*2.6. Analysis of E*ff*ects of Applying Di*ff*erent Doses of TA and TS to Tomatoes: Experiment 2*

The experimental procedure followed for experiment 2 was similar to that described for experiment 1, although conducted in winter. Again, propagated in substrate appropriately irrigated according to climate and crop necessity under commercial plant nursery conditions and supplemented with a commercial complex nutrient fertiliser, 96 tomato seedlings per replicate of four were treated with three solutions of spore suspension, each with 5 mL of TA, TS conidia and TA + TS (M) (TA D1, TS D1 and M D1: 105 spores mL<sup>−</sup>1; TA D2, TS D2 and M D2: 106 spores mL<sup>−</sup>1; and TA D3, TS D3 and M D3: 10<sup>7</sup> spores mL<sup>−</sup>1). After 30 days of sowing, twenty plants from each of the three treatment batches and control were randomly selected for harvest. The plants were measured, and data were recorded for the same parameters described in experiment 1. In mid-February a further 25 plants were transplanted into a sandy soil and analysed in mid-May.

In all tests, roots inoculated with *Trichoderma* isolates were collected at the end of the tests. Roots were surface sterilized in 0.1% sodium hypochlorite and washed with sterilised water. Root fragments were placed in PDA medium to determine root colonisation by the fungal isolate.

#### *2.7. Statistical Analysis*

The experimental results are presented as the means and standard error (± SE) for the different replicates. Mean separation was carried out using Fisher's Least Significant Difference (LSD) test. The data were tested by one-way analysis of variance (ANOVA) or Student's *t*-test with significance defined as *p*-values less than 0.05 (*p* < 0.05). Statgraphics Centurion 18 Software was utilised for statistical analysis.

#### **3. Results**

#### *3.1. Mass Production of Trichoderma Isolates on Solid Substrates*

The results are outlined in Table 1. Both isolates grew and sporulated well in all mixtures tested. The proportion of 70 + 30% for buckwheat husk and oats (Figure 1), respectively, and 80 + 20% for buckwheat husk and rice, resulted in significantly higher spore production for both species, followed by

90 + 10% and 70 + 30% of BH + 10% R (Table 1). The lowest spore production rate was observed for 80 BH + 20% O.


**Table 1.** Mass production of spores on solid substrates (CFU g<sup>−</sup>1).

BH: buckwheat husk; O: oat; R: rice; CFU: colony forming unit. Data were analysed by ANOVA and treatment means were compared according to Fisher's Least Significant Difference (LSD) statistical procedure (*F*-test at *p* < 0.05). Different letters indicate significant differences according to the one-way ANOVA test (*p* = 0.05).

**Figure 1.** Mass production of (**A**) *Trichoderma aggressivum* f*. europaeum* and (**B**) *Trichoderma saturnisporum* on 70 + 30% for buckwheat husk and oats.
