The present study was undertaken to find the most suitable organic substrates for the biomass production, viability and efficacy of the biocontrol strain Trichoderma harzianum AS12-2 in the solid-state fermentation system. In total, 13 inexpensive, locally available substrates (agricultural wastes or by-products) were inoculated with the antagonist, and following one month of incubation at room temperature, all colonized substrates were air dried and ground to powder. The shelf life and viability of the Trichoderma strain were assessed as colony-forming units per gram (CFUs g⁻¹) of each substrate on a monthly basis for up to one year at room temperature (25 ± 2 °C) and in the refrigerator (4 °C). In order to find out the effect of the substrate on the bioefficacy of T. harzianum AS12-2, the biocontrol potential of the formulations was evaluated against rice sheath blight disease caused by Rhizoctonia solani. The results showed that the fungus colonized more or less all substrates after one month, although the degree of colonization and conidiation was different among the substrates, being especially high in broom sorghum grain, rice husk, rice straw, rice bran and sugar beet pulp. Analysis of variance (ANOVA) of the population in the substrates in “Month 0” showed that the effect of treatment was significant, and the means were significantly different. The maximum population was recorded for broom sorghum grain and rice straw (6.4 × 10¹⁰ and 5.3 × 10¹⁰ CFUs g⁻¹, respectively). The population declined in all substrates after one year of incubation at room temperature. This decline was relatively smaller in broom sorghum grain, rice straw and rice husk. On the other hand, the population in the same substrate incubated in the refrigerator was decreased in a mild slope, and the final population was high. In addition, the results of greenhouse assay showed that all bioformulations were effective in controlling the disease, and there were no significant differences among the substrates. According to the results of this study, broom sorghum grain, rice husk, rice straw, sugar beet pulp and cow dung could be recommended as suitable fermentation media for the industrial-scale production of T. harzianum strains.

Brown spot caused by Bipolaris oryzae is an important rice disease in Southern coast of Caspian Sea, the major rice growing region in Iran. A total of 45 Trichoderma isolates were obtained from rice paddy fields in Golestan and Mazandaran provinces which belonged to Trichoderma harzianum, T. virens and T. atroviride species. Initially, they were screened against B. oryzae by antagonism tests including dual culture, volatile and nonvolatile metabolites and hyperparasitism. Results showed that Trichoderma isolates can significantly inhibit mycelium growth of pathogen in vitro by producing volatile and nonvolatile metabolites Light microscopic observations showed no evidence of mycoparasitic behaviour of the tested isolates of Trichoderma spp. such as coiling around the B. oryzae. According to in vitro experiments, Trichoderma isolates were selected in order to evaluate their efficacy in controlling brown spot in glasshouse using seed treatment and foliar spray methods. Concerning the glasshouse tests, two strains of T. harzianum significantly controlled the disease and one strain of T. atroviride increased the seedling growth. It is the first time that the biological control of rice brown spot and increase of seedling growth with Trichoderma species have been studied in Iran

In order to identify a specific marker for T. harzianum AS12-2, a strain capable of controlling rice sheath blight caused by Rhizoctonia solani, UP-PCR was performed using five universal primers (UP) both separately and in pairwise combinations. The application of two UP primers resulted in the amplification of unique fragments from the genomic DNA of T. harzianum AS12-2, clearly distinguishing it from other Trichoderma strains. The unique fragments had no significant sequence homology with any other known sequence available in databases. Based on the sequences of the unique fragments, 14 oligonucleotide primers were designed. Two primer sets amplified a fragment of expected size from the DNA of strain T. harzianum AS12-2 but not from any other examined strains belonging to T. harzianum, to other Trichoderma species assayed, or to other common fungi present in paddy fields of Mazandaran province, Iran. In conclusion, SCAR (sequence characterized amplified regions) markers were successfully identified and rapid, reliable tools were provided for the detection of an effective biocontrol Trichoderma strain, which can facilitate studies of its population dynamics and establishment after release into the natural environment.

Tomato Fusarium wilt, caused by Fusarium oxysporum f. sp. lycopersici (Fol), is a destructive disease that threatens the agricultural production of tomatoes. In the present study, the biocontrol potential of strain KR2-7 against Fol was investigated through integrated genome mining and chemical analysis. Strain KR2-7 was identified as B. inaquosorum based on phylogenetic analysis. Through the genome mining of strain KR2-7, we identified nine antifungal and antibacterial compound biosynthetic gene clusters (BGCs) including fengycin, surfactin and Bacillomycin F, bacillaene, macrolactin, sporulation killing factor (skf), subtilosin A, bacilysin, and bacillibactin. The corresponding compounds were confirmed through MALDI-TOF-MS chemical analysis. The gene/gene clusters involved in plant colonization, plant growth promotion, and induced systemic resistance were also identified in the KR2-7 genome, and their related secondary metabolites were detected. In light of these results, the biocontrol potential of strain KR2-7 against tomato Fusarium wilt was identified. This study highlights the potential to use strain KR2-7 as a plant-growth promotion agent.