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Construction of a YAC library from a Beta vulgaris fragment addition and isolation of a major satellite DNA cluster linked to the beet cyst nematode resistance locus Hs1 pat−1

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Abstract

A YAC library was constructed from the Beta vulgaris fragment addition AN5-203b. This monosomic fragment addition harbors an approximate 12-Mbp fragment of B.patellaris chromosome 1 accomodating the Hs1 pat-1conferring resistance to the beet cyst nematode (Heterodera schachtii). The YAC library consists of 20,000 YAC clones having an average size of 140 kb. Screening with organelle-specific probes showed that 12% of the clones contain chloroplast DNA while only 0.2% of the clones hybridizes with a mitochondrial specific probe. On the basis of a sugar beet haploid genome size of 750 Mbp this library represents 3.3 haploid genome equivalents. The addition fragment present in AN5-203b harbors a major satellite DNA cluster that is tightly linked to the Hs1 pat-1locus. The cluster is located on a single 250-kb EcoRI restriction fragment and consists of an estimated 700–800 copies of a 159-bp core sequence, most of which are arranged in tandem. Using this core sequence as a probe, we were able to isolate 1 YAC clone from the library that contains the entire 250-kb satellite DNA cluster.

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Abbreviations

YAC :

Yeast artificial chromosome

BCN :

beet cyst nematode

RAPD :

random amplified polymorphic DNA

RFLP :

restriction fragment length polymorphism

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Authors and Affiliations

  1. Department of Molecular Biology, DLO-Center for Plant Breeding and Reproduction Research (CPRO-DLO), P.O. Box 16, NL-6700 AA, Wageningen, The Netherlands

    R. M. Klein-Lankhorst, E. M. J. Salentijn, W. G. Dirkse, M. Arens-de Reuver & W. J. Stiekema

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  1. R. M. Klein-Lankhorst
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  2. E. M. J. Salentijn
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  3. W. G. Dirkse
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  4. M. Arens-de Reuver
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  5. W. J. Stiekema
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Communicated by F. Salamini

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Klein-Lankhorst, R.M., Salentijn, E.M.J., Dirkse, W.G. et al. Construction of a YAC library from a Beta vulgaris fragment addition and isolation of a major satellite DNA cluster linked to the beet cyst nematode resistance locus Hs1 pat−1 . Theoret. Appl. Genetics 89, 426–434 (1994). https://doi.org/10.1007/BF00225377

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