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Plant Molecular Biology Reporter

, Volume 27, Issue 2, pp 117–125 | Cite as

Construction, Characterization, and Expressed Sequence Tag (EST) Analysis of Normalized cDNA Library of Thermo-Photoperiod-Sensitive Genic Male Sterile (TPGMS) Wheat from Spike Developmental Stages

  • D. Yang
  • Z. H. Tang
  • L. P. Zhang
  • C. P. ZhaoEmail author
  • Y. L. ZhengEmail author
Article

Abstract

Thermo-photoperiod-sensitive genic male sterile (TPGMS) wheat is important in utilization of heterosis. To facilitate the use of such wheat line in agriculture, more knowledge about molecular mechanisms of TPGMS genes is required. In this study, we set up a normalized complementary DNA (cDNA) library based on the strategy of saturation hybridization with genomic DNA using TPGMS wheat line. This normalized cDNA library consists of cDNA from six directionally cloned cDNA libraries constructed with spike and anther tissues from spike developmental stages. From the normalized cDNA library, 3,264 single-pass expressed sequence tag (EST) were obtained. Exclusion of sequences shorter than 100 bp resulted in 3,223 vector-trimmed ESTs with a mean length of 926 bp. Clustering and assembly analysis resulted in 2,175 unique ESTs from 423 contigs and 1,752 singletons. Taking advantage of various tools and database, gene function classification showed that 60% of the ESTs were predicted to have putative gene function. Of the 2,175 unique ESTs, 264 (12%) displayed significant homology (BlastX E values <10−5) to genes previously reported to be involved in cold-response related processes. Among these, sequences encoding activities related to primary metabolism, signal transduction, and transcriptional regulation were observed. Finally, in the total EST sequences, 108 potential SSRs were found. The unigene dataset will now be used to fabricate biochips carrying all identified genes for TPGMS wheat functional genomic research.

Keywords

EST Gene ontology Normalized cDNA library TPGMS Wheat 

Abbreviations

CHA

chemical hybridizing agents

CMS

cytoplasmic male sterility

contigs

contiguous consensus sequences

CSRD

cold-stress-related database

EST

expressed sequence tag

GO

gene ontology

IP3

inositol-1,4,5-triphosphate

MAP kinase

mitogen- activated protein kinase

nr

non-redundant

ROS

reactive oxygen species

SSRs

simple sequence repeats

TPGMS

thermo-photoperiod-sensitive genic male sterile

TPGMSSUniGene

TPGMS wheat Spike development related UniGene

TPP

trehalose-6-phosphate phosphatase

Notes

Acknowledgments

This study was supported by the Program of Beijing Basic Research and Innovation Platform for Agricultural Breeding (No. YZPT01-04), National “863” Program (no. 2006AA100102), and Excellence Scholar Fostered Program of Beijing Government.

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Copyright information

© Springer-Verlag 2008

Authors and Affiliations

  1. 1.National Key Laboratory of Crop Genetic ImprovementHuazhong Agricultural UniversityWuhanChina
  2. 2.Beijing Engineering and Technique Research Center of Hybrid WheatBeijing Academy of Agricultural and Forestry SciencesBeijingChina

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