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Journal of Plant Health Issues DOI:XXXXXXXXXXXXXX Print ISSN XXXX-XXXX- December 2020 Online ISSN XXXX-XXXX JPHI 2020 1(3):061-068 Article JPHI0015 Research Article Native Diversity of Endotrophic Mycorrhizal Fungi of Forage Grass Species Occurring in Asan River Basin, Mussoorie Hills, Dehradun, Uttarakhand Megha1, Vipin Parkash1*, Ramesh Chhetri2, Akshita Gaur1 and Rahul Agnihotri1 1Forest Pathology Discipline, Forest Protection Division, Forest Research Institute (Indian Council Forestry Research & Education, Autonomous Council under Ministry of Environment & Forests, Government of India), Dehradun, Uttarakhand (248 006), India 2Kali Gandaki Polytechnic Institute, CTEVT, Ghiring - 1, Tanahun, Gandaki Pradesh, Nepal Open Access Abstract Corresponding Author Arbuscular Mycorrhizal (AM) fungi colonize more than 80% of plants on land in which grasses are known to have higher endomycorrhizal colonization. AM Vipin Parkash fungi are a common group of symbiotic fungi in the order, Glomale of Division, e-mail: [email protected] Zygomycota. These fungi are known to benefit growth through increased nutrient Keywords uptake especially phosphorus. In this study, a total of 21 grass species collected from the Asan river basin, Mussoorie hills, Dehradun, Uttarakhand were screened Abiotic stress, Arbuscular mycorrhiza, Mycorrhizal for AM fungal root colonization and their mycorrhizal diversity. Traditional method symbiosis, Plant microbe interaction, Root of sieving and decanting was used for isolating mycorrhizal spores whereas for colonization, Spore count studying colonization rapid staining and clearing method was used. The highest Conflict of interests: The author has declared that root colonization (95 ± 2.9) and AM spore count (234 ± 3.56) were observed in no conflict of interest exists. Phalaris minor whereas Saccharum spontaneum exhibited least colonization (30 ± 0.53) and AM spore count (46.7 ± 14.5) respectively. The Andropogoneae How to cite this article? (Sorghum tribe) was observed to be the most diverse tribe in association with Megha et al., 2020. Native Diversity of Endotrophic endomycorrhizal fungi among the studied grasses. This study confirms that the Mycorrhizal Fungi of Forage Grass Species grass species are highly colonized and dependent on endomycorrhizal association. Occurring in Asan River Basin, Mussoorie Hills, The diversity and colonization patterns of endotrophic mycorrhizal fungi are Dehradun, Uttarakhand. Journal of Plant Health described in details in this research paper. The AM fungal association with grass Issues 1(3): 0061-068. species provides new vistas and insight on the functioning of any grass ecosystem and also helps in harnessing the benefits of AM fungi through their usage in waste and abundant land reclamation programmes. Introduction extensively in terms of improving soil structure and its water holding capacity (Candido et al., 2015). AM fungi are Microorganisms are present in great numbers near the well known to improve plant growth and production under feeder roots of plants and they play vital roles in numerous abiotic stress conditions e.g. salinity, drought, cold, heat, physiological processes (Marx, 1975). Arbuscular mycorrhizal mineral deficiency and metals (Daei et al., 2009; Chen et (AM) fungi are microscopic fungi that colonize the roots al and their rhizosphere at the same time and spread out over ., 2015). These fungi act as bio fertilizers, bio protectors numerous centimeters (Bonfante and Genre, 2010). AM and biocontrol agents and significantly have benefits to the fungi are primarily involved in nutrient transfer from soil plants (Sreenivasa and Bagyaraj, 1989; Hoeksema et al., 2010; particularly phosphorus and other poorly mobile nutrients Jeffries and Barea, 2012). The term ‘endomycorrhiza’ is used (Sanders and Tinker, 1973; Rhodes and Gerdemann, 1975; to describe mutualistic symbiotic associations between certain O’Keefe and Sylvia, 1991). AM fungal hyphae contribute fungi and plant roots in which the fungal partner grows mainly Article History RECEIVED in 20th September 2020 RECEIVED in revised form 23rd October 2020 ACCEPTED in final form 25th October 2020 Journal Home: www.bioticainternational.com © 2020 061 Megha et al., 2020 inside the root cortex and penetrates the cells of the host This study was undertaken with the objective of studying plant root. The symbioses between arbuscular mycorrhizal AMF association and diversity among different grass species. fungi and plants could also be important because 80% of The survey and collection of different grass species samples/ all terrestrial plant species form this type of symbiosis in rhizospheric root samples was done from different study areas nature (Van der Heijden et al., 1998; Hart and Reader, 2002). in the Asan river basin, Mussoorie hills, Dehradun. Laboratory Knowledge and understanding of endotrophic mycorrhizal experiments on endotrophic mycorrhizal association were status on grass species is necessary for perceptive relation carried out in the Forest Pathology Division, Forest Research of eco-system functioning. According to (Read, 1991) who Institute, Dehradun, Uttarakhand. The collected samples suggested that in the situation of the low mean annual were studied for root colonization, AM spore count in temperatures, rates of mineralization of nutrients are slow its rhizosphere and their mycorrhizal diversity. Further and result in lack of the availability of major nutrients like biochemical analysis and forage quality analysis experiments nitrogen and phosphorus. The endotrophic mycorrhizal are still under investigation. associations have many benefits to plant communities in an A total of 21 samples of grass species were collected from ecosystem. Mycorrhizal associations have direct correlations different sites and their specimens were preserved in with the major climatic factors i.e. water, temperature, etc., herbarium sheets for identification at Botany Division, Forest which regulate the distribution of plants as well as more Research Institute, Dehradun. Rhizospheric soil samples were localized edaphic conditions (Brundrett, 1991; Smith and taken by digging out a small amount of soil (200 g) close to Read, 2008). There is a very limited number of research studies plant roots up to the depth of 5-30 cm and these samples were concerned with correlation of mycorrhizal association with taken into a polythene bag for further study. To study the root different grass species. Most endotrophic mycorrhizae are colonization of arbuscular mycorrhizae, the rapid clearing and ‘balanced’ mutualistic associations in which the fungus and staining method was employed (Phillips and Hayman, 1970). plant exchange commodities are required for their growth The percentage mycorrhizal root colonization was calculated and survival (Brundrett, 2004). The aim of this study is to using the following formula. investigate whether grass species, which naturally coexist, respond differently to AMF association and secondly how Percentage mycorrhizal root colonization=(Total no. of infected AMF communities vary with plant to plant species in a group root segments/total no. of root segments examined)×100 or family? AM spores were isolated using wet sieving and decanting technique (Gerdemann and Nicolson, 1963). The quantitative Materials and Methods estimation of AM spores was done by modified method of (Gaur and Adholeya, 1994). The AM fungal isolates with their Mussoorie, known as ‘Queen of hills’ is located 35 kilometres accession numbers are preserved, trapped on host plants from the state capital of Dehradun with the GPS coordinates and retained with Biofertilizers Research Laboratory, Forest of 30°27’35.6724’’ N and 78°3’59.0364’’ E. The mountain is Research Institute, Dehradun, Uttarakhand, India. The AM situated at an average altitude of 2,005 metres (6,578 ft). To fungi were identified by using the keys of (Taber and Trappe, the northeast of the hills are the Himalayan snow ranges and 1982; Walker, 1992; Pérez and Schenck, 1990; Morton and to the south are the Doon Valley and Shiwalik ranges with Benny, 1990; Srivastava et al., 1996; Morton and Redecker, Asan river basin. Doon Valley is situated in the Shiwalik ranges 2001; Sharma et al., 2008; Sharma et al., 2009). of the Himalayas between 78°00’ E to 78°10’ E and 30°15’ N to 30°25’ N (Jain, 2007). This valley is spread over 454.95 sq. Results and Discussion km. It is surrounded by river Song on the east, river Tons on Twenty one different grass species were collected during the west, Himalayan ranges on the north and Sal forests in the survey and a herbarium was made for their confirmed the south (Singh et al., 2013). The following map is showing identification at Botany Division, Forest Research Institute, the study area and collections points. Dehradun. The detailed list of the collected grass species along with common name and their uses are tabulated in following Table 1. Some of the species like Poa annua, Phalaris minor, Eragrostis tenella are utilized as good sources of food and fodder for cattle. Some of the species are used by locals as their food like Aegilops speltoides, Avena fatua, Paspalum scrobiculatum. The grass species i.e. Cynodon dactylon is a sacred grass, local name- Dhoob, which is used to feed the cows as well as dogs for their stomach ailment (Table 1). The collected rhizospheric samples were analyzed for the number of AM spores present in their rhizospheric soil. The results indicated great variation among these rhizospheric samples.