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A large insert Thellungiella halophila BIBAC library for genomics and identification of stress tolerance genes

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Abstract

Salt cress (Thellungiella halophila), a salt-tolerant relative of Arabidopsis, has turned to be an important model plant for studying abiotic stress tolerance. One binary bacterial artificial chromosome (BIBAC) library was constructed which represents the first plant-transformation-competent large-insert DNA library generated for Thellungiella halophila. The BIBAC library was constructed in BamHI site of binary vector pBIBAC2 by ligation of partial digested nuclear DNA of Thellungiella halophila. This library consists of 23,040 clones with an average insert size of 75 kb, and covers 4× Thellungiella halophila haploid genomes. BIBAC clones which contain inserts over 50 kb were selected and transformed into Arabidopsis for salt tolerant plant screening. One transgenic line was found to be more salt tolerant than wild type plants from the screen of 200 lines. It was demonstrated that the library contains candidates of stress tolerance genes and the approach is suitable for the transformation of stress susceptible plants for genetic improvement.

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Abbreviations

BIBAC:

Binary bacterial artificial chromosome

T. halophila :

Thellungiella halophila

MS:

Murashige and Skoog

PFGE:

Pulsed-field gel electrophoresis

GUS:

Glucronidase

HMW:

High-molecular-weight

LMP:

Low-melting-point

DTT:

Dithiothreitol

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Acknowledgments

This research was supported by grants by National Natural Science Foundation of China (No. 90717006/30670195) to Q. Xie and Y. Wu, the Chinese Academy of Science (KSCX2–YW–N–010 and CXTD–S2005–2).

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Authors and Affiliations

  1. State Key Laboratory for Biocontrol, Sun Yat-sen (Zhongshan) University, 510275, Guangzhou, China

    Weiquan Wang, Yin Li, Jiaying Xie, Zhonghui Zhang, Zhiyong Deng, Yiyue Zhang, Cuiping Yang & Chengwei Yang

  2. State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Datun Road, 100101, Beijing, China

    Yaorong Wu, Yiyue Zhang, Jianbin Lai, Huawei Zhang, Hongyan Bao, Sanyuan Tang & Qi Xie

  3. State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, 100080, Beijing, China

    Peng Gao, Guixian Xia & Huishan Guo

Authors
  1. Weiquan Wang
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  2. Yaorong Wu
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  3. Yin Li
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  4. Jiaying Xie
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  5. Zhonghui Zhang
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  6. Zhiyong Deng
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  7. Yiyue Zhang
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  9. Jianbin Lai
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  10. Huawei Zhang
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  11. Hongyan Bao
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  12. Sanyuan Tang
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  13. Chengwei Yang
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  14. Peng Gao
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  15. Guixian Xia
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  16. Huishan Guo
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  17. Qi Xie
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Corresponding author

Correspondence to Qi Xie.

Additional information

Weiquan Wang, Yaorong Wu, and Yin Li have contributed equally to this work.

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Wang, W., Wu, Y., Li, Y. et al. A large insert Thellungiella halophila BIBAC library for genomics and identification of stress tolerance genes. Plant Mol Biol 72, 91–99 (2010). https://doi.org/10.1007/s11103-009-9553-3

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  • DOI: https://doi.org/10.1007/s11103-009-9553-3

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