Identifikasi Ganoderma spp dan Trichoderma spp menggunakan marka molekuler ITS-RFLP

Abstract

Genetic informations of Ganoderma spp and Trichoderma spp are needed because Ganoderma spp is one of important fungi patogen in estate crops, whereas Trichoderma spp is fungi that can control Ganoderma spp. Research has been conducted to identify the fungi morphologically and physiologically. Due to the low level of accuracy, the use of ITS-RFLP method is expected to identify the species of Trichoderma spp and Ganoderma spp more accurate and specific. The method used are preparation of pure isolates, genomic DNA isolation, amplification by PCR using primers ITS1_ITS4 and ITS5_ITS4, digesting with restriction enzymes AluI, MspI, MluI, and MnlI, and sequencing amplicons. The size of genomic DNA isolated from six samples were about 3000 bp. Trichoderma spp amplicons size were 680 bp, whereas Ganoderma spp amplicons were ranged from 600 to 700 bp. Restriction enzymes that produce the most polymorphic bands pattern are MspI. Samples with different patterns and size of bands indicated a different species, whereas the similar pattern and size of bands shows close relations.