Crops ›› 2025, Vol. 41 ›› Issue (1): 83-88.doi: 10.16035/j.issn.1001-7283.2025.01.010

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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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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

Fig.1

PCR amplification results of mutation site detection primers"

Fig.2

Three genotypes of J11 lines Arrows indicate mutant base locations."

Table 1

Information of NtJAZ1 gene mutants screened"

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

Fig.3

Structural diagram of NtJAZ1 protein"

Fig.4

Nicotine contents in tobacco leaves of homozygous mutant strains with different loci “*”indicates significant difference at P < 0.05 level. The same below."

Fig.5

Nicotine content in two lines of J11 M4 generation in greenhouse before topping “**”indicates significant difference at P < 0.01 level. The same below."

Fig.6

Nicotine content in two lines of J11 M4 generation in greenhouse after topping"

Fig.7

Expression level of NtPMT1a and NtQPT genes in J11 I-1 line before topping"

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