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Construction of a binary BAC library for an apomictic monosomic addition line of Beta corolliflora in sugar beet and identification of the clones derived from the alien chromosome

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Abstract

A plant-transformation-competent binary BAC library was constructed from the genomic DNA of the chromosome 9 monosomic addition line of Beta corolliflora Zoss. in sugar beet (B. vulgaris. L). This monosomic addition line (designated M14) is characterized by diplosporic reproduction caused by the alien chromosome carrying the gene(s) responsible for diplospory. The library consists of 49,920 clones with an average insert size of 127 kb, representing approximately 7.5 haploid genome equivalents and providing a greater than 99% probability of isolating a single-copy DNA sequence from the library. To develop the scaffold of a physical map for the alien chromosome, B. corolliflora genome-specific dispersed repetitive DNA sequences were used as probes to isolate BAC clones derived from the alien chromosome in the library. A total of 2,365 positive clones were obtained and arrayed into a sublibrary specific for B. corolliflora chromosome 9 (designated bcBAC-IX). The bcBAC-IX sublibrary was further screened with a subtractive cDNA pool generated from the ovules of M14 and the floral buds of B. vulgaris by the suppression subtractive hybridization method. One hundred and three positive binary BACs were obtained, which potentially contain the genes of the alien chromosome specifically expressed during the ovule and embryo development of M14, and may be associated with apomictic reproduction. Thus, these binary BAC clones will be useful for identification of the genes for apomixis by genetic transformation.

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Acknowledgements

The authors thank Dr. Christian Jung for kindly providing the B. corolliflora genome-specific repetitive DNA sequences, and Dr. David M. Stelly for helpful suggestions on the experimental designs. This work was supported by National Plant Transgenic Research and Industrialization Special Foundation of China (J99-A-01), and in part by grants from National Scientific Funding Committee (39980015).

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Authors and Affiliations

  1. Institute of Genetics and Developmental Biology, No.3 Zhongguancun Nanyitiao, 100080, Beijing, China

    Xiaohua Fang, Suhai Gu, Fan Chen & Naihu Wu

  2. Department of Biology, Harbin, 150080, Heilongjiang , China

    Dedong Guo

  3. Department of Soil and Crop Sciences and Institute for Plant Genomics and Biotechnology, Texas A&M University, College Station, TX 77843, USA

    Zhanyou Xu & Hong-Bin Zhang

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  1. Xiaohua Fang
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  2. Suhai Gu
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  3. Zhanyou Xu
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  4. Fan Chen
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  5. Dedong Guo
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  6. Hong-Bin Zhang
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  7. Naihu Wu
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Correspondence to Naihu Wu.

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Communicated by H. C. Becker

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Fang, X., Gu, S., Xu, Z. et al. Construction of a binary BAC library for an apomictic monosomic addition line of Beta corolliflora in sugar beet and identification of the clones derived from the alien chromosome. Theor Appl Genet 108, 1420–1425 (2004). https://doi.org/10.1007/s00122-003-1566-8

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  • DOI: https://doi.org/10.1007/s00122-003-1566-8

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