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Fungal Assemblages in the Rhizosphere and Rhizoplane of Grasses of the Subfamily Panicoideae in the Lakkavalli Region of Karnataka, India

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Fungal Assemblages in the Rhizosphere and Rhizoplane of Grasses of the Subfamily Panicoideae in the Lakkavalli Region of Karnataka, India Microbes Environ. Vol. 26, No. 3, 228–236, 2011 http://wwwsoc.nii.ac.jp/jsme2/ doi:10.1264/jsme2.ME10163 Fungal Assemblages in the Rhizosphere and Rhizoplane of Grasses of the Subfamily Panicoideae in the Lakkavalli Region of Karnataka, India MADHUGIRI MALLAIAH VASANTHAKUMARI1, and MANCHANAHALLY BYRAPPA SHIVANNA1* 1Department of P.G. Studies and Research in Applied Botany, Kuvempu University, Shankaraghatta 577451, Shimoga District, Karnataka, India (Received September 3, 2010—Accepted April 24, 2011—Published online June 11, 2011) Fungal communities associated with roots play an important role in nutrient cycling, supporting plant growth and the biocontrol of plant diseases. Experiments were conducted in 2004–2006 to isolate and characterize, based on their morphological features, rhizosphere and rhizoplane fungi from perennial grasses of the subfamily Panicoideae growing in forests of the Western Ghats in India. Fungal species were isolated on potato dextrose, czapeck dox and water agar, in different locations and seasons. The results obtained on PDA were used for detailed analysis since most fungi occurred in high percentages. While certain grasses harbored diverse fungal species, others supported only a few species. Most fungi were isolated during winter followed by the rainy and summer seasons. The species richness, diversity and evenness of fungal assemblages in the rhizosphere and rhizoplane depended on the grass species and season. Ascomycetes were isolated in large numbers in most grass species. Species of Aspergillus, Chaetomium, Penicillium and Trichoderma occurred frequently. Certain others and non-sporulating fungi were grass species-specific. Most fungal species colonized the middle of the root more than the root tip or root base. Results suggest that perennial grasses harbor diverse fungal communities whose potential could be tapped for producing secondary metabolites and managing plant diseases. Key words: Fungal diversity, Western Ghats, perennial grass, seasonal variation, Ascomycetes Fungi colonize diverse habitats and substrates and play a 37, 40, 45). These studies prompted the authors to document substantial role in plant health and productivity as well as in fungal assemblages in the rhizosphere of perennial grasses disease. The rhizosphere is the zone of soil directly influenced of India and evaluate their biocontrol characteristics. Based by the roots of plants and provides a distinct biologically on this, a survey of rhizosphere fungi in certain perennial active micro-habitat within the terrestrial ecosystem (5, 11, grass species was undertaken which indicated the occurrence 30, 41). The organic compounds present in root exudates of anamorphic and teleomorphic ascomycetes, anamorphic make the rhizosphere environment very different from that basidiomycetes, zygomycetes and certain non-sporulating of the bulk soil and support a hoard of saprophytic fungi (Vasanthakumari et al. 2006; conference abstract). In microorganisms. These microorganisms are attracted to the this context, the isolation and characterization of rhizosphere release of ions, oxygen and water, in addition to carbon- and rhizoplane fungi from certain perennial grass species of containing compounds, amino acids, hormones and vitamins the subfamily Panicoideae were taken up. An attempt was (52). There is a considerable body of evidence which suggests also made to document variation in the diversity of fungal that the loss of soluble organic substances from roots is communities in relation to season, growing stage and location. significantly stimulated by the presence of microbes (33, 55). The present study is aimed at understanding the diversity, A search of the literature indicated that certain plant species evenness and richness of fungal communities in different sea- have been studied extensively for the occurrence of fungi in sons in the rhizosphere and rhizoplane of certain Panicoideae the rhizosphere and rhizoplane (34). A number of fungi have grasses growing in the area adjoining the Bhadra reservoir in been isolated and characterized from the rhizosphere of crops the Lakkavalli state forest of Karnataka, India. and to some extent, non-crop plants (1, 2, 29, 39). However, few reports are available on the occurrence of rhizosphere Materials and Methods fungi in grass species (23, 26, 42, 46). The diversity of rhizosphere fungi has been shown to vary depending on Study site season, location and soil type (7). Fungal species have been The study was carried out in the Bhadra Wildlife sanctuary, characterized mainly on a morphological basis; however, located in the Western Ghats region of Karnataka, India. The Western Ghats is recognized as a ‘hot spot’ of global biodiversity. some attempts have been made using molecular techniques The Lakkavalli state forest (13°34'–13°39' N latitude, 75°36'–75°39' (23, 42). Previous work on the grass rhizosphere indicated E longitude), an area adjoining the Bhadra reservoir situated in the the occurrence of a large number of fungal species, of which peripheral region of the Bhadra Wildlife sanctuary, was selected as a few were studied for their growth promotion and biocontrol the study area. Three study sites were established at a distance of potential and systemic resistance inducing ability (15, 27, 36, two kilometers from each other within the study area. Three quadrates (1×1 m) representing three replicates were established at each site. The data were collected in July, November and March, * Corresponding author. E-mail: [email protected]; representing the rainy, winter and summer seasons, of 2004–2006. Tel: +91–9448836859; Fax: +91–08282–256255. Fungal Assemblages in Panicoideae Grasses 229 Plant species and sampling design design (RCBD) and trials were conducted during 2004–05 and 2005– Five perennial grass species—Alloteropsis cimicina (L.) Stapf. 06. Fungal populations from the rhizosphere samples were quantified −1 (Bug seed-grass), Digitaria bicornis (Lam). Roem. & Schult as the number of colony forming units (cfu g ). The root colonization Beauv. (Asian crab grass), Heteropogon contortus (L.) P. Beauv. frequency (%) of rhizoplane fungi (20) and the predominantly (Tangle head), Ischaemum ciliare Retz. (Indian murianagrass) and occurring fungi (%) (31) were calculated. The diversity, evenness Paspalidium flavidum (Retz.) A. Camus (Shot grass) growing in the and richness of fungal species in the rhizosphere and rhizoplane of study area were selected. Initially, all species were identified to the grass species were analyzed using Simpson and Shannon indices. tribe level in the field based on vegetative and floral characteristics Jaccard’s similarity co-efficient and rarefaction indices were also (8) and characterized to the species level as per the descriptions in calculated (http://palaeo-electronica.org/2001. Accessed 20 Decem- manuals (10, 47, 57). The voucher specimens were collected at ber 2008). different growth stages. The identity of grasses was kindly confirmed by a grass specialist, Dr. G.K. Bhat, Retired Professor, Poornaprajna Results College, Udupi, Karnataka. Simultaneously, herbarium specimens were also prepared and deposited in the Department of Applied Study area, soil mineral nutrients and grass species Botany for future reference. Tests of homogeneity of two trials by ANOVA indicated Collection of soil samples no significant difference in the incidence of fungal commu- Soil samples were collected from soil adjacent to grass plants of nities isolated from rhizospheres and rhizoplanes of three each species up to a depth of 15–20 cm. Soil samples from each quadrate were pooled, air-dried and tested regarding soil pH, organic locations in the study area. Therefore, the data for the two carbon and available N, P and K (4, 28, 48). trials over three locations were averaged and subjected to ANOVAs and further analyses. Preparation of rhizosphere and rhizoplane samples The mineral nutrient composition of bulk soil (pH 5.3– Rhizosphere and rhizoplane samples were collected from five 5.6) collected from different locations did not vary much. plants (per quadrate) of each grass species. The zone of soil immediately adjacent to the roots of each plant (2 cm from the However, the N, P and K contents of soil samples varied in −1 culm) up to a depth of 15–20 cm was removed carefully using a different seasons. The N content was high (4.05 kg ha each) trowel. Samples consisting of soil as well as root systems were during winter and summer and very low (0.36 kg ha−1) during collected and brought to the laboratory in separate sterile polypro- the rainy season. However, the P and K contents were highest pylene bags and used within 4 h of collection. The root systems during winter (23.05 and 10.12 kg ha−1, respectively) followed were carefully removed from the sample and gently shaken off to by the rainy season (16.59 and 9.51 kg ha−1, respectively). remove superfluous soil. Soil particles closely adhering to the roots In summer, their availability was low (15.98 and 7.69 kg were collected by gentle scraping using a sterile spatula and brushing −1 with a camel hair brush. The soil so collected formed the rhizosphere ha , respectively). The organic carbon content remained the sample. The root system without soil particles was considered the same (0.99–1.00%). rhizoplane sample. Out of 20 perennial grass species in the study area, five Isolation and characterization of fungal species from the species of the sub family Panicoideae were selected for the rhizosphere and rhizoplane study
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