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Acta Agron Sin ›› 2011, Vol. 37 ›› Issue (05): 764-771.doi: 10.3724/SP.J.1006.2011.00764

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Isolation of NBS-LRR Late Blight Resistance Genes from Potato BAC Library by System of Magnetic Separation

XU Jian-Fei, JIN Li-Ping, PANG Wan-Fu, BIAN Chun-Song, DUAN Shao-Guang, LIU Jie, HUANG San-Wen, and QU Dong-Yu*   

  1. Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, China
  • Received:2010-11-18 Revised:2011-03-08 Online:2011-05-12 Published:2011-03-24
  • Contact: 屈冬玉, E-mail: dyqu@mail.caas.net.cn, Tel: 010-82105943

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Abstract: Cultivated potato (Solanum tuberosum) is an auto-tetraploid crop, which is very difficult to isolate late blight resistance genes. Nevertheless, it is discovered that there is conseved NBS-LRR domain among late blight resistance genes. In this study, conserved probes of R3a late blight resistance genes family were developed by alignment of sequences. And then, the DNA of BAC SH23G23 containingthe R3a gene was partially digested into 7–11 kb fragments. By magnetic separation system combined with conserved probes, the 7–11 kb fragments were enriched and cloned into binary vector pBINPLUS. Through identification of positive clones and colony PCR, the ratio of clones including R3a gene to all positive clones reached 82.76%, which was nearly 19 times higher than enrichment before. The system of magnetic separation for R genes and their analogs established in this study provides a new strategy for conserved domain genes cloning from polyploid crops.

Key words: Potato, Late blight, Resistance genes, Magnetic separation

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