Abstract
MicroRNAs (miRNAs) are approximately 22-nucleotide (nt)-long non-coding RNAs that play a key role in plant development and abiotic stresses. We have developed a simple but effective array platform for profiling plant miRNAs from various plant species. The array is composed of 188 non-redundant miRNA probes that can detect both conserved and species-specific miRNAs from most plant species, including Arabidopsis, rice, and poplar. In this chapter, we describe a protocol for developing the miRNA array platform, which can be used to identify stress-responsive miRNAs in diverse plant species. Profiling of miRNAs in tobacco seedlings exposed to different abiotic stress conditions has revealed that a number of miRNAs, miR398, miR399, miR408, miR156, miR164, and miR168, were responsive to stresses. This comprehensive and easy-to-follow protocol will be useful for studying roles of miRNAs in plant stress response.
The first two authors contribute equally to this work.
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Acknowledgments
G.T. is supported by the Kentucky Tobacco Research and Development Center (KTRDC), the USDA-NRI grants 2006-35301-17115 and 2006-35100-17433, the NSF MCB-0718029 (Subaward No. S-00000260), and an award from the Kentucky Science and Technology Corporation under Contract # KSTC-144-401-08-029.
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Jia, X., Mendu, V., Tang, G. (2010). An Array Platform for Identification of Stress-Responsive MicroRNAs in Plants. In: Sunkar, R. (eds) Plant Stress Tolerance. Methods in Molecular Biology, vol 639. Humana Press. https://doi.org/10.1007/978-1-60761-702-0_15
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DOI: https://doi.org/10.1007/978-1-60761-702-0_15
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