利用CRISPR/Cas9编辑Badh2基因改良迁粳184香味性状

doi:10.12190/j.issn.2096-1197.2026.03.01

北方农业学报 ›› 2026, Vol. 54 ›› Issue (3): 1-12.doi: 10.12190/j.issn.2096-1197.2026.03.01

• 作物遗传育种·分子生物学 • 下一篇

利用CRISPR/Cas9编辑Badh2基因改良迁粳184香味性状

李凡, 陈春, 郭新亚, 王飞飞, 史玉良, 陈卫军

江苏省农业科学院宿迁农科所,江苏宿迁 223800

收稿日期:2025-11-12 修回日期:2026-04-08 出版日期:2026-06-20 发布日期:2026-08-11
通讯作者: 陈卫军（1969—）,男,研究员,学士,主要从事水稻育种与栽培技术方面的研究工作。
作者简介:李凡（1992—）,男,助理研究员,博士,主要从事作物分子育种方面的研究工作。
基金资助:
宿迁市科技计划项目（L202301）

Improvement of aroma traits in Qianjing 184 via CRISPR/Cas9-mediated editing of Badh2 gene

LI Fan, CHEN Chun, GUO Xinya, WANG Feifei, SHI Yuliang, CHEN Weijun

Suqian Institute of Agricultural Sciences,Jiangsu Academy of Agricultural Sciences,Suqian 223800,China

Received:2025-11-12 Revised:2026-04-08 Online:2026-06-20 Published:2026-08-11

摘要/Abstract

摘要： 【目的】改良自主育成优质粳稻品种迁粳184的香味性状,进一步提升其食味品质。【方法】针对甜菜碱脱氢酶Badh2基因第3外显子序列确定编辑位点,构建CRISPR/Cas9元件后通过农杆菌转化至迁粳184愈伤组织中,获得不同突变类型的Badh2突变体株系,对纯合突变体的香味性状及各关键农艺性状进行检测分析。【结果】通过CRISPR/Cas9技术编辑迁粳184的Badh2基因,获得13株T₀代突变株系,纯化获得4种不携带基因编辑元件的T₁代纯合突变体株系Badh2-1、Badh2-2、Badh2-3、Badh2-4,4株突变体株系较野生型迁粳184Badh2基因分别缺失5 bp、插入1 bp、缺失1 bp、缺失2 bp碱基。农艺性状鉴定结果表明,除Badh2-3株高显著低于野生型外（P<0.05）,Badh2-1、Badh2-2、Badh2-4的株高与野生型相比均无显著差异（P>0.05）;4种突变体的粒长、粒宽、每穗粒数、千粒重、结实率均与野生型无显著差异（P>0.05）。经鉴定,4种突变体籽粒及叶片均有明显香味,且籽粒中2-乙酰-1-吡咯啉含量极显著提升（P<0.01）,其含量分别为1.52、1.40、1.41、1.49 mg/kg,而野生型籽粒及叶片均无香味。【结论】迁粳184的4个Badh2突变株系均实现明显增香,且除Badh2-3外,其余突变体的关键农艺性状未受到显著影响,表明利用CRISPR/Cas9技术对该位点进行靶向编辑可用于粳稻品种的香味品质提升。

关键词: 水稻, 迁粳184, CRISPR/Cas9, Badh2, 香味, 2-乙酰-1-吡咯啉

Abstract: 【Objective】To improve the aroma traits of self-bred high-quality japonica rice Qianjing 184 and further enhance its eating quality.【Methods】The editing site was identified in the third exon sequence of the betaine aldehyde dehydrogenase homologue 2（Badh2） gene. The CRISPR/Cas9 elements were constructed and introduced into the calli of Qianjing 184 via Agrobacterium-mediated transformation. The Badh2 mutant lines with different mutation types were obtained and the aroma traits and key agronomic traits of the homozygous mutants were detected and analyzed.【Results】A total of thirteen T₀ generation mutant lines were obtained by targeted editing of Badh2 in Qianjing 184 using CRISPR/Cas9 system. After genetic purification,four T₁ homozygous mutant lines（Badh2-1,Badh2-2,Badh2-3,Badh2-4） without CRISPR/Cas9 editing elements were obtained. Compared with wild-type Qianjing 184,the Badh2 gene of the four mutant lines carried a 5 bp deletion,1 bp insertion,1 bp deletion and 2 bp deletion,respectively. The results of agronomic trait phenotypic identification showed that except for plant height in Badh2-3,which was significantly lower than that of the wild-type（P<0.05）,the plant heights of Badh2-1,Badh2-2,and Badh2-4 showed no significant differences from that of the wild-type（P>0.05）;the grain length,grain width,grain number per panicle,1 000-grain weight and seed setting rate of the four mutants were not significantly different from those of the wild-type（P>0.05）. After identification,the grains and leaves of the four mutants had obvious aroma,and the content of 2-acetyl-1-pyrroline in the grains was extremely significantly increased（P<0.01）,reaching 1.52,1.40,1.41 and 1.49 mg/kg,respectively,while the wild-type grains and leaves had no aroma.【Conclusion】The four Badh2 mutant lines of Qianjing 184 all exhibit significant aroma enhancement,and the key agronomic traits of the other mutants are not significantly affected except Badh2-3,indicating that targeted editing at this site via CRISPR/Cas9 technology can be applied to improve aroma quality of japonica rice varieties.

Key words: Rice, Qianjing 184, CRISPR/Cas9, Badh2, Aroma, 2-acetyl-1-pyrroline

中图分类号:

S511.22

李凡, 陈春, 郭新亚, 王飞飞, 史玉良, 陈卫军. 利用CRISPR/Cas9编辑Badh2基因改良迁粳184香味性状[J]. 北方农业学报, 2026, 54(3): 1-12.

LI Fan, CHEN Chun, GUO Xinya, WANG Feifei, SHI Yuliang, CHEN Weijun. Improvement of aroma traits in Qianjing 184 via CRISPR/Cas9-mediated editing of Badh2 gene[J]. Journal of Northern Agriculture, 2026, 54(3): 1-12.

参考文献

[1] 韩建军,侯婧祎,郭志涛,等.复杂网络视角下的世界主粮贸易格局与趋势分析[J].河南农业大学学报,2023,57(2):342-351.
[2] 高维维,陈思平,王丽平,等.稻米蒸煮品质性状与分子标记关联研究[J].中国农业科学,2017,50(4):599-617.
[3] 余亚莹,邵高能,圣忠华,等.国内外香稻资源遗传多样性研究[J].植物分类与资源学报,2015,37(6):871-880.
[4] 吴钱蓉,吴娴,姜雪,等.利用CRISPR/Cas9技术编辑Badh2基因改良水稻优质恢复系QR79的香味[J/OL].作物杂志,2025:1-11(2025-04-24).https://kns.cnki.net/KCMS/detail/detail.aspx?Filename=ZWZZ20250423002&dbname=CJFD&dbcode=CJFQ.
[5] JEZUSSEK M,JULIANO B O,SCHIEBERLE P.Comparison of key aroma compounds in cooked brown rice varieties based on aroma extract dilution analyses[J].Journal of Agricultural and Food Chemistry,2002,50(5):1101-1105.
[6] KOVACH M J,CALINGACION M N,FITZGERALD M A,et al.The origin and evolution of fragrance in rice (Oryza sativa L.)[J].Proceedings of the National Academy of Sciences of the United States of America,2009,106(34):14444-14449.
[7] YANG D S,LEE K S,JEONG O Y,et al.Characterization of volatile aroma compounds in cooked black rice[J].Journal of Agricultural and Food Chemistry,2008,56(1):235-240.
[8] HINGE V R,PATIL H B,NADAF A B.Aroma volatile analyses and 2AP characterization at various developmental stages in Basmati and Non-Basmati scented rice (Oryza sativa L.) cultivars[J].Rice,2016,9(1):38.
[9] 张来桐,杨乐,刘洪,等.水稻香味物质的研究进展[J].中国水稻科学,2025,39(2):171-186.
[10] BRADBURY L M T,FITZGERALD T L,HENRY R J,et al.The gene for fragrance in rice[J].Plant Biotechnology Journal,2005,3(3):363-370.
[11] CHEN S H,YANG Y,SHI W W,et al.Badh2,encoding betaine aldehyde dehydrogenase,inhibits the biosynthesis of 2-acetyl-1-pyrroline,a major component in rice fragrance[J].The Plant Cell,2008,20(7):1850-1861.
[12] SHI W W,YANG Y,CHEN S H,et al.Discovery of a new fragrance allele and the development of functional markers for the breeding of fragrant rice varieties[J].Molecular Breeding,2008,22(2):185-192.
[13] SHI Y Q,ZHAO G C,XU X L,et al.Discovery of a new fragrance allele and development of functional markers for identifying diverse fragrant genotypes in rice[J].Molecular Breeding,2014,33(3):701-708.
[14] 邵高能,谢黎虹,焦桂爱,等.利用CRISPR/CAS9技术编辑水稻香味基因Badh2[J].中国水稻科学,2017,31(2):216-222.
[15] 朱永生,沈丽华,马静,等.利用CRISPR/Cas9技术编辑Badh2定向改良宁粳50号的香味[J].福建稻麦科技,2023,41(4):18-22.
[16] 张文婷,李阳,裘实,等.基于CRISPR/Cas9基因编辑技术研究Badh2基因对稻米品质的影响[J].中国农业科技导报(中英文),2025,27(5):39-48.
[17] GAO Y B,ZHAO Y D.Self-processing of ribozyme-flanked RNAs into guide RNAs in vitro and in vivo for CRISPR-mediated genome editing[J].Journal of Integrative Plant Biology,2014,56(4):343-349.
[18] 苏军,胡昌泉,翟红利,等.农杆菌介导籼稻明恢86高效稳定转化体系的建立[J].福建农业学报,2003,18(4):209-213.
[19] MÄDE D,DEGNER C,GROHMANN L.Detection of genetically modified rice:a construct-specific real-time PCR method based on DNA sequences from transgenic Bt rice[J].European Food Research and Technology,2006,224(2):271-278.
[20] SHAN Q W,ZHANG Y,CHEN K L,et al.Creation of fragrant rice by targeted knockout of the OsBADH2 gene using TALEN technology[J].Plant Biotechnology Journal,2015,13(6):791-800.
[21] 齐金岗,陈颖,刘开强,等.利用CRISPR-Cas9技术编辑OsBADH2基因改良优质恢复系桂恢852的香味品质[J].南方农业学报,2024,55(7):2058-2068.
[22] 胡明珠,杨瑰丽,刘洪,等.香型水稻种质资源筛选与Badh2等位变异分析[J].分子植物育种,2025,23(15):5028-5036.
[23] 韦新宇,曾跃辉,杨旺兴,等.利用CRISPR-Cas9技术编辑Badh2基因创制优质香型籼稻三系不育系[J].作物学报,2023,49(8):2144-2159.
[24] 邹德堂,韩政宏,郑洪亮,等.利用CRISPR/Cas9技术创制香糯型粳稻种质[J].东北农业大学学报,2022,53(5):1-7.
[25] 段敏,谢留杰,岳雅妮,等.基于CRISPR/Cas9技术创制优质香味粳稻品系[J/OL].中国水稻科学,2025:1-12(2025-09-03).https://kns.cnki.net/KCMS/detail/detail.aspx?filename=ZGSK20250902003&dbname=CJFD&dbcode=CJFQ.

利用CRISPR/Cas9编辑Badh2基因改良迁粳184香味性状

Improvement of aroma traits in Qianjing 184 via CRISPR/Cas9-mediated editing of Badh2 gene

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