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. 2025 Mar 5:16:1537924.
doi: 10.3389/fpls.2025.1537924. eCollection 2025.

Microstructure observation and flavor substances excavation of Yunyan 87 tobacco leaves with different oil contents

Xianfeng Hu #  1 , Wei Xu #  2 , Yubo Zhang  1 , Shouhui Pan  3 , Yanlan Xie  1 , Rui Liao  2 , Shenggang Yang  2 , Youxiang Wu  2 , Daomao Deng  3
Affiliations

Microstructure observation and flavor substances excavation of Yunyan 87 tobacco leaves with different oil contents

Xianfeng Hu et al. Front Plant Sci. .

Abstract

Introduction: The oil content of tobacco leaves is intimately associated with their aromatic characteristics. This study aims to explore the microstructure and distinctive flavor substances of Yunyan 87 high-oil-content tobacco leaves.

Methods: The microstructure and characteristic flavor substances of Yunyan 87 tobacco leaves with different oil contents were analyzed using scanning electron microscope (SEM) and comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry (GC×GC-TOF MS).

Results: The results indicate that the surface of high-oil tobacco leaves was characterized by a high density of glandular hairs, primarily composed of short-stalked glandular hairs featuring enlarged glandular heads. A total of 1551 flavor substances were detected in high-oil tobacco leaves, compared to 1500 metabolites were identified in low-oil tobacco leaves. Among these flavor substances, eight exhibited up-regulated, while three were down-regulated. Notably, the oil-related substances hexadecanoic acid methyl ester and the aroma-related substances nonanoic acid methyl ester and pseudoionone were found to be significantly more abundant in high-oil tobacco leaves compared to their low-oil counterparts. Consequently, hexadecanoic acid methyl ester may serve as a reliable indicator for evaluating the oil content in tobacco leaves, while nonanoic acid methyl ester and pseudoionone could play crucial roles as flavor substances influencing the aroma of tobacco leaves.

Discussion: These findings provide a theoretical foundation for future research on the regulatory mechanisms underlying the synthesis of aroma-producing flavor substances in tobacco leaves.

Keywords: flavor substances; hexadecanoic acid methyl ester; nonanoic acid methyl ester; pseudoionone; tobacco oil content.

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Conflict of interest statement

Authors WX, RL, SY, and YW were employed by the company China Tobacco Guizhou Industrial Co Ltd. SP and DD were employed by the company Guizhou Province Tobacco Company Anshun Company. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.

Figures

Figure 1
Figure 1
Agronomic characteristics of tobacco leaves. (A)Average leaf length, (B) average leaf width, (C) leaf shape index, and (D) single leaf weight. The letters ‘H’ and ‘L’ denote high oil content and low oil content tobacco leaves, respectively. In the bar chart, means sharing the same letter are not significantly different from each other as determined by ANOVA and Duncan’s test (p < 0. 05).
Figure 2
Figure 2
Appearance morphology of dried tobacco leaves with different oils contents. (A–C) high-oil tobacco leaves, (D–F) low-oil tobacco leaves.
Figure 3
Figure 3
Microscopic morphology of dried tobacco leaves with different oils contents. (A–D) high-oil dry tobacco leaves, (E–H) low-oil dry tobacco leaves.
Figure 4
Figure 4
Total ion flow chromatogram of tobacco leaves with different oil contents. (A–C) high-oil dry tobacco leaves, (D–F) low-oil dry tobacco leaves.
Figure 5
Figure 5
Principal component analysis (PCA) of tobacco leaves with different oil contents. (A) PCA Score Plot, (B) PLS-DA Score Plot, (C) OPLS-DA Score Plot. The letters ‘H’ and ‘L’ denote high oil content and low oil content tobacco leaves, respectively.
Figure 6
Figure 6
Statistical analysis of identification flavor substances of tobacco leaves with different oil contents. (A) Column chart of the number of identification substances; (B) Venn diagram of the number of identification substances. The letters ‘H’ and ‘L’ denote high oil content and low oil content tobacco leaves, respectively.
Figure 7
Figure 7
Analysis of flavor substances in tobacco leaves with different oils. The letters ‘H’ and ‘L’ denote high oil content and low oil content tobacco leaves, respectively.
Figure 8
Figure 8
Screening of various substances in tobacco leaves and analysis of their sensory flavor characteristics. (A) Statistical column chart of different flavor substances; (B) heat map of different flavor substances; (C) volcano plot of different flavor substances; (D) sensory flavor characteristic chart of tobacco leaves with different oil contents. The letters ‘H’ and ‘L’ denote high oil content and low oil content tobacco leaves, respectively.
Figure 9
Figure 9
Correlation network diagram of sensory flavor characteristics and flavor substances.
See this image and copyright information in PMC

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