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At each sampling time, six tobacco leaves with similar appearances were randomly selected to remove the main vein and branch vein and divided into three parts: one part was used to determine the moisture content of the tobacco leaves, another part was wrapped in tin foil and stored at 80C for analyses of the tobacco leaf microbiota, and the final part was dried and the water-soluble sugar, starch, total nitrogen, nicotine, and protein contents of the tobacco leaves were measured after removal of the main vein