eng Oriental Scientific Publishing Company Biosciences Biotechnology Research Asia 0973-1245 2023-12-31 20 4 1355 1363 10.13005/bbra/3181 48627 article Amit Kumar Sehgal 1 Anand Sagar 1 Department of Biosciences, Himachal Pradesh University, Summer Hill, Shimla (H.P.) , India The present investigations are aimed to synthesize in vitro ectomycorrhizae between Pinus gerardiana and two gilled ectomycorrhizal (ECM) mushrooms (Amanita ceciliae and Lactarius sanguifluus). To carry out in vitro synthesis, pure cultures of ECM mushrooms (A. ceciliae and L. sanguifluus) were isolated on Potato Dextrose Agar (PDA) and Modified Melin-Norkans (MMN) Medium respectively. The synthesis was achieved successfully in the surfaced sterilized seedlings of P. gerardiana geminated under aseptic conditions by using vermiculite, peat, medium for ECM fungi and inoculum of each fungus in the test tubes. Mycorrhization was checked periodically in the test tubes. P. gerardiana seedlings were lifted from test tubes after five months to observe ectomycorrhizae formation on the root system with A. ceciliae and L. sanguifluus. The synthesized ectomycorrhizae were dark brown in case of A. ceciliae whereas in case of L. sanguifluus the colour of ECM roots was yellowish brown. Anatomy of synthesized ectomycorrhizae with both ECM fungi showed fully developed fungal mantle and Hartig net.  The seedlings with ECM synthesis showed a significant effect on the growth and development. https://www.biotech-asia.org/vol20no4/aseptic-in-vitro-synthesis-of-pinus-gerardiana-ectomycorrhizae-with-amanita-ceciliae-and-lactarius-sanguifluus/ Amanita ceciliae; Ectomycorrhiza; In vitro synthesis; Lactarius sanguifluus; Pinus gerardiana