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Construction of an SSH Library of Pinellia ternata Under Heat Stress, and Expression Analysis of Four Transcripts

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Abstract

Suppression subtractive hybridization (SSH) was used to clone transcripts upregulated during heat stress in leaves of Pinellia ternata. The complementary structures of deoxyribonucleic acid (cDNAs) of heat-stressed plants were used as the “tester,” and cDNAs of unstressed plants were used as the “driver.” Sequencing the whole SSH library yielded 303 expressed sequence tags (ESTs). Among these, 61 showed insignificant homology to any previously identified genes. Forty four of the remaining 242 ESTs represented singletons. Real-time quantitative reverse transcriptase polymerase chain reaction analysis of the expression patterns of four transcripts showed that three of these were upregulated after heat shock, and one was downregulated. The sequences showed high homology to small heat shock protein (sHSP), heat shock protein 90 (HSP90), heat shock protein 70 (HSP70), and stearoyl-ACP-protein desaturase (SAD), respectively. Rapid amplification of cDNA ends (RACE) was used to obtain full-length cDNAs of PtSAD and PtsHSP, and characterization and phylogenetic analyses were carried out according to their sequences.

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Acknowledgments

This work was sponsored by the National Natural Science Foundation of China (No. 30973963), the Natural Science Foundation of Anhui Province for the Special Program (No. 090413252), and the Natural Science Foundation of Anhui Universities for the Key Project (No. KJ2009A160).

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Authors and Affiliations

  1. Huaibei Normal University, Huaibei, Anhui, China

    Hedong Lu, Tao Xue, Aimin Zhang, Wei Sheng, Yanfang Zhu, Li Chang & Yunxian Song

  2. College of Life Science, Huaibei Normal University, Huaibei, 235000, Anhui Province, People’ s Republic of China

    Jianping Xue

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  1. Hedong Lu
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  2. Tao Xue
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  3. Aimin Zhang
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  4. Wei Sheng
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  6. Li Chang
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  7. Yunxian Song
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  8. Jianping Xue
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Correspondence to Jianping Xue.

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Lu, H., Xue, T., Zhang, A. et al. Construction of an SSH Library of Pinellia ternata Under Heat Stress, and Expression Analysis of Four Transcripts. Plant Mol Biol Rep 31, 185–194 (2013). https://doi.org/10.1007/s11105-012-0488-5

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