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The maternally heritable endosymbiont provides many ecosystem functions.Antibiotic elimination of a specific symbiont and establishment of experimental host lines lacking certain symbionts enable the roles of a given symbiont to be explored.The whitefly Bemisia tabaci (Gennadius) in China harbors obligate symbiont Portiera infecting each individual,as well as facultative symbionts,such as Hamiltonella,Rickettsia and Cardinium,with co-infections occurring relatively frequently.So far no studies have evaluated the selectivity and efficacy of a specific symbiont elimination using antibiotics in whiteflies co-infected with different symbionts.Furthermore,no success has been achieved in establishing certain symbiont-free B.tabaci lines.In this study,we treated Hamiltonella-infected B.tabaci line,Hamiltonella-Rickettsia-co-infected line and Hamiltonella-Cardinium coinfected line by feeding B.tabaci adults with cotton plants cultured in water containing rifampicin,ampicillin or a mixture of them,aiming to selectively curing symbiont infections and establishing stable symbiont-free lines.We found ampiciilin selectively eliminated Cardinium without affecting Portiera,Hamiltonella and Rickettsia,although they coexisted in the same host body.Meanwhile,all of the symbionts considered in our study can be removed by rifampicin.The reduction of facultative symbionts occurred at a much quicker pace than obligate symbiont Portiera during rifampicin treatment.Also,we measured the stability of symbiont elimination in whitefly successive generations and established Rickettsia-infected and Cardinium-infected lines which are absent in natural populations.Our results provide new protocols for selective elimination of symbionts co-existing in a host and establishment of different symbiont-infected host lines.