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An anther-specific gene PhGRP is regulated by PhMYC2 and causes male sterility when overexpressed in petunia anthers

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An anther-specific GRP gene, regulated by PhMYC2 , causes a significant reduction of male fertility when overexpressed in petunia, and its promoter is efficient in genetic engineering of male-sterile lines.

Abstract

Glycine-rich proteins (GRPs) play important roles in plant anther development; however, the underlying mechanisms and related regulatory networks are poorly understood. In this study, a novel glycine-rich family gene designated as PhGRP was isolated from Petunia hybrida ‘Fantasy Red’. The qRT-PCR analysis showed that it expressed specifically in anthers, and its expression peaked earlier than those well-known tapetum-specific genes, such as TA29, and several genes with the classic cis-regulatory element ‘anther-box’ in petunia during its anther development. The male fertility was significantly reduced in PhGRP overexpression lines, due to the abnormal formation of pollen wall. The PhGRP promoter (pPhGRP) could drive the GUS genes expressing specifically in the anthers of the transgenic Arabidopsis plants, indicating that the anther-specific characteristic of this promoter was conserved. In addition, when pPhGRP was used to drive the expression of BARNASE, complete male-sterile petunia lines were created without changes in vegetative organs and floral parts other than anthers. Finally, when pPhGRP was used as the bait to screen a yeast-one-hybrid (Y1H) library, a transcription factor (PhMYC2) belonging to the bHLH family was successfully selected, and the binding between pPhGRP and PhMYC2 was validated both by Y1H and dual-luciferase reporter assay. Overall, these results suggest that PhGRP, which is a male fertility-related gene that expresses specifically in anthers, is regulated by PhMYC2 and whose promoter can be used as an effective tool in the creation of male-sterile lines.

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Acknowledgements

This research work was funded by the Fundamental Research Funds for the Central Universities (Project 2662016PY042), the National Natural Science Foundation of China (Project 31601785), the National Natural Science Foundation of China (Project 30972020) and the Natural Science Foundation of Hubei Province of China (Project 2014CFB926). We gratefully acknowledge valuable comments by Dr. Jihong Liu and Xuebo Shi.

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Authors and Affiliations

  1. Key Laboratory of Horticultural Plant Biology, Ministry of Education, College of Horticulture and Forestry Sciences, Huazhong Agricultural University, Wuhan, 430070, People’s Republic of China

    Yuanzheng Yue, Chaoqun Yin, Rui Guo, Hao Peng, Zhaonan Yang, Guofeng Liu, Manzhu Bao & Huirong Hu

  2. College of Landscape Architecture, Nanjing Forestry University, Nanjing, 210037, People’s Republic of China

    Yuanzheng Yue

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  1. Yuanzheng Yue
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  2. Chaoqun Yin
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Correspondence to Huirong Hu.

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Communicated by Da-Bing Zhang.

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Yue, Y., Yin, C., Guo, R. et al. An anther-specific gene PhGRP is regulated by PhMYC2 and causes male sterility when overexpressed in petunia anthers. Plant Cell Rep 36, 1401–1415 (2017). https://doi.org/10.1007/s00299-017-2163-7

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