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作物杂志,2026, 第4期: 36–45 doi: 10.16035/j.issn.1001-7283.2026.04.005

• 遗传育种·种质资源·生物技术 • 上一篇    下一篇

水稻OsDOG1-like基因家族突变体种子的萌发势及其GA敏感性的研究

黄新元1(), 张英2, 陶迎梅1, 江青山2, 魏丕伟1,3()   

  1. 1 四川轻化工大学食品与酿酒工程学院， 644005， 四川宜宾
    2 宜宾市农业科学院， 644000， 四川宜宾
    3 长江上游地区白酒数智化管理与生态决策优化四川省重点实验室， 644005， 四川宜宾
  • 收稿日期:2025-05-19 修回日期:2025-06-17 出版日期:2026-08-15 发布日期:2026-08-11
  • 通讯作者: 魏丕伟
  • 作者简介:黄新元，研究方向为水稻遗传育种，E-mail：482151489@qq.com
  • 基金资助:
    国家自然科学基金(31701513);宜宾五粮液股份有限公司资助课题(CXY2022ZR008)

Influence of OsDOG1-like Family Mutants on Seed Germination Potential and Gibberellin Sensitivity in Rice

Huang Xinyuan1(), Zhang Ying2, Tao Yingmei1, Jiang Qingshan2, Wei Piwei1,3()   

  1. 1 College of Food and Brewing Engineering, Sichuan University of Science & Engineering, Yibin 644005, Sichuan, China
    2 Yibin Academy of Agricultural Sciences, Yibin 644000, Sichuan, China
    3 Sichuan Provincial Key Laboratory of Digital-Intelligent Management and Ecological Decision-Making Optimization for Baijiu in the Upper Yangtze River Region, Yibin 644005, Sichuan, China
  • Received:2025-05-19 Revised:2025-06-17 Online:2026-08-15 Published:2026-08-11
  • Contact: Wei Piwei

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摘要： 拟南芥DOG1（DELAY OF GERMINATION 1）基因是控制种子休眠数量性状位点（QTL）的主效基因。水稻OsDOG1-like3基因可通过增强脱落酸（ABA）通路促进种子休眠，OsDOG1-like1和OsDOG1-like2基因的功能尚不明确。采用CRISPR-Cas9技术创制了osdog1l-2、osdog1l-3等单突变体，以及osdog1l-2/3双突变体和osdog1l-1/2/3三突变体。结果表明，osdog1l-2、osdog1l-3等单突变体的种子休眠表型变化很微弱。贮藏6个月后，与浸种后第10天野生型种子的萌发率（72.92%）相比，osdog1l-2/3和osdog1l-1/2/3的萌发率分别降至16.67%和8.33%。0.1和1.0 μmol/L低浓度赤霉素GA3处理使osdog1l-2/3的萌发率提高到45.83%，而对osdog1l-1/2/3的萌发率基本没有影响。osdog1l-1/2/3三突纯合子对低浓度GA3和GA4+7几乎不敏感，这说明OsDOG1-like1对水稻种子萌发在响应GA信号时至关重要，或者OsDOG1-L1、L2、L3三者存在功能上的冗余。高浓度GA3（10.0 μmol/L）处理一定程度上提高了osdog1l-1/2/3的萌发率，达27.08%，同种方法使用GA4+7处理则具有类似的规律。这些结果表明OsDOG1-like基因作用于GA信号的上游（或共同）调控水稻种子萌发。TTC染色证实了osdog1l-2/3和osdog1l-1/2/3突变体胚胎的高活力，可萌发的种子后期能正常生长。

关键词: 水稻, OsDOG1-like基因家族, 三重突变体, 种子萌发

Abstract:

The Arabidopsis thaliana DOG1 (DELAY OF GERMINATION 1) is a regulator gene responsible for the quantitative trait locus (QTL) controlling seed dormancy. In rice, the OsDOG1-like3 gene promotes seed dormancy by enhancing the abscisic acid (ABA) pathway, whereas the functions of OsDOG1-like1 and OsDOG1-like2 remain unclear. This study employed CRISPR-Cas9 technology together with conventional crossing to generate osdog1l-2 and osdog1l-3 single mutants, and osdog1l-2/3 double mutants and osdog1l-1/2/3 triple mutants. The results showed that there were few phenotypic changes in seed germination rates in osdog1l-2 and osdog1l-3 single mutants. However, the germination rates of osdog1l-2/3 and osdog1l-1/2/3 mutants decreased to 16.67% and 8.33% respectively, in contrast to the 72.92% germination rate of wild-type (WT) seeds at 10 days after imbibition. Treatment with low concentration of gibberellin (0.1 and 1.0 μmol/L GA3) increased the germination rate of osdog1l-2/3 to 45.83%, while the germination rate of osdog1l-1/2/3 remained largely unaffected. The homozygous osdog1l-1/2/3 mutant seeds exhibited insensitivity to GA3 and GA4+7, indicating that OsDOG1-like1 would be critical for rice seed germination in response to GA signal, or that OsDOG1-L1, OsDOG1-L2, and OsDOG1-L3 had functional redundancy in the regulation of seed germination. High- concentration GA3 (10.0 μmol/L) partially restored the germination rate of osdog1l-1/2/3 to 27.08%, with GA4+7 treatment showing similar effects. These results suggest that OsDOG1-like genes act upstream of (or in parallel with) GA signaling to regulate rice seed germination. Embryo viability in both osdog1l-2/3 and osdog1l-1/2/3 mutants was confirmed to be similar to that of WT by TTC staining, and normal post-germination growth occurred in the germinated seeds.

Key words: Rice, OsDOG1-like gene family, Triple mutant, Seed germination

表1

水稻OsDOG1-like家族基因编辑的靶点序列

靶点名称Target name 序列（5′-3′）Sequence (5′-3′)
OsDOG1L-1 target 1 TTGGCGAACGACTCGCCGGAGGG
OsDOG1L-2 target 1 CCTCTGGCTCCGCGGGCTGCGGG
OsDOG1L-3 target 1 GACGAACGGGCGGCGTATTATGG

表2

OsDOG1-like基因家族成员鉴定所用引物及序列

基因名称
Gene name		 引物名称
Primer name		 引物序列（5'-3'）
Primer sequence (5'-3')
OsDOG1L-1 DOG1L1-F1 GGCCACCAGTATTTTCTCTGC
DOG1L1-R1 CGCACCAGAGGTAGAGGTTCT
OsDOG1L-2 Sg7047-F1 CACCTATCGCCTCCTCAT
Sg7047-R1 GCCGACAGCGTCCACACC
OsDOG1L-3 GP7029-5796-F CTCGCCTCGTCTCTTCCTTC
GP7029-5796-R CACGCCCAGAGGAAGGTG

图1

OsDOG1-like基因家族成员突变类型的鉴定 OsDOG1L-1基因ID为LOC_Os01g06560、OsDOG1L-2基因ID为LOC_Os05g48650、OsDOG1L-3基因ID为LOC_Os01g20030。

图2

osdog1l-2单突变体种子萌发势的比较

图3

osdog1l-3单突变体种子萌发势的比较

图4

osdog1l-2/3双突变体与osdog1l-1/2/3三重突变体种子萌发势的比较

图5

GA处理下osdog1l-2/3双突变体与osdog1l-1/2/3三重突变体种子萌发率的比较

图6

不同GA浸泡处理下osdog1l-1/2/3三突变体萌发率在浸种第10天的变化 不同小写字母表示差异显著（P < 0.05）。

图7

osdog1-like突变体种子萌发后第10天的幼苗生长情况

图8

osdog1-like突变体的胚胎TTC染色结果

图9

OsDOG1-like基因家族成员互作蛋白预测 (a)、(b)、(c)分别为OsDOG1L-1、OsDOG1L-2、OsDOG1L-3基因互作蛋白预测。(d)和(e)为预测的OsDOG1L-2基因编码蛋白的三维结构。

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