Abstract:The tobacco β-1, 3-Glucanase gene, the alfalfa defensin gene alfAFP and their bivalent gene GLU-AFP were introduced into tomato line Micro-Tom via Agrobacterium-mediated method. The aim is to gain transgenic lines resistant to Botrytis cinerea, and test the synergistical effects of bivalent gene. Transgenic lines carrying each of the transgenes were confirmed for integration into the tomato genome using GFP, PCR and Southern blot hybridization. One to four copies of the transgene were integrated into the tomato genome. Performance test of resistance analyses to B.cinerea with T1 generation transgenic tomato lines showed the transgenic lines exhibited higher resistance to the pathogens infected than that of the non-transgenic plants, and resistance levels were related to expression levels of the transgene, showing dosage-effect. The transgenic tomato harboring GLU-AFP cassette showed the highest disease resistance. It suggested that the alfAFP and Glucanase gene have some synergistical effects on the resistance to B.cinerea. Performance test of resistance of T2 families with GLU-AFP gene showed that M3, M8 and M20 had a resistance index of 19 to 22, which showed a high resistance level. Some independent lines with high disease resistance, low variability and stable expression of transgenes could be selected for the further studies and molecular breeding.