Abstract

Microbiome plays an important role during the tobacco aging process which was an indispensable link in the production and processing of cigarettes. However, the structure and functions of microbiome have not been clarified during the tobacco aging process. In this study, 16S rDNA and ITS amplicon sequencing techniques were used to analyze the core microbiome of 15 tobacco samples from five different aging stages. The whole bacterial microbiome was classified into 29 microbial phyla and 132 orders. Enterobacteriales (63%), Pseudomonadales (16%), Sphingomonadales (8%), Xanthomonadales (4%), Burkholderiales (4%), Rhizobiales (3%), and Bacillales (2%) comprised the core bacterial microbiome. The whole fungal microbiome was classified into five microbial phyla and 52 orders. Incertae_sedis_Eurotiomycetes (27%), Wallemiales (25%), Sporidiobolales (17%), Capnodiales (5%), Eurotiales (2%), an unclassified Ascomycota (12%), and an unidentified Eurotiomycetes (4%) comprised the core fungal microbiome. FAPROTAX function prediction suggested that the core microbiome has a substantial potential for the carbon cycle, nitrate metabolism, aromatic compound degradation, chitinolysis, cellulolysis, and xylanolysis, but simultaneously, the core microbiome is also a source of human pathogens. The dynamics of the bacterial community were primarily determined by the total nitrogen in tobacco leaves during the aging process, while those of the fungal microbiome were primarily determined by total organic carbon. This study indicated that the core microbiome activities may play an important role in regulating the loss of carbon organic compounds and enhancing the secondary metabolites during tobacco leaves aging process.

Highlights

  • Flue-cured tobacco is one of the important cash crops in the world, which requires high quality and security

  • The 16S sequencing results were selected for additional analysis of their functional prediction because the abundance of bacterial communities obtained from tobacco leaves was far greater than that of the fungal communities

  • We believed that the nutritional component had an important effect on the changes of microbiome in tobacco leaves during the aging process, in which total nitrogen (TN) and total organic carbon (TOC) played an important role in the change in the microbiome

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Summary

| INTRODUCTION

Flue-cured tobacco is one of the important cash crops in the world, which requires high quality and security. A process called natural aging is often used to improve the quality of flue-cured tobacco. Tobacco aging is a process that changes the physical and chemical characteristics and significantly improves the aroma and flavor of tobacco leaves under certain temperature and humidity conditions. This process typically requires 24 to 30 months (Dixon, Darkis, Wolf, & Hall, 1936), following microbial (or/and enzymatic) actions and other chemical interactions in the leaves (Huang et al, 2010). Microbial communities in nature are usually complex, and this complexity is exacerbated by interactions among the ecosystem members (Butler & O’Dwyer, 2018). We predicted the bacterial functional profiles of the core microbiome in the tobacco leaves using Functional Annotation of Prokaryotic Taxa (FAPROTAX) software

| MATERIALS AND METHODS
Findings
| DISCUSSION
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