Re-evaluation of transcription factor function in tomato fruit development and ripening with CRISPR/Cas9-mutagenesis

Rufang Wang, Eveline Carla da Rocha Tavano, Michiel Lammers, Adriana Pinheiro Martinelli, Gerco C. Angenent, Ruud A. de Maagd*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

8 Citations (Scopus)

Abstract

Tomato (Solanum lycopersicum) is a model for climacteric fleshy fruit ripening studies. Tomato ripening is regulated by multiple transcription factors together with the plant hormone ethylene and their downstream effector genes. Transcription Factors APETALA2a (AP2a), NON-RIPENING (NOR) and FRUITFULL (FUL1/TDR4 and FUL2/MBP7) were reported as master regulators controlling tomato fruit ripening. Their proposed functions were derived from studies of the phenotype of spontaneous mutants or RNAi knock-down lines rather than, as it appears now, actual null mutants. To study TF function in tomato fruit ripening in more detail, we used CRISPR/Cas9-mediated mutagenesis to knock out the encoding genes, and phenotypes of these mutants are reported for the first time. While the earlier ripening, orange-ripe phenotype of ap2a mutants was confirmed, the nor null mutant exhibited a much milder phenotype than the spontaneous nor mutant. Additional analyses revealed that the severe phenotype in the spontaneous mutant is caused by a dominant-negative allele. Our approach also provides new insight into the independent and overlapping functions of FUL1 and FUL2. Single and combined null alleles of FUL1 and FUL2 illustrate that these two genes have partially redundant functions in fruit ripening, but also unveil an additional role for FUL2 in early fruit development.

Original languageEnglish
Article number1696
JournalScientific Reports
Volume9
Issue number1
DOIs
Publication statusPublished - 8 Feb 2019

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Clustered Regularly Interspaced Short Palindromic Repeats
Lycopersicon esculentum
Mutagenesis
Fruit
Transcription Factors
Phenotype
Alleles
Gene Knockout Techniques
Plant Growth Regulators
RNA Interference
Genes

Cite this

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title = "Re-evaluation of transcription factor function in tomato fruit development and ripening with CRISPR/Cas9-mutagenesis",
abstract = "Tomato (Solanum lycopersicum) is a model for climacteric fleshy fruit ripening studies. Tomato ripening is regulated by multiple transcription factors together with the plant hormone ethylene and their downstream effector genes. Transcription Factors APETALA2a (AP2a), NON-RIPENING (NOR) and FRUITFULL (FUL1/TDR4 and FUL2/MBP7) were reported as master regulators controlling tomato fruit ripening. Their proposed functions were derived from studies of the phenotype of spontaneous mutants or RNAi knock-down lines rather than, as it appears now, actual null mutants. To study TF function in tomato fruit ripening in more detail, we used CRISPR/Cas9-mediated mutagenesis to knock out the encoding genes, and phenotypes of these mutants are reported for the first time. While the earlier ripening, orange-ripe phenotype of ap2a mutants was confirmed, the nor null mutant exhibited a much milder phenotype than the spontaneous nor mutant. Additional analyses revealed that the severe phenotype in the spontaneous mutant is caused by a dominant-negative allele. Our approach also provides new insight into the independent and overlapping functions of FUL1 and FUL2. Single and combined null alleles of FUL1 and FUL2 illustrate that these two genes have partially redundant functions in fruit ripening, but also unveil an additional role for FUL2 in early fruit development.",
author = "Rufang Wang and {da Rocha Tavano}, {Eveline Carla} and Michiel Lammers and Martinelli, {Adriana Pinheiro} and Angenent, {Gerco C.} and {de Maagd}, {Ruud A.}",
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Re-evaluation of transcription factor function in tomato fruit development and ripening with CRISPR/Cas9-mutagenesis. / Wang, Rufang; da Rocha Tavano, Eveline Carla; Lammers, Michiel; Martinelli, Adriana Pinheiro; Angenent, Gerco C.; de Maagd, Ruud A.

In: Scientific Reports, Vol. 9, No. 1, 1696, 08.02.2019.

Research output: Contribution to journalArticleAcademicpeer-review

TY - JOUR

T1 - Re-evaluation of transcription factor function in tomato fruit development and ripening with CRISPR/Cas9-mutagenesis

AU - Wang, Rufang

AU - da Rocha Tavano, Eveline Carla

AU - Lammers, Michiel

AU - Martinelli, Adriana Pinheiro

AU - Angenent, Gerco C.

AU - de Maagd, Ruud A.

PY - 2019/2/8

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