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Identification of transcription factors interacting with a 1274 bp promoter of MaPIP1;1 which confers high-level gene expression and drought stress Inducibility in transgenic Arabidopsis thaliana

Authors
  • Xu, Yi1
  • Jin, Zhiqiang2
  • Xu, Biyu2
  • Li, Jingyang1
  • Li, Yujia1
  • Wang, Xiaoyi1
  • Wang, Anbang1
  • Hu, Wei2
  • Huang, Dongmei1
  • Wei, Qing1
  • Xu, Zhuye3
  • Song, Shun1
  • 1 Haikou Experimental Station, Chinese Academy of Tropical Agricultural Sciences, Haikou, China , Haikou (China)
  • 2 Institute of Tropical Bioscience and Biotechnology, Chinese Academy of Tropical Agricultural Sciences, Haikou, China , Haikou (China)
  • 3 Hainan University, Haikou, China , Haikou (China)
Type
Published Article
Journal
BMC Plant Biology
Publisher
Springer (Biomed Central Ltd.)
Publication Date
Jun 16, 2020
Volume
20
Issue
1
Identifiers
DOI: 10.1186/s12870-020-02472-7
Source
Springer Nature
Keywords
License
Green

Abstract

BackgroundDrought stress can severely affect plant growth and crop yield. The cloning and identification of drought-inducible promoters would be of value for genetically-based strategies to improve resistance of crops to drought.ResultsPrevious studies showed that the MaPIP1;1 gene encoding an aquaporin is involved in the plant drought stress response. In this study, the promoter pMaPIP1;1, which lies 1362 bp upstream of the MaPIP1;1 transcriptional initiation site, was isolated from the banana genome..And the transcription start site(A) is 47 bp before the ATG. To functionally validate the promoter, various lengths of pMaPIP1;1 were deleted and fused to GUS to generate pMaPIP1;1::GUS fusion constructs that were then transformed into Arabidopsis to generate four transformants termed M-P1, M-P2, M-P3 and M-P4.Mannitol treatment was used to simulate drought conditions. All four transformants reacted well to mannitol treatment. M-P2 (− 1274 bp to − 1) showed the highest transcriptional activity among all transgenic Arabidopsis tissues, indicating that M-P2 was the core region of pMaPIP1;1. This region of the promoter also confers high levels of gene expression in response to mannitol treatment. Using M-P2 as a yeast one-hybrid bait, 23 different transcription factors or genes that interacted with MaPIP1;1 were screened. In an dual luciferase assay for complementarity verification, the transcription factor MADS3 positively regulated MaPIP1;1 transcription when combined with the banana promoter. qRT-PCR showed that MADS3 expression was similar in banana leaves and roots under drought stress. In banana plants grown in 45% soil moisture to mimic drought stress, MaPIP1;1 expression was maximized, which further demonstrated that the MADS3 transcription factor can synergize with MaPIP1;1.ConclusionsTogether our results revealed that MaPIP1;1 mediates molecular mechanisms associated with drought responses in banana, and will expand our understanding of how AQP gene expression is regulated. The findings lay a foundation for genetic improvement of banana drought resistance.

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