Cargando…

CRISRP/Cas9-Mediated Targeted Mutagenesis of Tomato Polygalacturonase Gene (SlPG) Delays Fruit Softening

Polygalacturonase (PG) gene has been documented as a key candidate for the improvement of fruit firmness, which is a target trait for tomato production because it facilitates transportation and storage. To reduce the expression of the PG gene, most of the elite commercial tomato varieties were obtai...

Descripción completa

Detalles Bibliográficos
Autores principales: Nie, Hongmei, Shi, Yu, Geng, Xueqing, Xing, Guoming
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9162796/
https://www.ncbi.nlm.nih.gov/pubmed/35665160
http://dx.doi.org/10.3389/fpls.2022.729128
_version_ 1784719786108780544
author Nie, Hongmei
Shi, Yu
Geng, Xueqing
Xing, Guoming
author_facet Nie, Hongmei
Shi, Yu
Geng, Xueqing
Xing, Guoming
author_sort Nie, Hongmei
collection PubMed
description Polygalacturonase (PG) gene has been documented as a key candidate for the improvement of fruit firmness, which is a target trait for tomato production because it facilitates transportation and storage. To reduce the expression of the PG gene, most of the elite commercial tomato varieties were obtained by RNA interference technology. However, this approach of producing commercialized tomatoes by integration of the exogenous gene is controversial. In this work, CRISPR/Cas9 technology was used to induce the targeted mutagenesis of the SlPG gene to delay the softening of tomato fruit. Results showed that the SlPG gene was frameshift mutated by 4 bp deletion, 10 bp deletion, and 1 bp insertion, which generated premature translation termination codons. Compared with wild-type (WT), homozygous T(1)-generation tomato plants exhibited late fruit softening under natural conditions. Consistent with this phenomenon, the firmness value of WT fruit was lower in slpg mutant fruit, and the physiological loss of water was higher. Collectively, these data demonstrate that the mutation of the SlPG gene delays tomato fruit softening. More importantly, 8 out of 20 transgene-free tomato plants, which were homozygous for null alleles of SlPG, were separated in the T(3)-generation of line slpgT(2)-#2. This transgene-free slpg may provide materials for more in-depth research of SlPG functions and the molecular mechanism of fruit softening in tomatoes.
format Online
Article
Text
id pubmed-9162796
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-91627962022-06-03 CRISRP/Cas9-Mediated Targeted Mutagenesis of Tomato Polygalacturonase Gene (SlPG) Delays Fruit Softening Nie, Hongmei Shi, Yu Geng, Xueqing Xing, Guoming Front Plant Sci Plant Science Polygalacturonase (PG) gene has been documented as a key candidate for the improvement of fruit firmness, which is a target trait for tomato production because it facilitates transportation and storage. To reduce the expression of the PG gene, most of the elite commercial tomato varieties were obtained by RNA interference technology. However, this approach of producing commercialized tomatoes by integration of the exogenous gene is controversial. In this work, CRISPR/Cas9 technology was used to induce the targeted mutagenesis of the SlPG gene to delay the softening of tomato fruit. Results showed that the SlPG gene was frameshift mutated by 4 bp deletion, 10 bp deletion, and 1 bp insertion, which generated premature translation termination codons. Compared with wild-type (WT), homozygous T(1)-generation tomato plants exhibited late fruit softening under natural conditions. Consistent with this phenomenon, the firmness value of WT fruit was lower in slpg mutant fruit, and the physiological loss of water was higher. Collectively, these data demonstrate that the mutation of the SlPG gene delays tomato fruit softening. More importantly, 8 out of 20 transgene-free tomato plants, which were homozygous for null alleles of SlPG, were separated in the T(3)-generation of line slpgT(2)-#2. This transgene-free slpg may provide materials for more in-depth research of SlPG functions and the molecular mechanism of fruit softening in tomatoes. Frontiers Media S.A. 2022-05-19 /pmc/articles/PMC9162796/ /pubmed/35665160 http://dx.doi.org/10.3389/fpls.2022.729128 Text en Copyright © 2022 Nie, Shi, Geng and Xing. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Plant Science
Nie, Hongmei
Shi, Yu
Geng, Xueqing
Xing, Guoming
CRISRP/Cas9-Mediated Targeted Mutagenesis of Tomato Polygalacturonase Gene (SlPG) Delays Fruit Softening
title CRISRP/Cas9-Mediated Targeted Mutagenesis of Tomato Polygalacturonase Gene (SlPG) Delays Fruit Softening
title_full CRISRP/Cas9-Mediated Targeted Mutagenesis of Tomato Polygalacturonase Gene (SlPG) Delays Fruit Softening
title_fullStr CRISRP/Cas9-Mediated Targeted Mutagenesis of Tomato Polygalacturonase Gene (SlPG) Delays Fruit Softening
title_full_unstemmed CRISRP/Cas9-Mediated Targeted Mutagenesis of Tomato Polygalacturonase Gene (SlPG) Delays Fruit Softening
title_short CRISRP/Cas9-Mediated Targeted Mutagenesis of Tomato Polygalacturonase Gene (SlPG) Delays Fruit Softening
title_sort crisrp/cas9-mediated targeted mutagenesis of tomato polygalacturonase gene (slpg) delays fruit softening
topic Plant Science
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9162796/
https://www.ncbi.nlm.nih.gov/pubmed/35665160
http://dx.doi.org/10.3389/fpls.2022.729128
work_keys_str_mv AT niehongmei crisrpcas9mediatedtargetedmutagenesisoftomatopolygalacturonasegeneslpgdelaysfruitsoftening
AT shiyu crisrpcas9mediatedtargetedmutagenesisoftomatopolygalacturonasegeneslpgdelaysfruitsoftening
AT gengxueqing crisrpcas9mediatedtargetedmutagenesisoftomatopolygalacturonasegeneslpgdelaysfruitsoftening
AT xingguoming crisrpcas9mediatedtargetedmutagenesisoftomatopolygalacturonasegeneslpgdelaysfruitsoftening