For isolation of endophytic fungi four districts of Assam,
North East India were selected. Isolation was done from
healthy and mature leaves of C. sinensis. The samples were
processed within 24 h of collection and isolation was done
according to Petrini and Dreyfuss [27,28]. The leaves were
cut into small pieces (1 cm2
), washed in running tap water followed
by 96% alcohol treatment for 30 s, washed with sterile
distilled water for 30 s followed by sodium hypochloride
NaOCl (15%)–distilled water (1:3) treatment for 5 min, treated
with 96% alcohol for 30 s and finally washed with sterile water
for 1–2 min followed by drying. Surface sterilization was followed
as per the protocol described by Petrini and Dreyfuss
[27,28]. It was then kept in Petriplates containing PDA media
and kept in incubator at 28 C at inverted position for 5–
6 days. As the hyphal growth appeared, it was finally transferred
to PDA slant for further processing and PDA slant is
prepared in test tube which helps to store the fungi for a long
period.