Curing cytoplasmic male sterility via TALEN-mediated mitochondrial genome editing

Tomohiko Kazama, Miki Okuno, Yuta Watari, Shungo Yanase, Chie Koizuka, Yu Tsuruta, Hajime Sugaya, Atsushi Toyoda, Takehiko Itoh, Nobuhiro Tsutsumi, Kinya Toriyama, Nobuya Koizuka, Shin ichi Arimura

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Sequence-specific nucleases are commonly used to modify the nuclear genome of plants. However, targeted modification of the mitochondrial genome of land plants has not yet been achieved. In plants, a type of male sterility called cytoplasmic male sterility (CMS) has been attributed to certain mitochondrial genes, but none of these genes has been validated by direct mitochondrial gene-targeted modification. Here, we knocked out CMS-associated genes (orf79 and orf125) of CMS varieties of rice and rapeseed, respectively, using transcription activator-like effector nucleases (TALENs) with mitochondria localization signals (mitoTALENs). We demonstrate that knocking out these genes cures male sterility, strongly suggesting that these genes are causes of CMS. Sequencing revealed that double-strand breaks induced by mitoTALENs were repaired by homologous recombination, and that during this process, the target genes and surrounding sequences were deleted. Our results show that mitoTALENs can be used to stably and heritably modify the mitochondrial genome in plants.

Original languageEnglish
Pages (from-to)722-730
Number of pages9
JournalNature Plants
Volume5
Issue number7
DOIs
Publication statusPublished - Jul 1 2019
Externally publishedYes

Fingerprint

cytoplasmic male sterility
nucleases
transcription (genetics)
genes
male fertility
homologous recombination
embryophytes
rapeseed
mitochondrial genome
nuclear genome
mitochondria
rice

All Science Journal Classification (ASJC) codes

  • Plant Science

Cite this

Kazama, T., Okuno, M., Watari, Y., Yanase, S., Koizuka, C., Tsuruta, Y., ... Arimura, S. I. (2019). Curing cytoplasmic male sterility via TALEN-mediated mitochondrial genome editing. Nature Plants, 5(7), 722-730. https://doi.org/10.1038/s41477-019-0459-z

Curing cytoplasmic male sterility via TALEN-mediated mitochondrial genome editing. / Kazama, Tomohiko; Okuno, Miki; Watari, Yuta; Yanase, Shungo; Koizuka, Chie; Tsuruta, Yu; Sugaya, Hajime; Toyoda, Atsushi; Itoh, Takehiko; Tsutsumi, Nobuhiro; Toriyama, Kinya; Koizuka, Nobuya; Arimura, Shin ichi.

In: Nature Plants, Vol. 5, No. 7, 01.07.2019, p. 722-730.

Research output: Contribution to journalArticle

Kazama, T, Okuno, M, Watari, Y, Yanase, S, Koizuka, C, Tsuruta, Y, Sugaya, H, Toyoda, A, Itoh, T, Tsutsumi, N, Toriyama, K, Koizuka, N & Arimura, SI 2019, 'Curing cytoplasmic male sterility via TALEN-mediated mitochondrial genome editing', Nature Plants, vol. 5, no. 7, pp. 722-730. https://doi.org/10.1038/s41477-019-0459-z
Kazama, Tomohiko ; Okuno, Miki ; Watari, Yuta ; Yanase, Shungo ; Koizuka, Chie ; Tsuruta, Yu ; Sugaya, Hajime ; Toyoda, Atsushi ; Itoh, Takehiko ; Tsutsumi, Nobuhiro ; Toriyama, Kinya ; Koizuka, Nobuya ; Arimura, Shin ichi. / Curing cytoplasmic male sterility via TALEN-mediated mitochondrial genome editing. In: Nature Plants. 2019 ; Vol. 5, No. 7. pp. 722-730.
@article{87961bddf30543f6b44bfe4fd152a7fe,
title = "Curing cytoplasmic male sterility via TALEN-mediated mitochondrial genome editing",
abstract = "Sequence-specific nucleases are commonly used to modify the nuclear genome of plants. However, targeted modification of the mitochondrial genome of land plants has not yet been achieved. In plants, a type of male sterility called cytoplasmic male sterility (CMS) has been attributed to certain mitochondrial genes, but none of these genes has been validated by direct mitochondrial gene-targeted modification. Here, we knocked out CMS-associated genes (orf79 and orf125) of CMS varieties of rice and rapeseed, respectively, using transcription activator-like effector nucleases (TALENs) with mitochondria localization signals (mitoTALENs). We demonstrate that knocking out these genes cures male sterility, strongly suggesting that these genes are causes of CMS. Sequencing revealed that double-strand breaks induced by mitoTALENs were repaired by homologous recombination, and that during this process, the target genes and surrounding sequences were deleted. Our results show that mitoTALENs can be used to stably and heritably modify the mitochondrial genome in plants.",
author = "Tomohiko Kazama and Miki Okuno and Yuta Watari and Shungo Yanase and Chie Koizuka and Yu Tsuruta and Hajime Sugaya and Atsushi Toyoda and Takehiko Itoh and Nobuhiro Tsutsumi and Kinya Toriyama and Nobuya Koizuka and Arimura, {Shin ichi}",
year = "2019",
month = "7",
day = "1",
doi = "10.1038/s41477-019-0459-z",
language = "English",
volume = "5",
pages = "722--730",
journal = "Nature Plants",
issn = "2055-026X",
publisher = "Palgrave Macmillan Ltd.",
number = "7",

}

TY - JOUR

T1 - Curing cytoplasmic male sterility via TALEN-mediated mitochondrial genome editing

AU - Kazama, Tomohiko

AU - Okuno, Miki

AU - Watari, Yuta

AU - Yanase, Shungo

AU - Koizuka, Chie

AU - Tsuruta, Yu

AU - Sugaya, Hajime

AU - Toyoda, Atsushi

AU - Itoh, Takehiko

AU - Tsutsumi, Nobuhiro

AU - Toriyama, Kinya

AU - Koizuka, Nobuya

AU - Arimura, Shin ichi

PY - 2019/7/1

Y1 - 2019/7/1

N2 - Sequence-specific nucleases are commonly used to modify the nuclear genome of plants. However, targeted modification of the mitochondrial genome of land plants has not yet been achieved. In plants, a type of male sterility called cytoplasmic male sterility (CMS) has been attributed to certain mitochondrial genes, but none of these genes has been validated by direct mitochondrial gene-targeted modification. Here, we knocked out CMS-associated genes (orf79 and orf125) of CMS varieties of rice and rapeseed, respectively, using transcription activator-like effector nucleases (TALENs) with mitochondria localization signals (mitoTALENs). We demonstrate that knocking out these genes cures male sterility, strongly suggesting that these genes are causes of CMS. Sequencing revealed that double-strand breaks induced by mitoTALENs were repaired by homologous recombination, and that during this process, the target genes and surrounding sequences were deleted. Our results show that mitoTALENs can be used to stably and heritably modify the mitochondrial genome in plants.

AB - Sequence-specific nucleases are commonly used to modify the nuclear genome of plants. However, targeted modification of the mitochondrial genome of land plants has not yet been achieved. In plants, a type of male sterility called cytoplasmic male sterility (CMS) has been attributed to certain mitochondrial genes, but none of these genes has been validated by direct mitochondrial gene-targeted modification. Here, we knocked out CMS-associated genes (orf79 and orf125) of CMS varieties of rice and rapeseed, respectively, using transcription activator-like effector nucleases (TALENs) with mitochondria localization signals (mitoTALENs). We demonstrate that knocking out these genes cures male sterility, strongly suggesting that these genes are causes of CMS. Sequencing revealed that double-strand breaks induced by mitoTALENs were repaired by homologous recombination, and that during this process, the target genes and surrounding sequences were deleted. Our results show that mitoTALENs can be used to stably and heritably modify the mitochondrial genome in plants.

UR - http://www.scopus.com/inward/record.url?scp=85068594260&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85068594260&partnerID=8YFLogxK

U2 - 10.1038/s41477-019-0459-z

DO - 10.1038/s41477-019-0459-z

M3 - Article

C2 - 31285556

AN - SCOPUS:85068594260

VL - 5

SP - 722

EP - 730

JO - Nature Plants

JF - Nature Plants

SN - 2055-026X

IS - 7

ER -