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

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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

Qifeng Wang, Qiong Yi, Kunzhi Li, Limei Chen, Yongxiong Yu . 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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