Trends in Biosciences 3 (1): 60-62, 2010 Influence of Ectomycorrhizal Inoculation on Blue Pine (Pinus wallchiania) and Deodar (Cedrus deodara) Seedlings GH. HASSAN DAR, MUZAFER, A. BEIG AND NADEEM, A. GANAI Division of Plant Pathology, S.K. University of Agricultural Sciences & Technology of Kashmir, Shalimar, Srinagar (J&K) 191 12 e-mail: [email protected] ABSTRACT MATERIALS AND METHODS The impact of five locally isolated mycorrhizae (Boletus edulis, Five pure cultures of mycorrhizal fungi viz., Boletus Suillus granulatus, Tricholoma album, Hygrophorus edulis (Be-2), Suillus granulatus (Sg-1), Tricholoma album camrophyllus and Scleroderma sp.) was assessed individually (Ta-2), Hygrophorous camrophyllus (Hy-2) and Scleroderma for mycorrhizal colonization and plant growth in blue pine sp. (Sc-7), were isolated from the fruiting bodies from Gulmarg (Pinus wallchiania) and deodar (Cedrus deodara). Typical and Bandipora forest areas in Kashmir valley as per the method ectomycorrhizae were established within 3-4 weeks of of Molina and Palmer, 1982. Better acid phosphatase activity inoculation. Acid phosphatase activities of these mycorrhizae varied from 197.0 to 323.5 m moles p-nitrophenol phosphate of these isolates, assessed as per the method of Ho, 1989, g-1. Mycorrhizal inoculation in blue pine resulted in 7-27, 22- served as the basis for their selection in this study. The 46, 16-48% increase in plant height and shoot and root biomass, performance of above mycorrhizal fungi was evaluated under respectively, in comparison to uninoculated control with in vitro conditions. The details of sites, where from these maximum increase in S. granulatus inoculated plants followed isolation were made, with respect to soil texture, organic carbon by B. edulis and T. album. Highest root colonization of 36.0% content, soil pH and dominant plant species were recorded. was recorded in S. granulatus followed by 35.4 % in B. edulis The surface sterilized seeds of kail pine and deodar and 33.5% in T. album. In deodar seedlings, mycorrhizal inoculation caused 15-38, 21-60 and 16-48% increase in plant were separately stratified in water and kept at 4+1ºC for five height and shoot and root biomass, respectively. Maximum days, and germinated on 1% water agar in petriplates. After increase was in B. edulis inoculated plants followed by S. 10-12 days the healthy germinated seedling were transferred granulatus and Scleroderma sp. inoculated plants. These fungi to test tubes (200x30 mm) containing vermiculite-peat moss also exhibited mycorrhizal colonization of 22.5 to 30.5% with medium and kept in growth chamber set at temperature of 25/ maximum colonization in B. edulis. 18+1ÚC under 16 h light and 8 h darkness. Each treatment was replicated five times in CRD. After 30 days of seedling growth, Key words Deodar, ectomycorrhizae, phosphatase activity, 2 mm dia. mycelial discs were placed near the root zone plant growth, pine separately and 2-3 ml modified Melin Norkran’s (MMN) liquid medium poured to initiate the fungal growth. A control without Jammu and Kashmir State has only 14% of its area under mycorrhizal inoculation was also maintained. Mycorrhizal forest cover; of which conifers occupy 41% area and rest are rootlets and total number of rootlets were counted under covered by broad leaves forest plants. Conifers, especially stereoscopic microscope for the establishment of mycorrhiza pine and deodar, have great potential as valuable timber and (Daughtridge, et al., 1986, Grand and Harvey, 1982). The as a source of fiber material for pulp and paper industry. Pine observations were made from 4th week of fungal inoculation and deodar show complete mycotrophic dependence in early and continued upto 10 weeks. Biomass of shoot and root was sapling establishment and survival. Mycorrhizal inoculation assessed after the plants were oven-dried at 110oC for 24 h. generally leads to the plant growth stimulation and better nutrient and water uptake as well as imparts resistance in RESULTS AND DISCUSSION plants to overcome the adversity of nature (Ekwebelam, 1979; Lans and Richards, 1971; Ruehle, et al., 1981). However, the The selected mycorrhizae were isolated from Gulmarg degree of tree dependence on fungal symbionts varies with and Bandipora forests of Kashmir valley. These forests are tree species and environment (Grove and Le Tacon, 1993). rich in coniferous trees (Cedrus deodara, Pinus wallichiana, The performance of eco-specific and specie-specific P. roxburghii and Abies pindrow). The soil texture at the indigenous isolates is always considered better than the selected sites was either clay loam or loam with varied organic introduced ones owing to their established acclimatization carbon content (0.99 to 1.11%) and soil pH (6.1 to 6.9). The under the prevailing conditions. The present study was aimed acid phosphatase activities of mycorrhizae varied from 197.0 to evaluate the performance of some locally isolated to 323.3 m moles p-nitrophenol phosphate g-1, with maximum indigenous mycorrhizae in plant growth stimulation in kail activity observed in Suillus granulatus and minimum in pine and deodar. Scleroderma sp. (Table 1). DAR, et al., : Influence of Ectomycorrhizal Inoculation on Blue Pine (Pinus wallchiania) and Deodar (Cedrus deodara) Seedlings 6 1 Table 1. Isolation of mycorrhizae and their acid phosphatase activity S. Species Acid Soil texture Soil Soil Dominant forest plant species No. phosphatase organic pH present activity (m carbon -1 moles PNP g (%) fungal biomass) 1. Tricholoma album (Ta-2) 291.2 Loam 1.01 6.4 Pinus wallichiana, Abies pindrow 2. Hygrophorus camrophyllus (Hy-2) 288.7 Clay loam 1.11 6.8 A .pindrow, Cedrus deodara 3. Scleroderma sp. (Sc-7) 197.0 Clay loam 1.08 6.9 A. pindrow; P. roxburghii 4. Boletus edulis (Be-2) 311.7 Clay loam 1.03 6.3 C. deodara; P. wallichiana 5. Suillus granulatus (Sg-1) 323.3 loam 0.99 6.1 C. deodara; P. wallichiana The plants showed differential response towards locally plant-1 in B. edulis and 190.2 and 24.2 mg plant-1 in T. album. isolated mycorrhizae with respect to their growth viz., root Highest root colonization of 36.0% was recorded in S. and shoot biomass as well as mycorrhizal root colonization. granulatus followed by 35.4 % in B. edulis and 33.5% in T. Typical ectomycorrhizae were established on inoculated pine album. The findings are in agreement with Riffle, 1973 who and deodar plants within 3-4 weeks of inoculation. These observed variable mycorrhization in Pinus ponderosa with findings corroborate with those of Kottke, et al., 1987 who the inoculation of Suillus granulatus, Lactarius delicious noticed mycorrhization within 2-3 weeks of inoculation and and Amanita sp. Similarly, Raj Kumar, et al., 1990 observed within 2 months of seedling germination in case of Larix positive correlation between mycorrhizal infection of decidua, Picea abies, Pseudotusga menzesii and Pinus Scleroderma aurantium, Boletus edulis and P. tinctorius with sylvestris. seedling growth, dry weight and phosphatase activity in The in vitro assessment of above mycorrhizae for root Khasi pine plant and soil. Wang, et al., 1985 reported that colonization and their influence on plant growth in kail pine ectomycorrhizae increased the biomass production of pine revealed an increase of 7-27, 22-46, 16-48% in plant height and seedlings (Pinus tabulaeformis) by about 40% with the shoot and root biomass, respectively, in comparison to inoculation of Boletus sp., Suillus greivilli and P. tinctorius. uninoculated control. Maximum plant height and higher root Like our observation in the present study, they also found colonization was observed in Suillus granulatus inoculated significant differences within the isolates and between plants followed by Boletus edulis and Tricholoma album inoculated and uninoculated saplings, with most effective (Table 2). The plants inoculated with mycorrhizae showed colonization by P. tinctorius, Rhizopogon sp. and Tricholoma more increase in height and biomass with age as compared to sp. control. The pine plants attained maximum growth of 11.3 cm In deodar seedlings, mycorrhizal inoculation resulted in after 70 days of inoculation in case of S. granulatus which 15-38, 21-60 and 16-48% increase in plant height and shoot was 27% higher than control. This was followed by 11.0 and and root biomass, respectively, in comparison to uninoculated 10.1 cm height observed in plants inoculated with B. edulis control. Maximum increase was noticed in B. edulis inoculated and T. album, respectively. Highest shoot and root dry biomass plants followed by S. granulatus and Scleroderma sp. of 204.2 and 27.0 mg plant-1, respectively, was observed in inoculated plants which attained a height of 9.5, 9.2 and 9.0 plants inoculated with S. granulatus which were 46 and 48% cm, respectively, after 70 days of inoculation in comparison to higher than control. This was followed by 202.4 and 26.8 mg the lowest of 6.9 cm in control (Table 3). Maximum shoot and Table 2. In vitro effect of mycorrhizal inoculation on plant height, biomass and root colonization inkail pine, Pinus wallchiania Mycorrhizal fungi Plant height (cm) Dry biomass Mycorrhizal root (mg)** colonization (%) 28d* 42d 56d 70d Shoot Root 28d 42d 56d 70d Tricholoma album (Ta-2) 5.0 7.3 8.4 10.1 190.2 24.2 12.8 16.2 28.7 33.5 Hygrophorus camrophyllus (Hy-2) 5.1 6.7 8.0 9.5 170.4 21.2 10.8 15.0 20.4 25.5 Scleroderma sp. (Sc-7) 4.2 6.5 9.0 9.9 173.2 21.4 11.9 15.4 25.2 28.4 Boletus edulis (Be-2) 6.2 8.5 9.2 11.0 202.4 26.8 13.9 20.8 28.7 35.4 Suillus granulatus (Sg-1) 6.7 8.9 9.5 11.3 204.2 27.0 14.0 21.2 28.0 36.0 Control 3.5 5.8 6.9 8.9 140.1 18.3 00.0 00.0 00.0 00.0 *d denotes the age of plant after inoculation; ** Dry matter taken at 70 days after inoculation 6 2 Trends in Biosciences 3 (1), 2010 Table 3.