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...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.展开更多
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 villo...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 susceptibility.Microscopic 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.展开更多
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 ...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.展开更多
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 provid...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.展开更多
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...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 mutant.The 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.展开更多
一年生二倍体簇毛麦携带多种抗性基因,被认为是小麦抗性改良的重要基因源。本研究前期创制了一个高抗小麦条锈病的普通小麦(中国春,CS)-簇毛麦3V(3D)代换系CD-3,并将该条锈病抗性基因初步定位于簇毛麦3V染色体上。本研究以川麦60作为父...一年生二倍体簇毛麦携带多种抗性基因,被认为是小麦抗性改良的重要基因源。本研究前期创制了一个高抗小麦条锈病的普通小麦(中国春,CS)-簇毛麦3V(3D)代换系CD-3,并将该条锈病抗性基因初步定位于簇毛麦3V染色体上。本研究以川麦60作为父本,以CD-3为母本,在回交自交系BC,F2群体中筛选到一个高抗条锈病的株系,命名为1901-245,并进一步对该株系进行分子标记、焚光原位杂交(FISH)及田间条绣病抗性鉴定。分子标记和熒光原位杂交(fluorescence in situ hybridization,FISH)结果显示,1901-245携带一对簇毛麦3V染色体,丢失了一对小麦3D染色体;1901-245花粉母细胞减数分裂中染色体配对正常,单价体出现频率较低,且3V染色体未出现单价体。田间条锈病抗性鉴定结果显示,1901-245及其亲本CD-3均表现为高抗小麦条锈病,而亲本川麦60则表现为高感小麦条锈病。以上结果表明,簇毛麦3V染色体上的条锈病抗性基因在1901-245的遗传背景下能正常表达;1901-245为普通小麦-簇毛麦3V(3D)代换系材料,高抗小麦条锈病且遗传稳定,可进一步在小麦遗传改良中加以应用。另外,FISH鉴定及植株形态观察结果显示,1901-245仍含有较多亲本CD-3(CS背景)的遗传背景,因此后期可继续将1901-245与川麦60进行回交,并在后代中加大簇毛麦3V染色体的追踪力度。展开更多
基金supported by the National Key Research and Develpment Program of China (2021YFD1200600)the National Natural Science Foundation of China (31871619, 32101703, and 32101800)+4 种基金the Natural Science Foundation of Jiangsu Province (BK20210152)the Jiangsu Seed Industry Revitalization Project (JBGS (2021) 013)the Key Research and Development Program of Jiangsu Province(BE2022346)Jiangsu Agricultural Science and Technology Innovation Fund of China (CX (20) 3029)supported by the European Regional Development Fund (CZ.02.1.01/0.0/0.0/16_019/0000827)。
文摘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.
基金supported by the National Natural Science Foundation of China(31971938)the Natural Science Foundation of Jiangsu Province(BK20181316)+1 种基金the Special Fund for Independent Innovation of Agricultural Science and Technology in Jiangsu,China(CX(19)1001)Fundamental Research Funds for the Central Universities(KYZ201809)。
文摘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 susceptibility.Microscopic 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.
基金financially supported by the National Key Research and Development Program,China (2016YFD0102000) the Agricultural Science and Technology Innovation Program (ASTIP) of the CAAS
文摘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.
基金supported by the National Key Research and Development Program of China (2016YFD0102001)the National Natural Science Foundation of China (31571653, 31771782, 31201204)+5 种基金the Technology Support Program of Jiangsu Province, China (BE2013439)the Fundamental Research Funds for the Central Universities, China (KYZ201403)the Jiangsu Agricultural Science and Technology Innovation Fund, China (CX151001)the Program of Introducing Talents of Discipline to Universities, China (B08025)the Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, China (PAPD)the Six Talent Peaks Project in Jiangsu Province, China
文摘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.
文摘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 mutant.The 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.
文摘一年生二倍体簇毛麦携带多种抗性基因,被认为是小麦抗性改良的重要基因源。本研究前期创制了一个高抗小麦条锈病的普通小麦(中国春,CS)-簇毛麦3V(3D)代换系CD-3,并将该条锈病抗性基因初步定位于簇毛麦3V染色体上。本研究以川麦60作为父本,以CD-3为母本,在回交自交系BC,F2群体中筛选到一个高抗条锈病的株系,命名为1901-245,并进一步对该株系进行分子标记、焚光原位杂交(FISH)及田间条绣病抗性鉴定。分子标记和熒光原位杂交(fluorescence in situ hybridization,FISH)结果显示,1901-245携带一对簇毛麦3V染色体,丢失了一对小麦3D染色体;1901-245花粉母细胞减数分裂中染色体配对正常,单价体出现频率较低,且3V染色体未出现单价体。田间条锈病抗性鉴定结果显示,1901-245及其亲本CD-3均表现为高抗小麦条锈病,而亲本川麦60则表现为高感小麦条锈病。以上结果表明,簇毛麦3V染色体上的条锈病抗性基因在1901-245的遗传背景下能正常表达;1901-245为普通小麦-簇毛麦3V(3D)代换系材料,高抗小麦条锈病且遗传稳定,可进一步在小麦遗传改良中加以应用。另外,FISH鉴定及植株形态观察结果显示,1901-245仍含有较多亲本CD-3(CS背景)的遗传背景,因此后期可继续将1901-245与川麦60进行回交,并在后代中加大簇毛麦3V染色体的追踪力度。
