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Characterization of abiotic stress-responsive Arabidopsis thaliana RD29A and RD29B genes and evaluation of transgenes

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Abstract

Abiotic stresses have adverse effects on plant growth and productivity. The homologous RD29A and RD29B genes are exquisitely sensitive to various abiotic stressors. Therefore, RD29A and RD29B gene sequences have potential to confer abiotic stress resistance in crop species grown in arid and semi-arid regions. To our knowledge, no information on the physiological roles of the proteins encoded by RD29A and RD29B are available in the literature. To understand how these proteins function, we used reverse genetic approaches, including identifying rd29a and rd29b T-DNA knockout mutants, and examining the effects of complementing transgenes with the genes under control of their native promoters and chimeric genes with the native promoters swapped. Four binary vectors with the RD29A and RD29B promoters upstream of the cognate RD29A and RD29B cDNAs and as chimeric genes with noncognate promoters were used to transform rd29a and rd29b plants. Cold, drought, and salt induced both genes; the promoter of RD29A was found to be more responsive to drought and cold stresses, whereas the promoter of RD29B was highly responsive to salt stress. Morphological and physiological responses of rd29a and rd29b plants to salt stress were further investigated. Root growth, and photosynthetic properties declined significantly, while solute concentration (Ψπ), water use efficiency (WUE) and δ13C ratio increased under salt stress. Unexpectedly, the rd29a and rd29b knockout mutant lines maintained greater root growth, photosynthesis, and WUE under salt stress relative to control. We conclude that the RD29A and RD29B proteins are unlikely to serve directly as protective molecules.

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Abbreviations

A :

Photosynthesis (μmol m−2 s−1)

ABA:

Abscisic acid

ABRE:

Abscisic acid-responsive element

AREB:

Abscisic acid-responsive element-binding protein

CaMV:

Cauliflower mosaic virus

CDPK:

Calcium-dependent protein kinase

COR:

Cold-regulated

DRE:

Dehydration-responsive element

DREB:

Dehydration-responsive element-binding protein

E :

Transpiration (mmol m−2 s−1)

GFP:

Green fluorescent protein

GUS:

Beta-glucuronidase

LEA:

Late embryogenesis abundant

LTI:

Low temperature induced

MAP:

Mitogen activated protein

MS:

Murashige–Skoog

PAR:

Photosynthetically active radiation (μmol m−2 s−1)

RD :

Responsive to desiccation

RH:

Relative humidity

rd29a :

RD29A knockout mutant

rd29b :

RD29B knockout mutant

RT-PCR:

Reverse transcriptase polymerase chain reaction

T :

Temperature

TF:

Transcription factor

WUE:

Water use efficiency

δ13C:

Carbon isotope ratio (‰)

Ψπ :

Osmotic potential (MPa)

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Acknowledgments

We are grateful to the LICOR Inc. (Lincoln, NE) for providing the LI-6400-17 WPA Arabidopsis gas exchange chamber, Dr. S. Madhavan for the carbon isotope ratio determinations, and Christian Elowsky for his microscopy expertise. This work was supported by an Interdisciplinary Agriculture Research grant, ARD-University of Nebraska-Lincoln. J. Msanne would like to acknowledge the graduate scholarship from the School of Natural Resources, University of Nebraska-Lincoln.

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  1. School of Natural Resources, University of Nebraska-Lincoln, 3310 Holdrege Street, Lincoln, NE, 68583, USA

    Joseph Msanne & Tala Awada

  2. Department of Biochemistry, University of Nebraska-Lincoln, 1901 Vine Street, Lincoln, NE, 68588, USA

    Jiusheng Lin & Julie M. Stone

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  1. Joseph Msanne
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Correspondence to Julie M. Stone.

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Msanne, J., Lin, J., Stone, J.M. et al. Characterization of abiotic stress-responsive Arabidopsis thaliana RD29A and RD29B genes and evaluation of transgenes. Planta 234, 97–107 (2011). https://doi.org/10.1007/s00425-011-1387-y

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