In vivo and in vitro histological localization of endophytic fungi in Tinospora cordifolia (Willd.) Miers ex Hook F. & Thomas

• Localization of endophytic fungi inside the healthy leaves of Tinospora cordifolia.
• Staining with lactophenol cotton blue showed the presence of endophytic fungi.
• Endophytic fungi inhabitat in in vivo and in in vitro conditions.
• Preliminary test to verify the presence of endophytic fungi and save time.
• Enhance the level of understanding about the interaction of endophytes.

The present research protocol was developed to locate endophytic fungi present inside the healthy leaves of Tinospora cordifolia. The surface sterilized leaf explants after staining with lactophenol cotton blue showed the presence of endophytic fungi in intercellular spaces of ground, dermal tissue especially in midrib region. Leaves have shown dense colonies of endophytic fungi. Microscopic examination of in vitro and in vivo leaves has shown mycelium which was dense, branched and septate. Hence, the comparative study was conducted in both in vitro and in vivo leaves of T. cordifolia to locate endophytic fungi. The present study revealed that the fungi are not only developed in plants under natural environment but also under controlled conditions i.e. tissue culture of plants. Therefore, the present research work favoured the view that endophytes can infect nutrient medium of in vitro culture of T. cordifolia and diversity of endophytic fungi increase after hardening and acclimatization of T. cordifolia plantlets. The presence of endophytes in in vitro cultures depends both on age of tissue and host genotype. Further studies are required to isolate the endophytic fungi, pure culture on PDA plates and their identification to explore their biosynthetic pathway.