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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

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Genetic behavior of Triticum aestivum–dasypyrum villosum translocation chromosomes T6V#4S·6DL and T6V#2S·6AL carrying powdery mildew resistance

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Journal of Integrative Agriculture 2017年第10期16卷 2136-2144页

作者： LIU Chang YE Xing-guo WANG Mei-jiao LI Shi-jin LIN Zhi-shan National Key Facility of Crop Gene Resources and Genetic Improvement Institute of Crop Sciences Chinese Academy of Agricultural Sciences Beijing 100081 P.R. China

T6V#2S·6AL and T6V#4S·6DL translocation chromosomes developed from the cross of wheat and different dasypyrum villosum accessions have good powdery mildew （PM） resistance, but their pairing and pyramiding behavior remains unclear. Results in this study indicated that the pairing frequency rate of the two differently originated 6VS chromosomes in their F1 hybrid was 18.9% according to genomic in situ hybridization （GISH）; the PM resistance plants in the F2 generation from the cross between T6V#4S·6DL translocation line Pm97033 and its PM susceptible wheat variety Wan7107 was fewer than expected. However, the ratio of the resistant vs. the susceptible plants of 15：1 in the F2 generation derived from the cross between the two translocation lines of T6V#2S·6AL and T6V#4S·6DL fitted well. Plants segregation ratio （homozygous：heterozygous：lacking） revealed by molecular marker for T6V#4S·6DL or T6V#2S·6AL in their F2 populations fitted the expected values of 1：2：1 well, inferring that the pairing of the two alien chromosome arms facilitates the transmission of T6V#4S·6DL from the F1 to the F2 generation. A quadrivalent was also observed in 21% of pollen mother cells （PMCs） of homozygote plants containing the two pairs of translocated chromosomes. The chromosome pairing between 6V#2S and 6V#4S indicates that it will be possible to obtain recombinants and clarify if the PM resistance determinant on one alien chromosome arm is different from that on the other.

关键词： Trificum aestivum dasypyrum villosum translocation genetic behavior powdery mildew resistance GISH molecular marker

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Development of EST-PCR markers specific to the long arm of chromosome 6V of dasypyrum villosum

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Journal of Integrative Agriculture 2018年第8期17卷 1720-1726页

作者： SUN Hao-jie SONG Jing-jing XIAO Jin XU Tao WEI Xing YUAN Chun-xia CAO Ai-zhong XING Li-ping WANG Hai-yan WANG Xiu-e State Key Laboratory of Crop Genetics and Germplasm Enhancement Ministry of Science and Technology/Cytogenetics InstituteNanjing Agricultural University/Jiangsu Collaborative Innovation Center for Modern Crop Production(JCIC-MCP)

Expressed sequence tags-derived polymerase chain reaction(EST-PCR) molecular markers specific for alien chromosomes can be used to not only monitor the introgressed alien chromatin in wheat background, but also provide the evidence of the syntenic relationship between homoeologous chromosomes. In the present study, in order to develop high density and evenly distributed molecular markers specific for chromosome 6 VL of dasypyrum villosum, 297 primer pairs were designed based on the expressed sequence tags(EST) sequences, which were previously mapped in different bins of the long arms of wheat homoeologous 6 AL, 6 BL, and 6 DL. By using the Triticum aestivum, D. villosum, T. durum-D. villosum amphiploid, and T. aestivum-D. villosum alien chromosome lines involving chromosome 6 V, it was found that 32(10.77%) primers could amplify specific bands for chromosome 6 V, and 31 could be allocated to chromosome arm 6 VL. These 6 VL specific markers provided efficient tools for the characterization of structural variation involving the chromosome 6 VL in common wheat background as well as for the selection of useful genes located on 6 VL in breeding programs.

关键词： dasypyrum villosum molecular marker common wheat alien chromosome line

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Introgression of sharp eyespot resistance from dasypyrum villosum chromosome 2VL into bread wheat

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The Crop Journal 2023年第5期11卷 1512-1520页

作者： Caiyun Liu Wei Guo Yang Wang Bisheng Fu Jaroslav Doležel Ying Liu Wenling Zhai Mahmoud Said István Molnár Kateřina Holušová Ruiqi Zhang Jizhong Wu Institute of Germplasm Resources and Biotechnology/Jiangsu Provincial Key Laboratory of Agrobiology Jiangsu Academy of Agricultural SciencesNanjing 210014JiangsuChina College of Agronomy/JCIC-MCP/National Key Lab of Crop Genetics and Germplasm Enhancement Nanjing Agricultural UniversityNanjing 210095JiangsuChina Institute of Experimental Botany Centre of the HanáRegion for Biotechnological and Agricultural ResearchŠlechtitelů31CZ-783671 Olomoucthe Czech Republic Field Crops Research Institute Agricultural Research Centre9 Gamma StreetGiz112619 CairoEgypt Agricultural Institute Centre for Agricultural ResearchELKHMartonvásár 2462Hungary Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops Yangzhou UniversityYangzhou 225009JiangsuChina Zhongshan Biological Breeding Laboratory Nanjing 210014JiangsuChina

Wheat sharp eyespot, a stem disease caused by the soilborne fungus Rhizoctonia cerealis van der Hoeven,has become a threat to wheat production worldwide. Exploiting resistance resources from wild relatives of wheat is a promising strategy for controlling this disease. In this study, a new wheat–dasypyrum villosum T2DS·2V#4L translocation line in the background of Chinese Spring(CS) showed stable resistance to R. cerealis. Introgression of the T2DS·2V#4L chromosome into wheat cultivar Aikang 58 by backcrossing produced a marked increase in sharp eyespot resistance in NIL-T2DS·2V#4L in comparison with NILT2DS·2DL, and no detrimental effects of 2V#4L on agronomic traits were observed in the BC2F2, BC2F2:3,and BC2F2:4generations. Flow-sorted sequencing of 2V#4L yielded 384.3 Mb of assembled sequence, and8836 genes were predicted of which 6154 had orthologs in at least one of the 2AL, 2BL, and 2DL arms of CS, whereas 1549 genes were unique to 2V#4L. About 100,000 SNPs were detected in genes of 2V#4L and2DL in 10 sequenced bread wheat cultivars. A Kompetitive Allele Specific Polymerase chain reaction and30 conserved ortholog sequence markers were developed to trace the 2V#4L chromatin in wheat backgrounds. T2DS·2V#4L compensating translocation lines represent novel germplasm with sharp eyespot resistance and the markers will allow rapid detection in breeding programs.

关键词： Triticum aestivum Rhizoctonia cerealis dasypyrum villosum Flow sorting KASP

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Characterization and Expression Profiling of Genes Encoding the Gibberellin 3-oxidase in dasypyrum villosum Dwarf Mutant

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Agricultural Science & Technology 2017年第9期18卷 1585-1589页

作者： Peng CAI Zaijun YANG Zhengsong PENG Maoqun YU Horticulture Institute Sichuan Academy of Agricultural Sciences College of Life Science China West Normal University Xichang College Chengdu Institute of Biology Chinese Academy of Sciences

Gibberellin 3-oxidase catalyzes the conversion of inactive gibberellin(GA) species into GAs with biological activity and it is subjected to strict developmental controls in the life cycle of a plant. In this study, 33 gene sequences, encoding the gibberellin 3-oxidase(GA3ox) from dasypyrum villosum and its dwarf mutant, were obtained. Each contained a 1 107 bp coding sequence(CDS) that encoded a putative protein containing 369 amino acids. The GA3ox protein showed 77% to 97% homology and shared the major conserved structural domains of GA3ox proteins with rice, sorghum bicolor, oat, barley, and wheat. Sequence alignment showed that there were 20 single nucleotide polymorphisms(SNPs) and 22 Insertion/deletions(In Dels) among these sequences, which could be divided into 2 haplotypes, haplotypes Ⅰ and Ⅱ. Haplotype Ⅰ was found in the wild type and was1 495 bp in length, and haplotype Ⅱ was found in the dwarf mutant and was 1 485 bp in length. The Q-PCR results showed that GA3ox was expressed in the leaves, roots, internodes, and stem nodes, and that there was a significant difference in the transcript level of the GA3ox between the wild type and dwarf mutant. The transcript levels of GA3ox in the leaves at the seedling stage, stem elongation stage and the heading stage, in the root and stem nodes at the stem elongation stage and in the internodes at the heading stage of the wild type, were significantly higher than those in the dwarf mutant. However, GA3ox expression in the rest of the wild type tissues at the 3 stages was slightly higher than or not different from the dwarf *** results suggested that the wild type and mutant allele sequences of GA3ox in D. villosum showed 2 amino acid changes in exons and variations in the lengths of introns or the SNPs in introns, which most probably impaired the function of the enzyme,affected the GA3ox expression level, and eventually gave rise to dwarfing.

关键词： dasypyrum villosum Gibberellin 3-oxidase Dwarf mutant Wheat SNP

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The gene Pm21,encoding a typical CC-NBS-LRR protein,confers broad spectrum resistance to wheat powdery mildew

The gene Pm21,encoding a typical CC-NBS-LRR protein,confers ...

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第八届全国小麦基因组学及分子育种大会

作者： Huagang He Shanying Zhu Yaoyong Ji Zhengning Jiang Renhui Zhao Tongde Bie Jiangsu University Yangzhou Academy of Agricultural Sciences

Common wheat is the most widely grown cereal crop providing 0% of the calories consumed by ***,wheat production is challenged constantly by devastating powdery mildew caused by Blumeria graminis ***(Bgt).Utilization of powdery mildew resistance(Pm) genes is an effective and economical way to reduce yield losses caused by *** to now,more than 100 Pm genes in wheat and its wild relatives have been documented,among which,the Pm21 gene,originating from dasypyrum villosum,confers high resistance to all known Bgt *** is important to clarify the genetic basis and functional mechanism of *** this study,fine genetic mapping of Pm21 was conducted on an F population(harboring 10,536 individuals)derived from the cross between the resistant *** line DvRes-1 and the susceptible line DvSus-1,and a high-density map was ***21 was mapped to a 0.01-cM interval flanked by the markers 6 VS-08.4 b and 6 VS-10 *** on comparative mapping,a conserved CC-NBS-LRR-encoding resistance gene analog(RGA) locus was found between wheat and *** to the conserved RGA locus,a large genomic sequence harboring three complete genes DvPP2 C,encoding a protein phosphatase,DvRGA2 and DvRGA1,as well as a separated gene DvRGA3,were then obtained by PCR *** analysis showed that all these genes co-segregate *** analysis demonstrated both DvRGA1 and DvRGA2 responded to Bgt invasion while DvRGA3 had no ***,the silencing effects of DvRGA1 and DvRGA2 in Yangmai 18 carrying Pm21 were analyzed by using the BSMV-VIGS *** silencing of DvRGA2 could cause the normal devebpment of Bgt spores and macroscopic disease symptom on ***,no obvious effects were observed after silencing of *** 6,408 M families of Yangmai 18 treated with EMS,a total of 58 independent lines highly susceptible to powdery mildew during the whole growth stage were screened *** for Y18-S16

关键词： dasypyrum villosum wheat powdery mildew resistance DvRGA2 Pm21

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