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Expression of Chlorovirus MT325 aquaglyceroporin (aqpv1) in tobacco and its role in mitigating drought stress

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Abstract

Main conclusions

A Chlorovirus aquaglyceroporin expressed in tobacco is localized to the plastid and plasma membranes. Transgenic events display improved response to water deficit. Necrosis in adult stage plants is observed.

Aquaglyceroporins are a subclass of the water channel aquaporin proteins (AQPs) that transport glycerol along with other small molecules transcellular in addition to water. In the studies communicated herein, we analyzed the expression of the aquaglyceroporin gene designated, aqpv1, from Chlorovirus MT325, in tobacco (Nicotiana tabacum), along with phenotypic changes induced by aqpv1 expression in planta. Interestingly, aqpv1 expression under control of either a constitutive or a root-preferred promoter, triggered local lesion formation in older leaves, which progressed significantly after induction of flowering. Fusion of aqpv1 with GFP suggests that the protein localized to the plasmalemma, and potentially with plastid and endoplasmic reticulum membranes. Physiological characterizations of transgenic plants during juvenile stage growth were monitored for potential mitigation to water dry-down (i.e., drought) and recovery. Phenotypic analyses on drought mimic/recovery of juvenile transgenic plants that expressed a functional aqpv1 transgene had higher photosynthetic rates, stomatal conductance, and water use efficiency, along with maximum carboxylation and electron transport rates when compared to control plants. These physiological attributes permitted the juvenile aqpv1 transgenic plants to perform better under drought-mimicked conditions and hastened recovery following re-watering. This drought mitigation effect is linked to the ability of the transgenic plants to maintain cell turgor.

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Abbreviations

AQPs:

Aquaporins

AQPV1:

Aquaglyceroporin protein virus1

MIPs:

Major intrinsic proteins

ORF:

Open reading frame

A net :

Net photosynthesis (µmol m−2 s−1)

g s :

Stomatal conductance (mol m−2 s−1)

E :

Transpiration (mmol m−2 s−1)

PAR:

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

iWUE:

Instantaneous water use efficiency (µmol mol−1)

J max :

Maximum election transport rate (µmol m−2 s−1)

V cmax :

Maximum carboxylation efficiency (µmol m−2 s−1)

Fv/Fm:

Maximum photochemical efficiency of photosystem II

F o :

Minimal or dark-adapted chlorophyll fluorescence

F m :

Maximal fluorescence

F v :

Variable fluorescence

\({\Psi}_{{\Pi }}\) :

Osmotic potential (MPa)

Ψw :

Water potential (MPa)

NL:

Total number of leaves

DWTL :

Total leaf dry weight (g)

DWAG :

Above-ground dry weight (g)

LAt :

Total leaf area (cm2)

SLA:

Specific leaf area (cm2 g−1)

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Acknowledgments

This investigation was supported in part by National Institutes of Health Grant GM32241 (JLVE) and grant P20-RR15635 from the COBRE program of the National Center for Research Resources (JLVE). Funds to support this study were also provided by the Nebraska Research Initiative through the University of Nebraska’s Center for Biotechnology. The authors would like to thank Christian Elowsky for the help with the confocal imaging, Dr. Mathew Giovanni for his assistance with the statistical analysis and Amy Hilske from Beadle Greenhouse Facility for care of plants.

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Authors and Affiliations

  1. School of Natural Resources, University of Nebraska-Lincoln, Lincoln, NE, 68583-0968, USA

    Saadia Bihmidine & Tala Awada

  2. Department of Agronomy and Horticulture, Center for Plant Science Innovation, University of Nebraska-Lincoln, Lincoln, NE, 68588-0660, USA

    Mingxia Cao & Tom E. Clemente

  3. Department of Plant Pathology, Nebraska Center for Virology, University of Nebraska-Lincoln, Lincoln, NE, 68583-0900, USA

    Ming Kang, James L. Van Etten & David D. Dunigan

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  1. Saadia Bihmidine
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  2. Mingxia Cao
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  3. Ming Kang
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  7. Tom E. Clemente
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Correspondence to Tom E. Clemente.

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Bihmidine, S., Cao, M., Kang, M. et al. Expression of Chlorovirus MT325 aquaglyceroporin (aqpv1) in tobacco and its role in mitigating drought stress. Planta 240, 209–221 (2014). https://doi.org/10.1007/s00425-014-2075-5

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