Phansomba: An Indian Herbal, Folk Medicine

Vaidya J.G.*, Bhosle S.R.**, Bapat Gauri*, Garad S.A.*, Sonawane H.B.*,
Sharifi Ashghar***, Ali Mirzae***, Lamrood Prasad*****, Aparna Sawant****,
Mathew Poomala*
*Department of Botany, University of Pune, Pune – 411 007. India.
**Science and Technology Park, University of Pune, Pune-411 007. India
*** Medical Science University, Yasuoj, Iran.
**** J.P. Naik College, Aurangabad. M.S., India
***** Ahmadnagar College, A, Nagar. M.S., India
Email: jitendra@unipune.ernet.in

Abstract:

“Phansomba”, a folk medicine used by tribes in the Western Ghats of Maharashtra,

comprises of many species of Phellinus occuring on Artocarpus heterophyllus

(vernacular name: Phanas). The present article gives total scenario of the research work

carried out to explore this medicine, which could prove to be a future valuable resource.

A thorough survey carried out, in the “Western Ghats of Maharashtra” yielded twenty-six

species of Phellinus after critical taxonomic identification. Out of these, five were

selected for further studies based on their frequency of occurance, namely, Phellinus

badius, P. merrillii, P. sublinteus, P. fastuosus and P. lloydii.

Initially Physico-chemical anaysis and biochemical analysis were carried out to get an

idea about the composition of the saples. Assays for antioxidant, antimicrobial, anticancer

and antidiabetic activities were performed, for the selected samples. They suggest

significant bioactivity. Toxicity testing of the samples done, using Allium root tip assay

which suggests that the samples are nontoxic and could be further exploited.

Key words: Phansomba, Ayurveda, Folk medicine, Phellinus, Pharmacognosy,

Bioactivity.
Introduction

There is always a erge to search for newer and more effective medicines arround the

globe. The resources could be uncountable but hidden in nature. The traditional medicinal

systems all over the world, could serve as templates for identifying new drugs. Fungi

especially macro-fungi (regarded as mushrooms) are long known to have medicinal

properties (Hobbs 1995, Denisova 2002, Smith et.al. 2002)

Mushrooms are traditionally used in the Chinese medicine and commonly used for the

pharmaceutical purposes and health foods. Increasing experimental evidence indicates

that mushrooms contain a large number of biologically active components that offer

health benefits and protection against degenerative diseases.

Many pharmaceutical substances with potent and unique health enhancing properties

were isolated from various mushrooms (Wasser and Weis 1999, Smith et. al. 2002). For

thousands of years different communities are making use of many mushrooms as a

medicine (Blanchette et. al. 1992; Vaidya 1995 ).

i) Phansomba

Phanasomba is a folk medicine used in the Western Ghats of Maharashtra. It is common

bracket mushroom causing heart rot disease of Artocarpus heterophyllus (vernacular

name : Phanas). Phansomba it is also known as Phanas-alambi, Phanas banda (Khory

1887, Dymok et.al. 1890, Andalkar 1988, Vaidya and Bhor 1991; Vaidya and Lamrood

2000).
According to Dymock et.al. (1980), the medicinal use of mushrooms was introduced by

Portugese in India. They used a kind of mushroom as a substitute for European Tinder

fungus for wound healing, the local name given to this mushroom lateron was

Phansomba, the description of the mushroom given resembles that of Phellinus.

Khory (1887) named Phanasamba as Phanas-alombe and considered it to be belonging to

Agaricus. The description of therapeutic use of this fungus is same as that of Phansamba

described by Dymock et. al. (1980). He described several other genera under the same

trade name. Nadkarni (1954) considered Phanasamba as Agaricus ostreatus and also

considered it to be similar to Boletus crocatus. The use of this is exactly similar to the

previous references.

Kirtikar (1918) noted that, Phansomba samples were sometimes dry and sometimes moist

and when they were moist, showed oozing of reddish juice. He further stated that,

Phansomba is yellow – orange in colour when fresh and dull brown, when old.

Dymock et.al. (1980) regard Phansomba as Agaricus ostreatus whereas Kirtikar 1918

identified it as Boletus nitus (Shah 1928). After genuine and critical examination of

samples sold under the trade name Phansomba resemble Polyporus and not Agaricus.

On the basis of knowledge acquired lateron, Phansomba comprises of many species of

Phellinus, a medicinal fungus from Hymenochaetaceae,such as Phellinus adamantinus

(Berk.)Ryv., P. carteri (Berk.et. Cke.)Ryv., P. conchatus (Pers.: Fr.) Quel., P. fastuosus,

P. linteus, P. merrilli.(Murr.)Ryv., P. pectinatus (Klotzsch) Quel., P. sublinteus

Murr.)Ryv. And lot many (Vaidya and Bhor 1991).

Much advancement in studies of Phellinus spp. has revealed newer dimentions of their

medicinal properties, some of which are noted below alongwith the international senario

of the medicinal use of Phellinus.

The extracts of P.linteus has anti-oxidation effect,whitening effect,moisture retentive

effect,anti-acne effect and it improves immunity(Kim et.al .2000a).P.L. extracts along

with additives serves as hair tonic and is effective in the regeneration of mother hair cell

and the promotion of blood circulation(Kim et.al. 2000b)

The Khanty people of west Siberia use the ash of Phellinus nigricans to cure toothache

and tooth decay by mixing in low-grade tobacco(Saar,1991).Same practice of using ash

of Phellinus igniarius with tobacco was also practiced in other tribes like Blackfoot,

Labrador Eskimo, Micmac, Kwakiutl, Inuit, Yup’ik and Inupiaq Eskimos of western

Alaska.The mixture was either smoked or chewed.This mixture was supposed to be more

strong than only tobacco.(Blanchette 2001; Blanchette et.al.2002).

P.igniarius is taken internally as a bitter tonic and laxative, externally as a

styptic(Nadkarni,1954;Chopra et.al.,1956) Several studies have showed that Phellinus

species can produce substances with cytotoxic (Atsumi, et. al; 1990, 1993, Withers, &

Umezawa, 1991, Han, et. al; 1999). Phellinus linteus (Bhendiomba) is used for the

urification of blood in skin diseases (Vaidya and Rabba 1991). Polysaccharides-

spachyman and pachymaran extracted from Phellinus exhibited strong anti-cancer and

immunomodulatory activities.(Hobbs, 1995).

Phellinus also shows antioxidant activity that highly regarded in traditional medicine and

is widely consumed in the belief that it promotes health and longevity lowers the risk of

cancer and heart disease and boosts the immune system. GL is widely consumed
throughout the world as a health food, being commonly and regularly used by many

Asian communities for the promotion of health and longevity, and as a remedy for illness

(Jong & Birmingham, 1992; Rajarathnam et al. 1998;Wachtel-Galor et al. 2003).

ii) Distribution: Phansomba is amply found in the Western Ghats of India. The whole

basidiocarp of this mushroom is powdered and used in many commercial ayurvedic

preparations. (Andalkar,1988). Phansomba from Western Ghats of Maharashtra,

Karnataka and Kerala has been well-studied and many ayurvedic preparations are known

from the region. In kerala Phellinus is known as ‘Pila mangal’ (pila=jack fruit tree,

mangal=turmeric). The name suggests that it is as medicinal as turmeric.

As for today the drug being limitedly explored is not cultivated, but it is possible to

cultivate it on mass scale for commercial production through technologies like “Solid

State Fermentation.”

iii) Names of species: With further studies, carried out in Western ghats of India, it

became clear that total 26 species of Phellinus (noted below) are found on Jackfruit trees.

Total 400 Phansomba samples were collected from Artocarpus heterophyllus and studied

for their micro amd macromorphological characters. Their correct taxonomic

identification was done by Larson and Cobb-Poulle (1990) and total 24 different species

of Phellinus were identified. (Lamrood 2004, Rabba 1994).

Table 1. List of Phellinus species identified on Artocarpus heterophyllus

Name of species No. of Accessions

1 Phellinus adamantinus 8
2 P. badius 9
3 P. calcitratus 3
4 P.caryophylleus 1
5 P. coffeatoporus 5
6 P. conchatus 4
7 P. dependens 1
8 P. durissimus 1
 9 P. fastuosus 20
10 P. grenadensis 2
11 P. linteus 8
12 P. lloydii 10
13 P. megregori 1
14 P. melanodermus 1
15 P. merrillii 26
16 P. rhytiphloeus 3
17 P. robustus 1
18 P. sanctigeorgii 1
19 P. setulosus 1
20 P. stratosus 3
21 P.chinensis 3
22 P.rimosus 1
23 P. nilgheriensis 3
24 P. allardii 3
25 P. acontextus 1
26 P. stratosus 1

Besides the above-mentioned species there are many samples, which cannot be acertained

to any known species, and the work is in progress for the confirmation of new species.

The table depics that the major species of Phellinus occuring on Artocrpus heterophyllus

are P. fastosus, P. lloydii and P. merrillii.

iv) Chemical constitution:

Phansomba shows the moisture range of 5-8% and the colour of powder was basic red

with different brown shades. The extractive value is 12-18% whereas pH of all the

samples is acidic. The crude fibre content varies in the range of 8% to 33%, the total ash

content is 2-2.5% (Lamrood 2004).

Besides the routine physicochemical parameters the tested Phansomba samples show

presence of Germanium in the range of 0.2-0.5 ppm. and Selenium 0.1-0.5 ppm which is

medicinally important (Lamrood 2004).

The carbohydrate content is in the range of 17-34%, total proteins are 7-21% while amino

acids contents are 0.56 to 3.38%. The total lipid contents are in a range of 0.01-0.02%

(Lamrood 2004).

The exact chemical constitution of the bioactive compound in Phansomba has not yet

been cracked but the work is still in progress. However it is proved that polysaccharides

and terpenoids are responsible for bioactivity in many mushrooms (Wasser and Weis

1999). In the study Ergosterol is also considered being a fungal specific bioactive

compound.

vi) Bioactivity:

Antibacterial activity: Polysachharides fractions were extracted from the powdered

basidiocarps of Phansomba (Mizuno 1999) and also terpenoids were extrated from the

same starting material (Harborne 1984). All four polysaccharide fractions were screened

for antimicrobial activity against seven human pathogenic microorganisms. Out of four

fractions, fraction- I and II were inactive while fraction-IIIa and IIIb showed moderate

activity. The present study, for the first time reports the antimicrobial assay of

polysaccharides. Fraction-I (water soluble polysaccharides) have been showed to be

potent antitumor agents, but were seen inactive against the microorganisms similar to

fraction-II, while fraction-IIIa, IIIb, which are also reported to exihibit strong antitumor

activity, showed moderate antimicrobial activity. Among the terpenoids, sesquiterpenes

exhibited the strongest activity, while diterpene and triterpene extracts showed moderate

activity.(Asghar sharifi, 2006)

Antioxidant activity: Aerobic organisms produce reactive oxygen species such as super

oxide anion, hydrogen peroxide and hydroxyl radicals through the chain reactions that
start with the reduction of molecular oxygen. These free radicals attack polyunsaturated

fatty acids in cell membranes and give rise to lipid peroxidation in living systems

(Cochrane 1991).Lipid peroxidation is also strongly associated with aging and

carcinogenesis (Yagi 1987). However, living systems are protected from active oxygen

species by enzymes such as superoxide dismutase, glutathione peroxidase and catalase.

These living systems have also been reported to receive non-enzymatic protection by

endogenous antioxidants such as -tocopherol, ascorbic acid, -carotene, and uric acid by

food (Ames et al. 1981). It has been suggested that antioxidants found in large quantities

in herbs and mushrooms may be responsible for this protective effect (Halliwell 1994).

Antioxidant activities of methanolic extracts from Phellinus species were in the

descending order of P. adamantinus (86.3%) > P. sublinteus (84.15%) > P. fastuosus

(81.83%) > P. merrillii (79.36%) > P. lloydii (76.46%) at 5 mg/ml. (Ali Mirzael 2006,

Vaidya et.al., 2007)

Antidiabetic activity : The medicinal mushroom Phellinus badius shows hypoglycemic

activity in alloxan induced diabetic rats. Hot water extract of fruit body of Phellinus

species shows 63% reduction of blood glucose and significant reduction in triglyceride,

cholesterol, AST, ALT and mycelial biomass of the Phellinus species shows 68%

reduction of glucose. Extracellular polysaccharide shows body weight reduced the

plasma glucose level of the diabetic animals by 49 %(Kim et.al.,2001). The

administration of Phellinus species exopolysacharide significantly lowered the plasma

glucose level by 52.3% as compared to control (Personal communication).

Anticancer activity: Anticarcinogenic activity of two species of Phansomba (P. merrillii

and P. fastuosus) on SiHa cell lines was done. In this experiment hot, cold water and 50%
ethanol extracts were used for the activity. These all extracts showed high cytotoxic

effect on SiHa cells (Bonde et. al. 2002)

Mizuno,(2000) reported the antitumor activity of five different polysaccharide fractions

of Phellinus linteus on Sarcoma 180.Studies by Ajith and Janardhan (2003) on cytotoxic

and anti-tumor activities of ethyl acetate methanol and aqueous extracts of Phellinus

rimosus revealed that ethyl acetate and methnol extracts show in vitro cytotoxic activity

against Dalton’s Lymphoma Ascites(DLA) and Ehrlich’s Ascites Carcino(EAC) cell

lines.The aqueous extracts did not show cytotoxicity.All The three extracts were highly

effective in inhibiting growth of solid tumor induced by DLA cell line in mice.But the

activity of ethyl acetate extract was higher than that of methnol and aqueous extracts.

Anti-arthritic property

Collagen Induced Arthritic (CIA)mice were treated with a proteoglycan isolated from

P.linteus.Both pre and post treated mice showed decrease in CIA as well as in anti-CII

IgG and IgG2a antibodies and varying kinds of cytokines including IL-12,TNF-, and

IFN-  (Kim et. al.2003).

Antiviral activity property

Aqueous extracts from some aphyllophorales including Phellinus rhabarbarinus (S 0389-

7) exhibited strong anti-HIV-1 activity, without toxicity for Molt-4 lymphocytic cells.

The test extracts specifically act at the level of CD4-gp120 binding. (Walder

et.al.,1995.a) : Aqueous extracts from P. rhabarbarinus, are reported to have potent in

vitro anti-HIV-1 activity and to inhibit HIV-1 reverse transcriptase (R T), active against

HIV-1.Test extracts induced an IFN type sharing with native human leukocyte interferon
(NHLe-IFN) both biological and antigenic properties as well as the conventional acid

stability (Walder, et.al.1995b, Kalvatchev et.al. 1995).

vii)&viii) Uses:

Many species of Phellinus are being regularly used under the trade name Phansomba, by

the tribals, local people of western Maharashtra, learned Ayurvedic doctors (Vaidus) of

Western Maharashtra (i.e. Konkan region) not only in several ailments in children but

also for throat problems (antitumor), antimicrobial and antiinflamatroy agent in

rheumatoid arthritis as well as a general health enhancer along with other ayurvedic drugs

(Andhalkar, 1988; Vaidya, 1991; Vaidya & Bhor, 1991; Vaidya & Lamrood, 2000,

Khory, 1887; Dymock et.al., 1890; Kirtikar, 1918; Desai, 1927, Shah, 1928; Nadkarni,

1954; Chopra et. al. 1956, Sawant, 1974 and Sathe, 1982).

The whole basidiocarp of Phellinus is ground in water to make paste and then applied to

affected area, like gums in case of excessive salivation in kids, for wound healing etc. this

paste may be administered internally for diarrhoea, dysentery (Desai 1927, Vaidya 1991,

Vaidya and Bhor 1991).

The dosage suggested by local ayurvedic doctors is as folows, take Phansomba (50g) in

2,000cc of cold water. Heat it over high heat until begins to boil then reduce the heat to

low. Wait until the water concentrates down to 1,400cc. Remove the liquid from the heat

and pour into another container.

Add 2,000cc cold water to the ingredients and boil them again. Repeat the procedure

three times then mix all soups. Pour the mixed mushroom soup (3,000~4,000cc) into a

glass container and keep it in the refrigerator. Drink 100-150ml. of it (about the size of a

cup of coffee) three times a day. You can use glassware for boiling or simply use a herb
extractor. Do not dispose of the remaining mushrooms boil it 2 or 3 times to drink as a

substitute for mineral water or barley tea.

In Kerala Phellinus is used for many ailments some of them are as follows.

1) For pitta- pitta gulika (tablet)

Phellinus is used along with “Kantham” ( a mineral taken from the earth) and

“annupedi”(an ayurvedic medicine). These three are ground together for seven days

in lemon juice and then taken and used for ailments that come out of pitta.

2) Phellinus is a constituent of the “Marmanithailam”, an ayurvedic oil used for curing

major and minor fractures.

3) Phellinus is a constituent of the “Narayanithailam”, an ayurvedic oil used for various

massages. This is also used to reduce burning sensations and severe joint pains.

4) Phellinus is used to cure cancers in the buccal cavity (mouth) by making an ayurvedic

decoction called “kashayam”

5) Phellinus is used to cure rectal and intestinal cancers. It is ground together with cattle

urine and use.

6) Phellinus is used also in cases of brain tumour and severe headache by doing “Nasya”

through the nose. Here it is mixed along with other ayurveic medicines.

For all the above uses Phellinus is collected from jack fruit tree and also from

Strychnos nux-vomica. It is first soaked in rice juice to release all the toxic chemicals

and then ground to prepare various combinations.

In Karnataka is commonly used against throat cancer, mums, and for children ailments

for apothae and excessive salivation

ix) Genotoxicity testing: It was done by Allium root tip assay. Root and shoot length

went on decreasing as concentration increased. Onion bulb showed good root growth at

0.1% concentration, but from 0.3% concentration onwards root and shoot showed scanty

growth. When bulbs treated with Ec50 concentration and nearby concentrations and

tested for genotoxicity, it did not show any chromosomal or mitotic aberrations This

experimentation indicates that Phansomba maybe nontoxic to the living systems (Sawant,

2004).

x) Contraindications: After the administration the drug, the first sign will be the

expression of the unknown ailment, which remains suppressed because of allopathic

drugs. This is followed by urination, slowly controlling the disorders. This can be used

effectively as immunomodulator which is similar to that of Ganotherapy (use of

Ganodema lucidum). This obsevation is based on animal trials conducted at NTC

(Personal communication).

xi) Precautions for uses: The medicine is mainly used as a food supplement or

neutraceuticals. When it is used as alternate medicine, firstly it is administered in adiition

to allopathic medicine and later slowly, allopathic drug is withdrawn stepwise till the

patient achieves normal health. The dose of Phansomba may be continued as an immune

enhancer.

xiii) Adulterants: Our survey from the ayurvedic medicine stores reported that Phellinus

fastuosus and Ganoderma lucidum were sold under the name phansomba. The work done

by our group revealed that in addition to above species, species like P. adamantinus, P.

carteri, P. conchatus, P. merrillii, P. melanodermus, P. pectinatus., P. sublinteus,

Ganoderma applanatum, Ganoderma philippi, G. testaceum, G. applanatum and G.

lipsiense, G. lucidum var. lucidum, G. lucidum var. capense, G. perzonatum, G. curtisii,

G. multiplicatum, G. praelongum, G. orbiformum and G. stipitatum were also sold as

samples of phansomba. Even a bark of Jackfruit tree (Artocarpus integrifolia L.) was sold

under this name as adulterated sample.

We therefore feel that, majority of the heartwood causing species of Phellinus

having significant thick conk and shining hymenial surface is knowingly or unknowingly

collected and sold as a sample of phansomba in the market.

xiv) Commercial products available already: Like Phansomba, Sang-hwang is used in

Korea and “Song gen” in China and "Mesimakobu" in Japan which comprises of

Phellinus Linteus. as "Song gen" in China, "Sang-hwang" in Korea and "Mesimakobu" in

Japan, are well known in the chronicles of the Oriental medicine.

Phellinus linteus have been used for chinese medicine, traditionally. The functional

compounds in phellinus linteus have been highlighted recently, as these substances be

able to lower cholesterolemia, modulate immune system, inhibit tumoral growth.

Scientists found extracts of the medicinal fungi called Phellinus linteus boosts the effects

of a particular anti cancer drug.

Researchers based at the Boston University School of Medicine in the USA studied

extracts of Phellinus linteus They tested its effects on prostate cancer cells and found that

when it was combined with the common chemotherapy treatment doxorubicin it

increased the number of prostate cancer cells killed by the drug.

There are many marketted drugs available which use Phellinus linteus as one of the

constituents. Several companies manufacture and market drug in various forms, to name

few; Ryu Chung Hyun Mushroom, Hangzhou New Asia International Co., Ltd etc.
Hangzhou New Asia International Co., Ltd has prepared Sang-Hwang mushroom extract

of Phellinus linteus polysaccharide.Sang-Hwang mushrooms are effective for cancer

arrest, stomach ailments, arthritis of joints, and other ailments. It contains properties that

improves immune system and acts as anti-cancer, tumor and diabetic effects.

Phellinus linteus Polysaccharide Capsule of Dabieshan Brand is also available in the

market.

Lintenol is a product in dried powder form of Phellinus linteus IJ209 strain

manufactured by Mushville Co. South Korea.It is used in functional food and general

food in preparation of bread noodles and medicine.

Organic Meshima

Mycosoft Gold Capsules: Capsules for immunotherapy prepared from P.linteus

Mycosoft Gold Extract

Meshima coffee mix

Natural Meshima Mushroom slice: Beverage prepared from Phellinus

Host Defense 60C Mesima capsules

Host Defense® has been shown to increase human Natural Killer (NK) Cell activity by

up to 300%. The immune-enhancing power of Host Defense® comes from two sources:

the unsurpassed quality of the MycoMedicinals® mushrooms

Native Women 2 Vcaps

Mushroom Complex to Enliven A Woman's Physical and Sexual Energy

Sanghwang 100 (K.B. Co. Ltd. Korea) is made with Phellinus linteus and other herbal

ingredients. Phellinus linteus has a powerful anti-cancerous effect. At present KBCL is

under development of several other functional food products, which are good for dieting,

and for preventing obesity, osteoporosis and alopecia (hair loss).

Phellinus tea is prepared by boiling the pieces of fruit body.

Sang-Hwang One: Health supplement from P.linteus mushroom manufactured by

Pulmone health and living company,Ltd.

Sang-Hwang Rice(150 g) manufactured by Gyeongbuk Institute of Bio Industry.

Sang-Hwang Pepper Paste: Paste prepared from Phellinus

Phellinus linteus X-P: Product from Phellinus

Ginseng Phellinus linteus liquid powder Gold: Phellinus and Red Ginseng in liquid

form
Activity of Phellinus extracts on various human pathogenic microorganisms.

Table 4-3: Activity of the Phellinus extracts against A.c (NCIB 2086)

Sample Zone of inhibition in response to various

extract (mm)
Poly Poly Sesqui- Tri- Diterpene
IIIa IIIb terpene terpene
Ph-1 16.33 17 26 20.67 21.33
Ph-2 15.33 15 25 21.33 21.33
Ph-3 14.67 16 21 19.33 18.67
Ph-4 14.33 14.67 16 14.33 14.67
Ph-5 9.33 10.33 15 10.33 10.67
Ph-6 13.67 14 18 15.33 16.33
Ph-7 13 13.67 21 17.67 18
Ph-8 11.33 11.67 17 12.67 13.67
Ph-9 13.67 14.67 19 16 17
Ph-10 12 12.67 15 10 11.33
Ph-11 11.67 12.33 16 12.67 13.67
Ph-12 13.67 14.33 17 14.67 14.33
Ph-13 14.67 15 20 15.33 16.67
Ph-14 14 15.33 19 16 17
Ph-15 13.33 14.33 21 15 17.33
Ph-16 14 14.33 17 15 16
Control 7 7 7 7 7
SE 0.381 0.399 0.717 0.343 0.382
CD 1.099 1.150 2.067 0.989 1.101
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Solid state culture and cultivation of Phellinus and Ganoderma:
Phellinus and Ganoderma fresh fruit body were collected. Cultures were isolated
by tissue culture method as per Stalper, 1978. Fresh marginal tissue was taken and
untouched tissue was cut into small pieces and transferred to PDA under sterile
conditions and cultures were obtained.
Five different species of Phellinus named as P. badius (Berk. Ex Cke) Cunn. P.
sublinteus (Murr.)Ryv., P. merrillii (Murr.)Ryv , P. fastuosus (Lev.Ryv.), P. lloydii
(Clel.) Cunn., and Ganoderma sitpitatum (Murrill) Murrill., G. multiplicatum (Mont.)
Pat., G. lipisiense(Batsch), G. resinaceum Boudier., G. multicornum Ryv. were screened
for their growth rate and Phellinus badius and G. sitpitatum were selected for further
work as they showed higher growth rate and biomass production as compared to other
species.
For the comparison G. lucidum produced from NRCM (Solan H. P.) and P.
linteus from FRI (Dehradun) were taken.
Zea mays (Maize), Triticum aestivum (Wheat), Sorghum bicolor Jowar,
Pennisetum americanum Bajara, and Eleusine coracana (Nachani) cereal grains were
screend for biomass production. Colonization by Phellinus was fast on and where as
Ganoderma showed fast colonization on Bajara grains.
Spawn were prepared by the method given by Stamets, 2000. Grains were washed
and cooked until they became soft and then autoclaved at 20 lbs pressure at 121 0 C for 30
min. and inoculated from culture plate. Mass multiplication was done by grain to grain
culture method. (Stamets, 2000).
For the solid state culture two basic substrates were selected. Saw dust and
sugarcane baggasse and three supplements were screened for higher biomass production
i.e. Rice hull, Rice bran and Peanut shell.
In case of supplement Rice bran was gave the best result so was selected for
further work. Different combinations of saw dust: Rice bran, Sugarcane baggasse: Rice
bran, Saw dust: Rice bran: Sugarcane molasses were done.
Substrates soaked overnight, excess water drained off. Basic substrates and
supplements were mixed in different combinations ( 100 : 0, 90 : 10, 80 : 20,
…….0:100 ) 100 gm of each mixer was filled in autoclavable bottles and autoclaved it at
15 lb pressure 121 0 C for 1 hour. Inoculated with grain spawn and incubated at 28 0 C ±
2. Colonization rate was observed after 3 days. Bottles were shaked daily.
In case of solid state culture it is very difficult to measure the quantitative
biomass. One of the easiest method is by estimation of percent ergosterol. Ergosterol is a
fungal membrane compound. It is fungal specific sterol. So it is directly proportional to
biomass produced. This method was suggested by Arthington and Skagg’s(1999).
Fermented biomass is crushed in 25% alcoholic KOH and kept it in waterbath for 4 hours
then 800 µl Distilled water was added and 5 ml of n-heptane was added. Vortexed it
properly for 3 min. and allow to stand for a while. Upper n-heptane layer contains
ergosterol, which is estimated spectrophotometrically. Control was pure mycelial
biomass.
In P. badius Sugarcane baggasse: rice bran : sugarcane molasses (88:10:2) was
the highest biomass producing combination. In this case as the percentage or rice bran
goes on increasing the biomass yield decreased. Polysaccharide production was also done
by the method suggested by Mizuno 2000. Polysaccharide extraction done by
fractionation method. Fr-I was highest in 88; 0:2 and Fr-IIIa and IIIb in 84:15:2 was
highest.Fr-II was found was very less so not recorded. P. linteus gave the highest biomass
production in 88:10:2 (SB:RB:SC)
 Cultivation trial was done for both Ganoderma and Phellinus. Both the species of
Ganoderma and Phellinus was successfully done but in case of Phellinus only precocious
fruit body was induced.