The nematicidal potential of three native biological agents containing the fungus Trichoderma harzianum strain YZL229, the bacteria Pseudomonas fluorescens strain P-72-10 and Bacillus subtilis strain Itb162 which collected from Meloidogyne spp. infested tobacco fields and infected roots in Chongqing, against the root-knot nematode, Meloidogyne incognita, infecting tobacco, was assessed in vitro and under greenhouse. All treatments displayed nematicidal potentials through ovicidal and larvicidal assays in vitro and resulted in significant improvement in tobacco growth in greenhouse. The significant suppression of gall index (GI) and eggmass index (EMI), the reduction of nematode population in soil and eggs per eggmass were obviously observed following the application of these biocontrol agents, especially Trichoderma harzianum strain YZL229. Generally, the results of this study indicated that the tested native biological agents could prove to play a significant role in integrated root-knot nematode management on tobaccos.