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Occurrence, diversity and morphology of poroid wood decay by Ganoderma spp. from tropical moist deciduous forest region of Bangladesh

There are many hot spots in tropical moist deciduous forest region where wood decay fungi grow naturally. An investigation was carried out to collect, identify and preserve wood decay Ganoderma spp. on the basis of morphological characteristics from Pabna, Dhaka and Rajshahi under tropical moist deciduous forest region in Bangladesh during June to September 2016 and July to October 2017. This study was conducted to record the morphological variability, distribution, habitat and diversity of the Ganoderma population. A total of 40 Ganoderma samples were collected and identified to nine species under Ganodermataceae family. The samples were collected from the sites by walking through the area following standard method. The field data and laboratory analytical data was recorded during sample collection and in the laboratory, respectively.The highest density of occurrence (23%) was recorded for Ganoderma lucidum and Ganoderma oregonense followed by Ganoderma applanatum (20%), Ganoderma praelongum (19%), Ganoderma lesklokorka (18.5%), Ganoderma pfeifferi (17%), Ganoderma boninense (15%), Ganoderma lipsiense (13%) and Ganoderma tsugae (11%). The highest frequency of occurrence (10%) was recorded for Ganoderma applanatum and Ganoderma oregonense followed by Ganoderma lucidum (9%), Ganoderma pfeifferi (8%), Ganoderma boninense (8%), Ganoderma praelongum (7%), Ganoderma lesklokorka (7%),Ganoderma tsugae (6%) and Ganoderma lipsiense (5%). During survey, Koroi (Albizia procera), Aurjun (Terminalia arjuna), Sisso plant (Dalbergia sissoo), Neem (Azadirachta indica), Golden shower (Acacia auriculiformis) and Rain tree (Albizia lebbeck) were found as hosts of Ganoderma spp. The specimens were preserved in the SAU Herbarium of Macro Fungi (SHMF) and might be useful in mushroom breeding and development program for medicine and food industry sector in future.


INTRODUCTION
The term "Mushroom" is used mainly for the fruiting body produced by the macro-fungi :Ascomycota or Basidiomycota, of which some are edible, many are poisonous and non-edible. More than 10,000 species of mushrooms are present in and around the world and about 2000 of them are considered as edible (Rumainul et al., 2015). These edible fungi are more important for a tropical/subtropical country like Bangladesh, which has a climate, most congenial for the natural growth of such fungi (Purakasthya & Chandra, 1985). Mushroom have high nutritional value along with high proteins, vitamins, minerals, fibers, trace elements content and even low or more or less no calories and cholesterol contents. The number of recognized mushroom species has been reported to be 14000, which is about 10% of the total estimated mushroom species on the earth (Cheung, 2008). Mushrooms have been found in fossilized wood that are estimated to be 300 million years old and almost certainly, prehistoric man has used mushroom collected in the wild as food. Edible mushrooms are considered as healthy food because their mineral content is higher than that of meat or fish and most vegetables, apart from their nutritional value mushrooms have potential medicinal values (Chan, 1981;Chang & Miles, 1988).
Ganoderma is regarded as the ''King of all herbs''all around the world. Gano means shine, Derma means skinit has shining skin which is of red color. Ganoderma is the greatest herb on earth which can enhance immunity to optimum levels. Ganoderma is a genus of polypore mushrooms that grow on wood, and include about 80 species, many from tropical regions (Kirk et al., 2010). Because of their extensive use in traditional Asian medicines, and their potential in bioremediation, they are a very important genus economically. Ganoderma can be differentiated from other polypores because they have a double-walled basidiospore. They are popularly referred to as shelf mushrooms or bracket fungi. Ganoderma are characterized by basidiocarps that are large, perennial, woody brackets also called "conks". They are lignicolous and leathery either with or without a stem. The fruit bodies typically grow in a fan-like or hoof-like form on the trunks of living or dead trees. They have double-walled, truncate spores with yellow to brown ornamented inner layers. Ganoderma, more than 5000 years old, has various names. In China, it is known as Lingzhi, Japan -Reishi, Arab -Al Kam'ah, Indonesia -Jamur. It has more than 200 natural active elements, divided into three categories 30% water soluble elements, 65% organic soluble elements, 5% volatile elements. Another study  recorded several Ganoderma species as Ganoderma lucidum, Ganoderma boninense (Growing stage); Ganoderma tsugae; Ganoderma zonatum (Young stage); Ganoderma applanatum and Ganoderma resinaceum from Bangladesh. They have high nutritive and medicinal values and contribute to a healthy diet, because of their rich source of vitamins, minerals and proteins (Garcha et al., 1993). There are 38,000 varieties of mushroom found on earth. 36,000 are poisonous, 2,000 are edible, 200 are having special medicinal properties. Out of these 200top 6 have greatest medicinal properties . Therefore, it is essential to give efforts in introducing new mushrooms as a source of food and medicinal interest. The Ganoderma red mushroom has exceptional anti-stress qualities that can help improve memory, reduce tension and sharpen your concentration. The knowledge on biodiversity at the community and species level is more important for monitoring the effectiveness and effects of natural and artificial disturbances (Packham et al., 2002). Data on their diversity in different vegetation types is important for planning and managing ecosystem biodiversity (Engola et al., 2007). The biodiversity of mushroom is recently reported from the tropical moist deciduous forest region of Bangladesh (Rumainul et al., 2015; 162 monsoon season extending from June to September. Collection and preservation of Ganoderma spp. is utmost important to improve the study of genetic diversity of collected specimen for future utilization. The survey was conducted to collect and identify Ganoderma spp. from Dhaka, Pabna and Rajshahi under tropical moist deciduous forest region of Bangladesh and to study distribution, diversity and habitat of collected specimen.

Collection site
The survey was conducted in Dhaka, Pabna, Rajshahi biosphere reserved areas, which are situated in tropical moist deciduous forest region of Bangladesh (Figure 1). Survey and collection of Ganoderma spp. was conducted during rainy season starting from June to October, 2016 and 2017 because maximum macro fungi bloom when there is plenty of rainfall occurs. The collection sites were forests, residential area, botanical garden, parks and nearby villages of those above mentioned areas of tropical moist deciduous forest region where woody plants are available.

Sampling of Ganoderma spp.
The collection was made following a standard method of Hailing (Hailing, 1996). Systematic and periodical survey of different locations and other habitats were done in the tropical moist deciduous forest region of the country. Necessary materials and equipment such as isolation kit, slants, isolation chamber, data recording sheet, digital camera for photography, digging equipment, were arranged. following the method of Molina (Molina et al., 1995). Spotted samples were minutely inspected in their natural habitats and brought to laboratory for detailed study.

Morphological and analytical experiment
The collected specimens were brought to the laboratory of the Department of Plant Pathology, Sher-e-Bangla Agricultural University (SAU), Dhaka, Bangladesh and the analytical experiments were done on the recording of data on morphological parameters used for identification such as size of the basidiocarp, umbo, scale, gills, color, gills edges, gill attachment, gill spacing, stipes, length, width, shape, type of veil, annulus (position), volva, carpophores shape, cap color, cap surface, cap margin, cap diameter, and spore diameter (Srivastova & Bano, 2010). The basidiocarps were rehydrated by soaking in water for few minutes before analyzing their morphology. Qualitative characters such as color, shape, and presence of hymenia were evaluated by eye observation while texture was determined by feeling the back and top surfaces using fingers (Svrcek, 2000). Most of the morphological data were recorded during collection period that is when the mushroom was in fresh form. For microscopic characters, permanent glass slides were made from rehydrated basidiocarps with the aid of a sharp surgical blade. Basidiocarps were immersed in cotton blue stain and glycerin and placed on glass slides and covered with cover slips. Motic compound microscope (40x) were used to observe the slides. Spore size was measured by Motic Images plus 2.0 software (Rumainul et al., 2015). Final identification and classification were done by comparing recorded characteristics of mushrooms with the color dictionary of mushroom given by Dickinson & Lucus, (1982); the mushroom guide and followed by the reference of Jorden, (2000) and Pegler & Spooner, (1997).

Drying of samples
Collected samples were cleaned and dried by using electrical air flow drier at the 1000 voltage, which can easily remove the moisture content from the collected mushrooms within 3-7 controlling hours with a regular interval basis power supply (15 minutes switch off and 30 minutes switch on) depending on the structure and texture of the species (Kim, 2004).

Preservation of samples
Dried mushrooms were preserved into a zip-lock type polybag for further studies. Silica gels were used at the rate of 10% of dry basis during the storage period (Kim, 2004).

Habitat, distribution and diversity analysis
The specimens were found attached to various substrata. The surrounding environment temperature, moisture condition, vegetation and soil type was recorded for biodiversity of macro fungi.
Number of site in which the species is present Frequency of fungal species Total number of sites Total number of species Collected samples were wrapped in polybag and brought to the laboratory for their further study. The frequency and density of different species has been determined by using the above formulas (Zoberi, 1973).

RESULTS AND DISCUSSION
Biodiversity, distribution, morphological and ecological characterization of collected Ganoderma spp. are presented in Table 1 and Table 2. Photographs of the collected specimens were presented in Plate 1 and Plate 2. Ganoderma spp. are wood decay poroid macro fungi using for long time in industry and medicinal purposes. In the present study a total of 40 Ganoderma samples were collected and identified to nine species under Ganodermataceae family. The species were Ganoderma applanatum, Ganoderma lipsiense,Ganoderma pfeifferi, Ganoderma tsugae, Ganoderma boninense, Ganoderma lucidum, Ganoderma oregonense, Ganoderma lesklokorka and Ganoderma praelongum.
Ganoderma applanatum was recorded in association with Terminalia arjuna (Aurjun) where the recorded temperature was 28℃ in the surveyed area. For the similarity in temperature, this species was also found in Chittagong and Rangamati Hill Tracts as well as in Sylhet and Moulvibazar under Tropical ever-green and semi-evergreen forest region. But the species was previously reported on Acacia auriculiformis (Rumainul et al., 2015;Rubina et al., 2017) and Swietinia mahagoni ) and on Dalbergia sissoo  also. The density of its presence was 20%. Ganoderma applanatum was found in National Botanical Garden, Dhanmondi Lake and in National Zoo also in association with Azadirachta indica (Neem) with the frequency of 1.35-4.05% and density 1.5-3.76% (Tanni et al., 2020). In a study Moncalvo & Ryvarden, (1997) published a world list of Ganoderma species. The study considered the species described in last 200 years listing 386 names for Ganodermataceae as whole. Taxonomy and 58 diversity of Ganoderma lipsiense and Ganoderma applanatum was also reported in India (Bhosle et al., 2010).
Ganoderma lipsiense was recorded in association with Neem (Azadirachta indica) (Rubina et al., 2017).The density of its presence was 13% and frequency was 5%. This species was recorded from national botanical garden, Dhaka with a density of 5% (Rubina et al., 2017). Ganoderma lipsiense was found in National Botanical Garden and in Romna Park also with the frequency and density 2.70% and 1.5%, respectively (Tanni et al., 2020). On the other hand, Ganoderma lipsiense was also reported from India in association with Dalbergia sissoo plant (Bhosle et al., 2010;Dwivedi et al., 2012) and in China (Wang et al., 2012). In present experiment, this species was collected when the recorded temperature was 29℃. This species were also described from Modhupur under tropical moist deciduous forest and southern region of Bangladesh due to climatic similarity (Rashid et al., 2017).  In current study, Ganoderma pfeifferi was found in association with Acacia auriculiformis with recorded temperature 32℃. This genus was also reported from Southern region and tropical ever-green and semi-evergreen forest region of Bangladesh due to common climatic conditions. The average temperature is 12.1-35.1℃, average annual rainfall is 195 cm in Southern region and in Tropical ever-green and semi-evergreen forest region (Rashid et al., 2016;Tanjim et al., 2019). The average temperature is 12.1-35.1℃, average annual rainfall is 195 cm and average relative humidity is 70-84% of the collection site in the present study.
Ganoderma tsuage was recorded in association with Terminalia bellirica (Bohera tree) Rubina et al., 2017). But this species was previously found in association with Acacia auriculiformis (Golden shower) (Rumainul et al., 2015) under tropical moist deciduous forest region of Bangladesh. The frequency and density of it's presence was 6-8% and 11% respectively. Another survey found the density of this species 12.5% and frequency 50% under social forest region of Bogra . Ganoderma tsugae was found in National Botanical Garden, Romna Park, Dhanmondi Lake and in National Zoo associated with Swietenia macrophylla (Mehogani) tree (Tanni et al., 2020). In other study (Rubina et al., 2017) recorded it's density 10% in national botanical garden, Dhaka. Ganoderma tsuage was also reported from India (Vyas et al., 2014). This species was also found in southern forest region of Bangladesh for similarity in required climatic condition.
Ganoderma boninense was recorded in association with Dalbergia sissoo (Sisso) tree . But this species was also found in association with Mangifera indica from national botanical garden (Rubina et al., 2017) and Bambusa sp. from tropical moist deciduous forest region of Bangladesh (Rumainul et al., 2015). The frequency and density of its presence was 8% and15%, respectively. Another survey found this species in social forest region of Bogra, Bangladesh with frequency of 75% and density of 43.5% . The pathogenicity of Ganoderma boninense was examined (Cooper et al., 2011). This species was also seen in Chittagong and Rangamati Hill Tracts as well as in Sylhet and Moulvibazar forest region (Tanjim et al., 2019). In tropical ever-green and semi-evergreen forest region, the average temperature is 12.1-35.1℃, average annual rainfall is 195 cm, average relative humidity is 70-84% recorded.
In the present study Ganoderma lucidum was recorded in association with Albizia procera (Koroi), Terminalia arjuna (Aurjun) and Dalbergia sissoo (Sisso plant) and which was supported by Aminuzzaman and Das (2017) and Rubina et al. (2017). Another study (Marzana et al., 2018) found Ganoderma lucidum in association with Tectona grandis (Shegun) in Kaptai, Rangamati of Chittagong Hill tracts under tropical evergreen and semievergreen forest of Bangladesh. Other study (Rumainul et al., 2015) found this species in association with Leucaena leucocephala (Ipil-Ipil) and Acacia auriculiformis (Golder shower) under tropical moist deciduous forest region of Bangladesh. The density of its presence was 19% and 23%. Another survey (Marzana et al., 2018) found this species from Chittagong Hill tracts with a frequency and density of 11.11% and 2.78% respectively. There are three species were collected during the investigation, viz-Ganoderma applanatum, Ganoderma tsugae and Ganoderma lucidum on the bark of tree and on dead plant, Ganoderma sp. Among the species, the highest frequency of Ganoderma tsugae was 33.33% and the density was 95% (Rashid et al., 2017). Ganoderma lucidum was found in every survey area associated with Azadirachta indica (Neem) (Tanni et al., 2020). But another finding  described the species from Bogra district under social forest region of Bangladesh with a frequency and density of 75% and 25% respectively. This species was found from national botanical garden, Dhaka, Bangladesh with a density of 25%.The difference in frequency and density of the species might be due to the difference in geographic area of the collection sites (Rubina et al., 2017). The morphology of 53 specimens of Ganoderma lucidum was studied from Norway and found large variation among the species (Ryvarden, 1995). This species was also recorded in mangrove forest region and Tropical ever-green and semi-evergreen forest region of Bangladesh due to resemblance in climatic requirements. In Mangrove forest region, the average temperature is 25.5℃, average annual rainfall is 164-200cm, average relative humidity is 80-90%. Ganoderma oregonense was recorded in association with Terminalia arjuna (Aurjun). The Journal of Agriculture and Natural Resources (2020) (Allen et al., 2009). This was also reported in Germany (Passie et al., 2002). Ganoderma praelongum was recorded in association with the Terminalia arjuna (Aurjun). But the species was previously reported on Diospyros ebony .Ganoderma spp. were recorded in association with the Dalbergia sissoo (Sisso plant) with the frequency and density of 9%, 8%, 18% and 16% respectively. Four species of Ganoderma were found during collection such as-Ganoderma spp., Ganoderma tsugae, Ganoderma applanatum and Ganoderma boninense from Sylhet division. The frequencies of collected of Ganoderma were 37.5, 12.5, 12.5% and the densities were 12, 24, 4%, respectively. The color of Ganoderma tsugae was dark brown and white, Ganoderma applanatum was dark brown and Ganoderma boninense was brick red. The spore size of Ganoderma spp. were 13.2×8.9μm, 11.8×8.7μm and 9.2×6.8μm, accordingly. These Ganoderma spp. were collected from soil, Mehogani (Swietenia macrophylla) and Shimul (Bombax ceiba) tree, respectively (Tanjim et al., 2019). Ganoderma spp. were also found at Rajshahi, Pabna, and Dhaka, and Bogra districts of Bangladesh in tropical moist deciduous forest (Rumainul et al., 2015). It was also reported in China (Wang et al., 2012) and in India (Dwivedi et al., 2012;Thiribhuvanamala et al., 2011;Ram et al., 2010).