Symbiotic Propagation of the Wet Meadow Orchid Anacamptis Laxiflora for Conservation and Reintroduction

Abstract

The study was designed to determine the diversity of mycorrhizal fungi associated with Anacamptis laxiflora, and to determine the symbiotic seed germination and the seedling development under in vitro and in situ conditions for reintroduction and conservation. Mycorrhizal fungi were isolated monthly from the roots ail. laxiflora adult individuals during the vegetation period. In addition, fungi were isolated from protocorms through in situ seed baiting. The isolates were identified as the members of Tulasnella, Ceratobasidium, Fusarium, Photna and Pezizales through the sequences of ITS. Non-Rhizoctonia isolates did not promote germination. The isolates, AL2 and AL3 did not affect germination while AL5, AL8, AL 11, AL12 and AL6 supported germination and seedling growth at different levels. Tubers were formed in symbiotic seedlings transferred to their natural habitats. The seeds inoculated with AL5 and AL6 germinated also and leaf primordium occurred in their natural habitats. Asymbiotic germination occurred in MS and VWD media with rates of 28.92% and 42.42%, respectively and no seedling development occurred. These results indicate that A. laxiflora has high mycorrhizal fungi diversity and the seeds have a high chance of germinating and becoming a new individual after forming a successful symbiotic association with compatible fungi in the soil.