Figure 4
Optimization and validation of LAMP assay. Yellow colour indicated a positive reaction while red/pink colour indicated no reaction. (A) LAMP assay optimized with pure fungal isolates with no template control. M: 100 bp (Promega); 1: R. solani AG-1 IA isolate PURS1; 2: R. solani AG-1 IA isolate PURS2 3: R. solani AG-1 IA isolate PURS3; 4: No Template control; L: 1 kb (Generuler). [upper panel: gel photograph; lower panel: colorimetric reactions in PCR tubes] (B) Specificity assay of the LAMP assay. L: 1 kb (Generuler); 1: Colletotrichum capsici isolate CABI-063597; 2: Sclerotium rolfsii; 3: Trichoderma asperellum; 4: Fusarium oxysporum; 5: Alternaria alternata; 6: Ustilaginoidea virens; 7: Curvularia prasadii; 8: Cochiliobolus tuberculatus; 9: Pseudomonas plecoglossicida 10: R. solani AG-1 IA isolate PURS1; 11: R. solani AG-1 IA isolate PURS2; 12: No Template control; M: 100 bp (Promega). [upper panel: gel photograph; lower panel: colorimetric reactions in PCR tubes] (C) M: 100 bp (Promega); 1: R. solani AG-1 IA isolate PURS1; 2: R. solani AG-1 IB 3: R. solani AG 2–2 IIIB; 4: R. solani AG-8; 5: R. solani AG-7; 6: R. oryzae-sativae 7: R. solani AG-3; 8: R. solani AG-3; 9: Sarocladium oryzae; 10: Curvularia oryzae; 11: Curvularia lunata; 12: Magnaporthe oryzae; 13: No template control; 14: 1 kb ladder (Promega) [upper panel: colorimetric reactions in PCR tubes; lower panel: gel photograph] (D) M: 100 bp (Promega); 1: R. solani AG-1 IA isolate PURS1; 2: R. solani AG-1 IA isolate PU RS14; 3: Fusarium fujikuroi isolate RPF19; 4: No template control [upper panel: colorimetric reactions in PCR tubes; lower panel: gel photograph].
