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黑龙江省是我国最大的水稻种植区,近年来水稻种植面积不断增加,为加快高纬度、低积温的地区品种选育进程,本研究利用CRISPR/Cas9基因编辑技术对第二积温带主栽品种绥粳14的抽穗期基因Hd2、香味基因Badh2进行编辑。通过T_1代的分子鉴定,Hd2和Badh2基因被敲除而且基因型纯合。田间鉴定表明,转基因抗性标记也被成功剔除的两个株系,绥粳14-d1和绥粳14-d2,比绥粳14提早13天左右。利用咀嚼法、氢氧化钾浸泡法和气相色谱-质谱联用技术(GCMS)测定香味,结果显示绥粳14-d1和绥粳14-d2中香味物质2-乙酰-1-吡咯啉(2-AP)含量显著高于绥粳14。综合以上结果,本研究利用基因编辑技术,成功获得了改良的抽穗期缩短香米品种,为早熟、优质水稻品种的选育提供了理论指导,加速了育种进程。
Heilongjiang Province is the largest rice planting area in China. In recent years, the area under rice cultivation has been continuously increasing. In order to speed up the breeding process of high latitudes and low accumulations, Heilongjiang Province, using the CRISPR / Cas9 gene editing technology, Japonica 14 heading date gene Hd2, fragrance gene Badh2 for editing. By T 1 generation of molecular identification, Hd2 and Badh2 gene knockout and genotype homozygous. Field identification showed that the two lines with transgenic resistance markers also successfully knocked out, SuiJing 14-d1 and SuiJing 14-d2, were 13 days earlier than SuiJing 14. The flavor was determined by chewing method, potassium hydroxide soaking method and gas chromatography-mass spectrometry (GCMS). The results showed that the flavor substance 2-acetyl-1-pyrroline (2- AP) content was significantly higher than Suijing 14. Based on the above results, we successfully obtained the improved heading date to shorten the rice varieties by using gene editing technology, which provided theoretical guidance for the breeding of early maturity and high quality rice varieties and accelerated the breeding process.