Diversity and Ethno-Mycological Importance of Mushrooms from Western Himalayas, Kashmir


 Wild edible mushrooms (WEM) are economically significant and used in traditional medicines worldwide. The region of Jammu and Kashmir (Western Himalayas) is enriched with the diversity of edible mushrooms, collected by the rural people for food and income generation. This is the first detailed study on diversity and ethno-medicinal uses of mushrooms from the State of Jammu and Kashmir. Methods: Consecutive surveys were conducted to record ethno-mycological diversity and socio-economic importance of wild edible mushrooms value chain in rural areas of Azad Jammu and Kashmir during 2015-2020. Data were collected with a semi-structured questionnaire having a set of questions on indigenous mycological knowledge and collection and retailing of wild edible mushrooms. A total of 923 informants from the study area provided results identifying the gender, type of mushroom species, medicinal uses, and marketing of mushrooms. Principal component analysis (PCA) was also applied to the data set to analyse the relationship between species distribution, the underlying environmental factors and habitat types. PCA identified the major species specific to the sites and put them close to the sites of distribution.Results: A total of 131 mushroom species were collected and identified during 2015-2020 from the study area. One hundred and one species of mushrooms were reported new to the State of Jammu and Kashmir. The dominant mushroom family was Russullaceae with 23 species followed by Agaricaceae, 16 species. Major mushroom species identified and grouped by the PCA were Coprinus comatus, Lactarius sangufulus, Amanita fulva, Armillaria gallica, Lycoperdon perlatum, Lycoperdon pyriforme, and Russula creminicolor. Sparassis crispa, Pleurotus sp and Laetiporus sulphurous were recorded most edible and medicinally significant fungi. Morels were the most expensive and medicinally important among all harvested macro-fungal species. These were reported to use against the common ailments and various health problems. Conclusions: Collection and retailing of WEM contribute to improve the socioeconomic status, providing alternative employment and food security to rural people of the area. These mushrooms are used as a source of food and traditional medicines among the rural informants and could be used a potential source of antibacterial and anticancer drugs in future.

minerals, and other nutrients (24,3). They have important antioxidant and biologically active compounds and used worldwide from ancient times in traditional medicines due to health-promoting and immune-stimulating properties [25,26,27). Different functional bioactive compounds Cardiac glycosides Anthraquinones, avonoids, Tannins, Terpenoids and proteins were reported from mushrooms [28,29,30]. Water extract of Chaga mushrooms contains potent anti-cancer compounds [31]. Mushrooms are gaining importance by researchers worldwide due to their nutritional and pharmacological importance [32,33]. Biomolecules of mushrooms have good biological and medicinal potential against different diseases [34].
Morels (Morchella) have been used as food with various health bene ts [35] Different polysaccharides have been identi ed from Morchella sextelata with immune-modulating properties [36]. Morchella esculenta has a wide variety of antioxidant and antitumor compounds and is used as a source of food and traditional medicines [37]. It contains polyunsaturated fatty acids, carbohydrates, and bioactive compounds with antibacterial activity [38]. The whole region of AJK is blessed with diverse geographic and climatic conditions with a diversity of mushrooms but there are no comprehensive studies have been taken previously to explore such resources for human welfare. There is a lack of proper documentation on the diversity, speci c habitat, ethno-mycological uses, production, harvesting and export of mushrooms. Present research work will contribute towards a detailed overview of the species diversity of mushrooms in AJK, their ethno-mycological uses and commercial and economic importance.

Study area
The study area lies in the Western Himalayan regions of Azad Jammu and Kashmir between 32°-17 and 36°-58 North latitude and 73°-6 and 80°-30 longitude in the western part of the Indian sub-continent with an area of 13297 square Kilometres. The elevation from sea level ranges from three-sixty meters in the south to 6325 meters in the north. Average annual rainfall 1300 mm. The population is 4 million and the ratio between rural to urban populations is 88:12. Forestry, livestock and agriculture are major economic activities for rural income. The climate of the study area is Subtropical monsoon type in the lower range to moist Temperate in the middle and Subalpine to Alpine in upper regions. The summer is hot at lower altitudinal zones and pleasant in upper zones with very cold winters. The area above 1200 m altitude receives heavy snowfall from November to April. The average temperature recorded in summer remains 34°C to 25°C and in winters, 10°C to 4°C. Annual rainfall (average) in the monsoon region is 900-1300 mm and in monsoon free region it remains 35-140 mm [39].
All the major terrestrial ecological sites and hotspots for mushroom species from the state of Azad Jammu and Kashmir were selected for this study. Sampling sites were nalized through consecutive eld visits based on speci c geographic and ecological signi cance from representative vegetation zones of Azad Jammu and Kashmir. A total of 21 sites were selected from Neelum, Muzaffarabad, Hattian, Bagh, Heveli, Poonch and Kotli districts of Azad Jammu and Kashmir during 2015-19 to study mushroom diversity ( Figure 1 & Table 1).

Collection and preservation of sporophores
Sporophores of fungi were collected from Cedrus deodara and Pinus wallichiana forest communities. Sporocarps were collected by using standard methods (Gateri et al., 2014). Ethno-mycological knowledge was obtained from different eld visits and semi-structured questionnaires and interviews with rural people. A speci c collection number was assigned to each sample in triplicate. Speci c characters of habitat and associated plant species were also recorded. Sporophores were cleaned gently, soil particles were removed, and photographs were taken with a digital camera Nikon D5600. Morphological characters of the sporophores were recorded during collection in the eld. Fruiting bodies were left into the air for drying before packing for preservation. For easy drying, the larger Sporophores were cut down into many smaller pieces. Dried samples were packed and labelled with separate tag numbers for further analysis and future references. A sample of the selected type of mushroom was assigned a voucher number and carried to the laboratory of Botany, University of Punjab, Lahore, Pakistan for detailed morpho-anatomical examination. Specimens were nally cross-checked with the published material, literature at the morpho-anatomical level. Appropriate taxonomic literature was used for the proper identi cation of mushrooms [40,41,42,43,44). Further citations were checked on mycobank and the index fungorum database. Final identi cation was made from fungal biology and systematic research laboratory Department of the Botany University of the Punjab Lahore after studying detailed morpho-anatomical study (identi cation keys and published material). Voucher specimens were deposited with the accession numbers at the Herbarium of Botany, University of Azad Jammu and Kashmir Muzaffarabad.

Results
Demographic characteristics and community involvement Wild mushroom value chain is seen to be gender oriented dominated by women in collection (61.1%, n=564) while men occupy only 38%, n=359 out of the 923 respondents (Table 2). Women were found to participate in every mushroom activity such as collection to preservation while men contributed only to collection and selling. Similar ndings were reported by [6] where female was found dominant in WEM collection. However, it was found that men dominated in selling of mushrooms (70%) to local shops, restaurant, markets, and local mushrooms entrepreneurs. The preponderance of female collectors in present study is supported by another research [45,46,47). Every stage of mushroom activities from collection to processing and even marketing was led by women in this study. Poor involvement of men in mushroom activities might be due to the belief that mushroom collection is only art for remote areas of women. In remote areas of studied districts of AJK, women are mostly unemployed, dedicating themselves to household and subsistence activities. Mushroom collection and selling are one of their sources of food and income. The study revealed that collection activities are dominated by people of middle age (53.9%) especially those of 31-50 years old between the ages ranged 14-85, followed by 19-30 (25.8 %), by 14 and over (17.6 %), and by 50 and above (13.3 %) ( Table 2). Similar ndings were also reported from the Finland [48] where it was shown that middle aged people by 30 (96.6 %) or above involved in mushrooms collection activity. It revealed the participation of older, more experienced people in mushroom collection. Similar results on age distribution were also reported by [47]. Among 923 respondents, 25.8 % had an education level of primary school, 22.8 % middle school, 20.9% % secondary or high school, 17 % illiterate and 13.5 % higher secondary, university or colleges ( Table 2). There were 41 % housewives 39.7 % farmers & entrepreneur, 12.6% employed, 6.7 % retired from 923 respondents ( Table 2). Data on education in the present study revealed that almost 83 % informants had a middle school education in accordance with the ndings of [49] who indicated that mushroom collection or cultivation were mostly managed by less educated people in the rural areas.

Collection and identi cation mushrooms
A total of 131 mushroom species were collected and identi ed during the study. Out of 131 mushroom species, 101 species of mushrooms were recorded new to the state of Azad Jammu and Kashmir (Figure 2), however few of these species have been identi ed from different parts of Pakistan at molecular level previously. Already identi ed mushroom species were morphologically cross checked with published material. Many of these species were collected by the rural peoples based on folk taxonomy and only a few are considered edible. The dominant mushroom family was Russulaceae with 23 species followed by Agaricaceae, 16 [52,53] also contributed to mushroom ora of AJK. They reported 25 edible mushrooms from different sites of the state.

Mushrooms edibility in the study area
The state of Azad Jammu &Kashmir (AJ&K) has a land of rich diversity of wild mushrooms which might have been contributed by the tropical and moist temperate forests, mostly Quercus and coniferous woodlands, and higher rainfall and annual precipitation. Among the identi ed wild mushrooms, 54 (48%) were identi ed as edible only, 24 (21%) as inedible, 14 (12%) as edible and medicinal ( Figure 3). The detail of different categories of identi ed wild mushrooms with their percentage is given in gure (3).

Principal component analysis
PCA is used to determine and analyse the relationship between species distribution and the underlying environmental factors and habitat types. It is an advanced technique which maximizes the species scores with respect of sampling sites having linear and appropriate weights. PCA identi ed the major species speci c to the sites and put them close to the sites of distribution. The sites grouped together by the PCA based upon their species interrelationship are Pir Chinasi, Haji peer and Peer hasimar, Toolipeer, and Leepa. All these sites have little variations in the biotic factors including species composition and topography. These sites have some common geographic features which are responsible for the similar species composition.
Major mushroom species collected from these sites and grouped together by the PCA are Coprinus comatus, Lactarius sangufulus, Amanita fulva, Armillaria gallica, Lycoperdon perlatum, Lycoperdon pyriforme, and Russula creminicolor, these sites have shown a little correlation with a village Khawaja bandi kahuta Havali. The mushroom species grouped together by the PCA are the common fungi which are present in these sites.
On the other hand, Nagdar (Upper Neelum), Dawarian, Sharda, Taobut, Chakar (Noonbangla), Sudhan Gali and Banjosa are grouped near to each other. These sites are almost lying in the temperate forest of AJK and almost have same topography, Forest cover and precipitation pattern so their mushroom composition is nearly like each other. Major fungal species of these sites were Amanita muscaria, Lactarius deliciosus, Gyromitra esculenta, Armillaria sp, Agaricus campestris, Russula breviceps, Polyporus squamosus, Trametes versicolor and Laccaria sp. Other mushroom species grouped at the centre of the PCA axis show that these species are almost equally distributed and present in all the sites of the study area. These species have no speci c distribution pattern. These species are most common and grow almost equally in different geographic condition with slight changes in their growth period and maturation.
PCA identi ed ve major keystone species from the data matrix and separated them along X-axis. Lactarius piperatus, L. deliciosus, L. torminosus, Hygrocibe avesence and Russula delica were extracted as most signi cant vectors having maximum Eigen vale scores represented by their distinct placement on PCA biplot. These ve species were characterized by the higher IVI values in the species dataset and enjoyed abundance and broad distribution across the study area. The major bulk of the fungal elements were clustered in the centre of the PCA biplot showing their random distribution without speci c site or habitat preference (Fig. 4).

Discussion
Ethno-mycological and socio-economic importance of wild mushrooms A total of 923 informants from 22 sites of selected districts were interviewed based on the harvesting, selling and consumption of wild edible mushrooms. Consecutive eld visits were carried out to different villages, local markets, shops of the study area for gathering of information about mushroom collection and selling. A semi-structured questionnaire (Appendix.1) was used to collect the information on wild edible mushrooms value chain, hunting, collection, preservation, and retailing [54]. Primary and secondary information was collected from all the available resources. Primary information gathered by structured and semi structured interviews with collectors, consumers, and sellers. Secondary information was collected from different literature, thesis, maps, and web sites. Both formal and informal discussions with forestry professionals, key informants, village elders, farmers, women, schoolteachers, social workers, shopkeepers were carried out to identify and verify the facts. Information on edibility, medicinal uses, preservation methods and any other uses was also collected. Mushrooms play a signi cant role in rural development. Many species of edible mushrooms and morels have been collected by the poor rural for socioeconomic purpose [55] and rural livelihood in terms of economic development [56].
Morels are collected by the people of rural areas of AJK for medicinal and commercial purposes. Morchella conica, M. costata, Morchella esculanta, M. elata and M. tridentina were considered highly prized morel species. These morel species widely grow under the dense forest cover of Pinus wallichiana and Cedrus deodar in association with Viburnum grandi orum. Among morels, Morchella esculanta and M. tridentina were valuable morels and due to compact fruiting bodies, less moisture, and higher nutritional contents and considered good for export. M. conica has more water contents than the M. esculanta and turns dark black, which affects the preservation as well as its marketing. Edible fungi i.e., Cantharellus cibarius, Lactarius deliciosus, Russula sp were collected and sold in the market for food purposes [57]. Morchella species were collected mostly due to their commercial importance as one kilogram of dried morel was solid in the market up to 32 thousand PKR. One kilogram of dry morels can full ll the basic needs of a family of average size. Prices of dried morels vary from market to market. In a village (Neelum) average price of 1kg of dried morel was 30000 PKR. Other edible mushroom species Pleurotus ostreatus and Agaricus campestris were supplied to the famous hotels of the city. One Kg of dried mushroom was sold in 1500-2000PKR. These mushrooms are mostly used in dishes for foreign visitors. Mushrooms are collected worldwide as a source of income. More than 300 species of mushrooms were collected by different ethnic groups in Mexico for nutritional and medicinal purposes [58]. In China, local farmers earn up to 62% of their cash income through mushroom export [59].
Prices of dry mushrooms in this region were higher than fresh mushrooms. Similarly, those mushrooms which were exported showed higher prices. The most common species collected and used for trade-in neighbouring countries of Pakistan were e.g. Boletus spp. Lactarius spp., Thelephora ganbajun, Suillus bovinus, Russula sp. and Termitomyces spp. [59,60]. In the present investigation, the socio-economic data showed that a family collects an average of 3-4 kg morels with an average income of about Rs. 100000-120000 in a season. Mushrooms are collected and exported from Pakistan to the neighbour countries for revenue generation. Fifty-six (56) species of mushrooms were reported as edible previously from Pakistan and unfortunately because of over-collection, urbanization, and deforestation some species are threatened [52]. Mushrooms are natural sources of bioactive compounds used in alternative traditional medicines. Today, in parallel with the increase in the number of diseases, alternative medicine and their usage is increasing due to the insu ciency of synthetic medicines and their disadvantages or side effects. Mushrooms have compounds that decrease oxidative stress and improve health [61,62].
Many unexplored species of medicinally and commercially important mushrooms were widely distributed in the forests of Azad Jammu and Kashmir. Mushroom species growing naturally were collected by the rural people for food and medicines.
Previously we reported medicinally signi cant mushrooms from the Neelum Valley [78]. They are also collected in different advanced countries of the world like the United Kingdom, Sweden and France [64]. In the present study, Twenty-six (26) species of mushrooms were recorded as medicinally important which are used for the treatment of some common ailments.
Among these mushrooms Fistulina sp, Hericium erinaceous, Laetiporus sulphurous, Polyporus squamosus, Ramaria fennica, Sparassis crispa, Morchella elata, M. conica, M. tridentina and M. deliciosa were the most delicious and widely used species as a nutritive food by the rural people of Neelum valley and Hattian in Jhelum Valley. Morchella esculanta is reported to contain antioxidant, anticancer and anti-in ammatory properties and is used as delicious food [62]. Soup of dried fruiting bodies of Ramaria fennica is used by women during breastfeeding to improve lactation. Ramaria fennica and morel species were considered effective against common cough and cold. Many mushroom species are considered medicinally important and used against stomach problems, heart burning and wound healing without considering any side effects or toxicity.
Previously it is reported that extract and powder of mushrooms are used in traditional medicines and have reported uses as a liver tonic, blood puri ers, fertility issue and diabetes [65]. Fruiting bodies of Laetiporus sulphureus were dried into a ne powder and used with milk as a portion of healthy food and anti-seminal weakness. Previously it is reported that Laetiporus sulphureus is used against speedy recovery of wounds and common cold [6]. In another study, it is found that dry powder of this mushroom is helpful to expel a retained placenta in women and against stomach pain [66]. Use values of mushrooms are given in (Table. 3). In the present study, we have found the use of morels in different traditional home remedies against common ailments, fever, cough and cold. Soup of Morchella is considered nutritious and used to treat the common cold. Extract of many edible species of mushrooms is effective against different human diseases like coronary disorders, oxidative stress, and cancer and provides different physiological bene ts to consumers [67]. Sparassis crispa and Polyporus squamosus were used to treat stomach issues and considered healthy food. Old villagers prefer to use these mushrooms as a source of food and traditional medicines. People use Morchella species, Hydnum repandum, Sparassis crispa and Polyporus squamosus for stomach problems, Lycoperdon perlatum and Auricularia auricula in wound healing and as anti-hypertension. Armillaria mellea, Boletus badius, Cantharellus cibarius, Pleurotus ostreatus and Lactarius deliciosus contain bioactive organic contents, p-coumaric protocatechuic, ferulic, sinapic, p-hydroxybenzoic, vanillic and Cinnamic with reported uses in traditional medicines [68]. [69] reported that morels were utilized both for food as well as medicines to cure different diseases.
Ethno-mycological uses of mushrooms vary from region to region and even among the communities of the same area [70].
Extract of mushrooms can be used due to cosmeceutical and nutricosmetic ingredients to treat in ammatory skin disease and hyperpigmentation [71]. Aqueous Extracts of Polyporus squamosus, Morchella spp and Sparassis crispa are considered more effective against common diseases of the stomach by the rural informants of Kashmir. As it is reported that mushrooms are effective against different diseases, but chemical evaluation is very important before using an extract of mushroom species [72]. It is concluded that mushrooms, potentially can provide opportunities to rural communities to generate income for household's development in rural areas of Azad Jammu and Kashmir. Mushroom collection can provide opportunities to the low-income areas to improve their living standards in terms of income generation and socio-economic development. It is very important to raise awareness among the local communities/mushroom collectors, about the importance of mushrooms as food and medicines. Mushrooms, if well addressed in society, are a potential source of traditional medicines, anti-cancer compounds, food, and nutrition security speci cally in developing countries. Availability of data and materials.
Data sharing not applicable to this article as no datasets were generated or analysed during the current study.
Ethics approval and consent to participate. Not Applicable.
Consent for publication. Not applicable.
Competing interests. The authors declare that they have no competing interests   Appendix1.docx