Background: Grey mould caused by Botrytis cinerea Pers. (teleomorph Botryotinia fuckeliana (de Bary) Whetzel) is one of the most destructive fungal diseases of Mediterranean crops. In Algeria, few studies have been made on the economic impact of this disease. Nevertheless, it is practically present in all tomato and strawberry greenhouses, as well as in prospected vineyards in the north and south of the country. The complexity of chemical control of this disease has led to search for Trichoderma strains that are effective in biological control. Results: Fifteen isolates of Trichoderma spp. were obtained from vigorous and healthy plants (tomatoes, strawberries, and vines) rhizosphere, and from a commercial bio-compost (Bio-composte®), then identified as T. afroharzianum (four isolates), T. gamsii (four isolates), T. longibrachiatum (three isolates), T. atroviride (one isolate), T. brevicompactum (one isolate), T. breve (one isolate), and T. lixii (one isolate) on the basis of DNA sequence analysis of four genes (ITS, tef1, rpb2, and acl1). In vitro biocontrol tests revealed that four Algerian isolates of Trichoderma spp. (TAtC11, TGS7, TGS10, and TBS1) had a high antagonistic activity against B. cinerea, the mycelial growth has been reduced by 62 to 65% in dual-culture technique, by 62.31 to 64.49% in volatile compounds test, and a high inhibition of germling growth was recorded by TBS1 isolate with 90.68% in Culture filtrates test. Biocontrol tests carried out on tomato plants with T. brevicompactum (TBS1), T. atroviride (TAtC11), and T. lixii (TLiC8) against B. cinerea (BCT04) showed that TBS1 inoculation significantly reduced the incidence of disease by 64.43 and 51.35% in preventive and curative treatment, respectively. Conclusion: The present study revealed the first report of T. brevicompactum, T. breve, and T. lixii in Algeria, and it also contributes to the promotion of the use of native strains of Trichoderma in biological control leading to a better preservation of soil microbial diversity.