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Engineered biocontainable RNA virus vectors for non-transgenic genome editing across crop species and genotypes

Molecular Plant 2023年第3期16卷 616-631页

作者： Qian Liu Chenglu Zhao Kai Sun Yinlu Deng Zhenghe Li State key laboratory of Rice biology Institute of BiotechnologyZhejang UniversityHangzhouChina Ministry of Agricutture key laboratory of Molecular biology of crop pathogens and Insect Pests Zhejiang UniversityHangzhouChina key laboratory of biology of crop pathogens and insects of zhejang province Zhejang UniversityHangzhouChina

CRISPR/Cas genome-editing tools provide unprecedented opportunities for basic plant biology research and crop ***,the lack of robust delivery methods has limited the widespread adoption of these revolutionary technologies in plant ***,we report an efficient,non-transgenic CRISPR/Cas delivery platform based on the engineered tomato spotted wilt virus(TSWV),an RNA virus with a host range of over 1000 plant *** eliminated viral elements essential for insect transmission to liberate genome space for accommodating large genetic cargoes without sacrificing the ability to infect plant *** resulting non-insect-transmissible viral vectors enabled effective and stable in planta delivery of Cas12a and Cas9 nucleases as well as adenine and cytosine base *** systemically infected plant tissues,the deconstructed TSWV-derived vectors induced efficient somatic gene mutations and base conversions in multiple crop species with little genotype *** with heritable,bi-allelic mutations could be readily regenerated by culturing the virus-infected tissues in vitro without antibiotic ***,we showed that antiviral treatment with ribavirin during tissue culture cleared the viral vectors in 100%of regenerated plants and further augmented the recovery of heritable *** many plants are recalcitrant to stable transformation,the viral delivery system developed in this work provides a promising tool to overcome gene delivery bottlenecks for genome editing in various crop species and elite varieties.

关键词： genome editing CRISPR-Cas base editor RNA virus vector tomato spotted wilt virus bunyavirus delivery non-transgenic

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FoxO is required for optimal fitness of the migratory brown planthopper, Nilaparvata lugens (Hemiptera: Delphacidae)

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Insect Science 2023年第5期30卷 1352-1362页

作者： Jin-Li Zhang Ke-Liang Liu Xin-Yu Cai Xin-Yang Liu Hai-Jun Xu State key laboratory of Rice biology Key Laboratory of Biology of Crop Pathogens and Insects of Zhejiang ProvinceInstitute of Insect SciencesCollege of Agriculture and BiotechnologyZhejiang UniversityHangzhouChina

The forkhead box O(FoxO)protein is the main transcriptional effector downstream of the insulin/insulin-like signaling pathway and regulates many developmental and physiological *** insects with loss-of-function mutations in FoxO exhibit phenotypes distinct from those of hemimetabolous insects in which RNA interference was *** the functional importance of FoxO,whether hemimetabolous insects share an evolutionally conserved function of FoxO with holometabolous insects remains to be *** used the clustered regularly interspaced short palindromic repeats/CRISPR-associated 9(CRISPR/Cas9)genome editing-system to establish a homozygous FoxO-null mutant(NlFoxO^(4E))of the wing-dimorphic brown planthopper(BPH)Nilaparvata lugens,an economically important insect pest of rice *** phenotypes of NlFoxO^(4E) mutants included extended nymphal duration,shortened lifespan,reduced reproduction,and decreased stress *** addition,depletion of NlFoxO promoted cell proliferation in wing buds and led to 100%long-winged morphs,in stark contrast to short-winged wild-type *** findings indicate that NlFoxO is highly functionally conserved with its counterpart in holometabolous insects,and is required for optimal fitness of *** insights from FoxO studies may facilitate the identification of potential target genes for BPH control applications.

关键词： cell proliferation CRISPR/Cas9 FoxO longevity planthopper reproduction

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Moth sex pheromones affect interspecific competition among sympatric species and possibly population distribution by modulating pre-mating behavior

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Insect Science 2023年第2期30卷 501-516页

作者： Jianjun Cheng Qinghua Chen Qianshuang Guo Yongjun Du Ministry of Agriculture key Lab of Molecular biology of crop pathogens and insects Key Laboratory of Biology of Crop Pathogens and Insects of Zhejiang ProvinceZhejiang UniversityHangzhouChina Institute of Plant Protection Sichuan Academy of Agricultural Sciences/Key Laboratory of Integrated Pest Management on Crops in SouthwestMinistry of AgricultureChengduChina

Premating behaviors mediated by pheromones play pivotal roles in animal mating *** natural populations of the striped stem borer Chilo suppressalis and the rice leaf roller Cnaphalocrocis medinalis in the rice field habitat,we discovered that Z11-16:Ald,a major component of the *** pheromone,modulated the premating behavior of ***.Z11-16:Ald evoked a strong olfactory response in male antennae and strongly inhibited the sex pheromone trapping of male *** in the *** functions of three *** sex pheromone receptor genes(CmedPR1–3)were verified through heterologous expression in Xenopus ***1 responded to Z11-18:OH and Z11-18:Ald,as well as the interspecific pheromone compound Z11-16:Ac of sympatric species;CmedPR2 responded to Z13-18:OH and Z13-18:Ald,as well as the sex pheromone compounds Z11-16:Ald and Z9-16:Ald of sympatric species;and CmedPR3 responded to Z11-18:OH and Z13-18:OH,as well as the interspecific pheromones Z11-16:OH,Z9-16:Ald,Z11-16:Ac,and Z11-16:Ald of sympatric ***,CmedPR2 and CmedPR3 share the ligand Z11-16:Ald,which is not a component of the *** sex ***,the sex pheromones of interspecific species affected the input of neural signals by stimulating the sex pheromone receptors on the antennae of male *** moths,thereby inhibiting the olfactory responses of the male moths to the sex *** results demonstrate chemical communication among sympatric species in the rice field habitat,the recognition of intra-and interspecific sex pheromones by olfactory receptors,and how insect premating behaviors are modulated to possibly affect resource partitioning.

关键词： Chilo suppressalis Cnaphalocrocis medinalis premating sex pheromone sex pheromone receptors two-electrode voltage-clamp

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An endoparasitoid uses its egg surface proteins to regulate its host immune response

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Insect Science 2022年第4期29卷 1030-1046页

作者： Zi-Wen Teng Hui-Zi Wu Xin-Hai Ye Qi Fang Hong-Xu Zhou Gong-Yin Ye China-Australia Cooperation Base of crop Health and Invasive Species China-Australia Joint Institute of Agricultural and Environmental HealthCollege of Plant Health and MedicineQingdao Agricultural UniversityQingdaoShandong ProvinceChina State key laboratory of Rice biology Ministry of Agricultural and Rural Affairs Key Laboratory of Molecular Biology of Crop Pathogens and InsectsKey Laboratory of Biology of Crop Pathogens and Insects of Zhejiang ProvinceInstitute of Insect SciencesZhejiang UniversityHangzhouChina

With proteomic analysis,we identified 379 egg surface proteins from an endoparasitoid,Cotesia *** containing conserved enzymatic domains constitute a large proportion of egg surface *** proteins,such as superoxidase dismutase,homolog of *** 32-kDa protein,and immunoevasive protein-2A,are classical parasitism factors that have known functions in host immunity *** assays revealed that a novel egg surface protein,*** egg surface serpin domain-containing protein had the same function as a *** venom serpin,as both suppressed host melanization in a dose-dependent *** egg surface serpin domain-containing protein is mainly transcribed in *** oocytes with follicular cells,and it is located on both the anterior and posterior sides of the mature egg ***,we used LC-MS/MS to identify 586 binding proteins sourced from *** plasma located on the eggshell surface of ***,which included some immunity-related *** results not only indicate that *** uses its egg surface proteins to reduce the immune response of its host but also imply that endoparasitoid egg surface proteins might be a new parasitism factor involved in host immune regulation.

关键词： egg surface protein immune regulation melanization parasitism factor parasitoid wasp

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OsATL32 ubiquitinates the reactive oxygen species-producing OsRac5-OsRbohB module to suppress rice immunity

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Journal of Integrative Plant biology 2024年第7期66卷 1459-1480页

作者： Yuqing Yan Hui Wang Yan Bi Jiajing Wang Muhammad Noman Dayong Li Fengming Song National key laboratory for Rice biology and Breeding Institute of BiotechnologyZhejiang UniversityHangzhou 310058China Ministry of Agriculture key laboratory of Molecular biology of crop pathogens and Insect Pests Institute of BiotechnologyZhejiang UniversityHangzhou 310058China key laboratory of biology of crop pathogens and insects of Zhejiang province Institute of BiotechnologyZhejiang UniversityHangzhou 310058China

Ubiquitination-mediated protein degradation is integral to plant immunity,with E3 ubiquitin ligases acting as key factors in this ***,we report the functions of OsATL32,a plasma membrane-localized Arabidopsis Tóxicos En Levadura(ATL)-type E3 ubiquitin ligase,in rice(Oryza sativa)immunity and its associated regulatory *** found that the expression of OsATL32 is downregulated in both compatible and incompatible interactions between rice and the rice blast fungus Magnaporthe *** OsATL32 protein level declines in response to infection by a compatible *** strain or to chitin ***32 negatively regulates rice resistance to blast and bacterial leaf blight diseases,as well as chitin-triggered *** and genetic studies revealed that OsATL32 suppresses pathogen-induced reactive oxygen species(ROS)accumulation by mediating ubiquitination and degradation of the ROS-producing OsRac5–OsRbohB module,which enhances rice immunity against *** protein phosphatase PHOSPHATASE AND TENSIN HOMOLOG enhances rice blast resistance by dephosphorylating OsATL32 and promoting its degradation,preventing its negative effect on rice *** study provides insights into the molecular mechanism by which the E3 ligase OsATL32 targets a ROS-producing module to undermine rice immunity.

关键词： E3 ligase OsATL32 OsRac5-OsRbohB module phosphatase OsPTEN reactive oxygen species rice immunity

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Synthesis and antifungal activity of trichodermin derivatives

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Chinese Chemical Letters 2010年第9期21卷 1037-1040页

作者： Jing Li Cheng Yong Zhou Jin Hao Zhao Chu Long Zhang Fu Cheng Lin Ministry of Agriculture key laboratory of Molecular biology of crop pathogens and insects Institute of Pesticide and Environmental Toxicology Zhejiang University Hangzhou 310029 China

A series of derivatives were synthesized from trichodermin（1） which was an antifungal metabolite produced by Trichoderma taxi *** structures were confirmed by ~1H NMR,MS *** antifungal activities were evaluated in *** preliminary structure activity relationships（SAR） results indicated that the double bond,epoxide moiety and ester group were main pharmacophore elements,the stereochemistry of C4 position played a key role as well,and the compounds 1e-1g displayed stronger antifungal activity against Magnaporthe grisea than 1.

关键词： Trichodermin derivatives Synthesis Antifungal activity

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Potato insect pest management

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Journal of Integrative Agriculture 2020年第2期19卷 311-315页

作者： GAO Yu-lin ZHOU Wen-wu State key laboratory for biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193P.R.China key laboratory of Molecular biology of crop pathogens and insects Ministry of Agriculture/Key Laboratory of Biology of Crop Pathogens and Insects of Zhejiang Province/State Key Laboratory of Rice Biology Institute of Insect SciencesZhejiang UniversityHangzhou 310058P.R.China

Having an estimated production of 388 million tons in 2017(FAO 2019),potato is one of the world's most important food crops,along with rice,wheat,and *** plants can adapt to different agricultural environm ents and are widely grown over many latitudes and elevations(Douches et al.2004).In addition to its consumption in Europe,North America and countries of the former Soviet Union,potato is now also popularized in Africa and Asia,especially in the developing countries including China and *** 2015,China has been boosting potato production to become the fourth major crop produced in the country following rice,wheat and maize(Zhang etal.2017;Xu et al.2019).

关键词： crops boosting latitude

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Gene expression profiling of the whitefly （Bemisia tabaci） Middle East- Asia Minor 1 feeding on healthy and Tomato yellow leaf curl China virus-infected tobacco

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Insect Science 2011年第1期18卷 11-22页

作者： Jun-Min Li Yong-Ming Ruan Fang-Fang Li Shu-Sheng Liu Xiao-Wei Wang Ministry of Agriculture key laboratory of Molecular biology of crop pathogens and insects Institute of Insect Sciences Zhejiang University Hangzhou College of Chemistry and Life Sciences Zhejiang Normal University Jinhua Zhejiang Province China

Begomoviruses are exclusively transmitted by whitefly （Bemisia tabaci） in a circulative, non-propagative manner. The influences of begomoviruses on whitefly vector are complex with both direct and indirect effects. However, the molecular mechanisms underlying these effects are poorly understood and the transcriptional profiles of whitefly on healthy and virus-infected plants have not yet been studied. Using suppression subtractive hybridization, we investigated the differentially expressed genes in whitefly Middle East - Asia Minor 1 feeding on healthy and Tomato yellow leaf curl China virus （TYLCCNV） infected tobacco. From the forward cDNA library, 124 differentially expressed expression sequence tags （ESTs） were obtained which represent up-regulated genes in the whiteflies feeding on the infected tobacco. From the reverse library, 112 ESTs were isolated which represent down-regulated genes. Among the up-regulated genes, we identified several genes that are probably involved in direct interaction between whitefly and TYLCCN~, including a 26/29-kDa proteinase that appears to participate in the elimination of foreign proteins, heparan sulfate proteoglycan which mediates the entry of several viruses into host cells and two genes of Rickettsia-a secondary endosymbiotic bacterium of whitefly. In addition, we identified a number of genes involved in metabolism, transcription and translation which might be the result of indirect effects of TYLCCNV on the whitefly via host plants. Collectively, our results suggest that TYLCCNV-infected tobacco changes the gene expression profiles of whitefly via both direct and indirect interactions. This study revealed a number of genes involved in whitefly-TYLCCNV-tobacco interactions and provided useful information for future study on this complex system.

关键词： begomovirus Bemisia tabaci suppression subtractive hybridization Tomatoyellow leaf curl China virus whitefly

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Crossing experiments and behavioral observations reveal reproductive incompatibility among three putative species of the whitefly Bemisia tabaci

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Insect Science 2010年第6期17卷 508-516页

作者： Peng Wang Yong-Ming Ruan Shu-Sheng Liu Ministry of Agriculture key laboratory of Molecular biology of crop pathogens and insects Institute of Insect Sciences Zhejiang University Hangzhou College of Chemistry and Life Sciences Zhejiang Normal University Jinhua Zhejiang Province China

The whitefly Bemisia tabaci has a global distribution and extensive genetic diversity. Recent phylogenetic analyses as well as crossing experiments suggest that B. tabaci is a complex composed of 〉 20 cryptic species, but more crossing studies are required to examine the reproductive compatibility among the putative species and thus further clarify the systematics of this species complex. We conducted crossing experiments and behavioral observations to investigate the reproductive compatibility between the Mediterranean, Asia II 3, and Asia II 1 putative species ofB. tabaci collected from Zhejiang, China. Female progeny were never produced in inter-species crosses, demonstrating a lack of egg fertilization; while 55%-75% females were produced in all the intra-species treatments. Continuous behavioral observations showed that frequent courtship events occurred in both intra-species treatments and inter-putative species crosses. However, copulation events occurred only in the three intra-species treatments with one exception： that one copulation event occurred between Asia II 3 and Mediterranean in the crosses where two cohorts of females and males of different putative species were enclosed together in a small arena but were not allowed access to their intra-specific mates for a long period of time. These data demonstrated complete reproductive isolation between the Mediterranean, Asia II 3, and Asia II 1 putative species, and further showed that the isolation is due to lack of copulation. Demonstration of reproductive isolation between the Mediterranean and two indigenous putative species from China provides further evidence for the existence of cryptic species within the B. tabaci complex.

关键词： Bemisia tabaci behavioral observations crossing experiments genetic groups putative species reproductive incompatibility

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Role of aminopeptidase N-like in the acquisition of begomoviruses by Bemisia tabaci,the whitefly vector

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Insect Science 2024年第3期31卷 707-719页

作者： Yun-Yun Fan Yao Chi Na Chen Wilmer J.Cuellar Xiao-Wei Wang Ministry of Agriculture key Lab of Molecular biology of crop pathogens and insects Key Laboratory of Biology of Crop Pathogens and Insects of Zhejiang ProvinceInstitute of Insect SciencesZhejiang UniversityHangzhouChina Zhejiang Tianmushan National Nature Reserve Administration HangzhouChina Institute of Plant Protection Liaoning Academy of Agricultural SciencesShenyangChina Virology laboratory Cassava ProgramInternational Center for Tropical Agriculture(CIAT)PalmiraColombia

Sri Lankan cassava mosaic virus(SLCMV)is a prominent causative agent of cassava mosaic disease in Asia and relies on the whitefly Bemisia tabaci cryptic complex for its ***,the molecular mechanisms involved in SLCMV transmission by *** have yet to be *** this study,we identified an aminopeptidase N-like protein(BtAPN)in *** Asia II 1,an efficient vector of SLCMV,which is involved in the SLCMV transmission *** the use of glutathione S-transferase pull-down assay and LC-MS/MS analysis,we demonstrated the interaction between BtAPN and the coat protein(CP)of *** interaction was further confirmed in vitro,and we observed an induction of BtAPN gene expression following SLCMV *** interfering with the function of BtAPN,the quantities of SLCMV were significantly reduced in various parts of *** Asia II 1,including the whole body,midgut,hemolymph,and primary salivary ***,we discovered that BtAPN is conserved in *** Middle East-Asia Minor 1(MEAM1)and interacts with the CP of tomato yellow leaf curl virus(TYLCV),a begomovirus known to cause severe damage to tomato *** BtAPN with antibody led to a significant reduction in the quantities of TYLCV in whitefly whole body and organs/*** results demonstrate that BtAPN plays a generic role in interacting with the CP of begomoviruses and positively regulates their acquisition by the whitefly.

关键词： BtAPN coat protein SLCMV TYLCV virus acquisition whitefly

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