Browsing by Author "Valo, Karoliina"

Studies have revealed that members of Archaea exist in various environments such as human gut and the sporocarps of some fungi in addition to the extreme environments in which they were first discovered. It is yet unknown how some archaea end up in these environments and what is their role there. To study the archaea efficiently in different environments, enrichment or pure cultures are required. In this work liquid media were developed for cultivation and enrichment of archaea from the edible sporocarps of Suillus bovinus, Craterellus tubaeformis, Agaricus bisporus and Boletus pinophilus. The source of carbon in each medium was determined to be yeast extract, crushed and sterile filtered sporocarp of either Craterellus tubaeformis or Agaricus bisporus, or casaminoacids. The highest amount of prokaryotes grew on the casaminoacid medium but it was also a very good medium for the bacteria in sporocarps. The media were anaerobic to prevent the fastest aerobic bacteria from growing. Antibiotics kanamycin and ampicillin were added to reduce bacterial growth but with further inoculation resistant bacteria outgrew the archaea. Adding lysozyme with the antibiotics reduced bacterial growth also with later cultivations and the archaea-bacteria ratio turned favourable. The amount of both archaea and bacteria was determined with the quantitative PCR of the 16S rRNA gene. The most archaea was enriched from the sporocarps of Suillus bovinus. On the contrary, prokaryotes that grew from the sporocarp of Craterellus tubaeformis were mostly bacteria. The Sporocarps of both Boletus pinophilus and Agaricus bisporus didn t harbour many archaea or bacteria that could have been detected or enriched in this study. It was also noted that there was great variation between individual sporocarps of the same species in the amount of prokaryotes they originally harboured. Archaea were successfully enriched from the sporocarps of Suillus bovinus for the duration of three consecutive cultivations on the casaminoacid medium with the addition of kanamycin, ampicillin and lysozyme.