Allelic Mutations in the <i>Ripening</i><i>-Inhibitor</i> Locus Generate Extensive Variation in Tomato Ripening

Ito, Yasuhiro; Sekiyama, Yasuyo; Nakayama, Hiroko; Nishizawa‐Yokoi, Ayako; Endo, Masaki; Shima, Yoshihiro; Nakamura, Nobutaka; Kotake-Nara, Eiichi; Kawasaki, Susumu; Hirose, Sakiko; Toki, Seiichi

doi:10.1104/pp.20.00020

Cited by 44 publications

(58 citation statements)

References 73 publications

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“…The classic rin mutant allele encodes a fusion of RIN with Macrocalyx (MC), which is encoded at the genomic region adjacent to RIN [ 9 , 30 ] ( Fig 1C ). The rinG2 allele was developed by genome editing of the RIN gene [ 23 , 26 ]. Genome editing resulted in a one-base insertion at position 596 from the translation start position in the mRNA, which caused a frame-shift producing an early stop codon just after the insertion ( Fig 1B ).…”

Section: Resultsmentioning

confidence: 99%

“…The rin allele produces a protein encoded by a fusion mRNA composed of RIN lacking the last exon and MC lacking the first exon [ 9 ]. The rinG2 mutation generates an early stop codon, which removes the C-terminus including the transcriptional activator domain of RIN [ 23 ]. D. Accumulation of mutant proteins in heterozygous fruits.…”

Section: Introductionmentioning

confidence: 99%

“…In contrast to the non-ripening phenotype of the rin mutant, RIN knockout mutant fruits undergo initial ripening, accumulating small amounts of lycopene and increasing ethylene production at a low but detectable level, suggesting that RIN is required for full ripening but not for the initiation of ripening [ 22 , 24 ]. Surprisingly, RIN-knockout mutant fruits softened excessively, even compared to wild-type fruits, suggest that RIN does not simply accelerate ripening-associated changes but also represses excess softening [ 23 ].…”

Section: Introductionmentioning

confidence: 99%

“…Another allelic mutant in the RIN locus, rinG2 , was also developed by CRISPR/Cas9 and exhibited another specific ripening phenotype. Similar to rin , the rinG2 mutation confers delayed softening compared to the wild type, but unlike rin fruits, rinG2 fruits show ripening-specific changes in coloration, turning orange due to the accumulation of carotenoids ( Fig 1A ) [ 23 ]. The phenotype of rinG2 offers great promise for use in breeding tomato cultivars with extended shelf life; however, the low lycopene accumulation in these fruits must be improved.…”

Section: Introductionmentioning

confidence: 99%

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Semi-dominant effects of a novel ripening inhibitor (rin) locus allele on tomato fruit ripening

2021

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The tomato (Solanum lycopersicum) ripening inhibitor (rin) mutation completely represses fruit ripening, as rin fruits fail to express ripening-associated genes and remain green and firm. Moreover, heterozygous rin fruits (rin/+) ripen normally but have extended shelf life, an important consideration for this perishable fruit crop; therefore, heterozygous rin has been widely used to breed varieties that produce red tomatoes with improved shelf life. We previously used CRISPR/Cas9 to produce novel alleles at the rin locus. The wild-type allele RIN encodes a MADS-box transcription factor and the novel allele, named as rinG2, generates an early stop codon, resulting in C-terminal truncation of the transcription factor. Like rin fruits, rinG2 fruits exhibit extended shelf life, but unlike rin fruits, which remain yellow-green even after long-term storage, rinG2 fruits turn orange due to ripening-associated carotenoid production. Here, to explore the potential of the rinG2 mutation for breeding, we characterized the effects of rinG2 in the heterozygous state (rinG2/+) compared to the effects of rin/+. The softening of rinG2/+ fruits was delayed compared to the wild type but to a lesser degree than rin/+ fruits. Lycopene and β-carotene levels in rinG2/+ fruits were similar to those of the wild type, whereas rin/+ fruits accumulated half the amount of β-carotene compared to the wild type. The rinG2/+ fruits produced lower levels of ethylene than wild-type and rin/+ fruits. Expression analysis revealed that in rinG2/+ fruits, the rinG2 mutation (like rin) partially inhibited the expression of ripening-associated genes. The small differences in the inhibitory effects of rinG2 vs. rin coincided with small differences in phenotypes, such as ethylene production, softening, and carotenoid accumulation. Therefore, rinG2 represents a promising genetic resource for developing tomato cultivars with extended shelf life.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Semi-dominant effects of a novel ripening inhibitor (rin) locus allele on tomato fruit ripening

2021

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“…The knockout mutant of RIN shows induction of fruit ripening associated with physiological changes such as lycopene accumulation albeit much lower than wild type and expression of fruit softening enzymes such as pectate lyase and polygalacturonase. This study suggests that RIN is essential for fruit ripening but not for induction of fruit ripening [ 370 ]. Virus-induced gene silencing of 11 putative tomato fruit ripening-related factors indicated that organelle RNA recognition motif-containing protein4 ( ORRM4 ) positively regulates tomato fruit ripening.…”

Section: Genome Editing For Resistance Breeding In Tomatomentioning

confidence: 99%

Application of Genome Editing in Tomato Breeding: Mechanisms, Advances, and Prospects

Salava

Thula

Mohan

et al. 2021

IJMS

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Plants regularly face the changing climatic conditions that cause biotic and abiotic stress responses. The abiotic stresses are the primary constraints affecting crop yield and nutritional quality in many crop plants. The advances in genome sequencing and high-throughput approaches have enabled the researchers to use genome editing tools for the functional characterization of many genes useful for crop improvement. The present review focuses on the genome editing tools for improving many traits such as disease resistance, abiotic stress tolerance, yield, quality, and nutritional aspects of tomato. Many candidate genes conferring tolerance to abiotic stresses such as heat, cold, drought, and salinity stress have been successfully manipulated by gene modification and editing techniques such as RNA interference, insertional mutagenesis, and clustered regularly interspaced short palindromic repeat (CRISPR/Cas9). In this regard, the genome editing tools such as CRISPR/Cas9, which is a fast and efficient technology that can be exploited to explore the genetic resources for the improvement of tomato and other crop plants in terms of stress tolerance and nutritional quality. The review presents examples of gene editing responsible for conferring both biotic and abiotic stresses in tomato simultaneously. The literature on using this powerful technology to improve fruit quality, yield, and nutritional aspects in tomato is highlighted. Finally, the prospects and challenges of genome editing, public and political acceptance in tomato are discussed.

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Recent Trends in Targeting Genome Editing of Tomato for Abiotic and Biotic Stress Tolerance

et al. 2022

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Allelic Mutations in the Ripening-Inhibitor Locus Generate Extensive Variation in Tomato Ripening

Cited by 44 publications

References 73 publications

Semi-dominant effects of a novel ripening inhibitor (rin) locus allele on tomato fruit ripening

Semi-dominant effects of a novel ripening inhibitor (rin) locus allele on tomato fruit ripening

Application of Genome Editing in Tomato Breeding: Mechanisms, Advances, and Prospects

Recent Trends in Targeting Genome Editing of Tomato for Abiotic and Biotic Stress Tolerance

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