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Molecular cloning and expression analysis of SpWRKY6 gene from Solanum pimpinellifolium

  • Original papers
  • Published:
Biologia Plantarum

Abstract

Ttranscription factors WRKY play vital roles in response to biotic and abiotic stresses, and previous studies have predominantly focused on model plants and fairly limited research has been performed with tomato. In the present study, a novel pathogen-induced WRKY gene named SpWRKY6 was isolated from the late blight resistant tomato (Solanum pimpinellifolium) cultivar L3708 using in silico cloning and reverse transcription polymerase chain reaction (RT-PCR) methods. Multiple sequence alignment with other plant WRKYs indicates that SpWRKY6 contains two WRKY domains and belongs to group I WRKY transcription factors. Furthermore, some cis-acting elements associated with responses to environmental stresses were observed in the promoter region of this gene. Gene expression patterns were determined by analyzing microarray data of SpWRKY6 in tomato and of an orthologous gene from Arabidopsis thaliana using the Genevestigator tool. The results reveal a very strong biotic and abiotic stress responsive behaviour of this gene. Moreover, bioinformatics results were confirmed by real time quantitative polymerase chain reaction and show that SpWRKY6 expression was rapidly induced after infection with Phytophthora infestans and Botrytis cinerea, respectively. Expression of SpWRKY6 was up-regulated by application of various phytohormones including salicylic acid, methyl jasmonate, and abscisic acid. Likewise, the SpWRKY6 expression was induced by NaCl, drought, heat, cold, and HgCl2 treatments.

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Abbreviations

ABA:

abscisic acid

ABRE:

ABA responsive element

EIRE:

elicitor-responsive element

ETH:

ethephon

hpi:

hours post-inoculation

hpt:

hours post-treatment

MeJA:

methyl jasmonate

MRE:

MYB binding site

NLS:

nuclear localization signal

ORF:

open reading frame

PDA:

potato dextrose agar

PEG:

polyethyleneglycol

qPCR:

quantitative polymerase chain reaction

RT-PCR:

reverse transcriptase polymerase chain reaction

SA:

salicylic acid

TF:

transcription factor

WT:

wild-type

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Authors and Affiliations

  1. School of Life Science & Biotechnology, Dalian University of Technology, Dalian, 116024, P.R. China

    Z. Liu, Y. S. Luan & J. B. Li

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  1. Z. Liu
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  2. Y. S. Luan
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  3. J. B. Li
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Correspondence to Y. S. Luan.

Additional information

Acknowledgments: This work was supported by grants from the National Natural Science Foundation of China (Nos. 31272167 and 31471880).

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Liu, Z., Luan, Y.S. & Li, J.B. Molecular cloning and expression analysis of SpWRKY6 gene from Solanum pimpinellifolium . Biol Plant 60, 226–234 (2016). https://doi.org/10.1007/s10535-016-0582-x

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  • DOI: https://doi.org/10.1007/s10535-016-0582-x

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