Four Ganoderma species (G. tsugae, G. subamboinense var. laevisporium, A TCC 52419, G. tropicum

and G. lucidum) were incubated in liquid medium containing different concentrations of germanium (Ge) for up to

20 days at 28 C. Increasing the Ge concentration of the medium resulted in a gradual decrease in the growth of the

fungal mycelium. However, the Ge content in the mycelium increased with increasing Ge concentration. Different

species recorded different levels of tolerance towards the Ge. In each case, the optimum concentration of the

incorporated Ge in the medium was established as 100 mg/l for both optimal uptake of Ge by the fungal mycelium

and optimal mycelial growth

Chow, Chin Tong

Pei, Joo Chong

Universiti Putra Malaysia Institutional Repository

Pertanika J. Trop. Agric. Sci. 19(2/3): 171-174 (1996)ISSN: 0126-6128© Penerbit Universiti Pertanian MalaysiaMycelial Growth and GerlllaniulIl Uptake by Four Species ofGanoderrnaCHOW-CHIN TONG and PEI-JOO CHONGCenter for Research in En.<;Yme and Microbial Technology (CREAM)Faculty of Science and Environmental StudiesUniversiti Pertanian Malaysia43400 UPM Serdang, Selangor, MalaysiaKeywords: Ganoderma tsugae, G. subamboinense var. laevisporium, G. tropicum, G. lucidum,germaniwnABSTRAKMiselia dari empat spesis Ganoderma (G. tsugae, G. subamboinense var. laevisporium ATCC 52419, G.tropicum dan G. lucidum) telah ditumbuhkan dalam medium cecair yang mempunyai kepekatan germanium(Ge) yang berlainan selama 20 hari pada suhu 2frC. Didapati bahawa kadar pertumbuhan dan berat keringmiselia keempat-empat spesis semakin menurun dengan penambahan kepekatan Ge dalam medium walaupunperatusan pengambilan Ge oleh miselia semakin bertambah. Tahap pertahanan miselia terhadap Ge di dalammedium pertumbuhan berbeza untuk keempat-empat spesis Ganoderma. Untuk setiap spesis, kepekatan optima Gedi dalam medium telah ditentukan sebagai 100mgtl untuk mendapatkan keadaan optima bagi pengambilan Ge sertapertumbuhan miselia yang optimum.ABSTRACTFour Ganoderma species (G. tsugae, G. subamboinense var. laevisporium, A TCC 52419, G. tropicumand G. lucidum) were incubated in liquid medium containing different concentrations ofgermanium (Ge) for up to20 days at 2fr C. Increasing the Ge concentration of the medium resulted in a gradual decrease in the growth of thefungal mycelium. However, the Ge content in the mycelium increased with increasing Ge concentration. Differentspecies recorded different levels of tolerance towards the Ge. In each case, the optimum concentration of theincorporated Ge in the medium was established as 100 mgtl for both optimal uptake of Ge by the fungal myceliumand optimal mycelial growth.INTRODUCTIONTraditionally in the Orient, Ganoderma hasbeen considered an "elixir of life". Today,there is a considerable body of contempor-ary research that shows that certain speciesof Ganoderma are highly effective medicinalagents as a popular remedy to treathepatopathy, chronic hepatitis, nephritis,hypertension, hyperlipedemia, arthritis,neurasthenia, insomnia, bronchitis, asth-ma, gastric ulcer, arteriosclerosis. leukope-nia, diabetes, anorexia, mushroompoisoning and debility due to prolongedillness (Willard 1990). The medicinal valueof G. lucidum is closely linked to the pre-sence of the following compounds: organic-Ge, polysaccharides, triterpenoids andadenosine (Tong 1995).In Malaysia, studies on cultivationtechniques of a suitable strain of G. lucidumwell adapted to the local climatic condi-tions (Tong and Chen 1990), its growthcharacteristics and Ge uptake by themycelium (Tong et al. 1994a) as well asthe fruiting bodies of this fungus (Tong et al.1994b) have been carried out. Because ofthe paucity of information on the uptake ofGe by other species of Ganoderma of com-CHOW-CHIN TONG AND PEI-JOO CHONGmercial value, the present study was under-taken to ascertain differences, if any, intheir uptake of Ge.MATERIALS AND METHODSCulturesFour species, G. tsugae, G. subamboinense var.laevisporium (ATCC 52419), G. tropicum andG. lucidum were obtained from Taiwan andmaintained on potato dextrose agar (PDA)at 28°C and subcultured every 3 weeks.analysis were prepared by sequential dilu-tion from the stock solution. All reagentsused were analytical grade. A solution of20/0 sodium borohydride in 0.1 DID sodiumhydroxide was prepared daily from NaBH4pellets.The rate of uptake (010) of Ge is calcu-lated as:Amount of Ge in known dry weight of myceliumAmount of Ge in 100 m1 medium x 100RESULTS AND DISCUSSION100 200 300 400Amount of Ge02 in medium (mg/l)Fig 1. Mycelial growth ofGanoderrna species in liquidmedium containing different concentrations of Ge02incubatedfor 20 days at 2fr C. Results represent average oftriplicates.• G. tsugae • G. tropicurn 0 G. lucidurn• G. subarnboinense var. laevisporiurn1.2Effect of Ge on Mycelial Growth in LiquidMediumWith increasing amounts of Ge02 incorpo-rated into the medium, there was acorresponding decrease in the dry weightof the mycelium (Fig 1). The effect wasmost obvious for G. tsugae, which experi-enced a continuous sharp drop in themycelial dry weight at concentrationsabove 50 mgjl of Ge02' Similarly for G.lucidum, the growth of the mycelium was1.6i :~~.3: ---._-_...1~ 0.8 _____\J •i .~~ .~ 0.4 .---=::::::::Uptake of Ge by Fungal Mycelium Grown znLiquid MediumSamples (100 ml) of potato dextrose brothcontaining each of the following concentra-tions (50, 100,200,300,400 mgjl) ofGe02were poured into 1-1 flat bottles andsterilized at 121°C, 15 psi for 15 min.Each bottle was then inoculated with threeagar mycelium discs (1.3 em diam) of a ten-day-old culture and incubated in the darkat 28°C for 20 days. The mycelium waslater harvested through pre-weighed What-man No. 1 filter paper and washed withseveral changes of double distilled deion- ~­ised water (200 ml). The filter paper,together with the washed mycelium, wasdried at 60°C for 48 h. The dry weight ofthe mycelium was recorded.Anarysis of GermaniumFungal mycelium samples were ashed at700-800°C for 1 - 3 hours. The ash wasthen dissolved in 5M HCI. Germanium inthe solution was determined by hydridegeneration - inductively coupled plasmaatomic emission spectrometry (ICP-AES)using the Labtest Plasmascan 710 instru-ment. The method of analysis was adaptedfrom the method for arsenic analysis (Leeand Low 1987).A spectropure (Aldrich Chemical Co.Inc.) Ge atomic absorption standard solu-tion of 990 mgjl was used as the stocksolution. The standard solutions used in the172 PERTANlKA J. TRap. AGRIC. SCI. VOL. 19 NO. 2/3, 1996MYCELIAL GROWTH AND GERMANIUM UPTAKE BY FOUR SPECIES OF GANODERMATABLE 1Effect of Ge02 in liquid medium on the uptake of Ge by mycelium after 20 days ofincubation at 28°CSpeciesG. tsugaeG. subamboinensevar laevisporiumG. tropicumG. lucidumCone. of Ge02 Mycelial Amount ofGe Uptakein 100 ml dry in mycelium ofGeliquid medium weight (mg/kg) (%)(mg/l) (g)50 0.79 752 + 5 11.8100 0.70 1892 + 15 13.3200 0.68 2456 + 22 8.3300 0.54 3855 + 127 6.9400 0.38 4312 + 90 4.050 1.43 34 + 4 1.0100 1.10 99 + 3 1.1200 0.93 175 + 11 0.8300 0.87 260 + 9 0.7400 0.82 310 + 8 0.650 1.53 235 + 7 7.2100 0.94 1080 + 22 10.1200 0.52 2072 + 15 5.4300 0.46 2715 + 43 4.2400 0.37 4002 + 90 4.350 0.12 687 + 18 6.4100 0.11 1796 + 14 8.0200 0.)0 2845 + 31 6.0300 0.05 4251 + 112 3.2400 0.04 5431 + 87 2.6Results represent average of triplicatesretarded considerably. This may be due totoxicity at high concentrations. Onlyextremely sparse growth occurred at con-centrations of 250 mg!l of Ge02 and atconcentrations above 300 mg!l of Ge02there was no growth of the mycelium at alluntil the tenth day of incubation (visualobservation). The limited growth whichcommenced after this lag period rnayprobably be due to the induction of somedetoxification mechanism.The growth of the mycelium for G.subamboinense var. laevisporum was leastaffected by the amount of Ge02 in themedium and there was a more gradualdecline in the mycelial dry weight than inthe other species (Fig 1). Of the four speciestested, G. tropicum produced the mostabundant mycelial growth at Day 20 andthe effect of Ge02 was more gradual above200 mg!l of Ge02 but significantly de-creased between 50 and 200 mg!I.Uptake of Ge by Fungal Mycelium in LiquidMediumTable 1 shows that in general, Ge uptakeby the mycelium increased with increasingamounts of Ge02 in the growth medium.Of the four species tested, G. subamboinensevar. laevisporium assimilated least Ge intothe mycelium, which may explain why itsmycelial growth is least affected by increas-ing quantities of Ge02 in the medium. At400 mg!l of the Ge02 in the medium, G.PERTANlKA J. TROP. AGRIC. SCI. VOL. 19 NO. 2/3, 1996 173CHOW-CHIN TONG AND PEI-JOO CHONGtsugae and G. tropicum took up almost 13times, while G. lucidum took up 17 timesmore Ge than G. subamboinense var. laevis-porium. However, it was noted that the rateof uptake (0,10) of Ge by the myceliumincreased to a maximum at 100 mgJl ofGe02 in all species and then decreased withincreasing concentration of Ge02 in themedium. The highest rate of uptakerecorded was 13.3,10.1,8.0 and 1.10/0 forG. tsugae, G. tropicum, G. lucidum and G.subamboinense var. laevisporium, respectively.Thus, under the conditions studied, itwas deduced that for maximum productionof fungal mycelium with optimal Gecontent, the optimum concentration ofGe02 to be added to the medium was 100mgJI.REFERENCESLEE, C.K. and K.S. Low. 1987. Determination ofarsenic in cocoa beans by hydride generationwith inductively coupled plasma atomicemission spectrometry. Pertanika 10: 69-73.TONG, C.C. 1995. Nutraceutical properties of themushroom Ganoderma. Buletin Maklumat Perta-nian Malaysia 15(3): 1-13.TONG, C.C. and Z.C. CHEN. 1990. Cultivation ofGanoderma lucidum in Malaysia. MushroomJournalfor the Tropics 10: 27-30.TONG, C.C., S.L. KHOONG and C.K. LEE.1994a. Growth characteristics and germa-nium content of Ganodermq, lucidum growingin different substrates. Asia-Pacific Journal ofMolecular Biology and Biotechnology 2(4): 327-333.TONG, C.C., S.L. KHOONG and C.K. LEE.1994b. Germanium uptake by the fruitingbodies and mycelium of the fungus Ganodermalucidum. Asean Food Journal 9(2): 69-72.WILLARD, T. 1990. Reishi Mushroom-Herb ofSpiritual Potency and Medical Wonder. Issa-quah, Washington: Sylvan Press.(Received 2 May 1996)(Accepted 20 August 1996)174 PERTANlKA J. TROP. AGRIC. SCI. VOL. 19 NO. 2/3, 1996

Mycelial growth and germanium uptake by four species of

ganoderma

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Mycelial growth and germanium uptake by four species of ganoderma

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