Abstract
Suppression subtraction hybridization (SSH) libraries were constructed from RNA isolated from leaves of control and cold stress-induced Lepidium latifolium, a cold-tolerant plant species from high altitudes for isolation of cold-responsive genes. A total of 500 clones were obtained from the cold stress library. Dot blot expression analysis identified 157 clones that were upregulated and 75 that were downregulated during cold stress. These clones selected on the basis of their expression patterns on dot blot were sequenced. As much as 27 and 17 genes were identified from the forward and reverse libraries, respectively. The genes identified revealed homology with genes involved in diverse processes such as gene regulation/signaling, photosynthesis, DNA damage repair protein, pathogenesis-related protein, senescence-associated proteins and proteins with unknown functions.
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Acknowledgments
We thank Director, DIHAR (FRL), Leh for kind provision of seeds of Lepidium latifolium and Dr. Y.S. Shouche (National Centre for Cell Science, Pune, India) for help in sequencing. Financial support to MA, VBS and RKS by DRDO is gratefully acknowledged.
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Communicated by J. Sadowski.
M. Aslam and V. B. Sinha equally contributed to this work.
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Aslam, M., Sinha, V.B., Singh, R.K. et al. Isolation of cold stress-responsive genes from Lepidium latifolium by suppressive subtraction hybridization. Acta Physiol Plant 32, 205–210 (2010). https://doi.org/10.1007/s11738-009-0382-4
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DOI: https://doi.org/10.1007/s11738-009-0382-4