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Puccinia triticina effector protein Pt_21 interacts with wheat thaumatin-like protein TaTLP1 to inhibit its antifungal activity and suppress wheat apoplast immunity

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作物学报:英文版 2023年第5期11卷 1431-1440页

作者： Fei Wang Songsong Shen Zhongchi Cui Shitao Yuan Ping Qu Hui Jia Linshuo Meng Xiaoyu Hao Daqun Liu Lisong Ma Haiyan Wang College of plant Protection Hebei Agricultural University/Technological Innovation Center for Biological Control of Crop Diseases and Insect Pests of Hebei ProvinceBaoding 071001HebeiChina center of Agriculture Through Science and Education Hebei Agricultural UniversityBaoding 071001HebeiChina The State Key Laboratory of North China Crop Improvement and Regulation Hebei Agricultural UniversityBaoding 071001HebeiChina

Puccinia triticina(Pt), as the causal agent of wheat leaf rust, employs a plethora of effector proteins to modulate wheat immunity for successful colonization. Understanding the molecular mechanisms underlying Pt effector-mediated wheat susceptibility remains largely unexplored. In this study, an effector Pt_21 was identified to interact with the apoplast-localized wheat thaumatin-like protein TaTLP1 using a yeast two-hybrid assay and the Pt_21-TaTLP1 interaction was characterized. The interaction between Pt_21 and TaTLP1 was validated by in vivo co-immunoprecipitation assay. A TaTLP1 variant,TaTLP1C71A, that was identified by the site-directed mutagenesis failed to interact with Pt_21. Pt_21was able to suppress Bax-mediated cell death in leaves of Nicotiana benthamiana and inhibit TaTLP1-mediated antifungal activity. Furthermore, infiltration of recombinant protein Pt_21 into leaves of transgenic wheat line overexpressing TaTLP1 enhanced the disease development of leaf rust compared to that in wild-type leaves. These findings demonstrate that Pt_21 suppresses host defense response by directly targeting wheat TaTLP1 and inhibiting its antifungal activity, which broadens our understanding of the molecular mechanisms underlying Pt effector-mediated susceptibility in wheat.

关键词： Wheat Puccinia triticina Effector Thaumatin-like protein Anitifungal activity

Molecular Markers for Leaf Rust Resistance Gene Lr45 in Wheat Based on AFLP Analysis

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Agricultural Sciences in China 2006年第12期5卷 938-943页

作者： ZHANG Na YANG Wen-xiang YAN Hong-fei LIU Da-qun CHU Dong MENG Qing-fang ZHANG Ting Department of plant Pathology College of Plant Protection/Biological Control Center of Plant Diseases and Pests of Hebei Province Agricultural University of Hebei Baoding 071001 P.R.China Agricultural Department of hebei Province Shijiazhuang 050011 P.R. China

Amplified fragment length polymorphism （AFLP） analysis was carried out in Thatcher, near isogenic lines （NILs） canting different genes conferring resistance against wheat leaf rust, and TcLr45 × Thatcher F2 progenies were used to develop markers for Lr45 gene. Sixty AFLP primer combinations were screened and most of them provided clear amplification products, 31 primer combinations displayed polymorphism of TcLr45 in 23 NILs. Two AFLP markers closely linked to the gene Lr45 were acquired： P-AGG/M-GAG261bp, which was found closely linked to the Lr45 locus at a distance of 0.6 cM on one side, and P-ACA/M-GGT105bp, which was found at a distance of 1.3 cM on the other side. The specific hands were cloned and subsequently sequenced. The 261-bp fragment produced by P-AGG/M-GAG showed 86% similarity with the sequence of Vulgate Hort I gene; the 105-bp fragment produced by P-ACA/M-GGT showed 96% similarity with the phosphatidylserine decarboxylase gene of the Triticum monococcum. Both included an open reading frame （ORF）.

关键词： wheat leaf rust AFLP Lr45 gene localization

Species-specific COI primers for rapid identification of a globally significant invasive pest, the cassava mealybug Phenacoccus manihoti Matile-Ferrero

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Journal of Integrative Agriculture 2019年第5期18卷 1042-1049页

作者： WANG Yu-sheng TIAN Hu WAN Fang-hao ZHANG Gui-fen State Key Laboratory for Biology of plant diseases and Insect pests/Key Laboratory of Integrated Pest Management of Crop Ministry of Agriculture/Institute of Plant Protection Chinese Academy of Agricultural Sciences Caofeidian Sub-center of hebei Entry-Exit Inspection and Quarantine Technical center

The globally invasive cassava mealybug Phenacoccus manihoti Matile-Ferrero is a pernicious pest of cassava,and its recent introduction into Asia has raised considerable *** slow or prevent further invasion,an accurate,simple,and developmental-stage-independent detection method for *** is *** the present study,a PCR method based on a species-specific mitochondrial DNA cytochrome oxidase I(SS-COI)marker was developed for rapid identification of *** pair of SS-COI primers(PMSSZW-1F and PMSSZW-1R)was designed based on sequence variations in the COI gene among *** and related mealybug *** of the primer pair was validated on 21 closely related *** tests were performed on four immature developmental stages and female *** tests demonstrated that at the relatively low concentration of(135.2±14.7)pgresuspended DNA,the specific fragment was detected in all ***,the SS-COI primer pair was assayed on three populations of *** from major exporting countries of *** PCR assay was proved to be a rapid,simple,and reliable molecular measure for the identification of *** tool will be useful for quarantine,monitoring,and management of this invasive pest.

关键词： Phenacoccus manihoti cassava mealybug invasive pest molecular identification species-specific COI primers

Systemic acquired resistance, NPR1, and pathogenesis-related genes in wheat and barley

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Journal of Integrative Agriculture 2018年第11期17卷 2468-2477页

作者： WANG Xiao-dong BI Wei-shuai GAO Jing YU Xiu-mei WANG Hai-yan LIU Da-qun College of plant Protection/Biological Control center for plant diseases and plant pests of hebei Hebei Agricultural University Baoding 071000 P.R.China College of Life Sciences Hebei Agricultural University Baoding 071000 P.R.China Graduate School of Chinese Academy of Agricultural Sciences Beijing 100081 P.R.China

In Arabidopsis, systemic acquired resistance （SAR） is established beyond the initial infection by a pathogen or is directly induced by treatment with salicylic acid （SA） or its functional analogs, 2,6-dichloroisonicotinic acid （INA） and benzothiadiazole （BTH）. NPR1 protein is considered the master regulator of SAR in both SA signal sensing and transduction. In wheat （Triticum aesfivum） and barley （Hordeum vulgare）, both pathogen infection and BTH treatment can induce broad-spectrum resistance to various diseases, including powdery mildew, leaf rust, Fusarium head blight, etc. However, three different types of SAR-like responses including acquired resistance （AR）, systemic immunity （SI）, and BTH-induced resistance （BIR） seem to be achieved by activating different gene pathways. Recent research on wheat and barley NPR1 homologs in AR and SI has provided the initial clue for understanding the mechanism of SAR in these two plant species. In this review, the specific features ofAR, Si, and BIR in wheat and barley were summarized and compared with that of SAR in model plants of Arabidopsis and rice. Research updates on downstream genes of SAR, including pathogenesis-related （PR） and BTH-induced genes, were highlighted.

关键词： systemic acquired resistance NPR1 pathogenesis-related genes wheat barley

Identification of the pathogen causing white radish scab in China

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Identification of the pathogen causing white radish scab in ...

Tri-milR29 from Trichoderma breve Targets Chitin Synthase Gene in Botrytis cinerea and its Enhanced Disease Control Efficacy through Nanodelievery-base Application

中国植物病理学会2015年学术年会

作者： YANG Fang-yan YANG De-jie YU Xiu-mei ZHAO Wei-quan LIU Da-qun College of plant Protection Agricultural University of Hebei/Biocontrol Center of Plant Diseases and Plant Pests of Hebei Province

Corky scab-like lesions on white radish(Raphanus sativus) surfaces were observed during a survey of vegetable diseases in Qingyuan county,hebei province of China in September *** incidence was approximately 50 percent of the investigated field(667 m).The lesions occurred mainly on the root *** symptoms varied from slightly raised to pitted corky brown *** determine the causal agent,ten plants were sampled and their roots were surface-disinfected with 0.3%***(approximately 5 mm) were excised from the roots,sliced into 1 mm pieces,plated on oatmeal agar medium(OMA),and incubated at 281 for 5 *** individual actinomycete colonies were transferred onto fresh OMA medium and incubated at 28℃ for ten *** fulfill Koch’s postulates,the spores of the isolates were washed with sterile distilled water from ***(200 ml of 1×10 CFU/ml) was added to each pot(20 cm diameter) 20 days after white radish(***) had been *** isolate was tested in five replicate *** six weeks after inoculation,all roots of the inoculated plants showed scab symptoms,and no symptoms were observed on non-inoculated *** same pathogen was re-isolated from the scab lesions on the *** pathogen(typical strain SAT-1) was identified based on morphological and physiological characterization and 16 S rRNA gene *** strain SAT-1 produced rectiflexuous spore chains on oatmeal agar,had white,smooth surface spores that measured 0.45 to 0.5 x 1.0 to 1.1 μ*** produced diffusible yellow pigments,but did not produce melanin on tyrosine-peptone-yeast extract agar *** SAT-1 grew at pH 4.0 and did not produce hydrogen *** utilized all the International Streptomyces Project(ISP) sugars as single carbon sources,and was not sensitive to phenol(0.1%),crystal violet(0.5 μg/ml),penicillin(10 IU/ml) and streptomycin(20 μg/ml).The strain SAT-1 produced thaxtomin A in oatmeal liquid medium and the txtAB gene fragment was

关键词： white radish scab pathogen Identification Streptomyces acidiscabies

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Tri-milR29 from Trichoderma breve Targets Chitin Synthase Ge...

QTL mapping of adult plant resistance to stripe rust and leaf rust in a Fuyu 3/Zhengzhou 5389 wheat population

中国植物病理学会2024年学术年会

作者：刘震 Raja Asad Ali Khan WANG Rui LI Yuejiao 刘铜 School of Breeding and Multiplication (Sanya Institute of Breeding and Multiplication) Hainan University Key Laboratory of Green Prevention and Control of Tropical diseases and pests Ministry of Education (School of Tropical Agriculture and Forestry)Hainan University Engineering center of Agricultural Microbial Preparation Research and Development of Hainan (Hainan University)

In the process of mycoparasitism,a form of biological control that one fungus attacking another fungus,the exchange of biologically active molecules such as RNAs may occur,playing an important role in *** these molecules,microRNAs(miRNAs) can inhibit translation and regulate gene expression at the post-transcriptional level through complementary base pairing,thereby degrading target mRNAs to prevent ***,a well-known mycoparasitic fungus,is one of the most studied biocontrol *** recent years,small RNAs(named milRNA) have also been discovered in ***,the role of Trichoderma RNAs in their mycoparasitic activity and their targets in pathogenic fungi still needs further *** study verified the cross-species regulatory role of Trichoderma-T069 miRNA,named *** demonstrated that Tri-milR29 from T069 was transferred to ***,a pathogenic fungus,where it targeted the chitin synthase gene,BcCHS7,resulting in a marked decrease in its ***-species transportation of sRNAs may occur through extracellular vesicles(EVs) ***,more research is needed on EVs in *** a series of experiments,we verified that the Tri-milR29 from T069 was transferred to *** through the EVs *** sRNA can effectively prevent and control plant diseases by targeting specific genes in pathogenic microbes through RNA ***,their susceptibility to nuclease degradation,off-target effects,and their low absorption efficiency by target pathogens limit their practical ***,the development of suitable delivery systems to protect and deliver sRNA is *** developed a carbon nanomaterial-based delivery system that enhances the stability and absorption of TrimilR29 by ***,increasing antifungal *** nano-delivered Tri-milR29 significantly lowered the disease incidence caused by *** on various fruits and tomato plants.

关键词： microRNA extracellular vesicles biocontrol mycoparasitism pathogen

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The Crop Journal 2020年第4期8卷 655-665页

作者： Takele Weldu Gebrewahid Peipei Zhang Yue Zhou Xiaocui Yan Xianchun Xia Zhonghu He Daqun Liu Zaifeng Li Department of plant Pathology College of Plant ProtectionHebei Agricultural UniversityBiological Control Center for Plant Diseases and Plant Pests of HebeiBaoding 071001HebeiChina College of Agriculture Aksum University-Shire CampusTigrayEthiopia Baoding University Baoding 071001HebeiChina Institute of Crop Sciences National Wheat Improvement CenterChinese Academy of Agricultural SciencesBeijing 100081China International Maize and Wheat Improvement center(CIMMYT)China Office c/o CAASBeijing 100081China

Stripe or yellow rust(YR)and leaf rust(LR)cause large losses in wheat production *** cultivars curtail the levels of *** present study aimed to identify quantitative trait loci(QTL)for YR and LR resistance in 147 F2:6 recombinant inbred lines(RIL)derived from the cross Fuyu 3/Zhengzhou *** RIL population and parents were genotyped with the Wheat55 K single nucleotide polymorphism(SNP)array and simple sequence repeat(SSR)*** materials were also phenotyped for YR severity at Mianyang in Sichuan province and Baoding in hebei province in the 2015/2016,2016/2017,and 2017/2018 cropping seasons,and LR severity at Zhoukou in Henan province and at Baoding in 2017/*** QTL for YR resistance and five for LR resistance were detected using inclusive composite interval mapping(Ici Mapping).Four of these QTL on chromosomes 1 BL,2 BS,3 AL,and 5 AL conferred resistance to both YR and *** QTL on 1 BL was Lr46/Yr29,and that on 7 BL might be *** QTL on chromosome 2 BS was detected at a similar position to previously detected ***-3 AL/***-3 AL,***-5 AL/***-5 AL,***-7 DL,***-4 BS,***-6 DL,and ***-2 AS are likely to be *** SSR marker for ***-7 DL was developed and validated in a diverse wheat panel from China,suggesting effectiveness in different genetic *** QTL with closely linked SNP and SSR markers could be useful for marker-assisted selection in wheat breeding programs targeting durable resistance to both diseases.

关键词： mapping wheat breeding

Functional characterization of a Niemann-Pick type C2 protein in the parasitoid wasp Micropfitis mediator

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Insect Science 2018年第5期25卷 765-777页

作者： Yao Zheng Shan-Ning Wang Yong Peng Zi-Yun Lu Shuang Shan Ye-Qing Yang Rui-Jun Li Yong-Jun Zhang Yu-Yuan Guo State Key Laboratory for Biology of plant diseases and Insect pests Institute of Plant Protection Chinese Academy of Agricultural Sciences Beijing China College of plant Protection Agricultural University of Hebei Baoding Hebei Province China IPM center of hebei Province Key Laboratory of Integrated Pest Management on Crops in Northern Region of North China Ministry of Agriculture Plant Protection Institute Hebei Academy of Agricultural and Forestry Sciences Baoding Hebei Province China College of plant Protection China Agricultural University Beijing China

Niemann-Pick type C2 （NPC2） is a type of small soluble protein involved in lipid metabolism and triglyceride accumulation in vertebrates and arthropods. Recent stud- ies have determined that NPC2 also participates in chemical communication of arthropods. In this work, two novel NPC2 proteins （MmedNPC2a and MmedNPC2b） in Microplitis mediator were identified. Real-time quantitative PCR （qPCR） analysis revealed that Mmed- NPC2a was expressed higher in the antennae than in other tissues of adult wasps compared with MmedNPC2b. Subsequent immunolocalization results demonstrated that NPC2a was located in the lymph cavities of sensilla placodea. To further explore the binding charac- terization of recombinant MmedNPC2a to 54 candidate odor molecules, a fluorescence binding assay was performed. It was found MmedNPC2a could not bind with selected fatty acids, such as linoleic acid, palmitic acid, stearic acid and octadecenoic acid. How- ever, seven cotton volatiles, 4-ethylbenzaldehyde, 3,4-dimethylbenzaldehyde, fl-ionone, linalool, m-xylene, benzaldehyde and trans-2-hexen-l-al showed certain binding abilities with MmedNPC2a. Moreover, the predicted 3D model of MmedNPC2a was composed of seven r-sheets and three pairs of disulfide bridges. In this model, the key binding residues for oleic acid in CjapNPC2 of Camponotus japonicus, Lue68, Lys69, Lys70, Phe97, Thr103 and Phe127, are replaced with Phe85, Ser86, His87, Leu113, Tyr119 and Ile143 in MmedNPC2a, respectively. We proposed that MmedNPC2a in M. mediator may play roles in perception of plant volatiles.

关键词： binding ability chemical communication immunolocalization Microplitismediator NPC2 protein 3D model

Diagnosis and characterization of the ribosomal DNA-ITS of potato rot nematode(Ditylenchus destructor) populations from Chinese medicinal herbs

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Journal of Integrative Agriculture 2023年第6期22卷 1763-1781页

作者： NI Chun-hui HAN Bian LIU Yong-gang Maria MUNAWAR LIU Shi-ming LI Wen-hao SHI Ming-ming LI Hui-xia PENG De-liang College of plant Protection Gansu Agricultural University/Biocontrol Engineering Laboratory of Crop Diseases and Pests of Gansu ProvinceLanzhou 730070P.R.China Institute of plant Protection Gansu Academy of Agricultural SciencesLanzhou 730070P.R.China Department of Biological Sciences University of LethbridgeLethbridge 4401Canada State Key Laboratory for Biology of plant diseases and Insect pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193P.R.China

The potato rot nematode(Ditylenchus destructor) is a very economically important nematode in agronomic and horticultural plants worldwide. In this study, 43 populations of D. destructor were collected from different hosts across China, including 37 populations from Chinese herbal medicine plants. Obtained sequences of ITS-rDNA and D2–D3of 28S-rDNA genes of D. destructor were compared and analyzed. Nine types of significant length variations in ITS sequences were observed among all populations. The differences in ITS1 length were mainly caused by the presence of repetitive elements with substantial base substitutions. Reconstructions of ITS1 secondary structures showed that the minisatellites formed a stem structure. Ten haplotypes were observed in all populations based on mutations and variations of helix H9. Among them, 3 known haplotypes(A–C) were found in 7 populations isolated from potato,sweet potato, and Codonopsis pilosula, and 7 unique haplotypes were found in other 36 populations collected from C. pilosula and Angelica sinensis compared with 7 haplotypes(A–G) according to Subbotin' system. These unique haplotypes were different from haplotypes A–G, and we named them as haplotypes H–N. The present results showed that a total of 14 haplotypes(A–N) of ITS-rDNA have been found in D. destructor. Phylogenetic analyses of ITSrDNA and D2–D3 showed that all populations of D. destructor were clustered into two major clades: one clade only containing haplotype A from sweet potato and the other containing haplotypes B–N from other plants. For further verification, PCR-ITS-RFLP profiles were conducted on 7 new haplotypes. Collectively, our study suggests that D. destructor populations on Chinese medicinal materials are very different from those on other hosts and this work provides a paradigm for relevant researches.

关键词： Ditylenchus destructor minisatellites ITS-RFLP phylogeny RNA secondary structure