| dc.description.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. | en_US |
