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Molecular cytogenetic characterization of stem rust and stripe rust resistance in wheat-Thinopyrum bessarabicum–derived doubled haploid lines

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Abstract

Thinopyrum species are potential sources of disease resistance for wheat. Thinopyrum bessarabicum is a well-known source of salt tolerance, but there are very few reports of rust resistance being transferred from this species to wheat. Here, we report wheat-Th. bessarabicum–derived stem rust and stripe rust–resistant doubled haploid (DH) lines and characterized using molecular markers and cytogenetic analyses. We used sequential non-denaturing fluorescence in situ hybridization (ND-FISH) and genomic in situ hybridization (GISH) to determine their genomic compositions. Cytological results were validated by molecular analysis using genome-specific PCR markers. These lines were either alien additions, partial diploids, or partial amphidiploids. Line DH9 was a disomic addition line carrying chromosome 4Jb whereas DH1 was a double disomic addition line for chromosomes 6Jb and 7Jb. Lines DH7 and DH8 were partial diploids that were cytologically unstable due to having ph1ph1 genotype. DH7 also had a pair of T6BL-6JbL.6JbS translocated chromosomes and was monosomic for a T5DS-5JbS.5JbL translocation. The size of the translocated alien segment in chromosome T5DS-5JbS.5JbL was L+(0–0.42)S, whereas the size of wheat chromatin in chromosome T6BL-6JbL.6JbS was S+(0–0.85)L. Lines DH5 (2n = 50) and DH11 (2n = 54) were partial amphidiploids with eight and 12 Th. bessarabicum chromosomes, respectively. Lines DH7, DH8, and DH11 expressed an intermediate level of resistance to stem rust with infection types (ITs) varying from 1+2 to 23-, whereas they were immune or near-immune (IT 0 to 0) to stripe rust. Comparison of the rust reactions, genome compositions, and molecular markers of the DH lines along with their parents indicated that the Thinopyrum chromosome conferring resistance to both rusts was 2Jb.

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Acknowledgments

The authors express their sincere thanks to Professors Ian King and Julie King, University of Nottingham, UK, for sharing the seeds of the amphidiploid of Chinese Spring and Th. bessarabicum. AKS is grateful to the University of Sydney for awarding Ph.D. scholarship and for providing the platform to conduct the research activities. We thank Dr. Nizam Ahmed for generation of DH lines. The authors would like to extend sincere thanks to Prof. R. A. McIntosh for reviewing this manuscript.

Funding

The authors are thankful to BBSRC-UK for funding under the grant BB/100260X/1.

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Authors and Affiliations

  1. Plant Breeding Institute, and Sydney Institute of Agriculture, University of Sydney, Cobbitty, NSW, 2570, Australia

    Amit Kumar Singh, Peng Zhang, Chongmei Dong, Jianbo Li, Richard Trethowan & Peter Sharp

  2. School of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu, 611731, Sichuan, China

    Jianbo Li

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  1. Amit Kumar Singh
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  2. Peng Zhang
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  3. Chongmei Dong
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  4. Jianbo Li
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Contributions

AKS, PS, and PZ conceived the project. AKS conducted the experiments and drafted the manuscript. PZ performed GISH experiments and phenotyped the rust reactions. CD assisted in running of the capillary gel electrophoresis. JL assisted in the oligo-FISH experiments. PZ, CD, PS, and RT reviewed the manuscript.

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Correspondence to Peter Sharp.

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Singh, A.K., Zhang, P., Dong, C. et al. Molecular cytogenetic characterization of stem rust and stripe rust resistance in wheat-Thinopyrum bessarabicum–derived doubled haploid lines. Mol Breeding 39, 125 (2019). https://doi.org/10.1007/s11032-019-1034-z

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