protein phosphatase in the fungal development and pathogenesis of M. oryzae.
Keyword ：
Magnaporthe oryzae Magnaporthe oryzae pathogenesis pathogenesis protein phosphatases protein phosphatases stress tolerance stress tolerance synergistic synergistic
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| GB/T 7714 | Biregeya, Jules , Otieno, Frankline Jagero , Chen, Meilian et al. Protein Phosphatases MoPtc5, MoPtc1, and MoPtc2 Contribute to the Vegetative Growth, Stress Adaptation, and Virulence of Magnaporthe oryzae [J]. | JOURNAL OF FUNGI , 2025 , 11 (3) . |
| MLA | Biregeya, Jules et al. "Protein Phosphatases MoPtc5, MoPtc1, and MoPtc2 Contribute to the Vegetative Growth, Stress Adaptation, and Virulence of Magnaporthe oryzae" . | JOURNAL OF FUNGI 11 . 3 (2025) . |
| APA | Biregeya, Jules , Otieno, Frankline Jagero , Chen, Meilian , Mabeche, Anjago Wilfred , Felix, Abah , Aimable, Nsanzinshuti et al. Protein Phosphatases MoPtc5, MoPtc1, and MoPtc2 Contribute to the Vegetative Growth, Stress Adaptation, and Virulence of Magnaporthe oryzae . | JOURNAL OF FUNGI , 2025 , 11 (3) . |
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本发明属于生物基因工程领域，具体涉及一种稻瘟病菌MoPCP1基因及其应用。所述稻瘟病菌MoPCP1基因全长序列如SEQ ID NO.1所示，cDNA序列如SEQ ID NO.2所示，编码的蛋白的氨基酸序列如SEQ ID NO.3所示。实验表明，利用潮霉素抗性基因替换稻瘟病菌MoPCP1基因后，所得到的稻瘟病菌敲除突变体与野生型稻瘟病菌相比，营养生长能力、附着孢形成能力以及分生孢子形态及大小上有明显变化，在应对外界环境因子的应答上有明显的区别；致病性实验表明，MoPCP1基因的缺失使稻瘟病菌致病性显著下降，说明MoPCP1基因在调控稻瘟病菌致病力方面有显著作用。本发明所提供的MoPCP1基因及其应用在在稻瘟病菌致病性方面具有重要作用，为进一步利用该基因防治稻瘟病菌提供了新的方向。
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| GB/T 7714 | 汤蔚 , 黄琳婉 , 陈欣如 et al. 稻瘟病菌MoPCP1基因及其应用 : CN202410105769.3[P]. | 2024-01-25 . |
| MLA | 汤蔚 et al. "稻瘟病菌MoPCP1基因及其应用" : CN202410105769.3. | 2024-01-25 . |
| APA | 汤蔚 , 黄琳婉 , 陈欣如 , 陈小敏 , 杨子锋 , 翁书凝 et al. 稻瘟病菌MoPCP1基因及其应用 : CN202410105769.3. | 2024-01-25 . |
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The signal peptidase complex (SPC) is a crucial membrane enzyme complex involved in the process of protein secretion and maturation in both prokaryotic and eukaryotic cells. SPC is responsible for the cleavage of N-terminal signal sequences from nascent proteins, a sequence of amino acids that directs the newly synthesized protein to the secretory pathway. The yeast SPC is composed of four subunits: Spc1, Spc2, Spc3, and Sec11. To understand how SPC functions in the fungal plant pathogen, we identified the SPC component gene MoSPC2 and characterized its functions in M. oryzae. Through measuring the colony diameter of the Delta Mospc2 mutant and control strains on culture medium plates, quantifying conidia production, observing conidial morphology, and assessing pathogenicity on rice and barley plants, we found that MoSpc2 contributes to fungal growth, asexual development, and pathogenicity. Since host-derived reactive oxygen species (ROS) are crucial for rice to defend against M. oryzae, we further investigated the role of MoSpc2 in ROS modulation. Our results indicate that MoSpc2 plays a pivotal role in suppressing the accumulation of ROS and regulating the activities of extracellular peroxidases and laccases. Notably, MoSpc2 mediates the accumulation and secretion of the effector protein MoSlp1. Furthermore, using affinity purification, we discovered MoSpc2-interacting proteins and identified potential SPC interactors. These candidates provide a foundation for future mechanistic studies aimed at elucidating their functional roles in SPC complex assembly and pathogenic regulation. Our results highlight the significance of the SPC component gene MoSPC2 involvement in fungal development and pathogenicity and widen our understanding of the connections between the SPC and fungal pathogenesis.
Keyword ：
Fungal development Fungal development Magnaporthe oryzae Magnaporthe oryzae MoSpc2 MoSpc2 Pathogenicity Pathogenicity Signal peptidase Signal peptidase
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| GB/T 7714 | Tang, Wei , Wang, Min , Bao, Ying et al. The Signal Peptidase MoSpc2 Orchestrates Fungal Development, Protein Secretion and Pathogenicity in Magnaporthe oryzae [J]. | RICE , 2025 , 18 (1) . |
| MLA | Tang, Wei et al. "The Signal Peptidase MoSpc2 Orchestrates Fungal Development, Protein Secretion and Pathogenicity in Magnaporthe oryzae" . | RICE 18 . 1 (2025) . |
| APA | Tang, Wei , Wang, Min , Bao, Ying , Chen, Weiquan , Jiang, Yanfang , Chen, Meilian et al. The Signal Peptidase MoSpc2 Orchestrates Fungal Development, Protein Secretion and Pathogenicity in Magnaporthe oryzae . | RICE , 2025 , 18 (1) . |
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由稻瘟病菌引起的稻瘟病是世界水稻产区严重的真菌病害,威胁粮食生产安全。病原菌侵染寄主植物的早期大量表达效应蛋白,干扰寄主免疫反应,促进侵染。真核生物mRNA中5'端存在帽状结构,该结构由倒置的7-甲基鸟苷组成,其与新合成的第一个转录核苷酸相连,随后被帽结合蛋白复合物(The cap-binding complex)结合,参与转录因子的招募和转录的调节过程。在酿酒酵母中,CBC通过招募Ctk2或Bur2促进了延伸和加帽酶(Capping enzyme)释放,CBC还通过其与酵母体内Mot1p (Modifier of transcription)的相互作用促进预启动复合物的形成。而病原菌致病基因在特定阶段的时空表达对其成功致病具有重要意义。目前,在植物病原真菌中尚不清楚CBC复合物成员的生物学功能。本论文主要研究稻瘟病菌帽结合蛋白MoCbc2在稻瘟病菌生长发育及致病过程中所发挥的功能,生物信息学分析结果表明,该蛋白包含两个RRM结构域,为RNA识别基序。分析其在不同阶段的表达水平发现,MoCBC2基因在稻瘟病菌传染中期高表达,提示MoCBC2可能在稻瘟病菌侵染水稻中期具有重要作用。通过基因敲除得到稻瘟病菌的突变体ΔMocbc2,并以野生型菌株Guy11作为对照,分析突变体的生物学表型。研究表明,MoCBC2在MM培养基上其生长明显受到抑制。此外,对孢子产量的统计显示,突变体的产孢量显著减少,说明MoCBC2基因参与了菌丝的无性繁殖。另外,MoCBC2缺失会降低稻瘟病菌对内质网压力和氧化压力胁迫的耐受性,在致病性方面,突变体表现出显著减弱,表明MoCBC2在调控稻瘟病菌致病性中的重要作用。
Keyword ：
帽结合蛋白复合物 帽结合蛋白复合物 效应子 效应子 稻瘟病菌 稻瘟病菌
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| GB/T 7714 | 黄琳婉 , 陈欣如 , 何瑀欣 et al. 稻瘟病菌MoCbc2蛋白的生物学功能 [C] //中国植物病理学会2024年学术年会 . 2024 . |
| MLA | 黄琳婉 et al. "稻瘟病菌MoCbc2蛋白的生物学功能" 中国植物病理学会2024年学术年会 . (2024) . |
| APA | 黄琳婉 , 陈欣如 , 何瑀欣 , 萧道巧 , 王宗华 , 汤蔚 . 稻瘟病菌MoCbc2蛋白的生物学功能 中国植物病理学会2024年学术年会 . (2024) . |
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Endoplasmic reticulum (ER) is an enclosed three-dimensional eukaryotic membrane network composed of flattened sacs. Fusion of homologous membranes to the ER membrane is essential for the maintenance of this network structure. In yeast, ER membrane fusion is mediated by Sey1p, whose paralogues function distinctly in different species. In this study, we investigated the biological functions of MoSEY1 in the devastating rice blast fungus Magnaporthe oryzae by functional genomic approach. Compared to wild type, deletion of MoSEY1 considerably decreased the growth and conidia production of M. oryzae. Additionally, the absence of MoSEY1 delayed appressorium formation and invasive hyphae growth. The appressorium function was also impaired in Delta Mosey1 mutant. Subcellular localization analysis revealed that MoSey1 is localized at the ER. The Delta Mosey1 mutant showed augmented sensitivity to ER stress. Additionally, we found that MoSey1 regulated the unfolded protein response, autophagy, and protein secretion in M. oryzae. In conclusion, our study unveiled the involvement of MoSey1 in the development, pathogenesis, and ER functions in M. oryzae.
Keyword ：
Endoplasmic reticulum Endoplasmic reticulum Magnaporthe oryzae Magnaporthe oryzae MoSey1 MoSey1 Pathogenicity Pathogenicity Protein secretion Protein secretion
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| GB/T 7714 | Yang, Zifeng , Li, Meiqin , Huang, Linwan et al. MoSey1 regulates the unfolded protein response, appressorium development, and pathogenicity of Magnaporthe oryzae [J]. | PHYTOPATHOLOGY RESEARCH , 2024 , 6 (1) . |
| MLA | Yang, Zifeng et al. "MoSey1 regulates the unfolded protein response, appressorium development, and pathogenicity of Magnaporthe oryzae" . | PHYTOPATHOLOGY RESEARCH 6 . 1 (2024) . |
| APA | Yang, Zifeng , Li, Meiqin , Huang, Linwan , Chen, Xinru , Weng, Shuning , Jules, Biregeya et al. MoSey1 regulates the unfolded protein response, appressorium development, and pathogenicity of Magnaporthe oryzae . | PHYTOPATHOLOGY RESEARCH , 2024 , 6 (1) . |
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氨肽酶是一类可以选择性剪切氨基酸残基的蛋白酶，液泡氨肽酶Y能够调控碱性氨基酸残基的剪切以及液泡的活性。本研究通过同源性比对获得酵母液泡氨肽酶Y在稻瘟病菌中的同源基因PoAPE3,利用基因敲除法获得PoAPE3基因敲除突变体(ΔPoAPE3),并构建回补体菌株进行生物学功能分析。表型测定结果表明，PoAPE3基因敲除突变体在产孢量、孢子萌发率、致病性、侵染过程等方面与野生型均无明显差异。突变体在完全培养基(CM)和稻秆培养基(SDC)上生长速率下降，且在应对胁迫剂H_2O_2时表现为更耐受，应对刚果红时表现为更敏感。以上结果表明PoAPE3参与调控稻瘟病菌的营养生长和对外界环境胁迫的应答过程。
Keyword ：
功能分析 功能分析 基因敲除 基因敲除 氨肽酶Y 氨肽酶Y 稻瘟病菌 稻瘟病菌
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| GB/T 7714 | 杨子锋 , 黄琳婉 , 翁书凝 et al. 液泡氨肽酶Y编码基因PoAPE3在稻瘟病菌中的生物学功能研究 [J]. | 植物保护 , 2024 , 50 (03) : 37-45 . |
| MLA | 杨子锋 et al. "液泡氨肽酶Y编码基因PoAPE3在稻瘟病菌中的生物学功能研究" . | 植物保护 50 . 03 (2024) : 37-45 . |
| APA | 杨子锋 , 黄琳婉 , 翁书凝 , 徐虎啸 , 王聪勐 , 赵爱玉 et al. 液泡氨肽酶Y编码基因PoAPE3在稻瘟病菌中的生物学功能研究 . | 植物保护 , 2024 , 50 (03) , 37-45 . |
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本发明公开了一种水稻基因OsCAD8B，其核苷酸序列如SEQ ID NO.1所示，其编码的蛋白的氨基酸序列如SEQ ID NO.2所示，以水稻基因OsCAD8B开放阅读框和氨基酸序列为基础，利用CRISPR/Cas9基因编辑技术，构建水稻基因编辑载体OsCAD8B‑pYLCRISPR/Cas9Pubi‑H，通过农杆菌介导的转化方法，将OsCAD8B‑pYLCRISPR/Cas9Pubi‑H基因编辑载体导入水稻愈伤组织中，获得相对应的转基因株系。本发明的优点：通过OsCAD8B基因编辑进行作物遗传改良，显著提高了抗稻瘟的能力；进一步对野生型和Oscad8B‑ko水稻植株进行稻瘟病菌接种实验，结果显示敲除OsCAD8B提高了水稻植株对稻瘟病菌的抗性，且不影响水稻形态，对种子的千粒重影响较小。
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| GB/T 7714 | 陈小敏 , 樊嘉鑫 , 黄琳婉 et al. 一种水稻基因OsCAD8B及其应用 : CN202410278043.X[P]. | 2024-03-12 . |
| MLA | 陈小敏 et al. "一种水稻基因OsCAD8B及其应用" : CN202410278043.X. | 2024-03-12 . |
| APA | 陈小敏 , 樊嘉鑫 , 黄琳婉 , 张君 , 汤蔚 , 王宗华 . 一种水稻基因OsCAD8B及其应用 : CN202410278043.X. | 2024-03-12 . |
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