Construction of a Suppression Subtractive Hybridization Library for Stylosanthes guianensis Under Aluminum Stress and Expressed Sequence Tag Analysis

  • Kunzhi Li ,
  • Limei Chen ,
  • Yongxiong Yu ,
  • Qifeng Wang ,
  • Qiong Yi
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  • 1Bioscience and Biotechnology Research Center, Kunming University of Science and Technology, Kunming 650224, China

    2College of Zoological Science and Technology, Southwest University, Chongqing 400716, China

Received date: 2010-05-24

  Revised date: 2010-07-05

  Online published: 2010-09-20

Abstract

Stylo (Stylosanthes guianensis) is a forage grass grown worldwide. A cultivar of stylo (‘Reyan No.2’) was shown to be able to exudate citrate and tolerate aluminum (Al). To identify Al-induced genes in stylo, we constructed a forward cDNA library for Reyan No.2 under 300 μmol·L–1 Al stress by suppression subtractive hybridization. Approximately 600 clones with inserts > 300 bp were selected for sequence analysis. In total, 504 expressed sequence tags (ESTs) were obtained: 12.1% were singletons and 61.4% presented as 2–16 copies. The highly repetitive ESTs were 10.5% for cytochrome P450-like TBP protein, with 53 copies; 8.7% for pathogen-inducible trypsin-inhibitor-like protein, with 44 copies; and 7.3% for putative senescence-associated protein, with 37 copies. BLASTX analysis revealed that the 504 ESTs could be assembled into 97 non-redundant sequences, including 46 genes with known function. Of 46 genes with known function, 30 were previously reported to be Al-responsive genes and the other 16 ESTs are novel Al-associated genes. The information provides important clues to facilitate understanding the Al detoxification mechanism in stylo at the molecular level.

Cite this article

Kunzhi Li , Limei Chen , Yongxiong Yu , Qifeng Wang , Qiong Yi . Construction of a Suppression Subtractive Hybridization Library for Stylosanthes guianensis Under Aluminum Stress and Expressed Sequence Tag Analysis[J]. Chinese Bulletin of Botany, 2010 , 45(06) : 679 -688 . DOI: 10.3969/j.issn.1674-3466.2010.06.005

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