Perancangan Primer Spesifik untuk Mendeteksi Dini Jamur Pangan Ektomikoriza Pelawan

Abstract

Pelawan mushroom (Heimioporus sp.) is an edible ectomycorrhiza which has significant economical value. The fruiting bodies of the mushroom are firstly developed in the symbiosis system in the mature host plant. The existence of the mycelia in the rhizoplane of the host plant in seedling stage need to be detected to ensure successful colonization of targeted fungi in mature host plant. Therefore, specific primer is needed for early detection. In this experiment, five sequences of ITS rDNA of pelawan mushroom (JPA, JB, JPC, JPD, and JPE) were successfully obtained which size of 743-745 bp and their bases variation was below 3,3%. The five sequences however, had a significant variation when they were compared with other ITS rDNA sequences of pelawan mushroom collection of the Mycology Division and RCBB-IPB laboratory (JPPK, JPPM, and JPPX). Therefore, the primers were designed into two different groups. The first group was primers designed based on ITS rDNA sequences of fruiting body of JPA-JPE. They were PelAF1-Pel5.8SR1 (±230 bp), Pel5.8SF2-PelAR2 (±320 bp), and PelAF1-PelAR2 (±650 bp). The second group was primers designed based on ITS rDNA sequences of fruiting body of JPPK-JPPX. They were PelBF1-Pel5.8SR1 (±180 bp), Pel5.8SF2-PelBR2 (±260 bp), and PelBF1-PelBR2 (±560 bp). The specificity of the designed primers were tested against fungal DNA of Mortierella sp., Penicillium sp., Hypoxylon sp., Fusarium sp., Glomerella sp., Volvariella sp., and Ganoderma applanatum (IPBCC.10.658) as well as DNA from both groups of pelawan mushroom. The results indicated that primers PelAF1-PelAR2 was specific to the first group and primers PelBF1-PelBR2 was specific to the second group of pelawan mushroom, but the other primers could amplify all or some of the fungal DNA tested.