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Biotechnology and Biological Transformations
CRISPR/Cas9-Mediated SlCBF1 Mutagenesis Reduces Tomato Plant Chilling Tolerance Rui Li, Lixing Zhang, Liu Wang, Lin Chen, Ruirui Zhao, Jiping Sheng, and Lin Shen J. Agric. Food Chem., Just Accepted Manuscript • DOI: 10.1021/acs.jafc.8b02177 • Publication Date (Web): 10 Aug 2018 Downloaded from http://pubs.acs.org on August 11, 2018
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Journal of Agricultural and Food Chemistry
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CRISPR/Cas9-Mediated SlCBF1 Mutagenesis Reduces Tomato Plant Chilling
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Tolerance
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Rui Li,† Lixing Zhang,† Liu Wang,† Lin Chen,† Ruirui Zhao,† Jiping Sheng, ‡ and Lin
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Shen†,*
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† College of Food Science and Nutritional Engineering, China Agricultural University,
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Beijing 100083, China
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‡ School of Agricultural Economics and Rural Development, Renmin University of
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China, Beijing 100872, China
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Journal of Agricultural and Food Chemistry
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ABSTRACT
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Chilling stress is the main constraint in tomato (Solanum lycopersicum)
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production, as this is a chilling-sensitive horticultural crop. The highly conserved
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C-repeat binding factors (CBFs) are cold-response system components found in
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many species. In this study, we generated slcbf1 mutants using the CRISPR/Cas9
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system and investigated the role of SlCBF1 in tomato plant chilling tolerances. The
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slcbf1 mutants exhibited more severe chilling injury symptoms with higher
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electrolyte leakage and malondialdehyde levels than wild type (WT) plants.
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Additionally, slcbf1 mutants showed lower proline and protein content and higher
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hydrogen peroxide content and activities of antioxidant enzymes than WT plants.
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Knockout of SlCBF1 significantly increased indole acetic acid content but decreased
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methyl jasmonate, abscisic acid and zeatin riboside contents. The reduced chilling
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tolerance of slcbf1 mutants was further reflected by the down-regulation of
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CBF-related genes. These results contribute for better understanding the molecular
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basis underlying SlCBF1 mediation of tomato chilling sensitivity.
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Keywords: CRISPR/Cas9, SlCBF1, tomato plants, chilling stress, membrane damage
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Journal of Agricultural and Food Chemistry
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INTRODUCTION
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Cold stresses, including chilling (0–18 °C) and freezing (
