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Genome-wide characterization of polyphenol oxidase (PPO) gene family in tobacco and the molecular improvement of browning reaction for K326 tobacco leaves

文献类型： 外文期刊

作者： Zhang, Shixu 1 ; Lin, Yingchao 2 ; Zeng, Xinhai 1 ; Liu, Guangju 1 ; Ye, Guixiang 1 ; Du, Changyang 1 ; Li, Yanchun 3 ; Lin, Wenxiong 1 ; Wei, Kesu 2 ; Li, Zhaowei 1 ;

作者机构： 1.Fujian Agr & Forestry Univ, Coll Life Sci, Fujian Prov Key Lab Agroecol Proc & Safety Monitor, Fuzhou 350002, Peoples R China

2.Guizhou Acad Tobacco Sci, Key Lab Mol Genet China Tobacco, Guiyang 550081, Peoples R China

3.Fujian Acad Agr Sci, Inst Resources Environm & Soil Fertilizer, Fuzhou 350003, Peoples R China

关键词： Tobacco; CRISPR/Cas9; Browning of tobacco leaves; NtPPOs

期刊名称：INDUSTRIAL CROPS AND PRODUCTS （ 影响因子：6.2； 五年影响因子：6.2 ）

ISSN： 0926-6690

年卷期： 2025 年 235 卷

页码：

收录情况： SCI

摘要： Polyphenol oxidase (PPO) oxidizes polyphenols, causing browning in tobacco (Nicotiana tabacum L.) leaves during roasting, which reduces their quality and commercial value. In this study, 13 NtPPO genes were identified through genome mining, and their bioinformatic analyses were performed, including gene structure, protein structure, and physicochemical properties. Tissue-specific expression analysis found that the NtPPO genes were substantially expressed in leaves, whereas NtPPO9 and NtPPO10 were also highly expressed in flowers. During leaf growth, NtPPO1, 3, 5, 6, and 7 expression levels changed dramatically, whereas NtPPO2, 9, 10, and 13 expression patterns remained steady. Notably, NtPPO1 displayed the highest expression levels during the maturity stage of tobacco leaves. Subcellular localization revealed that NtPPO1, 6, 9, and 12 were primarily distributed in chloroplasts. KO-87 was generated through CRISPR/Cas9 technology, including mutations in the NtPPO1, 2, 4, 6, 8, 9, 11, 12, and 13 genes. KO-87 exhibits similar growth to the wild-type, but with decreased PPO activity in leaves and browning levels after roasting. Metabolomics analysis indicated that polyphenol metabolites were significantly upregulated, while quinone metabolites were significantly downregulated in the leaves of mutant plants. This study systematically explored the functions of NtPPO genes, providing preliminary insights into their key roles in the browning process of tobacco leaves. These findings offer new research directions and potential solutions for addressing the browning problem during tobacco leaf roasting.

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