Molecular Analysis Uncovers the Mechanism of Fertility Restoration in Temperature-Sensitive Polima Cytoplasmic Male-Sterile Brassica napus

Temperature-sensitive male sterility is a heritable agronomic trait affected by genotype-environment interactions. In rapeseed (Brassica napus), Polima (pol) temperature-sensitive cytoplasmic male sterility (TCMS) is commonly used for two-line breeding, as the f...

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Autores principales:	Qing Xiao, Huadong Wang, Hui Chen, Xiaohan Chen, Jing Wen, Cheng Dai, Chaozhi Ma, Jinxing Tu, Jinxiong Shen, Tingdong Fu, Bin Yi
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	ambient temperature temperature-sensitive cytoplasmic male sterility <i>Brassica napus</i> single-molecule long-read isoform sequencing Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/375c2473f1734d58b6cf11ec1a95cdec
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spelling	oai:doaj.org-article:375c2473f1734d58b6cf11ec1a95cdec2021-11-25T17:56:47ZMolecular Analysis Uncovers the Mechanism of Fertility Restoration in Temperature-Sensitive Polima Cytoplasmic Male-Sterile <i>Brassica napus</i>10.3390/ijms2222124501422-00671661-6596https://doaj.org/article/375c2473f1734d58b6cf11ec1a95cdec2021-11-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/22/12450https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Temperature-sensitive male sterility is a heritable agronomic trait affected by genotype-environment interactions. In rapeseed (<i>Brassica napus</i>), Polima (<i>pol</i>) temperature-sensitive cytoplasmic male sterility (TCMS) is commonly used for two-line breeding, as the fertility of <i>pol</i> TCMS lines can be partially restored at certain temperatures. However, little is known about the underlying molecular mechanism that controls fertility restoration. Therefore, we aimed to investigate the fertility conversion mechanism of the <i>pol</i> TCMS line at two different ambient temperatures (16 °C and 25 °C). Our results showed that the anthers developed and produced vigorous pollen at 16 °C but not at 25 °C. In addition, we identified a novel co-transcript of <i>orf224-atp6</i> in the mitochondria that might lead to fertility conversion of the <i>pol</i> TCMS line. RNA-seq analysis showed that 1637 genes were significantly differentially expressed in the fertile flowers of 596-L when compared to the sterile flower of 1318 and 596-H. Detailed analysis revealed that differentially expressed genes were involved in temperature response, ROS accumulation, anther development, and mitochondrial function. Single-molecule long-read isoform sequencing combined with RNA sequencing revealed numerous genes produce alternative splicing transcripts at high temperatures. Here, we also found that alternative oxidase, type II NAD(P)H dehydrogenases, and transcription factor <i>Hsfs</i> might play a crucial role in male fertility under the low-temperature condition. RNA sequencing and bulked segregant analysis coupled with whole-genome sequencing identified the candidate genes involved in the post-transcriptional modification of <i>orf224</i>. Overall, our study described a putative mechanism of fertility restoration in a <i>pol</i> TCMS line controlled by ambient temperature that might help utilise TCMS in the two-line breeding of <i>Brassica</i> crops.Qing XiaoHuadong WangHui ChenXiaohan ChenJing WenCheng DaiChaozhi MaJinxing TuJinxiong ShenTingdong FuBin YiMDPI AGarticleambient temperaturetemperature-sensitive cytoplasmic male sterility<i>Brassica napus</i>single-molecule long-read isoform sequencingBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 12450, p 12450 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	ambient temperature temperature-sensitive cytoplasmic male sterility <i>Brassica napus</i> single-molecule long-read isoform sequencing Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	ambient temperature temperature-sensitive cytoplasmic male sterility <i>Brassica napus</i> single-molecule long-read isoform sequencing Biology (General) QH301-705.5 Chemistry QD1-999 Qing Xiao Huadong Wang Hui Chen Xiaohan Chen Jing Wen Cheng Dai Chaozhi Ma Jinxing Tu Jinxiong Shen Tingdong Fu Bin Yi Molecular Analysis Uncovers the Mechanism of Fertility Restoration in Temperature-Sensitive Polima Cytoplasmic Male-Sterile <i>Brassica napus</i>
description	Temperature-sensitive male sterility is a heritable agronomic trait affected by genotype-environment interactions. In rapeseed (<i>Brassica napus</i>), Polima (<i>pol</i>) temperature-sensitive cytoplasmic male sterility (TCMS) is commonly used for two-line breeding, as the fertility of <i>pol</i> TCMS lines can be partially restored at certain temperatures. However, little is known about the underlying molecular mechanism that controls fertility restoration. Therefore, we aimed to investigate the fertility conversion mechanism of the <i>pol</i> TCMS line at two different ambient temperatures (16 °C and 25 °C). Our results showed that the anthers developed and produced vigorous pollen at 16 °C but not at 25 °C. In addition, we identified a novel co-transcript of <i>orf224-atp6</i> in the mitochondria that might lead to fertility conversion of the <i>pol</i> TCMS line. RNA-seq analysis showed that 1637 genes were significantly differentially expressed in the fertile flowers of 596-L when compared to the sterile flower of 1318 and 596-H. Detailed analysis revealed that differentially expressed genes were involved in temperature response, ROS accumulation, anther development, and mitochondrial function. Single-molecule long-read isoform sequencing combined with RNA sequencing revealed numerous genes produce alternative splicing transcripts at high temperatures. Here, we also found that alternative oxidase, type II NAD(P)H dehydrogenases, and transcription factor <i>Hsfs</i> might play a crucial role in male fertility under the low-temperature condition. RNA sequencing and bulked segregant analysis coupled with whole-genome sequencing identified the candidate genes involved in the post-transcriptional modification of <i>orf224</i>. Overall, our study described a putative mechanism of fertility restoration in a <i>pol</i> TCMS line controlled by ambient temperature that might help utilise TCMS in the two-line breeding of <i>Brassica</i> crops.
format	article
author	Qing Xiao Huadong Wang Hui Chen Xiaohan Chen Jing Wen Cheng Dai Chaozhi Ma Jinxing Tu Jinxiong Shen Tingdong Fu Bin Yi
author_facet	Qing Xiao Huadong Wang Hui Chen Xiaohan Chen Jing Wen Cheng Dai Chaozhi Ma Jinxing Tu Jinxiong Shen Tingdong Fu Bin Yi
author_sort	Qing Xiao
title	Molecular Analysis Uncovers the Mechanism of Fertility Restoration in Temperature-Sensitive Polima Cytoplasmic Male-Sterile <i>Brassica napus</i>
title_short	Molecular Analysis Uncovers the Mechanism of Fertility Restoration in Temperature-Sensitive Polima Cytoplasmic Male-Sterile <i>Brassica napus</i>
title_full	Molecular Analysis Uncovers the Mechanism of Fertility Restoration in Temperature-Sensitive Polima Cytoplasmic Male-Sterile <i>Brassica napus</i>
title_fullStr	Molecular Analysis Uncovers the Mechanism of Fertility Restoration in Temperature-Sensitive Polima Cytoplasmic Male-Sterile <i>Brassica napus</i>
title_full_unstemmed	Molecular Analysis Uncovers the Mechanism of Fertility Restoration in Temperature-Sensitive Polima Cytoplasmic Male-Sterile <i>Brassica napus</i>
title_sort	molecular analysis uncovers the mechanism of fertility restoration in temperature-sensitive polima cytoplasmic male-sterile <i>brassica napus</i>
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/375c2473f1734d58b6cf11ec1a95cdec
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Molecular Analysis Uncovers the Mechanism of Fertility Restoration in Temperature-Sensitive Polima Cytoplasmic Male-Sterile <i>Brassica napus</i>

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