Fig. S1 Nucleotide alignment of the ITS region in the genus Coniothyrium . a ITS sequences of C. sporulosum , C. fuckelii , and C. minitans were aligned with Clustal W. Identical nucleotide regions are highlighted in black. Arrows indicate C. minitansspecific primers.
The ITS region of three Coniothyrium species was sequenced; and nucleotide sequence size was 596 bp in C. minitans (identical to GenBank AJ293809), 598 bp in C. sporulosum
(similar [1 bp deletion] to GenBank GU566257), and 599 bp in C. fuckelii (similar [1 bp deletion] to GenBank JX496038). b C. minitansspecific primers exclusively amplified C. minitans genomic DNA. Cm; C. minitans , Cs; C. sporulosum , Cf; C. fuckelii , K; KOD FX
Neo DNA polymerase (Toyobo), Q; Quick Taq DNA polymerase (Toyobo), M 100-bp ladder.
Fig. S2 Field map of disease protection test against sclerotinia disease in lettuce in Hyogo
Prefecture. Each of three ridges was 14 m long with 140 cm inter-row spacing, with 90 lettuce seedlings (Santos No. 2) per ridge (2 plants 30 cm apart per row, 45 rows total). The ridges received no treatment, 100 L of 125-fold-diluted Minitan WG, or 200 L of 250-fold diluted Minitan WG.
Fig. S3 Effect of DNA extraction buffer on the amplification curve in qPCR. Amplification curve of soil samples from Gunma Prefecture. eDNAs were extracted with Lysis solution BB
( a ) or BB SP1 ( b ). Blue 2.0 mg/0.5 g soil, red 0.2 mg/0.5 g soil, purple 0.02 mg/0.5 g soil, green negative control.
Fig. S4 Quantification of BCA biomass in field of Gunma Prefecture. BCA biomass after each treatment was estimated from the standard curve in Fig. 1c. Field soils were sampled in
July and September. eDNA extraction was done in triplicate; standard deviations are presented.