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Papaya CpERF9 acts as a transcriptional repressor of cell-wall-modifying genes CpPME1/2 and CpPG5 involved in fruit ripening

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Abstract

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CpERF9 controls papaya fruit ripening through transcriptional repression of cell-wall-modifying genes CpPME1/2 and CpPG5 by directly binding to their promoters.

Abstract

Papaya fruit ripening is an intricate and highly coordinated developmental process which is controlled by the action of ethylene and expression of numerous ethylene-responsive genes. Ethylene response factors (ERFs) representing the last regulators of ethylene-signaling pathway determine the specificities of ethylene response. However, knowledge concerning the transcriptional controlling mechanism of ERF-mediated papaya fruit ripening is limited. In the present work, a gene-encoding AP2/ERF protein with two ERF-associated amphiphilic repression (EAR) motifs, named CpERF9, was characterized from papaya fruit. CpERF9 was found to localize in nucleus, and possess transcriptional repression ability. CpERF9 expression steadily decreased during papaya fruit ripening, while several genes encoding pectin methylesterases (PMEs) and polygalacturonases (PGs), such as CpPME1/2 and CpPG5, were gradually increased, paralleling the decline of fruit firmness. Electrophoretic mobility shift assay (EMSA) demonstrated a specific binding of CpERF9 to promoters of CpPME1/2 and CpPG5, via the GCC-box motif. Transient expression of CpERF9 in tobacco repressed CpPME1/2 and CpPG5 promoter activities, which was depended on two EAR motifs of CpERF9 protein. Taken together, these findings suggest that papaya CpERF9 may act as a transcriptional repressor of several cell-wall modifying genes, such as CpPME1/2 and CpPG5, via directly binding to their promoters.

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Abbreviations

EAR:

ERF-associated amphiphilic repression

EMSA:

Electrophoretic mobility shift assay

ERF:

Ethylene response factor

GFP:

Green fluorescence protein

PG:

Polygalacturonase

PME:

Pectin methylesterase

RT-qPCR:

Real-time quantitative polymerase chain reaction

TF:

Transcription factor

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Acknowledgments

The authors would like to thank Professor Junping Gao (Department of Ornamental Horticulture, China Agricultural University), Professor Shouyi Chen (Institute of Genetics and Developmental Biology, Chinese Academy of Sciences), and Professor George P. Lomonossoff (Department of Biological Chemistry, John Innes Centre, Norwich Research Park) for the generous gifts of the transient expression vectors and pEAQ vectors, respectively. Comments and suggestion from Dr. Prakash Lakshmanan during revision is gratefully acknowledged. This work was supported by the National Key Research and Development Program (No. 2016YFD0400103), National Science and Technology Pillar Program during the Twelfth Five-Year Plan Period of China (No. 2011BAD24B02-4).

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Authors and Affiliations

  1. State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources/Guangdong Key Laboratory for Postharvest Science, College of Horticulture, South China Agricultural University, Guangzhou, 510642, China

    Chang-Chun Fu, Yan-Chao Han, Xiu-Ye Qi, Wei Shan, Jian-Ye Chen, Wang-Jin Lu & Jian-Fei Kuang

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  1. Chang-Chun Fu
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  2. Yan-Chao Han
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Correspondence to Jian-Fei Kuang.

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Communicated by P. Lakshmanan.

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Fu, CC., Han, YC., Qi, XY. et al. Papaya CpERF9 acts as a transcriptional repressor of cell-wall-modifying genes CpPME1/2 and CpPG5 involved in fruit ripening. Plant Cell Rep 35, 2341–2352 (2016). https://doi.org/10.1007/s00299-016-2038-3

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