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Acta Agron Sin ›› 2013, Vol. 39 ›› Issue (04): 744-752.doi: 10.3724/SP.J.1006.2013.00744

• RESEARCH NOTES • Previous Articles     Next Articles

Construction and Analysis of SSH Library in Rapeseed (Brassica napus L.) under Drought Stress

XIE Xiao-Yu,ZHANG Bing,ZHANG Xia,MA Zhong-Lian,LI Jia-Na*   

  • Received:2012-08-04 Revised:2012-12-08 Online:2013-04-12 Published:2013-01-28
  • Contact: 李加纳, E-mail: ljn1950@swu.edu.cn E-mail:xiexy8009@163.com

Abstract:

The positive SSH library in leaves of rapeseed (Brassica napus L.) Holiday (with high drought resistance ability) was constructed using the sample under drought stress as Tester and the sample under normal water management as Driver. Twenty-four positive clones selected randomly were verified by using PCR. The result of electrophoresis showed that 23 clones contained inserts whose average length was 750 bp. Ninety-six positive clones were sequenced and analyzed by CAP3 Sequence Assembly Program, showing that four contigs and 82 singletons were contained and the average length of 86 ESTs was 542 bp. Blastx alignment with Nr database showed 11 ESTs without significant similarity and 75 ESTs with similarity in GenBank database. KOBAS suggested that 28 ESTs were mapped to 67 pathways. It could be predicted that the carbon fixation in photosynthetic organism, electron donor III of aerobic respiration, nitrogen metabolism, glyoxylate and dicarboxylate metabolism played a key role in drought tolerance ofrapeseed. The analysis of ESTs showed that a large share of function could be attributed to organelle (58.82%), binding (30.77%), and metabolic process (43.72%), which plays important roles in metabolism of rapeseed under drought stress.

Key words: Rapeseed (Brassica napus L.), Suppression subtractive hybridization (SSH), Drought stress, cDNA library, Express sequence tag

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