The great diversity in colors and forms become the orchids a business with high economic value. The habitat fragmentation contributes to the extinction of orchids. Inoculation of orchid with mycorrhizal fungi for seedlings can guarantee the success of reintroduction. For this purpose, seeds of Hadrolaelia jongheana were germinated using an isolate of Tulasnella sp. Seedlings were transferred to the natural field. Roots samples were collected before re-introduction, and 120th and 240th days. The diversity of mycorrhizal fungi was performed by ITS-PCR-DGGE. The ecological succession occurred in the field and the diversity was higher after 240th d. This work comprises the first study using tropical orchids for reintroduction for approaching to ecological aspects of mycorrhizal fungi association in Brazil with conservation purposes.