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Acta Agron Sin ›› 2009, Vol. 35 ›› Issue (6): 1156-1160.doi: 10.3724/SP.J.1006.2009.01156

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Expression of GMCHI gene，isolated from Soybean，Enhances the Survival in Prokaryotes to Low Temperature Stress

CHENG Li-Bao1,LI Shu-Yan2,HE Guang-Yuan1*   

  1. 1China-UK HUST-RRes Genetics Engineering and Genomics Joint Laboratory,Huazhong University of Science Technology,Wuhan 430074,China；2Institute of Botany，Chinese Academy of Sciences，Beijing 100093，China；3Life Science and Technique College of Xiaogan University，Xiaogan432000，China
  • Received:2008-11-05 Revised:2009-01-28 Online:2009-06-12 Published:2009-03-23
  • Contact: HE Guang-Yuan，E-mail:hegy@hust.edu.cn；Tel：027-87792271
  • About author:CHEN Li-Bao，E-mail:chenglibao453@163.com；Tel:027-87792271

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Abstract:

Low temperature during germination being a natural disaster usually occurs in North of China, which results in low germination rate and low seedling vigor, and even ultimately severe loss in yield. A gene was isolated from soybean Zhonghuang 22 cultivar (tolerant to low temperature in imbibition) for 24 h under 4℃ via cDNA-AFLP, name as GMCHI, GenBank accession No. EU699765. The full-length sequence of GMCHI which was 387 bp in nucleotide was obtained by RACE method. The sequence of gene shared low similarity with that of genes documented in NCBI, we inferred that GMCHI was a new discovered gene in soybean. Semi quantitative RT-PCR result revealed that GMCHI was induced by ABA and IPEG.. The gene ligated with Prokaryotic expression vector was transformed into prokaryotic cell, the result of 6% SDS-PAGE induced by IPTG was expressed in Prokaryotes. The E. coli carrying GMCHI survived and showed tolerance to low temperature under the treatment of –20℃ for 2 h. RT-PCR validated that GMCHI in Prokaryotes improved the adaptation to low temperature.

Key words: Soybean, GMCHI, E.coli, cDNA-AFLP, Gene


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