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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

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Population genetic structure of Chinese puccinia triticina races based on multi-locus sequences

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Journal of Integrative Agriculture 2018年第8期17卷 1779-1789页

作者： LIU Tai-guo GE Run-jing MA Yu-tong LIU Bo GAO Li CHEN Wan-quan State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant Protection Chinese Academy of Agricultural Sciences

puccinia triticina, the causal agent of wheat leaf rust, is one of the most devastating rust fungi attacking wheat worldwide. Seventy-six isolates of the wheat leaf rust pathogen from Yunnan, Sichuan, Gansu and Henan provinces, China, were tested on wheat leaf rust differentials and the population structure was analyzed using four presumably neutral partial sequence markers such as elongation factor-1α(EF-1α), glyceraldehyde-3-phosphate dehydrogenase(GAPDH), β-tubulin(TUB) and the second largest RNA polymerase subunit(RPB2). The phenotypic diversity of Yunnan and Sichuan populations was higher than that of Gansu and Henan populations. The four populations were separated into two clusters based on the pathogenic data. A total of 12 single nucleotide polymorphisms(SNPs) and 32 haplotypes were identified among the four sequences. The 32 haplotypes were divided into two clusters in a neighbor-joining tree. Bayesian analyses also identified two clusters. Pairwise FST between populations in different regions were significantly different(P<0.05). Analysis of molecular variance(AMOVA) indicated that 68% of the total genetic variation was within populations.

关键词： population structure polymorphism virulence wheat leaf rust puccinia triticina

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Screening and Analysis of Effector Proteins from puccinia triticina

Screening and Analysis of Effector Proteins from Puccinia tr...

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中国植物病理学会2017年学术年会

作者： Li Jianyuan Wei Jie An Zhe Zhang Yue Yang Wenxiang Liu Daqun Department of Plant Pathology Agricultural University of Hebei/Biological Control Center of Plant Diseases and Plant Pests of Hebei ProvinceNational Engineering Research Center for Agriculture in Northern Mountainous Areas

Leaf rust caused by the fungus puccinia triticina（Pt） is a major constraint to wheat production *** molecular events that underlie Pt pathogenicity are largely *** all rusts,Pt forms a specialized infection structure called haustoria to take nutrients and water from host plant tissue,and to secrete pathogenicity factors called effector *** this study,we extracted the total RNA of urediniospores,germinated urediniospores and infected wheat leaves harvest at 6 d from three different virulence strains of Pt and used Illumina Hiseq sequencing platforms to assemble the transcriptomes and then we got 46008 *** effectors were screened through Signal P 4.1,Target P 1.1,TMHMM 2.0 and Effector P,and 325 genes which constitute candidate effectors were ***-PCR analysis confirmed the expression patterns of 96 candidate effectors and 42 genes showed aspecial increase in various stages which may be involved in the pathogenic process.33 candidate effectors were cloned successfully and 17 were identified to be able to suppress the cell death caused by BAX in tobacco *** analysis of candidate effectors is an initial step towards functional research and established the foundation forclarify the pathogenic mechanism of leaf rust pathogens.

关键词： puccinia triticina Effector proteins Screening

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RNA-seq analysis reveals differentially expressed genes during the Lr47-mediated wheat resistance to puccinia triticina

RNA-seq analysis reveals differentially expressed genes duri...

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中国植物病理学会第十一届全国会员代表大会暨2018年学...

作者： Gao Jing Bi Weishuai Zhao Jiaojie Li Huanpeng Wu Jiaojiao Yu Xiumei Liu Daqun Wang Xiaodong College of Plant Protection Biological Control Center for Plant Diseases and Plant Pests of HebeiHebei Agricultural University College of Life Science Hebei Agricultural University Graduate School of Chinese Academy of Agricultural Sciences

Wheat leaf rust,caused by puccinia triticina(Pt),is one of the most severe fungal diseases threatening the global wheat *** use of leaf rust resistance(Lr) genes in wheat breeding program is the major solution to solve this *** resistance Lr47 gene was introgressed from chromosome 7 S of Aegilops speltoides to chromosome 7 AS of hexaploid wheat Triticum *** lines carrying Lr47 showed immune responses to all the Pt pathotypes detected in ***,this gene has not been cloned yet and its molecular mechanism remains *** the present study,RNA-seq analysis was applied on three different wheat lines carrying Lr47 inoculated with epidemic Pt pathotype THTT at 3 days post-inoculation,when a clear hypersensitive response(HR)was *** reads were assembled using reference genomes of Triticum uratu(AA),Aegilops tauschii(DD) and Triticumaestivum(AABBDD),respectively,to reveal differentially expressed genes(DEGs) during the Lr 47-mediated resistance.A total of eight Chromosome 7-located DEGs encoding NBS-LRR proteins were identified and selected for further functional *** with mock inoculation with water,a total of 152 up-regulated(higher than 4 fold) and 82 down-regulated(less than 0.25 fold) DEGs were *** expression profiles of several pathogenesis-related(PR) genes and transcription factor gene families were generated to reveal the regulatory network of *** identified in the current study might be valuable resources to improve wheat resistance to puccinia triticina.

关键词： RNA-seq Differentially expressed genes Leaf rust resistance Wheat puccinia triticina

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Screening and Analysis of Effector Proteins from puccinia triticina by Transcriptome Sequencing

Screening and Analysis of Effector Proteins from Puccinia tr...

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中国植物病理学会2016年学术年会

作者： LI Jian-yuan WANG Zhi ZHANG Yu-mei YANG Wen-xiang LIU Da-qun Department of Plant Pathology Agricultural University of Hebei/Biological Control Center of Plant Diseases and Plant Pests of Hebei Province/National Engineering Research Center for Agriculture in Northern Mountainous Areas

Leaf rust caused by the fungus puccinia triticina（Pt） is a major constraint to wheat production *** molecular events that underlie Pt pathogenicity are largely *** all rusts,Pt forms a specialized infection structure called haustoria to take nutrients and water from host plant tissue,and to secrete pathogenicity factors called effector *** extracted the total RNA of urediniospores,germinated urediniospores and infected wheat leaves harvest at 6d from three different virulence strains of Pt and used Illumina Hiseq sequencing platforms to assemble the transcriptomes and then we got 80392 *** effectors were screened through Signal P 4.1,Target P 1.1,TMHMM 2.0 and Effector P,and 325 genes which constitute candidate effectors were *** two thirds of these were up-regulated in the harvest leaves compared to urediniospores and germinated spores.42 of the genes were predicted to code for proteins of known function like a variety of metabolism processes,nutrient transport,and biosynthesis by using a BLAST2 GO platform and manual *** was used to predict the frameworks and 61 proteins were predicted with Pfam *** remaining 264 secreted proteins were screened and predicted the de novo structure by using MEME *** results showed that 3 conserved domains were found in 11 effector ***-PCR analysis confirmed the expression patterns of 52 effector candidates and four genes showed a special increase in infected leaves which may be associated with the formation of *** analysis of candidate effectors is an initial step towards functional research and established the foundation for clarify the pathogenic mechanism of leaf rust pathogens.

关键词： puccinia triticina Effector proteins Transcriptome sequencing

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Novel WRKY transcriptional factors associated with NPR1 during systemic acquired resistance in barley are potential resources to improve wheat resistance to puccinia triticina

Novel WRKY transcriptional factors associated with NPR1 duri...

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中国植物病理学会第十一届全国会员代表大会暨2018年学...

作者： Li Huanpeng Wu Jiaojiao Gao Jing Bi Weishuai Yu Xiumei Liu Daqun Wang Xiaodong College of Plant Protection Biological Control Center for Plant Diseases and Plant Pests of HebeiHebei Agricultural University College of Life Science Hebei Agricultural University Graduate School of Chinese Academy of Agricultural Sciences

In Arabidopsis,both initial pathogen infection and benzothiadiazole（BTH） treatment are sufficient to induce systemic acquired resistance（SAR）,which is an inducible form of enhanced resistance to secondary pathogen ***1 is reported to be the master regulator of *** Arabidopsis and rice,WRKY transcription factors are designated as key components during SAR in either NPR1-dependent or-independent *** several SAR-like responses discovered in wheat and barley can improve their resistance to various fungal diseases including Blumeria graminis and puccinia triticina（Pt）,roles of NPR1 and WRKY genes during SAR in these crop species are largely *** a series of transcriptome analysis on barley transgenic lines over-expressing wheat wNPR1（wNPR1-OE） and knocking down barley HvNPR1（HvNPR1-Kd）,the regulatory networks of NPR1 during SAR triggered by Pseudomonas syringae *** DC3000 and BTH were clarified.A total of 17 differentially expressed WRKY genes were selected for further functional ***,some of them may also function through physical interaction with NPR1 *** transient expression of three novel WRKY genes,including HvWRKY165,HvWRKY213,and HvWRKY248,in wheat leaves by Agrobacterium-mediated infiltration enhanced the resistance to *** transgenic lines over-expressing these WRKY genes may be potential resources to improve wheat resistance to various fungal diseases.

关键词： WRKY transcription factors NPR1 Systemic acquired resistance Barley Wheat puccinia triticina

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Three co-located resistance genes confer resistance to leaf rust and stripe rust in wheat variety Borlaug 100

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The Crop Journal 2022年第2期10卷 490-497页

作者： Bowei Ye Ravi P.Singh Chan Yuan Demei Liu Mandeep S.Randhawa Julio Huerta-Espino Sridhar Bhavani Evans Lagudah Caixia Lan College of Plant Science&Technology Huazhong Agricultural UniversityWuhan 430070HubeiChina International Maize and Wheat Improvement Center(CIMMYT) Apdo.Postal 6-64106600 Mexico D.F.Mexico Qinghai Provincial Key Laboratory of Crop Molecular Breeding Northwest Institute of Plateau BiologyChinese Academy of ScienceXining 810008QinghaiChina Campo Experimental Valle de México INIFAP Apdo.Postal 1056230ChapingoEdo.de MexicoMexico CSIRO Agriculture&Food GPO Box 1600CanberraACT 2601Australia

Leaf rust(LR) and stripe rust(YR) are important diseases in wheat producing areas worldwide and cause severe yield losses under favorable environmental conditions when susceptible varieties are grown. We determined the genetic basis of resistance to LR and YR in variety Borlaug 100 by developing and phenotyping a population of 198 F6 recombinant inbred lines derived from a cross with the susceptible parent Apav#1. LR and YR phenotyping were conducted for 4 and 3 seasons, respectively, at CIMMYT research stations in Mexico under artificial epidemics. Mendelian segregation analyses indicated that 3–5 LR and 2 YR genes conferred resistance in Borlaug 100. Lr46/Yr29(1 BL), Yr17(2 AS) and Yr30(3 BS) were present in the resistant parent and segregated in the RIL population based on characterization by molecular markers linked to these genes. When present alone, Lr46/Yr29 caused average 13% and 16% reductions in LR and YR severities, respectively, in RILs. Similarly, Yr17 and Yr30 reduced YR severities by 57% and 11%, respectively. The Yr30 and the Yr17 translocation were also associated with 27% and 14% reductions, respectively, in LR severity, indicating that the 3 BS and 2 AS chromosomal regions likely carry new slow rusting LR resistance genes, temporarily designated as Lr B1 and Lr B2, respectively. Additive effects of Yr30*Yr17, Yr29*Yr17 and Yr29*Yr30 on YR and LR were significant and reduced YR severities by 56%,55%, and 45%, respectively, and LR severities by 34%, 40%, and 45%, respectively. Furthermore, interaction between the three genes was also significant, with mean reductions of 70% for YR and 54% for LR severities. Borlaug 100, or any one of the 21 lines with variable agronomic traits but carrying all three colocated resistance loci, can be used as resistance sources in wheat breeding programs.

关键词： Co-located resistance loci Common wheat Gene interaction puccinia triticina puccinia striiformis Triticum aestivum

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Progress Towards Genetics and Breeding for Minor Genes Based Resistance to Ug99 and Other Rusts in CIMMYT High-Yielding Spring Wheat

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Journal of Integrative Agriculture 2014年第2期13卷 255-261页

作者： Ravi Prakash Singh Sybil Herrera-Foessel Julio Huerta-Espino Sukhwinder Singh Sridhar Bhavani Caixia Lan Bhoja Raj Basnet CIMMYT Apdo.Postal 6-641Mexico DF 06600Mexico INIFAP-CEVAMEX Apdo.Postal 10Chapingo 56230Mexico CIMMYT ICRAF BuildingNairobi 00621Kenya

Wheat rusts continue to cause significant losses worldwide despite major efforts given to their genetic control. This is due to frequent evolution and selection of virulence in pathogen overcoming the deployed race-specific resistance genes. Although the life of effective race-specific resistance genes can be prolonged by using gene combinations, an alternative approach being implemented at CIMMYT is to deploy varieties that posses adult plant resistance （APR） based on combinations of minor, slow rusting genes. When present alone, the APR genes do not confer adequate resistance especially under high disease pressure; however, combinations of 4 or 5 minor genes usually result in ＂near-immunity＂ or a high level of resistance. Although only a few APR genes are catalogued, various APR QTLs are now known and could lead to further characterization of additional genes. Four characterized genes have pleiotropic effects in conferring partial APR to all 3 rusts and powdery mildew, thus simplifying the task of breeding wheat varieties that are resistant to multiple diseases. Significant progress was made recently in developing high-yielding wheat germplasm that possesses high levels of APR to all three rusts by implementing a Mexico- Kenya shuttle breeding scheme. Parents with APR to Ug99 were hybridized with high-yielding parents that had adequate to high levels of APR to leaf rust and yellow rust. Segregating populations and advanced lines from these crosses were selected under high rust pressures in Mexico （leaf rust and yellow rust） and Kenya （Ug99 stem rust and yellow rust） to identify high- yielding progenies that possess high to adequate APR to all three rusts. International distribution of these high-yielding wheats is underway through CIMMYT intemational yield trials and screening nurseries. It is expected that several wheat varieties with APR to three rusts will be released and grown in various countries in the near-future that will allow determining the durability o

关键词： Triticum aestivum puccinia graminis puccinia triticina puccinia striiformis genetics breeding wheat rustresistance

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Postulation of Leaf Rust Resistance Genes in Seven Chinese Spring Wheat Cultivars

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Journal of Integrative Agriculture 2013年第9期12卷 1580-1588页

作者： SHI Li-hong ZHANG Na HU Ya-ya WEI Xue-jun YANG Wen-xiang LIU Da-qun Department of Plant Pathology College of Plant Protection/Biological Control Center for Plant Diseases and Plant Pests of HebeiAgricultural University of Hebei

To detect the leaf rust resistance genes in the 7 Chinese spring wheat clultivars Shenmian 99025, Shenmia 99042, Shenmian 85, Shenmian 91, Shenmian 96, Shenmian 1167 and Shenmian 962, Thatcher, Thatcher backgrounded near-isogenic lines and 15 pathotypes of P. triticina were used for gene postulate at the seedling stage, and 9 of the 15 pathotypes were used in the field tests. Molecular markers closely linked to, or co-segregated with resistance genes Lrl, Lr9, LrlO, Lrl9, Lr20, Lr21, Lr24, Lr26, Lr28, Lr29, Lr32, Lr34, Lr35, Lr37, Lr38, and Lr47 were screened to assist detection of the resistance genes. As results, 4 known resistance genes, including Lrl, Lr9, Lr26, and Lr34, and other unknown resistance genes were postulated singly or in combination in the tested cultivars. Shenmian 85, Shenmian 91, Shenmian 96, Shenmian 962, Shenmian 1167, and Shenmian 99042 are potentially useful for wheat production and breeding programs. The result suggested that combining gene postulation, molecular markers and pedigrees is effective and more accuracy method to know the resistance genes in cultivars.

关键词： gene postulation molecular marker Triticum aestivum puccinia triticina resistance gene

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Postulation of seedling leaf rust resistance genes in 84 Chinese winter wheat cultivars

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Journal of Integrative Agriculture 2015年第10期14卷 1992-2001页

作者： REN Xiao-li LIU Tai-guo LIU Bo GAO Li CHEN Wan-quan State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant Protection Chinese Academy of Agricultural Sciences

Wheat leaf rust（caused by puccinia triticina） is one of the most important fungal diseases in China. There are tens of winter wheat cultivars which are approved to be released by the government at a national level and more than 100 wheat cultivars at the provincial level. But there is no information about leaf rust（Lr） genes in these cultivars, which makes it difficult for farmers and breeders to select which cultivars they should plant in their fields and use in their breeding programs. The objective of this paper was to identify the leaf rust resistant genes at seedling stage present in the 84 commercial wheat cultivars from China that have been released in the past few years. A set of 20 near isogenic lines with Thatcher background and 6 lines with known Lr genes were used to test the virulence of 12 races of P. triticina（Pt）. By comparing the infection types（ITs） produced on the 84 cultivars by the 12 Pt races with the ITs on the differential sets, the Lr genes were postulated. In addition, 8 molecular markers of Lr genes such as Lr9, Lr10, Lr19, Lr20, Lr21, Lr24, Lr26 and Lr29, which are closely linked to or co-segregated with the Lr gene, were used for further validation of the genes in the 84 Chinese winter wheat cultivars. Twelve Lr genes, including Lr1, Lr3,（Lr3bg）,（Lr3ka）, Lr11, Lr13, Lr14 a, Lr16, Lr26, Lr27, Lr30 and Lr31 were postulated to be present either singly or in combinations in these Chinese wheat cultivars. Lr3 and Lr26 were detected most often in the tested cultivars, with frequencies of 51.2 and 38.1%, respectively. No wheat Lr genes were detected in 16 cultivars, and 4 cultivars may carry unknown Lr genes other than those used in this study. Lr9, Lr20, Lr21, Lr24, Lr25 and Lr29 were not present in any of the 84 tested accessions.

关键词： gene postulation molecular marker puccinia triticina wheat leaf rust

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