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学者姓名：宾羽
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Abstract ：
本发明属于植物病害防治技术领域，具体涉及一种柑橘木虱源的抗菌肽DcAMP2的制备和应用。所述抗菌肽DcAMP2的氨基酸序列为SEQ ID NO.1，编码所述抗菌肽DcAMP2的基因的核苷酸序列为SEQ ID NO.2。本发明采用原核表达质粒构建包含编码抗菌肽DcAMP2的基因的重组质粒，实现了抗菌肽DcAMP2的原核表达，方法简单易行。原核表达获得的抗菌肽DcAMP2在纯化后对柑橘黄龙病菌近缘细菌费氏中华根瘤菌具有明显的抑菌效果，可显著降低带有柑橘黄龙病菌的甜橙树中的柑橘黄龙病菌滴度。
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| GB/T 7714 | 魏太云 , 李猷 , 陈倩 et al. 一种柑橘木虱源的抗菌肽DcAMP2的制备和应用 : CN202510699670.5[P]. | 2025-05-28 . |
| MLA | 魏太云 et al. "一种柑橘木虱源的抗菌肽DcAMP2的制备和应用" : CN202510699670.5. | 2025-05-28 . |
| APA | 魏太云 , 李猷 , 陈倩 , 宾羽 , 杜宇 . 一种柑橘木虱源的抗菌肽DcAMP2的制备和应用 : CN202510699670.5. | 2025-05-28 . |
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本发明公开了一种抗病毒肽及其防治水稻病毒病的应用。本发明打破了从土壤微生物寻找抗病毒肽的传统思路，创新性地从动物生物体内寻找可能的抗病毒肽；经一系列前期研究，最终确定了8条抗病毒肽序列，经验证，本发明提供的天然来源的抗病毒肽均对水稻病毒病有显著的防治作用，为水稻病毒病的防治提供了一种新的农药，对环境无害，效果非常明显。
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| GB/T 7714 | 魏太云 , 杜宇 , 李猷 et al. 抗病毒肽及其防治水稻病毒病的应用 : CN202510341064.6[P]. | 2025-03-21 . |
| MLA | 魏太云 et al. "抗病毒肽及其防治水稻病毒病的应用" : CN202510341064.6. | 2025-03-21 . |
| APA | 魏太云 , 杜宇 , 李猷 , 宾羽 , 陈倩 . 抗病毒肽及其防治水稻病毒病的应用 : CN202510341064.6. | 2025-03-21 . |
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Phloem-inhabiting unculturable bacterial pathogens are persistently transmitted by insect vectors. However, how they evade insect immune responses to ensure persistent transmission remains unknown. The important melanization immune response in insects is triggered by cleavage of prophenoloxidase (PPO) into active phenoloxidase (PO) via clip-domain serine proteases (CLIPs). Here, we demonstrate that infection of Candidatus Liberibacter asiaticus (CLas) in psyllid vectors activates the peptidoglycan recognition protein (PGRP)-CLIP1-CLIP4-PPO-PO signaling cascade to induce a mild melanization response, ensuring persistent CLas infection without causing significant insect fitness costs to the insect. A CLas-encoded secretory protein, SDE3230, directly interacts with PGRP and suppresses its activity in transducing this signaling cascade. CLIP4 cleaves PPO between arginine 125 and methionine 126 residues to active PO to induce melanization, and this cleavage pattern in psyllid is distinct from other insects. However, SDE3230 competitively binds to this cleavage site of PPO with CLIP4, thereby suppressing PPO effective cleavage. Collectively, these findings reveal the dual role of SDE3230 in facilitating the mild melanization response, benefitting persistent CLas infection and insect fitness.IMPORTANCEPsyllid-borne huanglongbing is the most destructive citrus disease worldwide, causing billions of dollars in annual production losses and threatening the entire citrus industry. Currently, the mechanism by which the causal agent Candidatus Liberibacter asiaticus (CLas) antagonizes psyllid innate immune responses to facilitate its coexistence with psyllid vectors is still unknown. Here, we report that CLas exploits the highly expressed secretory protein SDE3230 in psyllids to suppress the important melanization immune response in hemolymph via inhibiting the pattern recognition receptor PGRP activity and the cleavage of prophenoloxidase into active phenoloxidase by clip-domain serine proteases. The pattern of PPO cleavage is novel, and this process ultimately ensures persistent CLas infection and insect fitness. Our findings provide insights into how CLas has evolved novel strategies to evade the insect melanization response, thereby facilitating persistent CLas transmission.
Keyword ：
Candidatus Liberibacter asiaticus Candidatus Liberibacter asiaticus huanglongbing huanglongbing melanization melanization persistent infection persistent infection psyllid psyllid secretory protein secretory protein
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| GB/T 7714 | Li, You , Du, Yu , Ren, Dongsheng et al. Unculturable bacteria exploit a secretory protein to antagonize insect melanization for persistent infection [J]. | MBIO , 2025 , 16 (10) . |
| MLA | Li, You et al. "Unculturable bacteria exploit a secretory protein to antagonize insect melanization for persistent infection" . | MBIO 16 . 10 (2025) . |
| APA | Li, You , Du, Yu , Ren, Dongsheng , Bin, Yu , Chen, Qian , Wei, Taiyun . Unculturable bacteria exploit a secretory protein to antagonize insect melanization for persistent infection . | MBIO , 2025 , 16 (10) . |
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The Asian citrus psyllid (Diaphorina citri) is the primary vector for Candidatus Liberibacter asiaticus (CLas), which is the causative agent of Huanglongbing (HLB). In recent years, increasing attention has been given to the endosymbiotic viruses harbored within D. citri, particularly Diaphorina citri picorna-like virus (DcPLV). However, the epidemiological distribution, biological effects, and role of DcPLV in pathogen transmission remain largely uncharacterized. This study investigates the field prevalence, transmission modes, and biological impacts of DcPLV in D. citri populations collected from 17 regions in China between 2023 and 2024. Detection of DcPLV revealed a widespread distribution, with an average infection rate of 35.18 %. Temporal monitoring in Gutian orchard further showed that DcPLV maintains consistently high infection levels throughout the year. The analysis of transmission showed that DcPLV is transmitted vertically from parents to offspring and horizontally from plant hosts. Biological assessments showed no significant effects of DcPLV on reproduction and adult longevity. Additionally, tissue-specific RT-qPCR revealed virus localization in the midgut and salivary glands, with viral accumulation progressively increasing during nymphal development and continuing into teneral adult. Furthermore, results demonstrated that DcPLV infection significantly enhanced CLas acquisition and accumulation in D. citri, which may contribute to increased vector competence. These findings provide novel insights on the interactions between DcPLV, the D. citri, and CLas.
Keyword ：
Citrus greening Citrus greening Insect-specific viruses Insect-specific viruses Vertical transmission Vertical transmission Viral symbionts Viral symbionts
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| GB/T 7714 | Wang, Shuai , Younas, Muhammad , Bao, Chengtao et al. Functional characteristics of endosymbiotic picornia-like virus in Diaphorina citri [J]. | VIRUS RESEARCH , 2025 , 360 . |
| MLA | Wang, Shuai et al. "Functional characteristics of endosymbiotic picornia-like virus in Diaphorina citri" . | VIRUS RESEARCH 360 (2025) . |
| APA | Wang, Shuai , Younas, Muhammad , Bao, Chengtao , Xiao, Yuxi , Ul Haq, Inzamam , Lu, Jia et al. Functional characteristics of endosymbiotic picornia-like virus in Diaphorina citri . | VIRUS RESEARCH , 2025 , 360 . |
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Huanglongbing (HLB) is a destructive disease that threatens the citrus industry worldwide. It is associated with "Candidatus Liberibacter asiaticus" (CLas) and vectored by Diaphorina citri, known as the Asian citrus psyllid (ACP). Australian finger lime (Citrus australasica), a promising candidate for HLB tolerance, was examined in response to CLas-infected ACPs through comprehensive transcriptomic profiling to identify key antimicrobial peptides (AMPs) and immune-related pathways. GO, KEGG, and genome enrichment analysis found that most DEGs were enriched in metabolic pathways, biosynthesis of secondary metabolites, and hormone signal transduction. Furthermore, immune pathways associated with pathogen recognition and defense mechanisms were characterized. DEGs were screened and confirmed through quantitative real-time qPCR (RT-qPCR). By analyzing gene expression in C. australasica, we identified several potential AMPs with significant antimicrobial properties. CaAMP1, CaAMP2, CaAMP4, and CaShepharin exhibited strong inhibitory effects against multiple bacterial suspensions with minimum EC50 values. Similarly, CaAMP4 also caused a significant reduction in CLas titer through Agrobacterium-mediated transient expression. This study provides a novel perspective for understanding the transcriptomic information on C. australasica in response to CLas-infected ACPs. These findings offer valuable insights into the development of AMP-based, environmentally sustainable strategies for managing HLB and enhancing citrus plant resilience.
Keyword ：
Asian citrus psyllid(Diaphorina citri) Asian citrus psyllid(Diaphorina citri) Australian finger lime Australian finger lime defense mechanisms defense mechanisms gene expression gene expression HLB HLB immune pathways immune pathways transcriptomicinformation transcriptomicinformation
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| GB/T 7714 | Younas, Muhammad , Wang, Chuang , Hassan, Mohamed Fathi et al. Comprehensive Transcriptomic Profiling of Citrus australasica Unveils Antimicrobial Peptides and Immune Pathways for Huanglongbing Tolerance [J]. | JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY , 2025 , 73 (27) : 16847-16859 . |
| MLA | Younas, Muhammad et al. "Comprehensive Transcriptomic Profiling of Citrus australasica Unveils Antimicrobial Peptides and Immune Pathways for Huanglongbing Tolerance" . | JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY 73 . 27 (2025) : 16847-16859 . |
| APA | Younas, Muhammad , Wang, Chuang , Hassan, Mohamed Fathi , Li, Wenqing , Zheng, Zheng , Bin, Yu et al. Comprehensive Transcriptomic Profiling of Citrus australasica Unveils Antimicrobial Peptides and Immune Pathways for Huanglongbing Tolerance . | JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY , 2025 , 73 (27) , 16847-16859 . |
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【目的】水稻矮缩病毒（rice dwarf virus,RDV）引起的水稻矮缩病是危害中国南方水稻生产的重要病害。结合前人的研究基础，利用分子生物学等手段，解析肽聚糖识别蛋白6(PGRP6)参与调控媒介昆虫黑尾叶蝉（Nephotettix cincticeps）体内共生菌携带RDV经卵垂直传播的机制，为水稻病毒病的生物防治提供理论依据。【方法】利用酵母双杂交技术筛选与经卵传播相关蛋白（prp、Nasuia porin、P8）均互作的PGRP，并通过GST pull-down进一步验证其互作关系；利用实时荧光定量PCR技术测定相关基因在无毒和带毒虫体内的表达差异，并分析干扰PGRP6、prp后对共生菌增殖的影响，同时通过Western blot检测对昆虫体内共生菌膜蛋白积累的影响；分析抑制PGRP6表达对叶蝉存活率、子代带毒率、带毒子代的病毒核酸水平的影响，并利用荧光原位杂交（FISH）和免疫荧光标记观察叶蝉卵巢内prp、PGRP6、Nasuia及RDV的分布情况；通过抑菌圈和肽聚糖降解实验验证PGRP6的功能。【结果】黑尾叶蝉的PGRP6与prp、Nasuia porin、P8均存在互作；PGRP6和prp在RDV侵染的黑尾叶蝉中的表达量均显著升高；抑制PGRP6表达可促进共生菌Nasuia和Sulcia的增殖，而抑制prp降低了共生菌的增殖，其功能与PGRP6相反，当两者同时被干扰后对共生菌的增殖无显著影响；ds PGRP6处理导致黑尾叶蝉存活率显著降低，子代带毒率增加，子代昆虫体内的病毒量明显高于对照组；免疫荧光标记发现PGRP6与prp、Nasuia、RDV均在卵巢组织中分布并发生共定位，prp和Nasuia是包裹的位置关系；PGPR6存在抑菌功能，可发挥抑制共生菌增殖的作用。【结论】PGRP6通过抑制黑尾叶蝉体内共生菌的增殖维持卵巢组织内的稳态，其与prp的功能相拮抗，其既确保了黑尾叶蝉的生命活力，又调控了共生菌携带RDV的垂直传播。
Keyword ：
共生菌 共生菌 卵巢 卵巢 富脯氨酸蛋白 富脯氨酸蛋白 抑菌 抑菌 水稻矮缩病毒 水稻矮缩病毒 经卵传播 经卵传播 肽聚糖识别蛋白 肽聚糖识别蛋白 黑尾叶蝉 黑尾叶蝉
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| GB/T 7714 | 徐元元 , 贾东升 , 宾羽 et al. 黑尾叶蝉PGRP6负调控共生菌抑制水稻矮缩病毒的经卵传播 [J]. | 中国农业科学 , 2025 , 58 (05) : 907-917 . |
| MLA | 徐元元 et al. "黑尾叶蝉PGRP6负调控共生菌抑制水稻矮缩病毒的经卵传播" . | 中国农业科学 58 . 05 (2025) : 907-917 . |
| APA | 徐元元 , 贾东升 , 宾羽 , 魏太云 . 黑尾叶蝉PGRP6负调控共生菌抑制水稻矮缩病毒的经卵传播 . | 中国农业科学 , 2025 , 58 (05) , 907-917 . |
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【目的】黏液样蛋白（mucin-like protein）参与半翅目昆虫取食过程中唾液鞘的形成。本研究旨在制备柑橘木虱（Diaphorina citri）黏液样蛋白DcMucin-like特异性抗体并对木虱取食孔进行免疫荧光标记，为DcMucin-like生物学功能研究提供依据。【方法】通过解剖柑橘木虱唾液腺、中肠、卵巢和精巢组织，根据木虱DcMcuin-like序列设计特异性引物，利用实时荧光定量PCR检测分析各组织中DcMcuin-like在转录水平的差异；扩增无信号肽的DcMucin-like序列并插入pET-28a中构建重组质粒，测序验证后转化Rosetta表达菌株，以终浓度0.5 mmol·L-1 IPTG在37℃下振荡培养8 h诱导重组蛋白表达，SDS-PAGE凝胶电泳检测重组蛋白表达情况；扩大培养后裂解菌体，裂解液上清经Ni-NTA纯化作为抗原免疫家兔5次，纯化血清IgG获得DcMucin-like多克隆抗体，Western blot检测抗体特异性；利用实时荧光定量PCR和Western blot比较健康木虱和感染黄龙病菌（Candidatus Liberibacter asiaticus,CLas）木虱中DcMucin-like在转录水平和蛋白水平的差异；利用交联荧光素（FITC）的DcMucin-like多克隆抗体标记木虱取食后的柑橘叶片，在共聚焦显微镜下观察柑橘木虱取食孔和唾液鞘。【结果】实时荧光定量PCR检测显示DcMcuin-like在木虱唾液腺中大量表达，且表达量显著高于中肠、卵巢和精巢组织。携带有pET28a-DcMucin的Rosetta表达菌株经IPTG诱导后在菌体裂解液的上清中获得大量表达的重组蛋白。重组蛋白免疫家兔后取抗血清纯化IgG,Western blot结果表明获得特异性好的DcMucin-like多克隆抗体。实时荧光定量PCR和Western blot分析结果显示DcMucin-like在木虱响应黄龙病菌侵染时上调表达。DcMucin-like(FITC)荧光抗体标记木虱取食后的柑橘叶片组织切片，共聚焦显微镜下观察到木虱的取食孔，并在取食孔周围检测到特异的FITC荧光信号，表明DcMucin-like随木虱取食释放到植株组织中参与唾液鞘形成。【结论】DcMucin-like在柑橘木虱唾液腺中高表达并响应黄龙病菌侵染上调表达；成功制备了特异性好的柑橘木虱DcMucin-l...
Keyword ：
DcMucin-like DcMucin-like 免疫荧光 免疫荧光 唾液鞘蛋白 唾液鞘蛋白 抗体制备 抗体制备 柑橘木虱 柑橘木虱
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| GB/T 7714 | 蒋丽琴 , 苏巧灵 , 李猷 et al. 柑橘木虱黏液样蛋白DcMucin-like抗体制备及应用 [J]. | 中国农业科学 , 2025 , 58 (02) : 281-290 . |
| MLA | 蒋丽琴 et al. "柑橘木虱黏液样蛋白DcMucin-like抗体制备及应用" . | 中国农业科学 58 . 02 (2025) : 281-290 . |
| APA | 蒋丽琴 , 苏巧灵 , 李猷 , 魏太云 , 宾羽 . 柑橘木虱黏液样蛋白DcMucin-like抗体制备及应用 . | 中国农业科学 , 2025 , 58 (02) , 281-290 . |
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本发明属于植物病害防治技术领域，具体涉及一种柑橘木虱源的抗菌肽DcAMP1的制备和应用。所述抗菌肽DcAMP1的氨基酸序列为SEQ ID NO.1，编码所述抗菌肽DcAMP1的基因的核苷酸序列为SEQ ID NO.2。本发明采用原核表达质粒构建包含编码抗菌肽DcAMP1的基因的重组质粒，实现了抗菌肽DcAMP1的原核表达，方法简单易行。原核表达获得的抗菌肽DcAMP1在纯化后对柑橘黄龙病菌近缘细菌费氏中华根瘤菌具有明显的抑菌效果，可显著降低带有柑橘黄龙病菌的甜橙树中的柑橘黄龙病菌滴度。
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| GB/T 7714 | 魏太云 , 李猷 , 宾羽 et al. 一种柑橘木虱源的抗菌肽DcAMP1的制备和应用 : CN202510702629.9[P]. | 2025-05-28 . |
| MLA | 魏太云 et al. "一种柑橘木虱源的抗菌肽DcAMP1的制备和应用" : CN202510702629.9. | 2025-05-28 . |
| APA | 魏太云 , 李猷 , 宾羽 , 陈倩 , 彭洪艳 . 一种柑橘木虱源的抗菌肽DcAMP1的制备和应用 : CN202510702629.9. | 2025-05-28 . |
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本发明公开了一种柑橘木虱唾液蛋白DcMucin的抗体及其制备方法和应用。本发明通过DcMucin‑like多克隆抗体的制备，首次验证了DcMucin‑like是柑橘木虱的唾液蛋白，并随木虱取食释放到植株组织中参与唾液鞘形成。本发明弥补了木虱唾液蛋白抗体制备和研究的空白，制备的柑橘木虱唾液蛋白多克隆抗体对DcMucin重组蛋白具有良好的特异性，基于该抗体免疫荧光标记观察木虱唾液鞘，为进一步研究DcMucin在黄龙病菌‑木虱‑柑橘三者互作中的生物学功能打下基础，为进一步获得柑橘木虱绿色防控特异性靶标奠定基础。
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| GB/T 7714 | 宾羽 , 魏太云 , 蒋丽琴 et al. 柑橘木虱唾液蛋白DcMucin的抗体及其制备方法和应用 : CN202411432625.5[P]. | 2024-10-14 . |
| MLA | 宾羽 et al. "柑橘木虱唾液蛋白DcMucin的抗体及其制备方法和应用" : CN202411432625.5. | 2024-10-14 . |
| APA | 宾羽 , 魏太云 , 蒋丽琴 , 李猷 . 柑橘木虱唾液蛋白DcMucin的抗体及其制备方法和应用 : CN202411432625.5. | 2024-10-14 . |
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