Abstract
Aging in crops like tea and tobacco involves the production of secondary metabolites, with symbiotic microbes playing a key role. However, their dynamic changes and correlation with metabolites during aging remain poorly understood. This study investigates changes in microbial communities, aroma compounds, and protein expression during tobacco leaf aging using artificial accelerated aging techniques, which combine GC-MS, metagenomics, and metaproteomics methods. We identified 62 aroma compounds with distinct change patterns and observed significant changes in the structure of symbiotic bacteria. Type one, represented by Wolbachia_endosymbiont_of_Diaphorina_citri, increased in abundance from the fourth month, correlating with compounds like 2-Furaldehyde. Type two, represented by Sphingomonas_sp_LK11, showed a bimodal abundance pattern, correlating with compounds like Tabanone. Metaproteomics revealed that protein functions were initially limited to cytoskeleton organization but diversified from the fourth month. Fungi also displayed two distinct clustering patterns, Rhizopus and Mortierella elongata were abundant early on, while Colletotrichum asianum and Trichophyton violaceum appeared later. Rhizopus and other fungi exhibited a significant positive correlation with 24 aroma compounds, including 5-Methylfuran-2(5 H)-one. Linderina pennispora and other fungi showed a significant positive correlation with 28 aroma compounds, including 2-Furaldehyde. The dynamic changes in microbial community structure during aging are closely related to the generation of aroma compounds. Overall, temporal shifts in microbial communities were closely linked to aroma formation. One set of microorganisms, such as Wolbachia_endosymbiont_of_Diaphorina_citri and Linderina pennispora, is positively correlated with 2-Furaldehyde, Isophorone, and 2-Methylbenzofuran. Another set, including Sphingomonas_sp_LK11 and Rhizopus, exhibits a positive correlation with 5-Methylfuran-2(5 H)-one and 1,2-Cyclohexanedione. These findings provide new insights into the biological mechanisms of tobacco leaf aging, and offer new research directions for the development and innovation of future tobacco products. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12870-025-07765-3.