Abstract: To improve the secretion mass fraction of tobacco glandular trichomes, a homozygous mutant plant TTB-1 (tobacco trichome bioreactor-1) was obtained by using gene editing technology to remove NtCycB2, a negative regulatory gene of glandular trichome development, in T.I.1068 germplasm. The results showed that TTB-1 plants grew vigorously and developed normally. Unlike the flue-cured tobacco cv. K326, the surface fur of TTB-1 and T.I.1068 was composed of glandular trichomes without protection. The glandular trichome density of TTB-1 was 53.41% higher than that of T.I.1068 and 7 times higher than that of K326. Compared to K326, the TTB-1 glandular head was fuller and Rhodamine staining was darker. The test results of the chemical components on the leaf surface of the middle leaves of tobacco plants at the bud stage indicated that the components in TTB-1 and T.I.1068 secretions were consistent, but the content of secretions was significantly higher in TTB-1, and its total secretion amount was 63.11% higher than that of T.I.1068. The secretions of TTB-1 differed from those of K326. In addition to cembranoid diterpene and type Ⅰ-Ⅱ sucrose esters, TTB-1 also contained labdane diterpene and type Ⅲ-Ⅴ sucrose esters, and its total secretion amount was 7.95 times of that of K326. The mature upper, middle and lower leaves were harvested in batches and the chemical components on the leaf surfaces were extracted. The concrete yield was 0.61%, 0.43%, and 0.30% of the fresh upper, middle, and lower leaf weights, respectively, and the average concrete yield per plant was 8.90 g. The concrete yield was 0.45% of the fresh leaf weight, and the average concrete yield per plant was 8.06 g when harvesting by one-priming at flowering stage.