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Integration of genetic and physical maps of the chickpea (Cicer arietinum L.) genome using flow-sorted chromosomes

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Abstract

Cultivated chickpea is the third most important legume after field bean and garden pea worldwide. Despite considerable breeding towards improved yield and resistance to biotic and abiotic stresses, the production of chickpea remained stagnant, but molecular tools are expected to increase the impact of current improvement programs. As a first step towards this goal, various genetic linkage maps have been established and markers linked to resistance genes been identified. However, until now, only one linkage group (LG) has been assigned to a specific chromosome. In the present work, mitotic chromosomes were sorted using flow cytometry and used as template for PCR with primers designed for genomic regions flanking microsatellites. These primers amplify sequence-tagged microsatellite site markers. This approach confirmed the assignment of LG8 to the smallest chromosome H. For the first time, LG5 was linked to the largest chromosome A, LG4 to a medium-sized chromosome E, while LG3 was anchored to the second largest chromosome B. Chromosomes C and D could not be flow-sorted separately and were jointly associated to LG6 and LG7. By the same token, chromosomes F and G were anchored to LG1 and LG2. To establish a set of preferably diagnostic cytogenetic markers, the genomic distribution of various probes was verified using FISH. Moreover, a partial genomic bacterial artificial chromosome (BAC) library was constructed and putative single/low-copy BAC clones were mapped cytogenetically. As a result, two clones were identified localizing specifically to chromosomes E and H, for which no cytogenetic markers were yet available.

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Abbreviations

BAC:

Bacterial artificial chromosome

FISH:

Fluorescence in situ hybridization

LG:

Linkage group

MAS:

Marker-assisted selection

NOR:

Nucleolus organizing region

rDNA:

Ribosomal DNA

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Acknowledgments

We are grateful to Dr. P. Winter (GenXPro GmbH, Frankfurt Innovation Centre, Frankfurt am Main, Germany) and Dr. B. Hüttel (MPI for Plant Breeding Research, Cologne, Germany) for providing primers for chromosome-specific microsatellite markers and valuable suggestions concerning PCR with these markers. We thank our colleagues Romana Šperková, Bc. and Jitka Weiserová, Bc. for excellent technical assistance. We appreciate valuable comments and suggestions by two anonymous reviewers. This work was supported by Ministry of Education, Youth, and Sports (grant award no. LC06004).

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  1. Pavlína Zatloukalová

    Present address: Department of Functional Genomics and Proteomics, Laboratory of Molecular Plant Physiology, Masaryk University, Kamenice 5, 625 00, Brno, Czech Republic

Authors and Affiliations

  1. Institute of Experimental Botany, Centre of the Region Haná for Biotechnological and Agricultural Research, Sokolovská 6, 772 00, Olomouc, Czech Republic

    Pavlína Zatloukalová, Eva Hřibová, Marie Kubaláková, Pavla Suchánková, Hana Šimková & Jaroslav Doležel

  2. Genetics Department, University of Córdoba, Campus Rabanales (C5), 14071, Córdoba, Spain

    Cabrera Adoración & Teresa Millán

  3. Molecular BioSciences, Biocenter, Max-von-Laue-Strasse 9, 60438, Frankfurt am Main, Germany

    Günter Kahl

  4. Institute of Experimental Botany, Sokolovská 6, 772 00, Olomouc, Czech Republic

    Jaroslav Doležel

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  1. Pavlína Zatloukalová
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Correspondence to Jaroslav Doležel.

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Zatloukalová, P., Hřibová, E., Kubaláková, M. et al. Integration of genetic and physical maps of the chickpea (Cicer arietinum L.) genome using flow-sorted chromosomes. Chromosome Res 19, 729–739 (2011). https://doi.org/10.1007/s10577-011-9235-2

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