Efficient Breeding of Early-Maturing Rice Cultivar by Editing Hd6 via CRISPR/Cas9

doi:10.1016/j.rsci.2024.06.007

Rice Science ›› 2024, Vol. 31 ›› Issue (6): 629-633.DOI: 10.1016/j.rsci.2024.06.007

Efficient Breeding of Early-Maturing Rice Cultivar by Editing Hd6 via CRISPR/Cas9

Chen Zhihui¹^,²^,³, Tao Yajun¹^,²^,³, Xu Yang¹^,²^,³^,⁴, Wu Jingjing¹^,²^,³, Wang Fangquan¹^,²^,³, Li Wenqi¹^,²^,³, Jiang Yanjie¹^,²^,³, Fan Fangjun¹^,²^,³, Li Xia¹^,²^,³, Zhu Jianping¹^,²^,³, Zhu Qian-Hao⁵(), Yang Jie¹^,²^,³^,⁴()

¹Key Laboratory of Germplasm Innovation in Downstream of Huaihe River, Ministry of Agriculture and Rural Affairs / Institute of Food Crops, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China
²Zhongshan Biological Breeding Laboratory, Nanjing 210014, China
³Jiangsu Co-Innovation Center for Modern Production Technology of Grain Crops, Yangzhou University, Yangzhou 225009, China
⁴Institute of Life Science, Jiangsu University, Zhenjiang 212013, China
⁵Commonwealth Scientific and Industrial Research Organisation, Agriculture and Food, GPO Box 1700, Canberra ACT 2601, Australia

Received:2024-03-29 Accepted:2024-06-12 Online:2024-11-28 Published:2024-12-10
Contact: Yang Jie (yangjie168@aliyun.com); Zhu Qian-Hao (qianhao.zhu@csiro.au)

Abstract

Chen Zhihui, Tao Yajun, Xu Yang, Wu Jingjing, Wang Fangquan, Li Wenqi, Jiang Yanjie, Fan Fangjun, Li Xia, Zhu Jianping, Zhu Qian-Hao, Yang Jie. Efficient Breeding of Early-Maturing Rice Cultivar by Editing Hd6 via CRISPR/Cas9[J]. Rice Science, 2024, 31(6): 629-633.

Figures/Tables 2

Fig. 1. Knockout of Hd6 by CRISPR/Cas9 method produces early heading phenotype. A, Gene structure of Hd6. The white boxes represent the untranslated regions, black boxes stand for the coding regions, and black lines represent the introns. B, DNA sequences flanking the two target sites of sgRNAs in Nanjing 46 (NJ46) and its three mutants (NJ46-1, NJ46-2, and NJ46-3). Inserted sequences are shown in red letter(s), deleted sequences are highlighted by red lines, and blue nucleotides indicate PAM (protospacer adjacent motif). On the right of the numbers, ‘-’ means deletion, and ‘+’ means insertion/substitution. WT, Wild type. C, Phenotypes of NJ46 and three mutants at the heading stage. Scale bars, 5 cm. D, Flowering time of NJ46 and three mutants under long-day conditions at Nanjing city, Jiangsu Province, China, in two years. E, qRT-PCR analysis of the transcription levels of Hd3a and RFT1 in NJ46 and three mutant lines. RNA was extracted from 30-day-old leaves. The rice ubiquitin (UBQ) gene was used as an internal control. Data are Mean ± SD (n = 3). ** represents significant differences at P ≤ 0.01.

Fig. 2. Agronomic traits of Nanjing 46 (NJ46) and its three mutants (NJ46-1, NJ46-2, and NJ46-3). A, Measurements of five agronomic traits in NJ46 and three mutants. Data are Mean ± SD (n = 5). * and ** represent significant differences at P ≤ 0.05 and P ≤ 0.01, respectively. ns, Not significant. B, Morphology of NJ46 and three mutants at the maturity stage. Scale bars, 5 cm. C, Morphology of milled rice grains of NJ46 and three mutants. Scale bar, 2 cm. D, Comparison of rapid viscosity analyzer spectra of rice flours between NJ46 and three mutants.

References 22

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Efficient Breeding of Early-Maturing Rice Cultivar by Editing Hd6 via CRISPR/Cas9

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