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Identification of differentially expressed transcripts involved in the salt-stress response of Salsola ferganica by suppression subtractive hybridization

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Abstract

To understand the molecular and cellular mechanisms underlying the salt response and to isolate differentially expressed genes, we constructed a forward subtractive cDNA library from Salsola ferganica treated with 600 mM NaCl for different lengths of time (0, 4, 8, 12, 16, 24, 36 h) using the suppression subtractive hybridization (SSH) technique. This SSH library contained approximately 2,200 positive clones, with the insert size ranging from 250 to 700 bp. Eighty clones were selected for further study based on results from reverse Northern blotting and sequencing and, these cDNAs are the first reported expressed sequence tags (ESTs) of Salsola ferganica listed in GenBank. Among these 80 differentially expressed genes, 32 are genes with known function that display a high homology (between 68 and 100% identity) to known sequences, 47 have an unknown function, showing no homology with known sequences, and one is a novel gene that does not match any previously reported genes. Five cDNA fragments, sf3d8, sf6d8, sf4a8, sf8d6, and sf6c7, were randomly chosen from the 80 differentially expressed genes for reverse transcription-PCR analysis. The expression of sf8d6 was entirely induced by salt stress and that of sf3d8, sf6d8, sf4a8, and sf6c7 was increased from a relatively lower level. The isolated ESTs reported here may enable us to find new genes involved in salt tolerance in Salsola ferganica.

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Abbreviations

DIG:

Digoxygenin

EST:

Expressed sequence tag

RT–PCR:

Reverse transcription PCR

SSH:

Suppression subtractive hybridization

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Acknowledgments

The research reported here was supported by the Educational Natural Science Foundation for Young teachers of China (No.XJEDU-2009S04). We would like to thank Dr Hai-yan Lan for her comments on this experiment. We also would like to acknowledge Dr. Ji Ma for her helpful suggestions on this paper.

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Authors and Affiliations

  1. Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, 14 Shengli Road, 830046, Urumqi, China

    Bo Guan, Ying-xin Wang, Zhi-cai Wang, Yakupjan Haxim, Qian Bao, You-zhen Hu, Fu-chun Zhang & Yan Wang

  2. Xinjiang Entry-Exit Inspection and Quarantine Bureau of the People’s Republic of China, 116 Nanhu Northern Road, 830063, Urumqi, China

    Gang-qiang Jiang

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  1. Bo Guan
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  9. Yan Wang
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Correspondence to Yan Wang.

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Guan, B., Jiang, Gq., Wang, Yx. et al. Identification of differentially expressed transcripts involved in the salt-stress response of Salsola ferganica by suppression subtractive hybridization. Plant Cell Tiss Organ Cult 103, 343–352 (2010). https://doi.org/10.1007/s11240-010-9787-4

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  • DOI: https://doi.org/10.1007/s11240-010-9787-4

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