4.7 Article

Fruit ripening specific expression of β-D-N-acetylhexosaminidase (β-Hex) gene in tomato is transcriptionally regulated by ethylene response factor SlERF.E4

Journal

PLANT SCIENCE
Volume 323, Issue -, Pages -

Publisher

ELSEVIER IRELAND LTD
DOI: 10.1016/j.plantsci.2022.111380

Keywords

beta-Hex; Ethylene Fruit Ripening; Promoter; RIN; SlASR1; SlERF.E4

Funding

  1. National Institute of Plant Genome Research
  2. Department of Biotechnology, Govt. of India
  3. CSIR, India [8862-A]

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N-glycans and N-glycan processing enzymes play important roles in regulating the ripening of tomato fruits. The study reveals that ethylene, together with RIN and SlASR1, transcriptionally regulates the expression of beta-Hex and is essential for fruit ripening in tomato.
N-glycans and N-glycan processing enzymes are key players in regulating the ripening of tomato (Solanum lycopersicum) fruits, a model for fleshy fruit ripening. beta-D-N acetylhexosaminidase (beta-Hex) is a N-glycan processing enzyme involved in fruit ripening. The suppression of beta-Hex results in enhanced fruit shelf life and firmness in both climacteric and non-climacteric fruits. Previously, we have shown that ripening specific expression of beta-Hex is regulated by RIPENING INHIBITOR (RIN), ABSCISIC ACID STRESS RIPENING 1 (SlASR1) and ethylene. However, the precise mechanism of ethylene-mediated regulation of beta-Hex remains elusive. To gain insights into this, we have performed 5' deletion mapping of tomato beta-Hex promoter and a shorter promoter fragment (pD-200, 200 bp upstream to translational start site) is identified, which was found critical for spatiotemporal transcriptional regulation of beta-Hex. Further, site specific mutagenesis in RIN and ASR1 binding sites in pD-200 provides key insights into ripening specific promoter activity. Furthermore, induction of GUS activity by ethylene, yeast one hybrid assay and EMSA identify Ethylene Response Factor SlERF.E4 as a positive regulator of beta-Hex. Taken together, our study suggest that SlERF.E4 together with RIN and SlASR1 transcriptionally regulates beta-Hex and all these three proteins are essential for fruit ripening specific expression of beta-Hex in tomato.

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