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Identification of chilling-responsive transcripts in peanut ( Arachis hypogaea  L.)
Tang, Yue Yi; Wang, Chuan Tang; Yang, Guan Pin; Feng, Tong; Gao, Hua Yuan; Wang, Xiu Zhen; Chi, Xiao Yuan; Xu, Ya Long; Wu, Qi & Chen, Dian Xu
Abstract
To isolate differentially expressed peanut genes responsive to chilling, a suppression subtractive hybridization (SSH) cDNA library was constructed for a chilling tolerant peanut cultivar A4 with mRNAs extracted from the seeds imbibed at 2°C and 15°C, respectively, for 24 hrs. A total of 466 cDNA clones were sequenced, from which 193 unique transcripts (73 contigs and 120 singlets) were assembled. Of these unique transcripts, 132 (68.4%) were significantly similar to the sequences in GenBank non-redundant (nr) protein database, which belonged to diverse functional categories including metabolism, signal transduction, stress response, cell defense and transcriptional regulation. The remaining 61 (31.6%) showed no similarity to either hypothetical or known proteins. Six differentially expressed transcripts were further confirmed with real-time quantitative PCR (RT-qPCR).
Keywords
chilling response, peanut, real-time quantitative PCR, suppression subtractive hybridization
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