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To clarify microorganisms resulted mildew on Hainan cigar tobacco leaves and screen their antagonistic strains to prevent tobacco leaves from mildew, mildew and antagonistic microorganisms were isolated from the surface of Hainan moldy cigar tobacco and purified. Morphological and molecular methods were combined to identify microbial species. The identification and screening results showed that the main microorganisms resulted mildew on Hainan cigar tobacco were Talaromyces funiculosus, Scopulariopsis brevicaulis and Aspergillus occultus. Two antagonistic strains with good antagonistic effects were screened out, namely Bacillus subtilis and Bacillus halotolerans. The main microorganisms resulted mildew on Hainan cigar tobacco were mainly Talaromycetes, Scopularia and Aspergillus strains. The screened two strains of Bacillus had good antagonism to mold, which provided a reference for the prevention and control of mildew on cigar tobacco. [ABSTRACT FROM AUTHOR]

In order to quantitatively detect Honghuadajinyuan in mixed tobacco leaves, cultivar-specific primers and TaqMan MGB probes were designed. The specificity and sensitivity of the primers and the probes were detected, and the reaction conditions of the fluorescent quantitative PCR were determined. On this basis, a regression equation was established to quantitatively detect the single cultivar of Honghuadajinyuan in mixed tobacco leaves. The results showed that the combination of the primers and the probes H1-R/F/T had satisfactory specificity. The regression equation was established and the determination coefficient R² was 0.997. When the template DNA (55 ng·µL-1) was 2 µL, the detection limit of Honghuadajinyuan was 0.11 ng. The error range of the content of Honghuadajinyuan in mixed tobacco leaves detected by this method was between 2.5% and 3.5%, suggesting that this method could be applied to the quantitative detection of Honghuadajinyuan in specific mixed tobacco leaves. [ABSTRACT FROM AUTHOR]