Acta Agriculturae Boreali-Sinica ›› 2022, Vol. 37 ›› Issue (2): 9-17. doi: 10.7668/hbnxb.20192762

Special Issue: Rice Biotechnology Hot Article

• Crop Genetics & Breeding·Germplasm Resources·Biotechnology • Previous Articles     Next Articles

Improvement of Grain Shape in Rice by Knocking GS3 and GS9 via CRISPR/Cas9 System

HAN Zhenghong, DUAN Yuxuan, XU Shanbin, WANG Jingguo, LIU Hualong, YANG Luomiao, JIA Yan, XIN Wei, ZHENG Hongliang, ZOU Detang   

  1. Key Laboratory of Germplasm Enhancement,Physiology and Ecology of Food Crops in Cold Region, Ministry of Education,Northeast Agricultural University,Harbin 150030,China
  • Received:2021-10-19 Published:2022-04-28

利用CRISPR/Cas9技术敲除GS3GS9基因改良水稻粒型性状

韩政宏, 段宇轩, 徐善斌, 王敬国, 刘化龙, 杨洛淼, 贾琰, 辛威, 郑洪亮, 邹德堂   

  1. 东北农业大学 寒地粮食作物种质创新与生理生态教育部重点实验室,黑龙江 哈尔滨 150030
  • 作者简介:
    作者简介:韩政宏(1997—),男,辽宁丹东人,在读硕士,主要从事水稻基因编辑育种研究。
  • 基金资助:
    黑龙江省“百千万”工程生物育种重大科技专项(2020ZX16B01)

Abstract:

In order to promote the breeding of long-grain japonica rice varieties,two knock-out vectors,pYLCRISPR/Cas9-GS3-RNA and pYLCRISPR/Cas9-GS3-GS9-RNA,were constructed with japonica rice varieties Dongfu 139,Longjing 31 and Dongnong 427,the genes of GS3 and GS9 were edited by Agrobacterium tumefaciens in vitro.In the end,GS3,GS9 and GS3,GS9 double-gene mutations without T-DNA elements were obtained in the T2.The results showed that the grain length and 1000-grain weight of gs3 mutant plants of three varieties were significantly increased compared with those of wild type,grain width,seed setting rate and grain number per panicle did not change significantly,grain length of gs9 mutant increased significantly,and grain width decreased significantly,while 1000-grain weight,seed setting rate and grain number per panicle did not change significantly,and grain length of gs3gs9 mutant increased more than gs3 and gs9,at the same time,grain width decreased significantly,1000-grain weight increased significantly,while seed setting rate and grain number per ear did not change significantly.To sum up,three japonica rice varieties,Dongfu 139,Longjing 31 and Dongnong 427 were improved by using CRISPR/Cas9 technique,which accelerated the breeding process of new long-grain japonica rice varieties.

Key words: Rice, Grain shape, GS3, GS9, CRISPR/Cas9

摘要:

为促进长粒型粳稻品种的选育,以粳稻品种东富139、龙粳31和东农427为试验材料,利用CRISPR/Cas9基因编辑技术构建了pYLCRISPR/Cas9-GS3-RNA和pYLCRISPR/Cas9-GS3-GS9-RNA 2个敲除载体,通过农杆菌转化法侵染受体材料的愈伤组织,对GS3GS9基因进行定点编辑。最终,3个品种在T2都获得了GS3单基因突变、GS9单基因突变和GS3GS9双基因突变,且无T-DNA元件的纯合植株。在成熟期对T2突变体及其野生型的农艺性状进行考察分析,结果表明,与野生型相比,3个品种的gs3突变植株的粒长、千粒质量均显著增加,粒宽、结实率和穗粒数无显著变化;gs9突变体粒长显著增加,粒宽显著减少,千粒质量、结实率和穗粒数无显著变化;gs3gs9突变体粒长增加,且增幅大于gs3gs9,同时粒宽显著减少,千粒质量显著增加,而结实率和穗粒数无显著变化。综上,利用CRISPR/Cas9技术对东富139、龙粳31和东农427等3个粳稻品种的粒型进行改良,加快了长粒型粳稻新品种的选育进程。

关键词: 水稻, 粒型, GS3, GS9, CRISPR/Cas9