title: Development of Commercial Thermo-sensitive Genic Male Sterile Rice Accelerates Hybrid Rice Breeding Using the CRISPR/Cas9-mediated TMS5 Editing System.
abstract: Hybrid rice breeding offers an important strategy to improve rice production, in which the cultivation of a male sterile line is the key to the success of cross-breeding. CRISPR/Cas9 systems have been widely used in target-site genome editing, whereas their application for crop genetic improvement has been rarely reported. Here, using the CRISPR/Cas9 system, we induced specific mutations in TMS5, which is the most widely applied thermo-sensitive genic male sterility (TGMS) gene in China, and developed new "transgene clean" TGMS lines. We designed 10 target sites in the coding region of TMS5 for targeted mutagenesis using the CRISPR/Cas9 system and assessed the potential rates of on- and off-target effects. Finally, we established the most efficient construct, the TMS5ab construct, for breeding potentially applicable "transgene clean" TGMS lines. We also discussed factors that affect the editing efficiency according to the characteristics of different target sequences. Notably, using the TMS5ab construct, we developed 11 new "transgene clean" TGMS lines with potential applications in hybrid breeding within only one year in both rice subspecies. The application of our system not only significantly accelerates the breeding of sterile lines but also facilitates the exploitation of heterosis.
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangzhou 510642, China.|Key Laboratory of Plant Functional Genomics and Biotechnology of Guangdong Provincial Higher Education Institutions, Guangzhou 510642, China.|College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Hai
authors.initials: H
authors.lastname: Zhou
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangzhou 510642, China.|Key Laboratory of Plant Functional Genomics and Biotechnology of Guangdong Provincial Higher Education Institutions, Guangzhou 510642, China.|College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Ming
authors.initials: M
authors.lastname: He
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangzhou 510642, China.|Key Laboratory of Plant Functional Genomics and Biotechnology of Guangdong Provincial Higher Education Institutions, Guangzhou 510642, China.|College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Li
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangzhou 510642, China.|Key Laboratory of Plant Functional Genomics and Biotechnology of Guangdong Provincial Higher Education Institutions, Guangzhou 510642, China.|College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Liang
authors.initials: L
authors.lastname: Chen
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangzhou 510642, China.|Key Laboratory of Plant Functional Genomics and Biotechnology of Guangdong Provincial Higher Education Institutions, Guangzhou 510642, China.|College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Zhifeng
authors.initials: Z
authors.lastname: Huang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangzhou 510642, China.|Key Laboratory of Plant Functional Genomics and Biotechnology of Guangdong Provincial Higher Education Institutions, Guangzhou 510642, China.|College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Shaoyan
authors.initials: S
authors.lastname: Zheng
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangzhou 510642, China.|Key Laboratory of Plant Functional Genomics and Biotechnology of Guangdong Provincial Higher Education Institutions, Guangzhou 510642, China.|College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Liya
authors.initials: L
authors.lastname: Zhu
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangzhou 510642, China.|Key Laboratory of Plant Functional Genomics and Biotechnology of Guangdong Provincial Higher Education Institutions, Guangzhou 510642, China.|College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Erdong
authors.initials: E
authors.lastname: Ni
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangzhou 510642, China.|Key Laboratory of Plant Functional Genomics and Biotechnology of Guangdong Provincial Higher Education Institutions, Guangzhou 510642, China.|College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Dagang
authors.initials: D
authors.lastname: Jiang
authors.affiliation: State Key Laboratory of Hybrid Rice, China National Hybrid Rice R&D Center, Changsha 410125, China.
authors.forename: Bingran
authors.initials: B
authors.lastname: Zhao
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangzhou 510642, China.|Key Laboratory of Plant Functional Genomics and Biotechnology of Guangdong Provincial Higher Education Institutions, Guangzhou 510642, China.|College of Life Sciences, South China Agricultural University, Guangzhou 510642, China.
authors.forename: Chuxiong
authors.initials: C
authors.lastname: Zhuang
mesh_terms: D001483:Base Sequence; D001947:Breeding / Q000379:methods*; D064113:CRISPR-Cas Systems / Q000235:genetics*; D000072669:Gene Editing; D006824:Hybridization, Genetic; D009154:Mutation / Q000235:genetics; D012275:Oryza / Q000033:anatomy & histology / Q000235:genetics*; D051479:Plant Infertility / Q000235:genetics*; D030821:Plants, Genetically Modified; D011058:Pollen / Q000235:genetics; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000378:metabolism; D013696:Temperature
chemical_list: D000094704:RNA, Guide, CRISPR-Cas Systems