Generation of a New Glutinous Photothermosensitive Genic-Male-Sterile (PTGMS) Line by CRISPR/Cas9-Directed Mutagenesis of Wx in Rice (<i>Oryza sativa</i> L.)

The Photothermosensitive Genic-Male-Sterile (PTGMS) line, Y58S, an indica rice variety, combines high-quality and high-light-efficiency use, disease and stress resistance, and excellent plant type and mating force. Y58S is widely used to assemble two-line hybrid rice varieties, especially super hybr...

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Autores principales:	Kaichong Teng, Xin Wang, Xinying Guo, Yaoguang Liu, Rongbai Li
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	rice genome editing crispr/cas9 waxy amylose content (ac) y58s Agriculture (General) S1-972
Acceso en línea:	https://doaj.org/article/4232785c74c44daa8399aaaf685a62ae
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spelling	oai:doaj.org-article:4232785c74c44daa8399aaaf685a62ae2021-11-25T15:58:10ZGeneration of a New Glutinous Photothermosensitive Genic-Male-Sterile (PTGMS) Line by CRISPR/Cas9-Directed Mutagenesis of Wx in Rice (<i>Oryza sativa</i> L.)10.3390/agriculture111110442077-0472https://doaj.org/article/4232785c74c44daa8399aaaf685a62ae2021-10-01T00:00:00Zhttps://www.mdpi.com/2077-0472/11/11/1044https://doaj.org/toc/2077-0472The Photothermosensitive Genic-Male-Sterile (PTGMS) line, Y58S, an indica rice variety, combines high-quality and high-light-efficiency use, disease and stress resistance, and excellent plant type and mating force. Y58S is widely used to assemble two-line hybrid rice varieties, especially super hybrids. The <i>Wx</i> gene is the main effector gene for controlling amylose synthesis, which determines the amylose content (AC) of rice grains. By editing this gene, a glutinous line with a low AC can be obtained. In this study, the CRISPR/Cas9 system was used to mediate the editing of the <i>Wx</i> gene, which caused ultra-low AC mutations that produced a PTGMS glutinous rice strain with excellent waxiness. The results showed that 18 positively transformed plants were obtained from the T<sub>0</sub> generation, with a mutation rate of 64.29%, of which six were homozygous mutant plants, indicating that the gene-editing target had a higher targeting efficiency and a higher homozygosity mutation rate. Compared to the wild type, the AC of the mutants was significantly lower. Through molecular marker detection and screening of T<sub>1</sub> and T<sub>2</sub> generations, five homozygous T-DNA-free mutant strains were identified that were consistent with Y58S in fertility and other agronomic traits except for AC. Among these, the AC of the W-1-B-5 homozygous mutant, the glutinous PTGMS line wx-Y58S, was as low as 0.6%. Our research revealed that the <i>Wx</i> gene of excellent PTGMS rice can be edited to generate a new waxy PTGMS line using the CRISPR/Cas9 system. This study provided a simple and effective strategy for breeding high-yield, high-quality, and glutinous two-line hybrid rice, and provided excellent sterile lines for their large-scale application. Once put into use, waxy hybrid rice will greatly improve the yield of glutinous rice and increase social benefits.Kaichong TengXin WangXinying GuoYaoguang LiuRongbai LiMDPI AGarticlericegenome editingcrispr/cas9waxyamylose content (ac)y58sAgriculture (General)S1-972ENAgriculture, Vol 11, Iss 1044, p 1044 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	rice genome editing crispr/cas9 waxy amylose content (ac) y58s Agriculture (General) S1-972
spellingShingle	rice genome editing crispr/cas9 waxy amylose content (ac) y58s Agriculture (General) S1-972 Kaichong Teng Xin Wang Xinying Guo Yaoguang Liu Rongbai Li Generation of a New Glutinous Photothermosensitive Genic-Male-Sterile (PTGMS) Line by CRISPR/Cas9-Directed Mutagenesis of Wx in Rice (<i>Oryza sativa</i> L.)
description	The Photothermosensitive Genic-Male-Sterile (PTGMS) line, Y58S, an indica rice variety, combines high-quality and high-light-efficiency use, disease and stress resistance, and excellent plant type and mating force. Y58S is widely used to assemble two-line hybrid rice varieties, especially super hybrids. The <i>Wx</i> gene is the main effector gene for controlling amylose synthesis, which determines the amylose content (AC) of rice grains. By editing this gene, a glutinous line with a low AC can be obtained. In this study, the CRISPR/Cas9 system was used to mediate the editing of the <i>Wx</i> gene, which caused ultra-low AC mutations that produced a PTGMS glutinous rice strain with excellent waxiness. The results showed that 18 positively transformed plants were obtained from the T<sub>0</sub> generation, with a mutation rate of 64.29%, of which six were homozygous mutant plants, indicating that the gene-editing target had a higher targeting efficiency and a higher homozygosity mutation rate. Compared to the wild type, the AC of the mutants was significantly lower. Through molecular marker detection and screening of T<sub>1</sub> and T<sub>2</sub> generations, five homozygous T-DNA-free mutant strains were identified that were consistent with Y58S in fertility and other agronomic traits except for AC. Among these, the AC of the W-1-B-5 homozygous mutant, the glutinous PTGMS line wx-Y58S, was as low as 0.6%. Our research revealed that the <i>Wx</i> gene of excellent PTGMS rice can be edited to generate a new waxy PTGMS line using the CRISPR/Cas9 system. This study provided a simple and effective strategy for breeding high-yield, high-quality, and glutinous two-line hybrid rice, and provided excellent sterile lines for their large-scale application. Once put into use, waxy hybrid rice will greatly improve the yield of glutinous rice and increase social benefits.
format	article
author	Kaichong Teng Xin Wang Xinying Guo Yaoguang Liu Rongbai Li
author_facet	Kaichong Teng Xin Wang Xinying Guo Yaoguang Liu Rongbai Li
author_sort	Kaichong Teng
title	Generation of a New Glutinous Photothermosensitive Genic-Male-Sterile (PTGMS) Line by CRISPR/Cas9-Directed Mutagenesis of Wx in Rice (<i>Oryza sativa</i> L.)
title_short	Generation of a New Glutinous Photothermosensitive Genic-Male-Sterile (PTGMS) Line by CRISPR/Cas9-Directed Mutagenesis of Wx in Rice (<i>Oryza sativa</i> L.)
title_full	Generation of a New Glutinous Photothermosensitive Genic-Male-Sterile (PTGMS) Line by CRISPR/Cas9-Directed Mutagenesis of Wx in Rice (<i>Oryza sativa</i> L.)
title_fullStr	Generation of a New Glutinous Photothermosensitive Genic-Male-Sterile (PTGMS) Line by CRISPR/Cas9-Directed Mutagenesis of Wx in Rice (<i>Oryza sativa</i> L.)
title_full_unstemmed	Generation of a New Glutinous Photothermosensitive Genic-Male-Sterile (PTGMS) Line by CRISPR/Cas9-Directed Mutagenesis of Wx in Rice (<i>Oryza sativa</i> L.)
title_sort	generation of a new glutinous photothermosensitive genic-male-sterile (ptgms) line by crispr/cas9-directed mutagenesis of wx in rice (<i>oryza sativa</i> l.)
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/4232785c74c44daa8399aaaf685a62ae
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Generation of a New Glutinous Photothermosensitive Genic-Male-Sterile (PTGMS) Line by CRISPR/Cas9-Directed Mutagenesis of Wx in Rice (<i>Oryza sativa</i> L.)

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