【Objective】Tobacco bacterial wilt is one of the most serious soil-borne pests affecting tobacco production in Southwest China. Biocontrol of tobacco bacterial wilt has been a hot research topic in recent years. For control of the pest, a kind of biomanure prepared out of microbes antagonistic to the pest is used together with some proper agronomic measures, so as to achieve sustainable and healthy development of tobacco production.【Method】In this study, a three-year long field experiment was conducted in a severely wilt infected tobacco field in Guizhou Province to explore effects of the pest control measures. The experiment was designed to have four treatments or measures, that is, T1 (Conventional fertilization), T2 (Conventional fertilization liming), T3 (Conventional fertilization L-25 Biomanure) and T4 (Conventional fertilization liming L-25 Biomanure). High-throughput sequencing was done of the microbial genome in the rhizosphere to explore effects of the integrated control measures on soil microflora.【Result】The experiment indicated that in 2013 Treatment T4 was significantly (p<0.05) higher than Treatments T2 and T3 in controlling effect during the entire tobacco growing period, and in 2014, Treatment T4 was significantly higher than Treatments T2 and T3 in controlling effect,60 days after transplanting and 6.26 and 1.99 times as high as Treatments T2 and T3, respectively, 90 days after transplanting. At the end of the experiment, the pest controlling effect reached up to 61.30%. In 2013, Treatment T4 was significantly higher than all the other treatments in tobacco yield or 3.71, 1.61 and 1.13 times as high as Treatment T1, T2 and T3, respectively. In 2014, Treatment 4 was the highest in tobacco yield and output value and Treatment T3 was significantly higher than the other two, which were more or less the same. In 2013, although the pathogen in the treatments amended with biomanure slowly increased in population, the counts of antagonistic bacteria in the treatments were higher than those of pathogen during the initial 70 days after transplanting, while in 2014, the population of pathogen in Treatment T4 was always kept below the level of 10⁶ cfu g^-1 soil, but the counts of pathogen in all the other treatments grew beyond the level of 10⁷ cfu g^-1 soil, 90 days after transplanting and on. Treatment T4 was higher than Treatment T1 in microbial species abundance, operational taxonomic unit (OTU) and Shannon index. Principal component and cluster analysis shows that Treatments T3 and T4 were quite similar, but significantly different from Treatment T1 in microfloral structure. 【Conclusion】The integrated measure, (like Treatment T4) can effectively control tobacco bacterial wilt and significantly improve yield and output value of cured-tobacco, and soil microflora as well, which demonstrates that the measure has a promising application and extension prospect.