Research on separation optimization of endophytic bacteria in tobacco
The best surface sterilization method and isolation medium of endophytic bacteria in tobacco were determined by grinding separation and surface sterilization efficacy check. Results showed that isolation and disinfection effect of soaking by 1%NaClO (5 min)+75%alcohol (2 min) was better in that surface epiphytic bacteria can be 100%eliminated and more bacteria can be separated. Four isolating media including Nutrient Agar (NA), Tryptic Soy Agar (TSA), Potato Dextrose Agar (PDA), Luria-Bertani medium (LB) were utlized to isolate endophytic bacteria from tobacco, aiming at comparing difference of isolation effect between them. Results showed that the isolation effect of TSA and NA was better than that of other media, while LB and PDA showed the lowest isolation effect. The amount of bacteria isolated from root, stem, leaf by TSA medium were 5.18 log cfu/g.fw, 2.17 log cfu/g.fw, 4.69 log cfu/g.fw respectively. The amount of bacteria isolated from root, stem, leaf by NA medium are 5.01 log cfu/g.fw, 1.81 log cfu/g.fw, 4.93 log cfu/g.fw respectively. The 16S rDNA of 127 endophytic strains were sequenced. Nine, seven and 13 different kinds of bacteria were isolated from root, stem, leaf by TSA medium respectively while only 8, 6 and 10 different kinds of bacteria isolated from root, stem, leaf by NA medium. These results indicated that TSA and NA medium were suitable for isolating endophytic bacteria in tobacco by which ideal separating effect can be obtained.
国家科技期刊平台
NSTL
万方数据
维普
