TOXICITY OF THE AQUEOUS EXTRACTS FROM THE FRUITING BODY OF GANODERMA SP. BY BRINE LETHALITY TEST OF Artemia salina Leach.

The level of toxicity of aqueous extract from Ganoderma sp. fruiting body isolated from different plant media was determined in this experiment using Brine shrimp lethality test (BST) on Artemia salina Leach. The influence of media on the toxicity level of aqueous extract from Ganoderma sp. fruiting body was determined in this experiment. Ganoderma sp. and G. Lucidum were grown on three flamboyant, (Delonix regia Bojer ex Hook Rafin), sengon (Paraserianthes falcataria (L) Nielsen) and coconut(Cocos nucifera Linn.) plant media. The aqueous extracts were prepared from fruiting body using 0.02 M phosphate buffer pH 7.2. The toxicity level of these aqueous extract were then screened using the BST method. The level of the toxicity was determined by LC50. The results indicated that the aqueous extracts of Ganoderma sp. and G. lucidum isolated from flamboyant, sengon and coconut plant media were influenced by plant media. Ganoderma sp. was isolated from flamboyant plant media possesed the highest toxicity (LC50 480 g aqueous extract/ml), then followed by Ganoderma sp. isolated from sengon plant media (LC50 770 g aqueous extract /ml) and coconut (LC50 1040 g aqueous extract /ml). G. lucidum from coconut plant media possesed the highest toxicity (LC50 660 g aqueous extract /ml), followed by G. lucidum were isolated from sengon plant media (LC50 1100 g aqueous extract /ml) and flamboyant (LC50 1970 g aqueous extract /ml). It was suggested that G.lucidum having highest toxicity might produce compounds having antitumor activity.Key words: Ganoderma sp., aqueous extracts, toxicity

Toksisitas Ekstrak Air Tubuh Buah ganoderma sp. terhadap Larva Udang Anemia salina Leach.: Toxicity of the aqueous extracts from the fruiting body of GANODERMA SP. By brine lethality test of Anemia salina Leach.

Penggunaan Ganoderma sp. sebagai obat perlu diuji toksisitasnya dengan metode kematian larva udang Anemia salina Leach. Diharapkan dari penelitian ini dapat diketahui tingkat â¢oksisitasnya dan kemungkinannya untuk dikembangkan sebagai penghasil obat antitumor. Tubuh buah Ganoderma sp. dan Ganoderma lucidum yang diperoleh dad media buatan kayu sengon (Paraserianthes falcataria (L) Nielsen), kelapa (Cocos nucifera Linn.) dan flamboyan (Delonix regia Bojer ex Hook Rafin) diekstraksi dengan dapar fosfat 0,02 M pH 7,2. Ekstrak yang diperoleh diuji toksisitasnya dengan metode kematian larva udang A. salina Leach. pada dosis 15030060012002400 dan 4800 itg ekstrak air/ml. Tingkat ketoksikan diukur dengan harga LC50. Hasil menunjukkan bahwa toksisitas ekstrak air tubuh buah Ganoderma sp. pada BST dipengaruhi oleh media penumbuh. Ganoderma sp. yang ditanam pada serbuk gergaji flamboyan menghasilkan senyawa yang paling toksik dengan harga LC50 480 11g ekstrak air/ml, diikuti dengan Ganoderma sp. yang ditanam pada media serbuk gergaji sengon (LC50 770 jag ekstrak air/m1) dan kelapa (LC50 1040 lig ekstrak air/m1). Ganoderma. lucid= dengan efek toksik paling tinggi adalah G. lucidum yang ditanam pada media serbuk kelipa dengan harga LC50 660 lig ekstrak air/ml, diikuti G. lucidum yang ditanam pada media sengon (LC50 1100 ug ekstrak air/ml) dan flamboyan (LC50 1970 jig ekstrak Ganoderma yang bersifat toksik terhadap BST ini kemungkinan besar berpotensi sebagai senyawa penghasil obat antitumor. Kata kunci: Ganoderma sp., ekstrak air, toksisitas N ABSTRACT The level of toxicity of aqueous extract from Ganoderma sp. fruiting body isolated from different plant media was determined in this experiment using Brine shrimp lethality test (BST) on Anemia salina Leach. The influence of media on the toxicity level of aqueous extract from Ganoderma sp. fruiting body was determined in this experiment. Ganoderma sp. and G. Lucidum were grown on three flamboyant, (Delonix regia Bojer ex Hook Rafin), sengon (Paraserianihes falcataria (L) Nielsen) and coconut(Cocos nucifera Linn.) plant media. The aqueous extracts were prepared from fruiting body using 0.02 M phosphate buffer pH 7.2. The toxicity level of these aqueous extract were then screened using the BST method. The level of the toxicity was determined by LC50. The results indicated that the aqueous extracts of Ganoderma sp. and G. lucidum isolated from flamboyant, sengon and coconut plant media were influenced by plant media. Ganoderma sp. was isolated from flamboyant plant media possesed the highest toxicity (LC50 480 1.tg aqueous extract/mi), then followed by Ganoderma sp. isolated from sengon plant media (LC50 770 1..tg aqueous extract /m1) and coconut (LC50 1040 pg aqueous extract /ml). G. lucidum from coconut plant media possesed the highest toxicity (LC50 660 pg aqueous extract /m1), followed by G. lucidum were isolated from sengon plant media (LC50 1100 pg aqueous extract /ml) and flamboyant (LC50 1970 pg aqueous extract /ml). It was suggested that G.lucidum having highest toxicity might produce compounds having antitumor activity. Key words: Ganoderma sp., aqueous extracts, toxicity

Pengaruh Trichoderma terhadap Perkembangan Mikoriza pada Akar Pinus Merkusii Jungh. et de Vriese = The Effect of Trichoderma to The Mycorrhizae Development of Pinus merkusii Jungh. et de Vriese Root

The objective of experiment was to determine the effect of introduction Trichoderma on development of mycorrhizae of seedling Pinus merkusii Jungh. et de Vriese. T. Koningii (T,), T. reesei (T]3), and T. harzianum (T27), were introduced into seedling of wood with different time of introduction. The different time was before, together and after inoculation of soil, which had mycorrhizae, or without mycorrhizae. The result of research showed that percentage of infection mycorrhizae, were created in variation according to the kind of Trichoderma and time of introduction. An introduction before inoculation mycorrhizae caused total infection 52,7% in the seedling of T,, 54,32% in T13, and 56,59% in T27. It was smaller than infection value that was introduced together with mycorrhizae (64,36% in T,, 71,03% in T]3, 57,35% on T^,). Introduction after inoculation of mycorrhizae caused infection on T, (67,17%), Tjj (72,88%) and T27 (63,92). Keywords: trichoderma -mycorrhizae -Pinus merkusi

Respon Jenis-jenis Akasia dan Eukaliptus Terhadap Patogen Akar Ganoderma

Many forest plantations in Indonesia were built on converted tropical forest. Minimal site preparation was often applied before planting, and the result was that various forest debris, particullarly dead stumps, were still remained in young plantation site. Since some root pathogens use the debris to survive in the absence of host, this might be potential as disease inoculum to the plantation establised. The experiment was aimed to evaluate the response of Acacia and Eucalyptus to root-rot pathogen. infection. Field study was made in some forest plantations to observe the root disease status. The root-rot pathogens were isolated and their pathogenicity were tested using Crotalaria juncea L. as indicator plant. In laboratory experiment, pieces of root of three species ofAcacia and four species of Eucalyptus were selected for root pathogen colonization test. Selected Ganoderma sp. isolated from A. mangium was inoculated to the root samples using modified soil block test. The attack ofGanodernia, on the root sample was evaluated based on fungal colonization on the root samples. The results indicated that root-rot disease was found to attack A. mangium and A. auriculifirmis, and 17 different isolates of Ganodernia were known associated with the disease. Laboratory experiment showed that theroot of A. mangium was totally colonized by Ganoderma on day 5 after inoculation, whereas root of A. auriculifirmis was colonized by the pathogen on day 8 after inoculation. On day 37 only 53.28% of the surface area of A. oraria\u27s root sample was colonized by the pathogen. E. pelita, E. alba, E. camaldulensis and E. urophylla were entirely colonized by Ganoderma on day 7, 8, 12 and 37 respectively, although for the last species the colonization was only 90% of total sample surface area. Key words: Ganoderma, root-rot disease, Eucalyptus, Acaci

Isolasi dan Karakterisasi B-1,3-glukanase Akar Semai Tusam (pinus merkusii jungh. Et de vriese) yang Berasosiasi dengan Fungi Ektomikorisa: Isolation and characterization -1,3-glucanase from tusam seedling roots (pinus

ABSTRACT Association between tusam (Pinus merkusii Jungh. et de Vriese) seedling roots with ectomycorrhiza fungi was expected to increase the activity f3-1,3-glucanase level in the plants. This enzyme has potency to protect seedling from soil borne fungal pathogens by degrading the fungal cell walls. The objectives of this research were to isolate and characterize 0-1,3-glucanase from tusam seedling roots associated with ectomycorrhiza fungi. Crude protein was isolated with ammonium sulfat precipitation and then purified by gel filtration chromatography and characterize molecular weight, temperature and pH for optimum the activity.- The enzym frdm tusam seedling roots associated with ectomycorrhizal fungi, designated as GLUC15 have 15 kD molecular weight, 30° â 40° C temperature and pH 5 â7 for optimum activity. Key words: f3-1,3-glucanase, tusam, fungi, ectomycorrhiza INTISARI Asosiasi antara akar tusam dengan fungi pembentuk ektomikorisa diduga mengimbas adanya enzim13-1,3-glukanase. Enzim ini memberikan kontribusi terhadap ketahanan semai tusam dalam menghadapi fungi patogen melalui aktivitas enzim tersebut dalam mendegradasi dinding sel fungi yang mengandung glukan. Penelitian ini bertujuan untuk melakukan isolasi dan karakterisasi 0-1,3-glukanase akar semai tusam yang berasosiasi dengan fungi pembentuk ektomikorisa. Crude protein diisolasi melalui tahapan pengendapan ammonium sulfat 70% dan kemudian dipisahkan menggunakan kromatografi gel filtrasi. 13 - 1 ,3-glukanase yang diperoleh dikarakterisasi berat molekul, pH, dan suhu optimal aktivitas enzim. Enzim 13-1,3-glukanase dari akar semai tusam yang berasosiasi dengan fungi pembentuk ektomikorisa, yang kemudian disingkat GLUC15 merniliki berat molekul 15 kD, kurang tahan pemanasan dengan suhu optimal aktivitas 30° -40° C, bekerja pada kondisi asam dengan aktivitas pH optimal 5 -7

Pengimbasan Ketahanan Pisang Terhadap Penyakit Layu Bakteri (Raistonia solanacearum) Dengan Pseudomonas Cepacia =Induced Resistance of Banana to Bacterial Wilt (Ralstonia solanacearum) using Fseudomonas cepacia

Abstract Bacterial wilt caused by Ralstonia solanacearum (synonym Pseudomonas solanacearum E.E Smith.), have been an important disease of banana plants. The objective of this study was to know the ability of P. cepacia in inducing resistance and salicylic acid content on phytoalexin extract of banana plants. The result showed that resistance of Ambon Kuning cultivar could be induced by viable and extraselulair protein of P. cepacia. The induced banana plants had lower disease intensity than uninduced plants. Salicylic acid content in phytoalexin extract from banana was induced by extraselulair protein of P. cepacia was 41,41 ppb. Keywords: induce resistance - salicylic acid

THE EFFECT OF ANTIFUNGAL ACTIVITY OF β-1,3-GLUCANASE Trichoderma reesei AGAINST ROOT FUNGI OF Ganoderma philippii

Purifi ed three isoform β-1,3-glucanases of 90 kDa β-1,3-glucanase-I, 75 kDa β-1,3-glucanase-II, and 64 kDa β-1,3-glucanase-III derived from Trichoderma reesei isolate T13were tested in vitro for its antifungal activity to the root fungi of Ganoderma philippii. Antifungal activity was tested by bioassay using a modifi ed cup plate method and microscopic observation to determine the presence of abnormal mycelia growth of G. philippii. The result showed that antifungal activity of those three isoform β-1 ,3-glucanase (β-1,3-glucanase-I, β-1,3 glucanase-II, and β-1,3-glucanase-III) in vitro could not form growth inhibition against the mycelia of G. philippii. Microscopic observation revealed that the enzyme caused necrotic lesion, whereas the combination of β-1,3-glucanase and chitinase from T. harzianum (Sigma) caused lyzed hyphae of G. philippii

Pengendalian Hayati Penyakit Akar Merah Pada Akasia Dengan Trichoderma = Biological Control Of Red Root-Rot Disease Of Acacia Using Trichoderma

The experiment was aimed to measure the distribution and intensity of root rot disease of Acacia spp. in the urban forest at the campus of Gadjah Mada University, to isolate and identify the causal organism and to select Trichoderma sp. as biological agent for controlling, the disease. The pathogenicity of the causal organisme was tested using Crotalaria juncea L. and trunk of A. mangium Willd. (10 cm diameter, 8 cm length) as indicator plant. The ability of Trichoderma sp. as antagonist was tested in vitro. It was concluded that the pathogen of the root rot disease was Ganoderma philippii. The pathogen attacked four species of Acacia spp. in the location namely A. auriculiformis, A. mangium, A. oraria and A. crassicarpa. Of the total individual trees of those species in the campus, as much as 38.6%, 22.2%, 28.9% and 66.7% were attacked by the root rot pathogen respectively. Of the 20 Trichoderma spp.isolates capable to inhibit the pathogen in vitro, three isolates were found as promising agents for biological control of the pathogen. The promising isolates were T. reseilT13, T. koningii/T/ and T. koningii/T/6 with inhibition effectivity of 94.58%93.66%and 91.76% respectively. Key word :biological control, red root-rot disease, Trichoderma, acaci

Isolasi dan Karakterisasi Kitinase Akar Tusam (Pinus merkusii Jungh. et de Vriese) yang Bersimbiosis dengan Fungi Ektomikorisa: Isolation and characterization chitinase in tusam (Pinus merkusii Jungh. et de Vriese) roots

ABSTRACT The experiment was aimed to detect the activity and characterize the chitinase of tusam root during ectomycorrhizal symbiosis. Tusam inoculated with tusam stands soil from Kaliurang as fungi inocula. Crude proteins were isolated from 4, 6, and 8 weeks age of Chitinase activity and isoform were detected using glycol chitin as a subtrat. The .enzyme was purified by ammonium sulfa( precipitation, dialysis, followed by gel filtration chromatography. The results showed that tusam produced chitinase with a molecular weight of approximately 52 kDa. The optimum activity was at pH 5 and temperature of 30°C. Key words: isolation, characterization, chitinase, ectomycorrhizal, tusam INTISARI Penelitian ini bertujuan untuk mengetahui akti vitas dan karakterisasi kitinase yang terjadi waktu akar tusam bersimbiosis dengan fungi ektomikorisa. Semai tusam diinokulasi dengan fungi ektomikorisa, yang berasal dari tanah dibawah tegakan di Kaliurang. Protein kasar diisolasi dari semai tusam pada umur 4, 6, dan 8 minggu. Aktivitas kitinase dan isoform dideteksi menggunakan substrat glikol kitin. Pemisahan protein diawali pengendapan dengan ammonium sulfat, dialisis, dilanjutkan dengan kromatografi gel filtrasi. Hasil penelitian menunjukkan bahwa tusam memproduksi kitinase yang mempunyai berat molekul 52 kDa, suhu 30°C, dan pH optimum 5. Kata kunci: isolasi, karakterisasi, kitinase, ektomikorisa, tusa

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