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Chinese Science Bulletin 2013年第33期58卷 4084-4091页

作者： YANG XueMing cao aizhong SUN YuLei CHEN PeiDu State Key Laboratory of Crop Genetics and Germplasm Enhancement Cytogenetics InstituteNanjing Agricultural University Provincial Key Laboratory of Agrobiology Institute of BiotechnologyJiangsu Academy of Agricultural Sciences

Bacterial artificial chromosomes(BACs)or yeast artificial chromosomes(YACs)containing large inserts as probes for fluorescence in situ hybridization(FISH)have been used in the physical mapping of specific DNA sequences,especially for single-or low-copy *** earlier study identified Stpk-V,a powdery mildew resistance-related gene located on the 6VS chromosome arm of the wild grass Haynaldia villosa(tribe Triticeae),and obtained several Triticum aestivum–*** alien chromosome lines carrying the Stpk-V ***,the precise physical location of the Stpk-V gene on chromosome 6VS is not *** this study,we used TAC-FISH with TAC15 as the probe coupled with sequential genomic in situ hybridization(GISH)to determine the physical location of the Stpk-V gene in different ***–*** 6V alien chromosome lines,including addition,substitution and translocation *** result indicated that the fraction length of the Stpk-V locus is 0.575±0.035 on the 6V chromosome short arm and this was confirmed by FISH using TAC15 as the probe for tracing the Stpk-V gene in other genetic *** cytological mapping strategies used in this study will be of benefit for tracing the alien gene location in the course of introducing desirable traits from wild species.

关键词：异源染色体相关基因物理位置抗白粉病簇毛麦小麦族 TAC 跟踪

Pm67, a new powdery mildew resistance gene transferred from Dasypyrum villosum chromosome 1V to common wheat(Triticum aestivum L.)

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The Crop Journal 2021年第4期9卷 882-888页

作者： Ruiqi Zhang Chuanxi Xiong Huanqing Mu Ruonan Yao Xiangru Meng Lingna Kong Liping Xing Jizhong Wu Yigao Feng aizhong cao College of Agronomy/JCIC-MCP/National Key Lab of Crop Genetics and Germplasm Enhancement Nanjing Agricultural UniversityNanjing 210095JiangsuChina Institute of Germplasm Resources and Biotechnology/Provincial Key Lab of Agrobiology Jiangsu Academy of Agricultural SciencesNanjing 210014JiangsuChina

Powdery mildew, caused by the biotrophic fungus Blumeria graminis f. sp. tritici(Bgt), is a global disease that poses a serious threat to wheat production. To explore additional resistance gene, a wheatDasypyrum villosum 1 V#5(1 D) disomic substitution line NAU1813(2 n = 42) with high level of seedling resistance to powdery mildew was used to generate the recombination between chromosomes 1 V#5 and1 D. Four introgression lines, including t1 VS#5 ditelosomic addition line NAU1815, t1 VL#5 ditelosomic addition line NAU1816, homozygous T1 DL·1 VS#5 translocation line NAU1817, and homozygous T1 DS·1 VL#5 translocation line NAU1818 were developed from the selfing progenies of 1 V#5 and 1 D double monosomic line that derived from F1 hybrids of NAU1813/NAU0686. All of them were characterized by fluorescence in situ hybridization, genomic in situ hybridization, 1 V-specific markers analysis, and powdery mildew tests at different developmental stages. A new powdery mildew resistance gene named Pm67 was physically located in the terminal bin(FL 0.70–1.00) of 1 VS#5. Lines with Pm67 exhibited seedling stage immunity and tissue-differentiated reactions at adult plant stage. The sheaths, stems, and spikes of the Pm67 line were still immune, but the leaves showed a low degree of *** observation showed that most penetration attempts were stopped in association with papillae on the sheath, and colonies cannot form conidia on the susceptible leaf of Pm67 line at adult plant stage, suggesting that the defence layers of the Pm67 line is tissue-differentiated. Thus, the T1 DL·1 VS#5 translocation line NAU1817 provides a new germplasm in wheat breeding for improvement of powdery mildew resistance.

关键词： Wheat Dasypyrum villosum Powdery mildew Pm67

Characterization of *** chromosome addition lines DA2H and MA5H

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Journal of Genetics and Genomics 2008年第11期35卷 673-678页

作者： Fang Kong Haiyan Wang aizhong cao Bi Qin Jianhui Ji Suling Wang Xiu-E Wang State Key Laboratory of Crop Genetics and Germplasm Enhancement Cytogenetics Institute Nanjing Agricultural University Nanjing 210095 China

In order to transfer useful genes of Hordeum californicum into common wheat （Triticum aestivum L.）, the T. aestivum c.v. Chinese Spring （CS）-H. californicum amphiploid was crossed to CS, and its backcrossing and self-fertilized progenies were analyzed by morpho- logical observation, cytological, biochemical and molecular marker techniques. Alien addition lines with two H. californicum chromo- somes were identified and their genetic constitution was characterized. STS-PCR analysis using chromosome 2B specific markers indi- cated that chromosome H3 of H. californicum belongs to homoeologous group 2, and was thus designated 2H. SDS-PAGE showed that chromosome H2 of H. californicum belongs to homoeologous group 5, and was designated 5H. The CS-H. californicum amphiploid and the chromosome addition lines （DA2H and MA5H） identified were evaluated for powdery mildew （Erysiphe graminis f. sp. triticii） resis- tance in field. The preliminary results indicated that the amphiploid showed higher powdery mildew resistance than CS. However, chro- mosome addition lines DA2H and MA5H were highly susceptible to powdery mildew, indicating that major powdery mildew resistant genes of H. californicum should be located on chromosomes other than 2H and 5H.

关键词： Hordeum californicum alien addition lines powdery mildew sequential C-banding/FISH sequence-tag-sites (STS)

Genetic Analysis of NAU9918 Mutant and Mapping of the Dwarf Gene

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Genetic Analysis of NAU9918 Mutant and Mapping of the Dwarf ...

江苏省遗传学会2013年学术研讨会

作者： Lu Yuan Wang Xiue cao aizhong National Key Laboratory of Crop Genetics and Germplasm Enhancement Cytogenetics InstituteNanjing Agricultural University

Plant height is an important agronomic trait in cereal crops. A dwarfing mutant NM9 is identified from a mutant library of the common wheat cultivar NAU9918 created by ethylmethyl sulfide（EMS） treatment of dry seeds. To identify this mutant, the phynotype, anatomical structure and endogenous hormones concentration of NM9 have been detected. The mutant plant showed obvious phenotypic differences in comparison with the wild type, especially the reduced height, increased tiller number, elongated spike and grain length. The mutant is insensitive to exogenous GA, IAA or BR. Differences of endogenous hormone concentration of ABA, IAA, GA, BR and ZR between NM9 and NAU9918 are confirmed in root, leaf and spike at different growth stages. The plant height of NM9 is dwarfed by reducing parenchyma cells elongation in internode. Allelism tests of the dwarfing gene in NM9 and varieties with different known dwarfing genes, indicating that the dwarfing gene in NM9 is non-allelic to Rht-B1 b, Rht-D1 b, Rht-B1 c, Rht-8 or Rht-D1 c. A genetic analysis using two F2 segregating populations indicated that a single recessive mutation on wheat chromosome 2 A, between two SSR markers gwm122 and gwm71. For the dwarfism phenotype and location, the dwarfing gene in NM9 is designed as Rht-x, temproarily.

关键词： Common wheat Plant height Mutant Genetic mapping Phytohormone

Characterization of a Putative New Semi-Dominant Reduced Height Gene, Rht_NM9, in Wheat(Triticum aestivum L.)

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Journal of Genetics and Genomics 2015年第12期42卷 685-698页

作者： Yuan Lu Liping Xing Shujuan Xing Ping Hu Chaofan Cui Mingyi Zhang Jin Xiao Haiyan Wang Ruiqi Zhang Xiue Wang Peidu Chen aizhong cao National Key Laboratory of Crop Genetics and Germplasm Enhancement Cytogenetics Institute Nanjing Agricultural University

Plant height is an important agronomic trait in cereal crops, and can affect both plant architecture and grain yield. New dwarfing genes are required for improving the genetic diversity of wheat. In this study, a novel dwarf mutant, NM9, was created by treating seeds of the wheat variety NAU9918 with ethyl methanesulfonate （EMS）. NM9 showed obvious phenotypic changes, which were distinct from those caused by other dwarfing genes, especially the reduced plant height, increased effective tiller number, and elongated spike and grain length. The reduced plant height in NM9 was attributable to a semi-dominant dwarfing gene Rht__NM9, which was flanked by two closely linked SNP markers, SNP34 and SNP41, covering an 8.86-Mb region on the chromosome arm 2AS. The results of gibberellic acid （GA） sensitivity evaluation, comparative genomics analysis and allelism test indicated that RhtNM9 was neither allelic to Rht7 and Rht21 nor homoe- oallelic to Rht8, so Rht_NM9 was proposed to be a new dwarfing locus on the homoeologous group 2 chromosomes of wheat. Rht_NM9 has a negative effect on plant height and positive effects on effective tiller number and grain size, thus, Rht_NM9 could be used for elucidating the mechanisms underlying plant architecture and grain development.

关键词： Wheat Mutant Plant architecture Gene mapping

Pm21 from Haynaldia villosa Encodes a CC-NBS- LRR Protein Conferring Powdery Mildew Resistance in Wheat

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Molecular Plant 2018年第6期11卷 874-878页

作者： Liping Xing Ping Hu Jiaqian Liu Kamil Witek Shuang Zhou Jiefei Xu Weihao Zhou Li Gao Zhenpu Huang Ruiqi Zhang Xiue Wang Peidu Chen Haiyan Wang Jonathan D.G. Jones Miroslava Karafiatova Jan Vrana Jan Baros Jaroslav Dolezel Yuanchun Tian Yufeng Wu aizhong cao National Key Laboratory of Crop Genetics and Germplasm Enhancement Cytogenetics Institute Nanjing Agricultural University/JGIC-MCP Nanjing 210095 China The Sainsbury Laboratory Norwich Research Park Norwich NR4 7UH UK Institute of Experimental Botany Centre of the Region Hana for Biotechnological and Agricultural Research Slechtitelu 31 78371 OlomoucCzech Republic

Dear Editor Wheat powdery mildew, caused by Blumeria graminis f. sp. tritici （Bgt）, is a destructive disease of wheat throughout the world. One of the most important environmental-friendly and economical methods to reduce wheat loss caused by Bgt is to develop highly resistant varieties （Kuraparthy et al., 2007）. Pm21 from the wild species Haynaldia villosa （also known as Dasypyrum villosum） confers high resistance to Bgt in wheat throughout all growth stages. It has now become one of the most highly effective genetic loci introgressed into wheat from wild species, and the commercial varieties harboring Pm21 have been widely used in wheat production with more than 4 million hectares in China.

关键词： Pm21 小麦粉蛋白质编码环境友好生长阶段小麦生产还原剂

NLR1-V,a CC-NBS-LRR encoding gene,is a potential candidate gene of the wheat powdery mildew resistance gene Pm21

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NLR1-V,a CC-NBS-LRR encoding gene,is a potential candidate g...

第八届全国小麦基因组学及分子育种大会

作者： Liping Xing Ping Hu Jiaqian Liu Yufeng Wu Jaroslav Dolezel Ruiqi Zhang Xiue Wang Peidu Chen aizhong cao National Key Laboratory of Crop Genetics and Germplasm Enhancement Cytogenetics InstituteNanjing Agricultural University/JCIC-MCP Institute of Experimental Botany

Wheat powdery mildew,caused by Blumeria graminis ***,is one of the most destructive diseases all over the ***21,transferred from the wild Haynaldia villosa to wheat,confers broad spectrum resistance throughout the whole growth stage,and this gene has been widely used in wheat production for more than 20 *** the candidate gene of Pm21 is the prerequisite for elucidating the resistance mechanism,and is a valuable attempt to clone the target genes from the evohitionarily distant wild *** this study,an innovative approach,which combined cytogenetic stocks development,mutagenesis,RenSeq(R gene enrichment sequencing) and PacBio approaches,was tried successfully to clone an NBS-LRR type gene NLR1-V from the Pm21 *** Stocks and Mutants ***,a new cryptic alien introgression line NAU427 with very small introgression segment of 6 VS but still containing Pm21 gene was identified by molecular markers analysis and resistance phenotype evaluation,and the line showed no fluorescent signal when detected by *** enables us to narrow the Pm21 locus into an extremely small region ***,6 independent susceptible M3 mutants of T6 VS·6 AL was identified from more than 20,000 M2 EMS treated lines,which could help us to anchoring the mutant ***-PacBio and NLR Gene ***,the transcriptome of *** was obtained by NGS and the full-length NBS-LRR gene database was constructed by *** the following study,45 individual NLR-carrying contigs showing >80%identity with 6 A/6 B/6 D/6 H NLR homologous were obtained which were identified as the putative NLRs located on the 6 V chromosome,but only two expressed NLR genes were located to the Pm21 locus using the NAU427 as the mapping material,and only NLR1-V showed polymorphism between the wild T6 VS-6 AL and its six ***21 Candidate Gene ***,both the genomic and cDNA sequences of NLR1-V were cloned and *** f

关键词： NLR1-V CC-NBS-LRR Pm21 Mutagenesis RenSeq-Pacbio Chromosome sorting Chicago Hi-C

HvFP3, a HMA domain protein interacted with the E3 ubiquitin ligase HvCMPG, is involved in wheat powdery mildew defense

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HvFP3, a HMA domain protein interacted with the E3 ubiquitin...

江苏省遗传学会2013年学术研讨会

作者： LI Yingbo WANG Wei FEI Fei cao aizhong WANG Haiyan XING Liping WANG Zongkuan CHENG Jiangyue ZHU Yanfei XIE Qi WANG Xiue National Key Laboratory of Crop Genetics and Germplasm Enhancement Cytogenetics InstituteNanjing Agricultural University Institute of Genetics and Developmental Biology Chinese Academy of Sciences

Powdery mildew is one of the most serious diseases threatened wheat production for last few decades. Haynaldia villosa L., species from wheat family, shows high powdery mildew resistance and confers the resistance gene Pm21. In our previous study, by using the Barley GeneChip microarray and cDNA library screening, a HvCMPG gene encoding E3 ligase has been cloned and was found to play an important role in the defense response to the powdery mildew disease. For further elucidating the molecular mechanism, a yeast two-hybrid library of H. villosa was constructed and used to screen the interact proteins with HvCMPG. A metal transporter gene was found to be interacted with the HvCMPG, and the full length was cloned from H. villosa and was designated as HvFP3. HvFP3 can interact with and be ubiquitinated by HvCMPG in vitro. Analysis by quantitative RT-PCR showed that HvFP3 was rapidly up-regulated in the leaf of *** after Blumeria graminis ***（Bgt） inoculation. The subcellular localization of the HvFP3 was in the nucleus and plasma membrane of the epidermal cell of onion. In the transient overexpression assay, it was found that the transient over-expression of HvFP3 led to significantly decreased in the haustoria index（HI） compared with the vector control in the epidermal cell of wheat variety Yangmai158, which is moderatly susceptible to powdery mildew disease. The HvFP3 transgenic plants over-expressed in Yangmai158 and the positive transgenic plants showed improved powdery mildew resistance than the receptor. These results indicated that the HvFP3 participated in the powdery mildew defense responses.

关键词： wheat powdery mildew yeast two hybrid metal transporter

A Disulphide Isomerase Gene（Pdi-V） From Haynaldia Villosa Contributes To Powdery Mildew Resistance In Common Wheat

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A Disulphide Isomerase Gene（Pdi-V） From Haynaldia Villosa ...

江苏省遗传学会第九次会员代表大会暨学术研讨会

作者： Muhammad Faheem Yingbo Li Muhammad Arshad Cheng Jiangyue Zhao Jia Xiao Jin Haiyan Wang Ruiqi Zhang Liping Xing aizhong cao Xiue Wang The State Key Laboratory of Crop Genetics and Germplasm Enhancement Nanjing Agricultural University

In this study, we reported the contribution of PDI-like gene from wheat wild relative Haynaldia villosa in combating against powdery mildew（Bgt）. PDI-V was identified by Yeast two hybrid as an interacting protein with CMPG1-V, which encodes an E3 ligase and is a positive regulator for powdery mildew resistance. The PDI-V contains two conserved thioredoxin active domains（a and a′） and an inactive domain（b）. Our results showed that PDI-V could be mono-ubiquitinated by CMPG1-V without detected degradation. The PDI-V was located on the 5 V chromosome in H. villosa genome and PDI-V is subcellularly localized in endoplasmic reticulum lumen. PDI-V was induced in leaves when challenged with Bgt. Visible Bgt spores could not be detected when silencing the PDIs in PM resistant T. durum-H. villosa amphiploid, but microscopically display increased fungal growth. Single cell transient over-expression of PDI-V decreased the haustorium index of a moderately PM susceptible wheat variety Yangmai-158. Stable transgenic lines over-expressing PDI-V displayed improved PM resistance at both seedling and adult stages. Over-expression of construct only containing the active thioredoxin domain a could also equally reduce the HI as the full length gene, indicating its critical role in defense responses. Over-expressed point mutated PDI-VC57 Ain Yangmai-158 could not improve PM resistance, further confirming the essential role of the conserved TRX-like domain a for proper functioning of PDI-V. Taken together, these evidences indicates that PDI-V plays positive role in PM resistance and represents a potential novel source for wheat improvement.

关键词： Protein disulphide isomerase Haynaldia villosa Tritucum aestivum Blumeria graminis disease resistance

The cloning of Rht23 revealed a point mutation in 5Dq leading to plant dwarfness and compact spike in common wheat

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The cloning of Rht23 revealed a point mutation in 5Dq leadin...

2017中国长三角遗传学大会

作者： Kaijun Zhao Jin Xiao Yu Liu Shulin Chen Chunxia Yuan aizhong cao Frank M.You Donglei Yang An Shengmin Haiyan Wang Xiue Wang State Key Laboratory of Crop Genetics and Germplasm Enhancement Cytogenetics InstituteNanjing Agricultural University/JCIC-MCP Morden Research and Development Centre Agriculture and Agri-Food Canada

The domesticated Q gene on wheat chromosome 5 A(5 AQ) encode an AP2 transcription factor,conferring characters of free-threshing and subcompact *** B and D homeoalleles of hexaploid bread wheat are thought to be either a pseudogene or expressed at a low level,*** function of the low expressed allele on chromosome 5 D(5 Dq) was still *** identified a compact spike and dwarf mutant of Sumai 3,NAUH164,by EMS mutagenesis,and the mutant gene was designated as ***23 was mapped to a physical interval of 8.5 Mb on chromosome 5 DL,in which the 5 Dq was *** present study,the 5 Dq/q’ were cloned from wild type Sumai 3 and its *** comparison showed that there occurred a SNP substitution(G3147 A) in the 5 Dq ORF-region,which has been proved as the binding site of *** SNP resulted in an Ala416 Thr mutation and changed the structure of 5 Dq *** expression change was observed for 5 AQ in the wild type and the *** the 5 Dq expression was significantly up-regulated in leaves,stems and spikes of NAUH164 than those in Sumai *** in the wild type Sumai 3,the Ta-miR172 expression was down regulated in leaf,stem and spike of the *** may due to the induced mismatch to *** further study the relationship of 5 Dq’ with Rht23,a SNP-based co-dominant marker,was *** analysis using a(NAUH164×Alondra’s) RIL population showed that the marker allele was co-segregated with the Rht23 mutant *** results suggest that Rht23 was a 5 Dq point mutation,which led to plant dwarf and compact spike *** results provided the evidence that the Q homolog on chromosome 5 D also has pleiotropic effect in regulation of plant architecture.

关键词： Common wheat Q gene Rht23 Mutagenesis Single nucleotide polymorphism(SNP)