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An experiment was conducted to study the influence of seed sources on seed germination and seedling vigour of Pinus gerardiana seeds collected from 23 seed sources during October, 2018 in Kinnaur and Chamba districts of Himachal Pradesh. The seeds were subjected to germination testing to identify the best seed source for raising Pinus gerardiana seedlings in the nursery. Significant differences were observed in germination percent, average seedling length and seedling vigour of seeds collected from different seed sources. The maximum germination of 83.00% was recorded in seeds collected from Jangi seed source followed by 74.00% germination in seeds collected from Purbani, 71.00% germination in seeds collected from Moorang in Kinnaur Forest Division whereas minimum germination of 36.00% germination was recorded in seeds collected from Luj in Pangi Forest Division. Similarly, maximum seedling length of 11.10 cm was recorded in seeds collected from Jangi followed by 10.50 cm in seeds collected from Purbani and 10.21 cm in seeds collected from Moorang in Kinnaur Forest Division whereas minimum seedling length of 7.70 cm was recorded from seeds collected from Luj in Pangi Forest Division. The maximum seedling vigour index of 921 was recorded in seeds collected from Jangi followed by seedling vigour index of 777 from Purbani and seedling vigour index of 724 from Moorang in a decreasing order whereas minimum seedling vigour index of 277 was recorded in seeds collected from Luj in Pangi Forest Division. It is recommended on the basis of present investigation that seeds of Pinus gerardiana should be collected from Jangi seed source in Kinnaur district for raising quality seedlings in the nursery.

In the present research, a chemical co-precipitation approach has been used to approach the synthesis, characterization, and photocatalytic applicability of Ni-doped α-Fe2O3 (hematite) nanoparticles. Biosynthesized iron oxide nanoparticles (IONPs) were successfully synthesized using a non-toxic leaf extract of the Azadirachta indica (AI) plant (neem) as a reducing and stabilizing agent. X-ray diffraction (XRD), transmission electron microscopy (TEM), scanning electron microscopy (SEM), Raman spectroscopy, FT-IR spectroscopy, ultraviolet-visible (UV-Vis) spectroscopy, and vibrating sample magnetometer (VSM) have all been used to examine the synthesized materials. All of the produced NPs contain only the nanocrystalline hematite phase, according to XRD measurements. The morphology studies of the Ni-doping hematite nanoparticles, as demonstrated by TEM and SEM. The phase purity and phonon modes of the prepared nanoparticles are confirmed by Raman spectroscopy. The UV-Vis absorption tests show also that value of the band gap increases together with the reduction in particle size, going from 2.26 eV for chemical α-Fe2O3 to 2.5 eV for green Ni-doped α-Fe2O3 nanoparticles. Additionally, it was clear from the magnetic characteristics that all of the samples behaved ferromagnetically at ambient temperatures. On the other side, malachite green (MG) dye was used as a surrogate industrial wastewater dye in order to study the photocatalytic efficiency of Ni-doped α-Fe2O3 particles. The pure/green Ni-doped α-Fe2O3 NPs showed that after 70 minutes of exposure, 92% of the MG had become discolored.

Mycorrhizal fungi boost host plant growth by improving roots' ability to absorb nutrients and water from the rhizosphere soil. In this study, a mass inoculum of the ectomycorrhizal fungus Scleroderma polyrhizum was produced on wheat grains and incorporated into polybags during seed sowing of Pinus gerardiana, with the expectation that the roots of the germinating seedlings would form a mycorrhizal association with S. polyrhizum. For 2 years, the seedlings' growth parameters were measured at 3-month intervals. The seedlings raised in inoculated bags exhibited ectomycorrhizal anatomy and higher growth indices like absolute growth rate, relative growth rate, sturdiness quotient, volume index, quality index etc. The growth parameters of 2-year-old inoculated seedlings were 44.36 to 94.36 percent higher than control. Shoot dry weight increased the most (94.36 percent), followed by root volume (93.59 percent), shoot fresh weight (91.42 percent), root fresh weight (79.46 percent), and collar diameter increased the least (49.30 percent). Two-year-old seedlings were outplanted in two locations: one within and one beyond its normal zone of occurrence. Inoculated seedlings fared better at both sites in terms of survival and growth. S. polyrhizum inoculation has accelerated the growth metrics of P. gerardiana seedlings. Thus, it can be recommended that the P. gerardiana nursery should be raised by artificial inoculation with S. polyrhizum to produce healthy, tall planting stock in a shorter nursery time while also lowering maintenance costs.

Rhizopogon himalayensis (Castellano, S.L. Miller, Singh & Lakhanpal) A.B. Mujic & M.E. Sm., comb. nov. -an edible truffle-like fungus- normally exists in a symbiotic association with Cedrus deodara (Roxb.) Loud. Because of this important association and the ecological significance as per the available literature attached to this fungus, the present study was undertaken with a specific objective to test the mycorrhizal viability of this mycobiont with this important tree species of Himalaya -the theme tree of Himachal Pradesh. An attempt to investigate and record morphological and anatomical characteristics and variations in this mycorrhizal association was also made in nursery conditions using pure culture (wheat grain spawn) of R. himalayensis. The seedlings grown and inoculated in the nursery were harvested after six months for further examination, including the microscopic details. The study and subsequent analysis revealed that that mycobiont had invaded most of the feeder roots, imparting a typical swollen appearance to the mycorrhizal root tips. These root tips were light ochre with moderately thick plectenchymatic fungal mantle with occasional clamp connections on the inner layer of the fungal mantle. The root surface, as seen, was found smooth and frequently covered with a loose aggregation of inter-woven hyphae that uniformly pervaded the epidermis extending into the inter-cortical spaces of outer cortical cells and formed a characteristic Hartig net. Thus, results obtained in fact for the first time, presented an in-depth analysis of the morphological and anatomical characteristics of R. himalayensis and C. deodara association.

Kumar A, Tapwal A. 2022. Diversity of arbuscular mycorrhizal fungi and root colonization in Polygonatum verticillatum. Nusantara Bioscience 14: 53-63. Diversity of arbuscular mycorrhizal fungi (AMF) associated with Polygonatum verticillatum (L.) All. was investigated in two sites of Himachal Himalaya, India. A total of 15 AMF species were isolated and identified from the rhizosphere soil of P. verticillatum. The spore density was 1.48 ± 1.91 and 3.99 ± 3.78 per 20 gram of rhizosphere soil at site-I and site-II, respectively. Mycorrhizal colonization in the roots of P. verticillatum was recorded 46.12 and 52.23 percent at site-I and site-II, respectively. The mycorrhizal structures like darkly stained endophytic hyphae, coiled intracellular hyphae, Y-shaped hyphae and ‘H’ connection (Arum type) were recorded.

Western Himalaya is endowed with rich treasure of medicinal plant wealth. Bergenia ciliata is one of the important ethnomedicinal plants collected from Western Himalaya and used for dissolving kidney stones by the local inhabitants. In present work ethanolic rhizome extract of plant was evaluated for its medicinal potential by assessing its antimicrobial and phytochemical properties. In antimicrobial studies the ethanolic rhizome extract of the plant was used against standard and clinical isolates of selected pathogenic bacterial strains. Maximum inhibition zone (12±1.0mm) was observed against clinical isolate of S. typhimurium followed by clinical isolate of E. coli with an inhibition zone of 11.33±0.57mm. The MIC of B. ciliata rhizome extract ranged from 100 to 200μg/100μl against different pathogenic strains. The rhizome extract showed presence of reducing sugars, alkaloids, saponins, tannins, flavonoids and cardiac glycosides. Further, GC-MS analysis of rhizome extract confirmed the presence of phytochemicals viz., beta sitasterol, ethyl iso-allocholate, hexadecanoic acid, cetene, monolinolein TMS, berginin, gallic acid, linolool, β-caryophyllene, calcitriol, carotene, oleic acid and astaxanthin with significant ethnomedicinal importance.

Sparassis crispa is an edible mushroom exhibiting a wide range of medicinal properties. It is recognized for therapeutic value because of the high β-glucan content in the basidiomes. The broad range of its reported curative effects include anti-tumour, anti-cancer, immune-enhancing, hematopoietic, anti-angiogenic, anti-inflammatory, anti-diabetic, wound-healing, antioxidant, anti-coagulant, and anti-hypertensive properties. However, most of the studies are conducted on immunomodulatory and anticancer activities. Besides this, it also exhibits anti-microbial properties due to the presence of sparassol. Technology is now available for the cultivation of S. crispa on coniferous sawdust. This review is an attempt to focus on its distribution, taxonomy, chemical composition, medicinal properties, potential applications, and artificial cultivation.

This chapter attempts to trace the developments in ectomycorrhizal (ECM) research in India. The research on ECM in India was initiated at erstwhile FRI (now ICFRE) at Dehradun after independence in the late fifties by Dr. B.K. Bakshi and collaborators. They described the ECM in some important forest trees, viz., Pines, Deodar and high-altitude conifers. They isolated the mycorrhizal associates in pure culture and artificially inoculated the seedlings in a few cases. More impetus to ECM research was provided in the late seventies and early eighties from South, North and Eastern India. Natarajan, Raaman, Reddy and Mohan and their associates intensely investigated the ECM and its implications in seedling regeneration in Pinus patula and other pines. Sharma, Mishra and their students carried out similar studies on P. kesiya in the eastern Himalayas. Lakhanpal and collaborators studied the various aspects of ECM in Chir Pine, Blue Pine, Deodar, Fir, Spruce, Chilgoza Pine, Yew and apple plants in N.W. Himalaya. All these studies concerned characterization, identification, mycobiont association, physical and chemical status and artificial inoculation with selected mycobionts. Recently (2019, 2020) Atri and his students have carried out similar studies on ECM of sal trees in H.P. Tapwal and associates at HFRI, Shimla, are working on the ECM relationship of some important Himalayan forest tree species. The emphasis has gradually shifted now towards evaluation of field performance of the inoculated seedlings which is the primary aim of studies on this symbiotic association.

Aging is a complicated biological process in which functional and structural alterations in a living organism take place over time. Reactive oxygen species is one of the main factors responsible for aging and is associated with several chronic pathologies. The relationship between aging and diet is quite interesting and has attained worldwide attention. Healthy food, in addition to dietary antioxidants, are required to delay the process of aging and improve the quality of life. Many healthy foods such as fruits are a good source of dietary nutrients and natural bioactive compounds which have antioxidant properties and are involved in preventing aging and other age-related disorders. Health benefits linked with healthy consumption of fruit have drawn increased interest. A significant number of studies have documented the advantages of fruit intake, as it suppresses free-radical development that further reduces the oxidative stress created in the body and protects against several types of diseases such as cancer, type 2 diabetes, inflammatory disorders, and other cardiovascular diseases that ultimately prevent aging. In addition, fruits have numerous other properties like anti-inflammatory, anti-cancerous, anti-diabetic, neuroprotective, and have health-promoting effects. Mechanisms of various bioactive compounds that aids in preventing various diseases and increases longevity are also described. This manuscript provides a summary of various bioactive components present in fruits along with their health-promoting and antiaging properties.

The ethnomedicinal plant extract of Trillium govanianum was assessed for its antimicrobial activity and for the presence of phytochemicals to provide scientific support for its ethnomedicinal use. Ethanolic rhizome extract of the plant was used for the antibacterial study and displayed a maximum inhibition zone against a standard isolate of K. pneumoniae (16.66 ± 0.57) followed by an inhibition zone against clinical isolate of K.pneumoniae (16.33 ± 0.57) at 200 μg/disc. The antimicrobial activity of plant rhizome extract was observed against five pathogenic microbial strains, with both standard strains and clinical isolates viz. Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Salmonella typhimurium and Pseudomonas aeruginosa. Standard analysis of phytochemicals in the plant showed the presence of flavonoids, cardiac glycosides, alkaloids, reducing sugars and saponins, however, tannins were found to be absent. In GC–MS profiling the plant rhizome extract showed the presence of important compounds possessing antibacterial, anticancer, antioxidant and antiinflammatory properties. DPPH and FRAP assays showed optimum antioxidant potential of the plant. Antimicrobial and phytochemical investigation of the plant rhizome extract revealed therapeutic potential of Trillium govanianum, thus scientifically supporting its ethnomedicinal use.

Biodeterioration of indoor wood and microbial pollution inside buildings is one of important problem in humid areas. Number of fungi are associated with indoor wood and many of them are responsible for its deterioration by causing decay and stain. Some of them may release mycotoxins, which have hazardous impact on human health. In present study, fifteen fungi associated with wood surface were isolated and out of which genus Aspergillus dominate with four species. A. flavus was recorded from all locations. The potential of leaf extracts of nine tree species on the growth of A. flavus was evaluated by amending culture media. Considerable growth inhibition of A. flavus in the range of 1.14-45.45% was recorded on solid media and 9.37-86.66% in liquid media. Amendment of culture media @ 30% concentration of the leaf extract of Corymbia torelliana have recorded maximum growth inhibition irrespective of the media used.

Tapwal A, Pradhan S, Chandra S, Rashmi. 2016. Antimycotic activity and phytochemical screening of fungal endophytes associated with Santalum album. Nusantara Bioscience 8: 14-17. The heartwood of Santalum album constitutes the central part of the tree is valued for its fragrance. The wood and oil are utilized in medicine. Sandalwood oil is extensively used in perfumery, cosmetics, aromatherapy, and pharmaceutical industry. The endophytic microorganisms inhabiting the plant tissues are expected to mimic some of the metabolites of its host. This study was aimed to isolate and screen the fungal endophytes inhabiting the Santalum album for antimicrobial activity and for the presence of important phytochemicals. Five fungal endophytes isolated from different parts of S. album have exhibited antimicrobial potential against Fusarium oxysporum in the range of 5.0-40.4%. The isolated endophytic fungi also indicated the presence of alkaloids, phenolics, and tannins, flavonoids, carbohydrates and glycosides, terpenoids, amino acids and proteins.

Tapwal A, Kumar R, Borah D. 2015. Effect of mycorrhizal inoculations on the growth of Shorea robusta seedlings. Nusantara Bioscience 7: 1-5. Shorea robusta is one of important timber yielding tree species of northeast India and known to have both ectomycorrhizal (EcM) and endomycorrhizal (AM) associations. It is hypothesized that under favorable conditions different mycorrhizal fungi present in soil develop symbiotic association with fine roots of trees. In present investigations, mycorrhizal inoculum of EcM and AM fungi applied to S. robusta seedlings raised in polyethylene bags in nursery. Observations on growth characters and mycorrhizal colonization were recorded at the interval of three months. The results revealed that irrespective of type of mycorrhizal inoculation, growth of the seedlings increased significantly in comparison to control. Maximum growth was observed for the seedlings inoculated with EcM alone, followed by dual inoculations (EcM+AM), seedlings inoculated with AM fungi and minimum in control.

Kumar R, Tapwal A, Pandey S, Raja-Rishi R, Mishra G, Giri K. 2014. Six unrecorded species of Russula (Russulales) from Nagaland, India and their nutrient composition. Nusantara Bioscience 6: 33-38. The young and matured carpophores of mushrooms were collected from the forests of Puliebzie, Zakhama, Pherma, Mankoi, Chungtia, and Tigit located in different districts of Nagaland, one of the north-eastern states of India. All the species were found associated with Pinus khasya, P. caribaea, P. patula, Cryptomeria japonica, Canarium resiniferum, Dipterocarpus macrocarpus, and Shorea assamica. The collected mushrooms were identified as Russula species on the basis of their macro and microscopic characteristics. The identified Russula species were also screened for their nutrient content. The protein and carbohydrate content was found to vary between 28.12 to 42.86, and 49.33 to 55%, respectively. These species [Russula aeruginea Lindblad; -Fr., R. alnetorum Romagnesi, R. brevipes Peck, R. fragrantissima Romagnesi, R. nobilis Velen. and R. ochroleuca (Pers.) Fr Gray.] are a new record from Nagaland state of India.

The antifungal activity of aqueous extract of Cannabis sativa, Parthenium hysterophorus, Urtica dioeca, Polystichum squarrosum and Adiantum venustum was investigated against Alternaria solani, Alternaria zinniae, Curvularia lunata, Rhizoctonia solani and Fusarium oxysporum at different concentrations (5, 10, 15 and 20%). At 20%, maximum antifungal potential was observed with the extracts of C. sativa, which recorded excellent inhibitory activity against C. lunata (100%), A. zinniae (59.68%), followed by leaf extract of P. hysterophorus (50%) against A. solani. The application of botanical extracts for disease management could be less expensive, easily available, non-polluting and eco-friendly.

In the present study diversity of arbuscular mycorrhizal fungi (AMF) was analyzed in seven important ethnomedicinal plants viz. Trillium govanianum Wall. ex D. Don, Bergenia ciliata (Haw.) Sternb., Leucas lanata Wall ex. Benth, Rumex obtusifolius L., Justicia adhatoda Nees, Perilla frutescens (L.) Britton and Acorus calamus L. The AMF diversity in rhizosphere and in the roots was investigated and reported for the first time in L. lanata, R. obtusifolius, T. govanianum and B. ciliata of district Shimla, Himachal Pradesh, India in the western Himalaya. Maximum mycorrhizal spore count was observed in the rhizosphere of P. frutescens. The most predominant genus was Glomus with fifteen species followed by Acaulospora, Gigaspora, Scutellospora, Entrophospora and Sclerocystis.

Sap-stain fungi attack the wood immediately after felling or sawing and cause reduction in the value of timber by discolouring sapwood. Wood blocks of three tree species (Populus deltoides, Syzygium cumini and Mangifera indica) were artificially inoculated with three sap-stain fungi (Lasiodiplodia theobromae, Trichoderma harzianum and Ceratocystis sp). After 5 weeks of incubation 1.97 to 9.68 per cent weight loss was recorded in wood blocks inoculated with sap-stain fungi. Maximum weight loss was recorded in the wood block of P deltoides followed by S cumini and M indica. The in vitro evaluation of selected fungicides on sap-stain fungi revealed that carbendazim and benomyl were quite effective while copper oxychloride was least effective.

Kumar R, Tapwal A, Pandey S, Rishi R. 2013. Fungal diversity associated with bamboo litter from Bambusetum of Rain Forest Research Institute, Northeast India. Biodiversitas 14: 79-88. Fungi play an important role in leaf litter decomposition due to their ability to break down the lignocelluloses matrix, which other organisms are unable to digest. Diversity of bamboo leaf litter fungi from fallen leaves and undergoing active decomposition leaves in different season and different depth was carried out in 2009-10. Twenty four samples were collected from Bambusetum of Rain Forest Research Institute (RFRI),  Northeast India. The moist chamber, direct isolation and dilution plate methods were used to assess the diversity of fungal species. Fungi were cultivated on 3% malt extract agar and half strength potato dextrose agar. The litter was divided into freshly fallen senescent leaves (grade 1) and leaves already undergoing active decomposition (grade 2). Moist chamber incubation of the litter revealed 45 fungal taxa belonging to 22 genera. fungal taxa were found on grade I and 39 fungal taxa found on grade II litter. Although 24 fungal taxa were common to both grades, Differences were observed in percentage occurrence of fugal species between the two grades of litter. Periodic surveys were carried out to collect macrofungi. Young and matured carpophores of 16 macro fungi species were collected in different seasons. Out of these macrofungi, 3 species belongs to family Entolomataceae and Agaricacea, two species belongs to Tricholomataceae and Geoglossaceae one species belongs to each family Dacrymycetaceae, Pluteaceae, Coprinaceae, Marasmiaceae Lycoperdaceae and Phallaceae. The bamboo leaf-litter was selected for the present syudy because of the dominance and great economic value of bamboo vegetation in North-east India.

Tapwal A, Kumar R, Pandey S. 2013. Diversity and frequency of macrofungi associated with wet ever green tropical forest in Assam, India. Biodiversitas 14: 73-78. A study was conducted in Jeypore Reserve Forest located in Assam, India to investigate the diversity of macrofungi associated with different tree species. The diversity of broad leaves trees and high humidity during monsoon period favours ideal growth of diverse group of macrofungal fruiting bodies. Thirty macrofungal species representing 26 genera belonging to 17 families were collected from six different sites in the study area. Out of these maximum six genera assignable to family Polyporaceae, five genera to Russulaceae, three genera to Agaricaceae, two genera to Ganodermataceae and Cantharellaceae each and rest of the families were represented by single genus only. The ecological preference of the species reveled that maximum (17) species were saprophyte, living on dead substrates or decaying wood debris, ten species were found associated with roots of higher trees, while three species were found parasitic. Overall 20 species were found edible including some species having medicinal utilization. The present study revealed that maximum frequency of occurrence was exhibited by Trametes versicolor and Schizophyllum commune (83.33%), followed by Microporus xanthopus, Pycnoporus sanguineus (66.67%) and Coprinus disseminates (50%). The rest of the species exhibited the frequency distribution ranging between 16.67-33.33%. The maximum density was recorded for Schizophyllum commune (126.67%) followed by Trametes versicolor (120%) and Xylaria polymorpha (93.33%). The density of rest of the species were ranged between 3.33- 6.67%. The key objective of the present study was to generate a database on macrofungal diversity of Jeypore Reserve Forest along with their ecological preferences and utilization, which is not earlier documented.

Kumar R, Tapwal A, Pandey S, Borah RK, Borah DP, Borgohain J. 2013. Macro-fungal diversity and nutrient content of some edible mushrooms of Nagaland, India. Nusantara Bioscience 5: 1-7. The northeast region of India abounds in forest wealth, including variety of flora and fauna. The high humidity during monsoon period provides ideal atmospheric conditions for the growth of diverse group of macrofungal fruit bodies. Nagaland, the northeastern state of India is rich in biodiversity and encompasses large numbers of edible and non-edible mushroom species. Young and matured carpophores of 15 wild edible mushroom species were collected from 12 locations in different districts of Nagaland. Out of these four species belongs to family Agaricaceae, two belongs to Tricholomataceae and rest belongs to Boletaceae, Cantharellaceae, Russulaceae, Sarcoscyphaceae, Auriculariaceae, Polyporaceae, Schizophyllaceae, Pleurotaceae, and Lyophyllaceae. The selected species were analyzed for proximate analysis of nutritional values. The protein content varies from 22.50-44.93% and carbohydrates were recorded 32.43-52.07% in selected species. The documentation of wild edible mushrooms is very scanty in Northeast India. The key objective of the present study was to generate a database on macrofungal diversity, ecology, ethnomycology, utilization and nutrient status of important wild edible mushroom species of Nagaland, which forms a part of the food culture of the native peoples.

Kumar R, Tapwal A, Pandey S, Rishi R, Borah D. 2013. Observations on arbuscular mycorrhiza associated with important edible tuberous plants grown in wet evergreen forest in Assam, India. Biodiversitas 14: 67-72. Non-timber forest products constitute an important source of livelihood for rural households from forest fringe communities across the world. Utilization of wild edible tuber plants is an integral component of their culture. Mycorrhizal associations influence the establishment and production of tuber plants under field conditions. The aim of present study is to explore the diversity and arbuscular mycorrhizal (AMF) colonization of wild edible tuber plants grown in wet evergreen forest of Assam, India. A survey was conducted in 2009-10 in Sunaikuchi, Khulahat, and Bura Mayong reserved forest of Morigaon district of Assam to determine the AMF spore population in rhizosphere soils and root colonization of 14 tuberous edible plants belonging to five families. The results revealed AMF colonization of all selected species in all seasons. The percent colonization and spore count was less in summer, moderate in winter and highest in rainy season. Seventeen species of arbuscular mycorrhizal fungi were recorded in four genera viz. Acaulospora (7 species), Glomus (5 species), Sclerocystis (3 species) and Gigaspora (2 species).