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作物杂志,2025, 第1期: 83–88 doi: 10.16035/j.issn.1001-7283.2025.01.010

• 遗传育种·种质资源·生物技术 • 上一篇    下一篇

突变NtJAZ1基因获得高烟碱烤烟新材料

高玉龙1(), 赵璐1, 王丙武1(), 孔光辉1, 王亚辉1, 刘剑金2, 段杰3, 吴兴富1, 李青2, 者开明3   

  1. 1云南省烟草农业科学研究院/国家烟草基因工程研究中心，650021，云南昆明
    2云南烟草公司普洱市公司，665000，云南普洱
    3云南省烟草公司临沧市公司耿马分公司，677500，云南临沧
  • 收稿日期:2023-02-27 修回日期:2024-05-14 出版日期:2025-02-15 发布日期:2025-02-12
  • 通讯作者: 王丙武，主要从事烟草功能基因研究，E-mail：bwwang76@hotmail.com
  • 作者简介:高玉龙，主要从事烟草育种研究，E-mail：gyl3000@163.com
  • 基金资助:
    中国烟草总公司云南省公司科技计划项目(2021530000242029)

Mutating NtJAZ1 Gene to Obtain High Nicotine Flue-Cured Tobacco New Material

Gao Yulong1(), Zhao Lu1, Wang Bingwu1(), Kong Guanghui1, Wang Yahui1, Liu Jianjin2, Duan Jie3, Wu Xingfu1, Li Qing2, Zhe Kaiming3   

  1. 1Yunnan Academy of Tobacco Agricultural Sciences / National Center for Tobacco Gene Engineering, Kunming 650021, Yunnan, China
    2Puer Branch of Yunnan Provincial Tobacco Company, Puer 665000, Yunnan, China
    3Lincang Company Gengma Branch of Yunnan Provincial Tobacco Company, Lincang 677500, Yunnan, China
  • Received:2023-02-27 Revised:2024-05-14 Online:2025-02-15 Published:2025-02-12

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摘要：

烟碱是栽培烟草中重要的化学成分之一，深刻影响烟叶质量。为了培育高烟碱含量的烟草新材料，在我国主栽品种云烟87的甲基磺酸乙酯（EMS）突变体库中筛选烟碱合成负调控因子基因NtJAZ1的突变体材料。结果表明，在突变体库中筛选获得了12个突变体材料，其中10个突变导致NtJAZ1氨基酸变化，1份突变位点位于内含子，1个无义突变。温室种植纯合突变体材料，现蕾期检测整株叶片烟碱含量，其中2个株系（J8和J11）的烟碱含量显著高于对照。J11 I-1和J11 Ⅳ-1打顶后其烟碱含量分别比对照提高48%和58%。qRT-PCR表明，NtPMT1a和NtQPT基因在突变体J11 I-1根中的表达水平显著升高。综上，筛选获得的高烟碱突变体材料J11不涉及转基因成分，可为培育高烟碱含量烤烟新品种提供种质资源。

关键词: 烤烟, 烟碱, 甲基磺酸乙酯（EMS）, NtJAZ1

Abstract:

Nicotine is one of the most important chemical compounds in cultivated tobacco, which largely determines the quality of tobacco leaves. In order to develop a new tobacco material with high nicotine content, the mutant material of NtJAZ1, a negative regulator of nicotine synthesis, was screened from the ethyl methyl sulfonate (EMS) mutant library of Yunyan 87, a major cultivar in China. The results showed that 12 mutant materials were selected from the mutant library, of which ten mutations resulted in the amino acid change of NtJAZ1, one mutation was located in the intron, and one nonsense mutation. The homozygous mutant material was planted in greenhouse. The nicotine contents in leaves of two strains (J8 and J11) were significantly higher than that of the control at bud stage. After topping, the nicotine content of J11 I-1 and J11 IⅤ-1 increased by 48% and 58%, respectively, compared with the control. qRT-PCR showed that the expression levels of NtPMT1a and NtQPT genes in mutant J11 I-1 were significantly increased. In conclusion, the high-nicotine mutant material J11 obtained by screening does not involve transgenic components, which can provide germplasm resources for breeding new varieties of flue-cured tobacco with high nicotine content.

Key words: Tobacco, Nicotine, Ethyl methyl sulfonate (EMS), NtJAZ1

图1

突变位点检测引物PCR扩增结果

图2

J11株系3个基因型 箭头所示为突变碱基位置。

表1

筛选获得的NtJAZ1基因突变体信息

突变体编号
Number of
mutants		 核苷酸突变
Nucleotide
mutation		 氨基酸突变体
Amino acid
mutation		 突变位点所在结构域
The domain of the
mutation site
J2 C305T T102I ZIM
J3 C346T P116S ZIM
J4 G584A R195K Jas
J5 C650T T217I C端
J6 G652A E218K C端
J8 G500A R167Q ZIM和Jas间隔区
J9 C708T A203V Jas
J10 C616T Q206* Jas
J11 C392T T131I ZIM和Jas间隔区
J14 C473T T158I ZIM和Jas间隔区
J15 G372A M124I ZIM
J16 内含子 内含子

图3

NtJAZ1蛋白结构示意图

图4

不同位点纯合突变体株系烟叶烟碱含量 “*”表示在P < 0.05水平差异显著。下同。

图5

打顶前温室J11 M4代2个株系烟碱含量 “**”表示在P < 0.01水平差异显著。下同。

图6

打顶后温室J11 M4代2个株系烟碱含量

图7

打顶前J11 I-1 NtPMT1a和NtQPT基因的表达水平

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