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Comparative functional analysis of six drought-responsive promoters in transgenic rice

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Abstract

Rice production is greatly affected by environmental stresses such as drought and high salinity. Transgenic rice plants tolerant to such stresses are expected to be produced. Stress-responsive promoters with low expression under normal growth conditions are needed to minimize the adverse effects of stress-tolerance genes on rice growth. We performed expression analyses of drought-responsive genes in rice plants using a microarray, and selected LIP9, OsNAC6, OsLEA14a, OsRAB16D, OsLEA3-1, and Oshox24 for promoter analysis. Transient assays using the promoters indicated that AREB/ABF (abscisic acid (ABA)-responsive element-binding protein/ABA-binding factor) transcription factors enhanced expressions of these genes. We generated transgenic rice plants containing each promoter and the β-glucuronidase (GUS) reporter gene. GUS assays revealed that the LIP9 and OsNAC6 promoters were induced by drought, high salinity, and ABA treatment, and both promoters showed strong activity under normal growth conditions in the root. The other promoters were strongly induced by stresses and ABA, but showed low activity under normal growth conditions. In seeds, GUS staining showed that Oshox24 expression was low and expressions of the other genes were high. Transgenic rice plants overexpressing OsNAC6 under the control of the Oshox24 promoter showed increased tolerance to drought and high salinity, and no growth defects. These data suggest that the Oshox24 promoter is useful to overexpress stress-tolerance genes without adversely affecting growth.

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Abbreviations

ABA:

Abscisic acid

ABRE:

ABA-responsive element

AREB:

ABRE-binding protein

CaMV:

Cauliflower mosaic virus

CRT:

C-repeat

CBF:

CRT-binding factor

DRE:

Dehydration-responsive element

DREB:

DRE-binding protein

GUS:

β-Glucuronidase

LEA:

Late embryogenesis abundant

TF:

Transcription factor

UBI:

Ubiquitin

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Acknowledgments

We are grateful for technical support provided by Kaori Amano, Emiko Kishi, Kyoko Murai, and Kyouko Yoshiwara of JIRCAS. We thank Masami Toyoshima for her excellent editorial support. This work was supported in part by the grants from the Ministry of Agriculture, Forestry and Fisheries (MAFF; in part by Genomics for Agricultural Innovation, Development of Abiotic Stress Tolerant Crops by DREB Genes), and the Programme for Promotion of Basic and Applied Researches for Innovations in Bio-oriented Industry (BRAIN) of Japan.

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  1. Asad Jan

    Present address: Institute of Biotechnology and Genetics Engineering, The University of Agriculture, Peshawar, Khyber Pakhtunkhwa, 25130, Pakistan

Authors and Affiliations

  1. Biological Resources and Post-harvest Division, Japan International Research Center for Agricultural Sciences (JIRCAS), Tsukuba, Ibaraki, 305-8686, Japan

    Kazuo Nakashima, Asad Jan, Daisuke Todaka, Kyonoshin Maruyama, Shingo Goto & Kazuko Yamaguchi-Shinozaki

  2. RIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa, 230-0045, Japan

    Kazuo Shinozaki

  3. Graduate School of Agricultural and Life Sciences, The University of Tokyo, Bunkyo-ku, Tokyo, 113-8657, Japan

    Daisuke Todaka & Kazuko Yamaguchi-Shinozaki

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  1. Kazuo Nakashima
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  2. Asad Jan
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Nakashima, K., Jan, A., Todaka, D. et al. Comparative functional analysis of six drought-responsive promoters in transgenic rice. Planta 239, 47–60 (2014). https://doi.org/10.1007/s00425-013-1960-7

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