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PsAP2 an AP2/ERF family transcription factor from Papaver somniferum enhances abiotic and biotic stress tolerance in transgenic tobacco

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Abstract

The AP2/ERFs are one of the most important family of transcription factors which regulate multiple responses like stress, metabolism and development in plants. We isolated PsAP2 a novel AP2/ERF from Papaver somniferum which was highly upregulated in response to wounding followed by ethylene, methyl jasmonate and ABA treatment. PsAP2 showed specific binding with both DRE and GCC box elements and it was able to transactivate the reporter genes in yeast. PsAP2 overexpressing transgenic tobacco plants exhibited enhanced tolerance towards both abiotic and biotic stresses . Real time transcript expression analysis showed constitutive upregulation of tobacco Alternative oxidase1a and Myo-inositol-1-phosphate synthase in PsAP2 overexpressing tobacco plants. Further, PsAP2 showed interaction with NtAOX1a promoter in vitro, it also specifically activated the NtAOX1a promoter in yeast and tobacco BY2 cells. The silencing of PsAP2 using VIGS lead to significant reduction in the AOX1 level in P. somniferum. Taken together PsAP2 can directly bind and transcriptionally activate NtAOX1a and its overexpression in tobacco imparted increased tolerance towards both abiotic and biotic stress.

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Abbreviations

3-AT:

3-Amino-1,2,4-triazole

AP2/ERF:

Apetela 2/ethylene response factor

bp:

Base pair

GRA:

Gel retardation assay

GST:

Glutathione S-transferase

IPTG:

Isopropyl β-d-1-thiogalactopyranoside

MeJA:

Methyl jasmonate

MQ:

Milli-Q water

MS media:

Murashige and Skoog media

TF:

Transcription factor

VIGS:

Virus-induced gene silencing

Y1H:

Yeast one hybrid

MIPS:

Myo-inositol-1-phosphate synthase

AOX1a:

Alternative oxidase1a

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Acknowledgments

The authors acknowledge Director, CSIR-Central Institute of Medicinal and Aromatic Plants, Lucknow, India for providing the necessary facilities. Author’s also acknowledge CIMAP-National Gene bank for providing Papaver somniferum cv. Sampada seeds and Arabidopsis Biological Resource Centre for providing pTRV1 and pTRV2 vectors. RKS acknowledge Department of Science and Technology, INDIA under Fast Track Project Number: SR/FT/LS-189/2009 for financial support. Sonal and Ujjal acknowledge UGC, New Delhi, India for providing fellowship.

Author contributions statement

SM, UJ and VT has performed most of the in vitro and in vivo experiments. DKS and SL has contributed towards antioxidant profiling of transgenic and vector transformed tobacco lines provided to them. RKS has designed and coordinated the work. RKS, SM, UJ, VT wrote the manuscript.

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Authors and Affiliations

  1. Biotechnology Division, Central Institute of Medicinal and Aromatic Plants (CSIR-CIMAP), P.O. CIMAP, Near Kukrail Picnic Spot, Lucknow, 226015, India

    Sonal Mishra, Ujjal J. Phukan, Dhananjay K. Singh, Suaib Luqman & Rakesh Kumar Shukla

  2. Botany Division, CSIR-Central Drug Research Institute, Sector 10, Jankipuram, Lucknow, 226031, India

    Vineeta Tripathi

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  1. Sonal Mishra
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  2. Ujjal J. Phukan
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  3. Vineeta Tripathi
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  4. Dhananjay K. Singh
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  5. Suaib Luqman
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  6. Rakesh Kumar Shukla
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Correspondence to Rakesh Kumar Shukla.

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Mishra, S., Phukan, U.J., Tripathi, V. et al. PsAP2 an AP2/ERF family transcription factor from Papaver somniferum enhances abiotic and biotic stress tolerance in transgenic tobacco. Plant Mol Biol 89, 173–186 (2015). https://doi.org/10.1007/s11103-015-0361-7

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