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Efficient chimeric plant promoters derived from plant infecting viral promoter sequences

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Abstract

In the present study, we developed a set of three chimeric/hybrid promoters namely FSgt-PFlt, PFlt-UAS-2X and MSgt-PFlt incorporating different important domains of Figwort Mosaic Virus sub-genomic transcript promoter (FSgt, −270 to −60), Mirabilis Mosaic Virus sub-genomic transcript promoter (MSgt, −306 to −125) and Peanut Chlorotic Streak Caulimovirus full-length transcript promoter (PFlt-, −353 to +24 and PFlt-UAS, −353 to −49). We demonstrated that these chimeric/hybrid promoters can drive the expression of reporter genes in different plant species including tobacco, Arabidopsis, petunia, tomato and spinach. FSgt-PFlt, PFlt-UAS-2X and MSgt-PFlt promoters showed 4.2, 1.5 and 1.2 times stronger GUS activities compared to the activity of the CaMV35S promoter, respectively, in tobacco protoplasts. Protoplast-derived recombinant promoter driven GFP showed enhanced accumulation compared to that obtained under the CaMV35S promoter. FSgt-PFlt, PFlt-UAS-2X and MSgt-PFlt promoters showed 3.0, 1.3 and 1.0 times stronger activities than the activity of the CaMV35S2 (a modified version of the CaMV35S promoter with double enhancer domain) promoter, respectively, in tobacco (Nicotiana tabacum, var. Samsun NN). Alongside, we observed a fair correlation between recombinant promoter-driven GUS accumulation with the corresponding uidA-mRNA level in transgenic tobacco. Histochemical (X-gluc) staining of whole transgenic seedlings and fluorescence images of ImaGene Green™ treated floral parts expressing the GUS under the control of recombinant promoters also support above findings. Furthermore, we confirmed that these chimeric promoters are inducible in the presence of 150 μM salicylic acid (SA) and abscisic acid (ABA). Taken altogether, we propose that SA/ABA inducible chimeric/recombinant promoters could be used for strong expression of gene(s) of interest in crop plants.

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Abbreviations

GUS:

β-Glucuronidase

GFP:

Green fluorescent protein

X-gluc:

5-bromo-4-chloro-3-indolyl β-d-glucopyranosiduronic acid

CLSM:

Confocal laser scanning microscope

Kanr :

Kanamycinresistant

Kans :

Kanamycinsusceptible

FMV:

Figwort mosaic virus

MMV:

Mirabilis mosaic virus

PClSV:

Peanut chlorotic streak virus

SA:

Salicylic acid

ABA:

Abscisic acid

ROI:

Region of interest

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Acknowledgments

We are greatly indebted to Institute of Life Sciences for providing funds and facilities. We are thankful to the Director, ILS, for his constructive suggestions and support. We sincerely acknowledge Mr. Abhimanyu Das, for his kind help and technical support. Mr. Bhabani S. Sahoo is gratefully acknowledged for confocal assistance. We also thank Ms. Adrita Roy, Bose Institute for her technical support in Arabidopsis protoplast culture maintenance and isolation. SA is thankful to University Grant Commission for fellowship and the two anonymous reviewers for critical reading and helpful comments in improving the manuscript.

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Authors and Affiliations

  1. Division of Gene Function and Regulation, Department of Biotechnology, Institute of Life Sciences, Government of India, Nalco Square, Chandrasekharpur, Bhubaneswar, 751 023, Odisha, India

    Sefali Acharya, Rajiv Ranjan & Nrisingha Dey

  2. Plant Genetic Engineering and Service, KTRDC, University of Kentucky, Lexington, KY, 40546-0236, USA

    Sitakanta Pattanaik & Indu B. Maiti

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  1. Sefali Acharya
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  2. Rajiv Ranjan
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  3. Sitakanta Pattanaik
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  4. Indu B. Maiti
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  5. Nrisingha Dey
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Correspondence to Nrisingha Dey.

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Acharya, S., Ranjan, R., Pattanaik, S. et al. Efficient chimeric plant promoters derived from plant infecting viral promoter sequences. Planta 239, 381–396 (2014). https://doi.org/10.1007/s00425-013-1973-2

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