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Chinese Journal of High Pressure Physics  > 2008  >  22(4): 370-376 .
LIU Xun-Cheng, ZHANG Mei, DUAN Jun. Construction and Analysis of SSH-cDNA Library Induced by High Pressure in Rice Seeds during Germination[J]. Chinese Journal of High Pressure Physics, 2008, 22(4): 370-376 . doi: 10.11858/gywlxb.2008.04.006
Citation: LIU Xun-Cheng, ZHANG Mei, DUAN Jun. Construction and Analysis of SSH-cDNA Library Induced by High Pressure in Rice Seeds during Germination[J]. Chinese Journal of High Pressure Physics, 2008, 22(4): 370-376 . doi: 10.11858/gywlxb.2008.04.006
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Construction and Analysis of SSH-cDNA Library Induced by High Pressure in Rice Seeds during Germination

doi: 10.11858/gywlxb.2008.04.006
  • 1.

    South China Botanic Garden, Chinese Academy of Sciences, Guangzhou 510650, China;

  • 2.

    Graduate University of Chinese Academy of Sciences, Beijing 100039, China

More Information
  • Corresponding author: DUAN Jun
  • Received Date: 28 Nov 2007
  • Rev Recd Date: 28 Mar 2008
  • Publish Date: 05 Dec 2008
    Abstract
  • In order to study the effect of expression pattern of high hydrostatic pressure on high plant, a SSH-cDNA library was constructed with rice germinating seeds in this paper. Suppression subtractive hybridization (SSH) in combination with mirror orientation selection (MOS) was performed using the mRNA from rice germinating seeds treated by 75 MPa high hydrostatic pressure as tester and the mRNA from control (seeds untreated by high hydrostatic pressure) as the driver. 580 recombinant clones were picked through blue/white screening. PCR detection of the recombinant clones showed that the size of inserts mainly ranged from 200 bp to 1000 bp, and the average size of these inserts were about 575 bp. Among the 580 recombinant clones, 542 clones (93.3%) were identified to be effective single band ones. After sequencing some of the recombinant clones, it was found that the genes induced by high hydrostatic pressure were mainly involved in metabolism, stress response and transcriptional regulation. The research paved the way for studying the gene expression patterns of rice seeds responding to high hydrostatic pressure.

     

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