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Stored and neosynthesized mRNA in Arabidopsis seeds: effects of cycloheximide and controlled deterioration treatment on the resumption of transcription during imbibition

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Abstract

Dry seeds accumulate translatable mRNAs as well as functional proteins for transcription and translation. They are possibly involved in early physiological responses after imbibition, however, their functions remain poorly understood. The aim of this study is to investigate the function of seed stored transcriptional machinery in resumption of gene expression after the onset of imbibition in Arabidopsis thaliana. First, we examined the characters of stored mRNAs in A. thaliana dry seeds using microarray data from non-dormant Columbia (Col) and dormant Cape Verde Islands (Cvi) accessions. Transcriptomes of Col and Cvi dry seeds resembled one another, suggesting that patterns of stored mRNA do not reflect either the degree of dormancy or germination potential, but rather reflect the developmental context, such as seed maturation. Upon imbibition, changes in mRNA abundance of many genes were initiated between 1- and 2-h after the onset of imbibition. RT–PCR expression analysis of imbibition-responsive genes indicates that early induction was not altered by treatment of cycloheximide. This suggests that de novo protein synthesis is not required for gene expression during early imbibition stages. Moreover, controlled deterioration treatment (CDT), which causes artificial damages on dry seeds, disrupted gene expression specifically during the first 3 h after the start of imbibition, suggesting that seed stored transcription factors play a pivotal role in gene expression during early imbibition periods. Furthermore, the negligible effect of CDT on germination indicates that early imbibition response is dispensable and de novo synthesized proteins compensate for the function of stored proteins for germination.

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Abbreviations

ABA:

Abscisic acid

ABI:

Abscisic acid-insensitive

ABRE:

Abscisic acid-responsive element

BME3:

Blue micropylar end 3

CDT:

Controlled deterioration treatment

CHX:

Cycloheximide

Col:

Columbia

Cvi:

Cape Verde Islands

GSTU22:

Glutathione S-transferase tau 22

GO:

Gene ontology

HSP:

Heat shock protein

LEA:

Late embryogenesis abundant

RT–PCR:

Reverse transcription-PCR

TT:

Transparent testa

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Acknowledgments

Authors acknowledge Drs. George Stamatiou and Danielle Vidaurre (University of Toronto) for their critical reading of this manuscript and Professor Naoto Kawakami (Meiji University) for providing the tt8 mutants. This work is supported by NSERC Discovery grant (to E.N.).

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  1. Department of Cell & Systems Biology, University of Toronto, 25 Willcocks Street, Toronto, ON, M5S 3B2, Canada

    Mitsuhiro Kimura & Eiji Nambara

  2. The Centre for the Analysis of Genome Evolution and Function (CAGEF), University of Toronto, Toronto, ON, M5S 3B2, Canada

    Eiji Nambara

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  1. Mitsuhiro Kimura
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Correspondence to Eiji Nambara.

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Kimura, M., Nambara, E. Stored and neosynthesized mRNA in Arabidopsis seeds: effects of cycloheximide and controlled deterioration treatment on the resumption of transcription during imbibition. Plant Mol Biol 73, 119–129 (2010). https://doi.org/10.1007/s11103-010-9603-x

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