Xia Xu ^1,2,3 Liang Zhao ^2,3* Yanfei Chen ^2,3 Hancheng Wang ^4* Liuti Cai ^4* Yanyan Wang ^2,3 Nalin N. Wijayawardene ⁵ Weihua Pan ⁶ Feng Wang ^4* Yingqian Kang ^2,3*

• ¹ Guizhou Medical University, Guiyang, China
• ² Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education of Guizhou & Institute of Health Research, Guizhou Medical University, Guiyang, P.R. China., Guiyang, Guizhou Province, China
• ³ Key Laboratory of Medical Microbiology and Parasitology, School of Basic Medical Sciences, Guizhou Medical University, Guiyang, P.R. China., Guiyang, Guizhou Province, China
• ⁴ Guizhou Provincial Academician Workstation of Microbiology and Health, Guizhou Academy of Tobacco Science, Guiyang, P.R. China, Guiyang, Guizhou Province, China
• ⁵ Center for Yunnan Plateau Biological Resources Protection and Utilization, College of Biological Resource and Food Engineering, Qujing Normal University, Qujing, Yunnan 655011, P.R. China, Yunnan, China
• ⁶ Department of Dermatology, Changzheng Hospital, Shanghai, P.R. China, Shanghai, China

Phyllospheric microbial composition of tobacco plants are influenced by multiple factors.Disease severity level is one of the main influencing factors. This study was designed to understand the microbial community in tobacco wildfire disease with different disease severity levels. Tobacco leaves at disease severity level of 1, 5, 7 and 9 (L1, L5, L7, and L9) were collected, both healthy and diseased leaf tissues for each level were collected. The community structure and diversity in tobacco leaves with different disease severity levels were compared using high-throughput technique and BIOLOG ECO. The results showed that in all healthy and diseased tobacco leaves, the most dominant bacterial phylum was Proteobacteria with a high prevalence of genus Pseudomonas, the relative abundance of Pseudomonas was most found at S9 diseased samples. Ascomycota represents the most prominent fungal phylum, with Blastobotrys as the predominant genus. In bacterial communities, the Alpha-diversity of healthy samples was higher than that of diseased samples. In fungal community, the difference in Alpha-diversity between healthy and diseased was not significant. LEfSe analysis showed that the most enriched bacterial biomarker was unclassified_Gammaproteobacteria in diseased samples, unclassified_Alcaligenaceae were the most enrich bacterial biomarker in healthy samples. FUNGuild analysis showed that Saprotroph was the dominated mode in health and lower disease samples, The abundance of Pathotroph-Saprotroph and Pathotroph-Saprotroph-Symbiotroph increases at high disease levels. PICRUSt analysis showed that the predominant pathway was metabolism function, and most bacterial gene sequences seem to be independent of the disease severity level. The BIOLOG ECO results showed that the utilization rates of carbon sources decrease with increasing disease severity level. The current study revealed the microbial community's characteristic of tobacco wildfire disease with different disease severity levels, providing scientific references for the control of tobacco wildfire disease.