Plant Molecular Biology

, Volume 75, Issue 1–2, pp 35–51

Identification and characterization of high temperature stress responsive genes in bread wheat (Triticum aestivum L.) and their regulation at various stages of development

  • Harsh Chauhan
  • Neetika Khurana
  • Akhilesh K. Tyagi
  • Jitendra P. Khurana
  • Paramjit Khurana
Article

Abstract

To elucidate the effect of high temperature, wheat plants (Triticum aestivum cv. CPAN 1676) were given heat shock at 37 and 42°C for 2 h, and responsive genes were identified through PCR-Select Subtraction technology. Four subtractive cDNA libraries, including three forward and one reverse subtraction, were constructed from three different developmental stages. A total of 5,500 ESTs were generated and 3,516 high quality ESTs submitted to Genbank. More than one-third of the ESTs generated fall in unknown/no hit category upon homology search through BLAST analysis. Differential expression was confirmed by cDNA macroarray and by northern/RT-PCR analysis. Expression analysis of wheat plants subjected to high temperature stress, after 1 and 4 days of recovery, showed fast recovery in seedling tissue. However, even after 4 days, recovery was negligible in the developing seed tissue after 2 h of heat stress. Ten selected genes were analyzed in further detail including one unknown protein and a new heat shock factor, by quantitative real-time PCR in an array of 35 different wheat tissues representing major developmental stages as well as different abiotic stresses. Tissue specificity was examined along with cross talk with other abiotic stresses and putative signalling molecules.

Keywords

Heat stress HSF HSPs Subtractive hybridization Transcriptome Wheat 

Supplementary material

11103_2010_9702_MOESM1_ESM.doc (55 kb)
Supplementary Table S1: List and sequences of primers used for semi-quantitative RT-PCR analysis along with accession numbers. (DOC 55 kb)
11103_2010_9702_MOESM2_ESM.doc (41 kb)
Supplementary Table S2: List and sequences of primers used for quantitative RT-PCR analysis along with accession numbers. (DOC 41 kb)
11103_2010_9702_MOESM3_ESM.doc (95 kb)
Supplementary Table S3: List of putative conserved domains in unknown/hypothetical proteins homologues of Rice. (DOC 95 kb)
11103_2010_9702_MOESM4_ESM.doc (392 kb)
Supplementary Table S4: Fold change of representative seedling FSH clones along with accession numbers. (DOC 392 kb)
11103_2010_9702_MOESM5_ESM.doc (125 kb)
Supplementary Table S5: Fold change of representative flower FSH library clones along with accession numbers. (DOC 125 kb)
11103_2010_9702_MOESM6_ESM.doc (126 kb)
Supplementary Table S6: Fold change of representative developing seed FSH library clones along with accession numbers. (DOC 126 kb)
11103_2010_9702_MOESM7_ESM.doc (143 kb)
Supplementary Table S7: Fold change of representative developing seed RSH library clones along with accession numbers. (DOC 143 kb)

Copyright information

© Springer Science+Business Media B.V. 2010

Authors and Affiliations

  • Harsh Chauhan
    • 1
  • Neetika Khurana
    • 1
  • Akhilesh K. Tyagi
    • 1
    • 2
  • Jitendra P. Khurana
    • 1
  • Paramjit Khurana
    • 1
  1. 1.Department of Plant Molecular BiologyUniversity of DelhiDhaula Kuan, New DelhiIndia
  2. 2.National Institute of Plant Genome ResearchNew DelhiIndia

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