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Heterologous expression of the haynaldia villosa pattern-recognition receptor CERK1-V in wheat increases resistance to three fungal diseases

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The Crop Journal 2022年第6期10卷 1733-1745页

作者： Anqi Fan Luyang Wei Xu Zhang Jia Liua Li Sun Jin Xiao Yajia Wang Haiyan Wang Jian Hua Ravi P.Singh Zongkuan Wang Xiue Wang State Key Lab of Crop Genetics and Germplasm Enhancement Cytogenetics InstituteNanjing Agricultural University/JCIC-MCPNanjing 210095JiangsuChina Department of Plant Biology Cornell UniversityIthacaNY 14853USA International Maize and Wheat Improvement Center(CIMMYT) El BatanTexcoco 56237Estado de MexicoMexico

Wheat production is under continuous threat by various fungal *** of multipledisease resistance genes may lead to effective disease control via the development of cultivars with broad-spectrum *** Lysin-motif(LysM)-type pattern-recognition receptors,which elicit innate immunity by recognizing fungal pathogen associated molecular patterns such as chitin,are potential candidates for such *** this study,we cloned a LysM receptor-like kinase gene,CERK1-V,from the diploid wheat relative haynaldia ***1-V expression was induced by chitin and Blumeria graminis ***,the causal agent of wheat powdery *** overexpression of CERK1-V in wheat inhibited the development of three fungal pathogens,thereby increased resistance to powdery mildew,yellow rust,and Fusarium head ***1-V physically interacted with the wheat Lys M protein Ta CEBi ***1-V/Ta CEBi Ps interaction promoted chitin recognition and activated chitin signal transduction in *** plants with excessively high CERK1-V expression showed high resistance but abnormal plant growth,whereas plants with moderate expression level showed adequate resistance level with no marked impairment of plant *** transgenic lines,RNA-seq showed that gene expression involved in plant innate immunity was *** of genes involved in photosynthesis,ER stress and multiple phytohormone pathways was also *** expression of CERK1-V in wheat can confer disease resistance without compromising growth or defense fitness.

关键词： Wheat Fungal disease LysM PRR haynaldia villosa CERK1-V

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Development of oligonucleotide probes for FISH karyotyping in haynaldia villosa,a wild relative of common wheat

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The Crop Journal 2020年第4期8卷 676-681页

作者： Jia Lei Jiawen Zhou Haojie Sun Wentao Wan Jin Xiao Chunxia Yuan Miroslava Karafiátová Jaroslav Dolezel Haiyan Wang Xiue Wang State Key Laboratory of Crop Genetics and Germplasm Enhancement Cytogenetics InstituteNanjing Agricultural University/JCIC-MCPNanjing 210095JiangsuChina Institute of Experimental Botany Czech Academy of SciencesCentre of the Region Hana for Biotechnological and Agricultural ResearchŠlechtitelů3177900 OlomoucCzech Republic

haynaldia villosa is a wild relative of wheat and a valuable gene resource for wheat *** to the limited number of probes available for fluorescence in situ hybridization(FISH),the resolution at which the karyotype of *** can be characterized is poor,hampering accurate characterization of small segmental alien *** designed ten oligonucleotide probes using tandem repeats in DNA sequences derived from the short arm of *** chromosome 6 V(6 VS).FISH with seven of them resulted in clear signals on *** *** these,we constructed FISH karyotypes for *** using oligo-6 VS-1 and oligo-6 VS-35 oligonucleotides and characterized the distribution of the two probes in five different *** *** new FISH probes can efficiently characterize *** introgressions into wheat.

关键词： Chromosome identification haynaldia villosa Oligonucleotide probes Tandem DNA repeats

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Genetic Analysis and Molecular Mapping of an All-Stage Stripe Rust Resistance Gene in Triticum aestivum-haynaldia villosa Translocation Line V3

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Journal of Integrative Agriculture 2013年第12期12卷 2197-2208页

作者： HOU Lu MA Dong-fang HU Mao-lin HE Miao-miao LU Yan JING Jin-xue State Key Laboratory of Crop Stress Biology for Arid Areas College of Plant ProtectionNorthwest A&F University Qinghai Academy of Agriculture and Forestry Science Department of Plant Pathology China Agricultural University/Key Laboratory of Plant PathologyMinistry of Agriculture

Triticum aestivum-Hayaldia villosa translocation line V3 has shown effective all-stage resistance to the seven dominant pathotypes of Puccinia striiforms *** prevalent in *** elucidate the genetic basis of the resistance,the segregating populations were developed from the cross between V3 and susceptible genotype Mingxian 169,seedlings of the parents and F 2 progeny were tested with six prevalent pathotypes,including CYR29,CYR31,CYR32-6,CYR33,Sun11-4,and Sun11-11,F 1 plants and F 3 lines were also inoculated with Sun11-11 to confirm the result *** genetic studied results showed that the resistance of V3 against CYR29 was conferred by two dominant genes,independently,one dominant gene and one recessive gene conferring independently or a single dominant gene to confer resistance to CYR31,two complementary dominant genes conferring resistance to both CYR32-6 and Sun11-4,two independently dominant genes or three dominant genes（two of the genes show cumulative effect） conferring resistance to CYR33,a single dominant gene for resistance to *** gene analog polymorphism（RGAP） and simple-sequence repeat（SSR） techniques were used to identify molecular markers linked to the single dominant gene（temporarily designated as YrV3） for resistance to Sun11-11.A linkage map of 2 RGAP and 7 SSR markers was constructed for the dominant gene using data from 221 F 2 plants and their derived F 2：3 lines tested with Sun11-11 in the *** of the complete set of nulli-tetrasomic lines of Chinese Spring with a RGAP marker RG1 mapped the gene on the chromosome 1B,and then the linked 7 SSR markers located this gene on the long arm of chromosome *** linkage map spanned a genetic distance of 25.0 cM,the SSR markers Xgwm124 and Xcfa2147 closely linked to YrV3 with genetic distances of 3.0 and 3.8 cM,*** on the linkage map,it concluded that the resistance gene YrV3 was located on chromosome arm *** chromosomal location,

关键词： haynaldia villosa translocation line stripe rust RGAP-SSR molecular mapping

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Microdissection of haynaldia villosa Telosome 6VS and Cloning of Species-specific DNA Sequences

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Acta Botanica Sinica 2002年第3期44卷 307-313页

作者：孔凡晶陈孝马有志辛志勇李连成张增艳林志姗中国农业科学院作物育种栽培研究所

The material T240_6 derived from SC 2 young embryo of the combination CA9211/RW15 (6D/6V alien substitution) was telosomic substitution line of 6VS identified by GISH (genomic in situ hybridization) analysis. The 6VS was microdissected with a needle and transferred into a 0.5 mL Ep tube. In the 'single tube', all the subsequence steps were conducted. After two round of LA (Linker adaptor)_PCR amplification, the size of PCR bands ranged from 100 to 3 000 bp, with predominate bands 600-1 500 bp. The products were confirmed by Southern blotting analysis using haynaldia villosa (L.) Schur. genomic DNA labeled with 32 P as probe. The PCR products were purified and ligated into clone vector-pGEM_T easy vector. Then, the plasmids were transformed into competence E. coli JM109 with cool CaCl 2. It was estimated that there were more than 17 000 white clones in the library. The size of insert fragments distributed from 100-1 500 bp, with average of 600 bp. Using H. villosa genomic DNA as probe, dot blotting results showed that 37% clones displayed strong and medium positive signals, and 63% clones had faint or no signals. It is demonstrated that there were about 37% repeat sequence clones and 67% single/unique sequence clones in the library. Eight H. villosa_specific clones were screened from the library, and two clones pHVMK22 and pHVMK134 were used for RFLP analysis and sequencing. Both of them were H. villosa specific clones. The pHVMK22 was a unique sequence clone, and the pHVMK134 was a repeat sequence clone. When the pHVMK22 was used as a probe for Southern hybridization, all the powdery mildew resistance materials showed a special band of 2 kb, while all the susceptible ones not. The pHVMK22 may be applied to detect the existence of Pm21.

关键词： microdissection and microcloning of chromosome haynaldia villosa genomic in situ hybridization alien substitution of telosome species_specific DNA sequences RFLP

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Development of EST-PCR Markers for the Chromosome 4V of haynaldia villosa and Their Application in Identification of 4V Chromosome Structural Aberrants

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Journal of Integrative Agriculture 2014年第2期13卷 282-289页

作者： ZHAO Ren-hui WANG Hai-yan JIA Qi XIAO Jin YUAN Chun-xia ZHANG Ya-jun HU Qing-shan WANG Xiu-e State Key Laboratory of Crop Genetics and Germplasm Enhancement Cytogenetics InstituteNanjing Agricultural University

EST-PCR based molecular markers specific for alien chromosomes are not only useful for the detection of the introgressed alien chromatin in the wheat background, but also provide evidence of the syntenic relationship between homoeologous chromosomes. In the present study, in order to develop high density and evenly distributed molecular markers on chromosome 4V of haynaldia villosa, a total of 607 primer pairs were designed according to the EST sequences, which were previously located in 23 different bins of wheat chromosomes 4A, 4B and 4D. By using the Triticum durum-H, villosa amphiploid and T. aestivum-H, villosa alien chromosome lines involving chromosome 4V, it was found that 9.23% of the tested primers could amplify specific bands for chromosome 4V. Thirty and twenty-six specific markers could be assigned to chromosome arms 4VS and 4VL, respectively. These 4V specific markers provided efficient tools for the characterization of structural variation involving the chromosome 4V as well as for the selection of useful genes located on chromosome 4V in breeding programs.

关键词： haynaldia villosa molecular marker Triticum aestivum chromosome engineering

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Establishment of 6VS Telocentric Lines of haynaldia villosa Resistant to Powdery Mildew Induced by Immature Embryo Culture

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Acta Botanica Sinica 2002年第2期44卷 127-131页

作者：李辉陈孝辛志勇徐惠君杜丽璞马有志中国农业科学院作物育种栽培研究所农业部作物遗传育种重点实验室

The line of T240-6 was selected from 32 SC 2 lines of immature embryo culture of T240 (common wheat (Triticum aestivum L.)× Wheat-haynaldia villosa (L.) Schur. 6D/6V substitution line) through powdery mildew characterization, glutamate oxaloacetate transaminase (GOT) enzyme and gliadin (Gli) analyses and in situ hybridization. All of the individual plants resistant to powdery mildew lacked the locus of GOT at 6VL arm (GOT-V 2) and had gliadin locus at 6VS arm (Gli-V 2) of haynaldia villosa. All the plants resistant to powdery mildew had one or two telocentric chromosomes that did not pair with wheat chromosomes but paired between themselves. T240-6 was identified as a telocentric line through in situ hybridization.

关键词： haynaldia villosa immature embryo culture telocentric chromosome glutamate oxaloacetate transaminase gliadin in situ hybridization

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Molecular Characterization of a Triticum durum-haynaldia villosa Amphiploid and Its Derivatives for Resistance to Gaeumannomyces graminis var. tritici

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Agricultural Sciences in China 2007年第5期6卷 513-521页

作者： HUANGDa-hui LIN Zhi-shan CHEN Xiao ZHANG Zeng-yan CHEN Cai-ceng CHENG Shun-he XIN Zhi-yong Key Laboratory of Crop Genetics and Breeding of Ministry of Agriculture~Institute of Crop Sciences Chinese Academy of Agricultural Sciences Beijing 100081 P.R.China Institute of Plant Protection Chinese Academy of Agricultural Sciences Beijing 100094 P.R. China Institute of Agricultural Science of Lixiahe District Yangzhou 225007 P.R.China

Take-all is a serious disease found in wheat across the world. haynaldia villosa is considered to be resistant to take-all at a high level. TH3 was an amphiploid （2n =42, AABBVV） between Triticum durum and haynaldia viUosa with significant resistance to take-all fungus isolated from China. In greenhouse experiment, the derivatives of the hybrid between wheat and TH3 showed better resistance to take-all than that of the wheat control. One of the derivatives named HW918-5 was selected for further analysis. Cytological and genomic in situ hybridization （GISH） analysis indicated that a monotelosome originated from H. villosa existed in the genome of the offspring of the line HW918-5. The monotelosome with promising resistant gene for take-all was located on the 3V chromosome of H. villosa in the further PCR-based molecular analysis.

关键词： AFLP Gaeumannomyces graminis var. tritici genomic in situ hybridization haynaldia villosa Triticum aestivum

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Structural Changes of 2V Chromosome of haynaldia villosa Induced by Gametocidal Chromosome 3C of Aegilops triuncialis

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Agricultural Sciences in China 2008年第7期7卷 804-811页

作者： CHEN Quan-zhan CAO Ai-zhong QI Zeng-jun ZHANG Wei CHEN Pei-du National Key Laboratory of Crop Genetics and Germplasm Enhancement/Nanjing Agricultural University Nanjing 210095 P.R. China Genetics Laboratory Life Science Department Nanjing Xiaozhuang University Nanjing 211171 P.R. China

haynaldia villosa （2n=2X= 14, VV）, a relative of wheat, plays important roles in wheat improvement mainly owing to its disease resistance. Powdery mildew resistance gene Pm21 has been successfully transferred into wheat by Cytogenetic Institute, Nanjing Agricultural University, China, and is widely used in the current wheat breeding programs. In this research, our objective is to further transfer and utilize the beneficial genes such as eye-spot resistance, yellow rust resistance, and gene of the tufted bristles on the glume ridge （a remarkable morphology） mapped on 2V of haynaldia villosa. A disomic addition line with gametocidal chromosome 3C ofAegilops triuncialis added in Norin-26 was crossed to the wheat-H, villosa disomic substitution 2V（2D） and the hybrid F1 was then self-crossed. Chromosome C-banding, genomic in situ hybridization （GISH）, and meiotic analysis in combination with molecular markers were applied to detect the chromosome variations derived from hybrids Fz and F3. To date, four translocations including one small segmental translocation T6BS·6BL-2VS, two whole arm translocations （preliminarily designed as T3DS·2VL and T2VS.7DL） and one intercalary translocation T2VS·2VL-W-2VL, one deletion Del. 2VS·2VL-, one monotelosomic Mt2VS, and one isochromosome 2VS·2VS line have been developed and characterized. One wheat SSR marker Xwmc25.120 tagging 2VS and one wheat STS marker NAU/STSBCD135-1 （2BL） tagging 2VL were successfully used to confirm the alien chromosome segments involved in the seven lines. The tufted bristles on the glume ridge appeared in lines T2VS-7DL, Mt2VS, 2VS-2VS as well as the parent DS2V（2D）, whereas in T3DS·2VL, this trait did not appear. The gene controlling the tufted bristles was located on 2VS. Gametocidal chromosome 3C ofAegilops triuncialis could successfully induce chromosome 2V structural changes.

关键词： Triticum aestivum haynaldia villosa gametocidal chromosome 3C chromosome aberration C-banding genomic in situ hybridization molecular marker

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The Meiotic Behavior of an Alien Chromosome in Triticum aestivum-haynaldia villosa Monosomic Addition Lines

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Agricultural Sciences in China 2002年第4期1卷 370-374页

作者： LI Rui-fen, LIANG Hong-xia and ZHAO Mao-lin(Beijing Agricultural Biotechnology Research Center , Beijing Academy of Agricultural and Forestry Sciences , Beijing 100089 , P.R. China) Beijing Agricultural Biotechnology Research Center Beijing Academy of Agricultural and Forestry Sciences Beijing 100089 P.R. China

By the combination of cytological analysis and using genomic in situ hybridization technique to identify an alien chromosome in wheat-haynaldia villosa monosomic addition lines, we studied the meiotic behavior of the alien chromosome. The results indicated that the frequency of bivalent pairing was lower than the value expected in PMCs of two monosomic addition lines, the frequency of wheat chromosomes unpairing increased, and the wheat homologous chromosome pairing was interfered with by the added chromosome 6V at metaphase I. The chromosome 6V lagged in 20.3% -29.3% of PMCs, sister chromatids 6V early divided in 29.0% - 34.1% of PMCs, the single chromosome 6V in 18.2% - 26.1% of PMCs went to a pole randomly, the breakage frequency of chromosome 6V was 1.2% - 2.9%. Meanwhile, it was also found that several wheat chromosomes showed earlier division, lagging and breakage in a few PMCs. It revealed that the added chromosome 6V influenced the behavior of wheat chromosomes at anaphase. It was also found that the translo-cation was produced between 6V and wheat chromosomes in 1.2% of PMCs. It offered evidence for transloca-tion between wheat and haynaldia villosa 6V chromosomes.

关键词： Wheat haynaldia villosa Monosomic addition Meiosis GISH Translocation

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Production of Somatic Hybrid Plants Between Two Types of Wheat Protoplasts and the Protoplasts of haynaldia villosa

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Acta Botanica Sinica 2002年第8期44卷 1004-1008页

作者：周爱芬夏光敏陈秀玲陈惠民山东大学生命科学学院济南250100

小麦 (TriticumaestivumL .)济南 177的两种原生质体 ,一种来自快速生长的悬浮细胞 ,它们因长期继代而丧失分化能力 ,其染色体只有 2n =2 4～ 2 8;另一种来自可以再生的愈伤组织 ,其原生质体不能持续分裂。它们中任一种与UV照射过的簇... 详细信息

小麦 (TriticumaestivumL .)济南 177的两种原生质体 ,一种来自快速生长的悬浮细胞 ,它们因长期继代而丧失分化能力 ,其染色体只有 2n =2 4～ 2 8;另一种来自可以再生的愈伤组织 ,其原生质体不能持续分裂。它们中任一种与UV照射过的簇毛麦原生质体融合均不能再生植株。然而当它们混合在一起作为受体时 ,能够获得再生绿色植株。细胞核基因和胞质基因的分析证明这些绿色植株是杂种。以上事实说明这两种原生质体在融合时存在某些互补的关系 ,讨论了这种融合方式的可能作用及重要性。

关键词： Triticum aestivum haynaldia villosa UV light somatic hybridization organellar genes

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