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Isolation of high quality RNA and construction of a suppression subtractive hybridization library from ramie (Boehmeria nivea L. Gaud.)

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Abstract

Isolation of high quality RNA from ramie (Boehmeria nivea L. Gaud.) is difficult due to its high levels of polyphenols, polysaccharides, pectin, fat, wax and other secondary metabolites. A modified procedure based on guanidinium isothiocyanate for RNA preparation of ramie was developed in this study. High concentrations (5%, v/v) of guanidinium isothiocyanate, PVP-4000, sodium citrate and sodium lauryl sarcosinate and β-mercaptoethanol were used in the extraction buffer, together with a low pH sodium acetate (pH 4.0) added to improve the RNA quality. The average yield was about 400 μg RNAg−1 fresh leaves. One SSH library which was induced by ramie anthracnose was constructed by utilizing the RNA extracted through the present method. These results showed that our protocol was applicable for RNA isolation from recalcitrant ramie tissues.

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Acknowledgments

This research was supported by National Natural Science Funds (30800696) and was also supported by Special Fund for Public Welfare Industry of China (nyhyzx07-018-16). We also wish to thank Pro. Yucheng Jie for providing ramie anthracnose diseased plants.

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Authors and Affiliations

  1. College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, 430070, Hubei Province, Poeple’s Republic of China

    Xu-xia Wang, Bo Wang, Li-jun Liu, Xiao-ping Cui, Jin-yu Yang, Hui Wang, Hui Jiang, Bei-bei Luo, Zhu Long, Wen-xiang Dou, Na Zhang & Ding-xiang Peng

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  1. Xu-xia Wang
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  2. Bo Wang
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Correspondence to Bo Wang.

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Wang, Xx., Wang, B., Liu, Lj. et al. Isolation of high quality RNA and construction of a suppression subtractive hybridization library from ramie (Boehmeria nivea L. Gaud.). Mol Biol Rep 37, 2099–2103 (2010). https://doi.org/10.1007/s11033-009-9671-7

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  • DOI: https://doi.org/10.1007/s11033-009-9671-7

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