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Acta Agron Sin ›› 2011, Vol. 37 ›› Issue (05): 918-923.doi: 10.3724/SP.J.1006.2011.00918

• RESEARCH NOTES • Previous Articles     Next Articles

Expression Analysis of Three Genes from SSH Library Constructed Using Tillering Nodes of Dongnongdongmai 1 under Low Temperature

MOU Yong-Chao, CUI Hong, YU Jing, ZENG Yan, MENG Jian-Nan,CANG Jing*   

  1. College of Life Science, Northeast Agricultural University, Harbin 150030, China
  • Received:2010-10-28 Revised:2011-03-08 Online:2011-05-12 Published:2011-03-24

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Abstract: To reveal the molecular mechanism of cold resistance of winter wheat variety Dongnongdongmai 1, under natural conditions in the north region of China, we collected tillering nodes before (5°C) and in winter season (-25°C) respectively, then constructed cDNA library of Dongnongdongmai 1 containing low temperature stress related genes by suppression subtractive hybridization. A total of 300 positive clones selected from cDNA library randomly were sequenced to obtain 230 high quality expressed sequence tags (EST). After BLAST and functional annotation, we found complex and diverse types of genes in the library, including stress response genes (such as heat shock protein, CS66, Wcor8 and other related genes) and genes encoding 40S and 60S ribosomal subunit. These genes with high frequency may play an important role in protecting Dongnongdongmai 1 from injuring under low temperature. Two unknown genes and Clp selected from the cDNA library were analyzed through quantitative PCR. The expression levels of these genes in Dongnongdongmai 1 were increased in various degrees with the decrease of temperature; however, their expression patterns were different in Jimai 22 (a winter wheat cultivar with low resistance to low temperature). This result suggests that these genes may play an important role in resistance to low temperature stress.

Key words: Dongnongdongmai 1, Gene for cold resistance, Suppression subtractive hybridization, Expressed sequence tag, Real-time PCR

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