Analisis Biomolekuler dan Sifat Patogenesitas Ganoderma Terhadap Penyakit Busuk Pangkal Batang pada Tanaman Kelapa Sawit (Elaeis Guinensis)

Abstract

Biomolekuler analysis and photogenesity test of Ganoderma, causal agent of basal stem rot (BSR) disease on oil palm (Elaeis Guinensis) have been carried out. The identification and characterization were preceded by extraction of purified mycelium DNA of 20 Ganoderma of arecanut (Areca catechu), 17 Ganoderma of coconut (Cocos nucifera), 4 Ganoderma of sugar palm (Arenga pinnata), 2 Ganoderma of sago palm (Metroxylon rumphii), 3 Ganoderma of rubber (Havea brasiliensis), 4 Ganoderma of acacia (Acacia mangium) and 3 Ganoderma of forest wood (Un known), that were suspected to be the source of transmission of basal stem rot. The purity of the extracted DNA was confirmed by an electrophoresis using the extracted DNA of 12 purified mycelium of Ganoderma phatogen on oil palm and the DNA marker (Gibco BRL Life Technologies Inc. Gaitherburg USA) AS a control and all produced a single band in accordance with the control. The extracted DNA was then amplified using a primer pair Internal Transcribed Spacer (ITS), ITS 1 - ITS 4 by Polymerase Chain Reaction (PCR) method and continued with the digestion of DNA result of amplification uses restriction enzyme Mlul specifically digest ITS DNA Ganoderma phatogen on oil palm. From the electrophoresis analysis, it has been found that : 1. 8 of 20 DNA Ganoderma of areacanut were digested by enzyme Miu l (40% test were+). 2. 4 of 17 DNA Ganoderma of coconut were digested by enzyme Mlul (50% test were+). 3. 2 of 4 DNA Ganoderma of sugar palm digested by enzyme Miu l (50% test were+). 4. 2 of2 DNA Ganoderma of sago palm were digested by enzyme Mlul ( 100% test were + ). 5. 11 of 12 DNA Ganoderma phatogen oil palm were digested by enzyme Miu 1 (91 % test were + ). 6. 3 of 3 DNA Ganoderma of rubber not digested by enzyme Miu 1 (100% test were -). 7. 4 of 4 DNA Ganoderma acacia plant were not digested by enzyme Mlul (100% test were-). 8. 3 of 3 DNA Ganoderma of forest wood not digested by enzyme Miu I (100% test were-). According to the literature, it was known all Ganoderma DNA digested by enzyme Miu 1 belong to Ganoderma phatogen on oil palm whereas all Ganoderma DNA not digested by enzyme Miu 1 belong to the Ganoderma non phatogen on oil palm. All the extracted DNA after amplified using primer pair ITS 1 - ITS 4 were sequeneed in Eijkman Institute Laboratory to determine the order of nucleotides direved in electrophenogram. The reading of electrophenogram showed that 5 of the total sequenced 22 sample were readable (23%) they are : 2 ITS l, 3 ITS 1, 3 ITS 4; 7 ITS 1 and 9 ITS 1 whereas the others were unreadable due to various N emerging so that they can not used in the classifying of Ganoderma samples. The use of the interpretation nucleotide DNA result of electrophenogram for each Ganoderma classification samples must be compared to DNA mark nucleotide in gen Bank reserves with acces number : http//www.ncbi.nlm.nih.gov/entrez/viwer:109390937 atau : NCBI : AJ.586520 Ganoderma sp kk ... [gi:109390937]. The phatogenicity test carried out by contact method between, the tested oil palm and the source of mycelium in greenhouse showed that all mycelium with the DNA digested by enzyme Mlul were phatogen on oil palm tested (agrees with the literature) unless of all purified mycelium of Ganoderma with DNA not digested by enzyme Miu 1 that were not phatogen against oil palm ( agrees with the literature) except the Ganoderma originated from forest wood which was phatogen.