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

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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.

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Acknowledgments

This work was financially supported by Department of Biotechnology, Government of India, and partially by Indo-Swiss Collaboration in Biotechnology (ISCB). HC thanks Council for Scientific and Industrial Research for Junior and Senior Research Fellowships.

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  1. Akhilesh K. Tyagi

    Present address: National Institute of Plant Genome Research, Aruna Asaf Ali Marg, New Delhi, 110067, India

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  1. Department of Plant Molecular Biology, University of Delhi, South Campus, Benito Juarez Road, Dhaula Kuan, New Delhi, 110 021, India

    Harsh Chauhan, Neetika Khurana, Akhilesh K. Tyagi, Jitendra P. Khurana & Paramjit Khurana

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Electronic supplementary material

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11103_2010_9702_MOESM1_ESM.doc

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

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

Supplementary Table S3: List of putative conserved domains in unknown/hypothetical proteins homologues of Rice. (DOC 95 kb)

Supplementary Table S4: Fold change of representative seedling FSH clones along with accession numbers. (DOC 392 kb)

11103_2010_9702_MOESM5_ESM.doc

Supplementary Table S5: Fold change of representative flower FSH library clones along with accession numbers. (DOC 125 kb)

11103_2010_9702_MOESM6_ESM.doc

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

Supplementary Table S7: Fold change of representative developing seed RSH library clones along with accession numbers. (DOC 143 kb)

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Chauhan, H., Khurana, N., Tyagi, A.K. et al. Identification and characterization of high temperature stress responsive genes in bread wheat (Triticum aestivum L.) and their regulation at various stages of development. Plant Mol Biol 75, 35–51 (2011). https://doi.org/10.1007/s11103-010-9702-8

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